CN114945665A - 用于增强地衣芽孢杆菌中蛋白产生的组合物和方法 - Google Patents

用于增强地衣芽孢杆菌中蛋白产生的组合物和方法 Download PDF

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CN114945665A
CN114945665A CN202180009565.4A CN202180009565A CN114945665A CN 114945665 A CN114945665 A CN 114945665A CN 202180009565 A CN202180009565 A CN 202180009565A CN 114945665 A CN114945665 A CN 114945665A
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S·D·多伊格
R·L·弗里希
H·贺
C·莱夫朗
马臻
B·J·保罗
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Danisco US Inc
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Abstract

本公开总体上涉及用于构建和/或获得包含增强的蛋白产生能力的地衣芽孢杆菌细胞(例如,蛋白产生型宿主)的组合物和方法。因此,某些实施例涉及衍生自亲本地衣芽孢杆菌菌株的经遗传修饰的地衣芽孢杆菌菌株,其产生增加量的一种或多种目的蛋白。

Description

用于增强地衣芽孢杆菌中蛋白产生的组合物和方法
技术领域
本公开总体上涉及细菌学、微生物学、遗传学、分子生物学、酶学、工业蛋白生产等领域。因此,本公开的某些实施例涉及用于构建具有增强的蛋白产生表型的地衣芽孢杆菌细胞/菌株的组合物和方法。
相关申请的交叉引用
本申请要求于2020年1月15日提交的美国临时专利申请号62/961,234的权益,将该申请通过引用以其整体并入本文。
序列表的引用
命名为“NB41684-WO-PCT_SequenceListing.txt”的文本文件序列表的电子提交的内容创建于2021年1月7日,并且大小为425KB,将其通过引用以其整体并入本文。
背景技术
***如枯草芽孢杆菌(Bacillus subtilis)、地衣芽孢杆菌和解淀粉芽孢杆菌(Bacillus amyloliquefaciens)由于其优异的发酵特性和高产率(例如,高达25克/升培养物;Van Dijl和Hecker,2013年),经常被用作微生物工厂,用于产生工业相关蛋白。例如,枯草芽孢杆菌因其产生食品、纺织品、洗衣、医疗器械清洁、制药工业等所必需的α-淀粉酶(Jensen等人,2000;Raul等人,2014)和蛋白酶(Brode等人,1996)而为人熟知(Westers等人,2004)。由于这些非致病性***产生完全不含毒性副产物(例如脂多糖;LPS,也称为内毒素)的蛋白,因此它们已经获得了欧洲食品***的“安全资格认定”(QPS)状态,并且其许多产品获得了美国食品和药品管理局的“通常认为安全”(GRAS)状态(Olempska-Beer等人,2006;Earl等人,2008;Caspers等人,2010)。
因此,微生物宿主细胞中蛋白(例如,酶、抗体、受体等)的产生在生物技术领域中是特别有意义的。同样,用于产生和分泌一种或多种目的蛋白的芽孢杆菌属宿主细胞的优化具有高度相关性,特别是在如下工业生物技术环境中,其中当蛋白以大的工业产量生产时该蛋白产率的微小改善具有重大意义。更特别地,地衣芽孢杆菌是具有高工业重要性的芽孢杆菌属物种宿主细胞,因此,对于构建新的和改善的地衣芽孢杆菌产生菌株,高度希望具有修饰和工程化地衣芽孢杆菌宿主细胞以获得增强的/增加的蛋白表达/产生的能力。因此,本公开涉及对于获得和构建具有增加的蛋白产生能力的地衣芽孢杆菌细胞(例如,蛋白产生型宿主细胞)的高度希望和未满足的需求。
发明内容
本公开总体上涉及用于获得包含增强的蛋白产生能力的地衣芽孢杆菌细胞(例如,蛋白产生型宿主)的组合物和方法。因此,本公开的某些实施例涉及用于构建此类经修饰的地衣芽孢杆菌细胞/菌株的方法,所述细胞/菌株产生增加量的一种或多种目的蛋白。
因此,本公开的某些实施例涉及在经修饰的地衣芽孢杆菌细胞中产生增加量的内源目的蛋白(POI)的方法,所述方法包括(a)获得表达POI的亲本地衣芽孢杆菌细胞并通过在亲本细胞中引入包含与天然prsA可读框(ORF)可操作地连接的天然prsA启动子的多核苷酸来修饰亲本细胞,和(b)在适于产生所述POI的条件下发酵步骤(a)的经修饰的细胞,其中当在相同条件下发酵时,经修饰的细胞相对于亲本细胞产生增加量的POI。在方法的特定实施例中,步骤(a)的引入的多核苷酸包含与SEQ ID NO:100具有至少95%序列同一性的天然prsA启动子。在方法的其他实施例中,步骤(a)的引入的多核苷酸包含与SEQ ID NO:101具有至少90%序列同一性的天然prsA ORF。在其他实施例中,引入的多核苷酸编码与SEQ IDNO:155具有约90%序列同一性的天然prsA蛋白。在某些优选的实施例中,亲本细胞包含编码天然prsA蛋白的内源(野生型)prsA基因,其中引入的多核苷酸由此编码与SEQ ID NO:155具有约90%序列同一性的prsA蛋白的第二(第2)拷贝。在其他实施例中,步骤(a)中引入的多核苷酸整合到经修饰的地衣芽孢杆菌细胞的基因组中。在方法的又其他实施例中,目的蛋白(POI)是蛋白酶或淀粉酶。在其他实施例中,经修饰的细胞包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏。在其他实施例中,经修饰的细胞包含与SEQ ID NO:121或SEQ ID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。在其他实施例中,经修饰的细胞包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏和与SEQ ID NO:121或SEQ ID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。
在某些其他实施例中,本公开涉及在经修饰的地衣芽孢杆菌细胞中产生增加量的异源目的蛋白(POI)的方法,所述方法包括(a)向亲本地衣芽孢杆菌细胞中引入(i)编码POI的表达盒和(ii)包含可操作地连接到天然prsA可读框(ORF)的天然prsA启动子的多核苷酸,和(b)在适于产生所述POI的条件下发酵步骤(a)的经修饰的细胞,其中当在相同条件下发酵时,经修饰的细胞相对于亲本细胞产生增加量的POI。在方法的特定实施例中,步骤(a)(ii)的引入的多核苷酸包含与SEQ ID NO:100具有至少95%序列同一性的天然prsA启动子。在某些其他实施例中,步骤(a)(ii)的引入的多核苷酸包含与SEQ ID NO:101具有至少90%序列同一性的天然prsA ORF。在方法的又其他实施例中,内源prsA基因编码与SEQ IDNO:155具有约90%序列同一性的天然prsA蛋白。在某些其他实施例中,步骤(a)(ii)中引入的多核苷酸整合到经修饰的地衣芽孢杆菌细胞的基因组中。在某些优选的实施例中,亲本细胞包含编码天然prsA蛋白的内源(野生型)prsA基因,其中引入的多核苷酸步骤(a)(ii)由此编码与SEQ ID NO:155具有约90%序列同一性的prsA蛋白的第二(第2)拷贝。在特定实施例中,目的蛋白(POI)是蛋白酶或淀粉酶。在其他实施例中,经修饰的细胞包含与SEQ IDNO:122具有至少90%序列同一性的dltA基因的缺失或破坏。在其他实施例中,经修饰的细胞包含与SEQ ID NO:121或SEQ ID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。在某些优选的实施例中,经修饰的细胞包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏和与SEQ ID NO:121或SEQ ID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。
本公开的其他实施例涉及经修饰的地衣芽孢杆菌细胞/菌株,其衍生自包含编码天然prsA蛋白的内源prsA基因的亲本地衣芽孢杆菌细胞/菌株。因此,在某些实施例中,本公开的经修饰的地衣芽孢杆菌细胞包含引入的多核苷酸,所述多核苷酸包含可操作地连接至天然prsA可读框(ORF)的天然prsA启动子。在特定实施例中,引入的多核苷酸包含与SEQID NO:100具有至少95%序列同一性的天然prsA启动子。在其他实施例中,引入的多核苷酸包含与SEQ ID NO:101具有至少90%序列同一性的天然prsA ORF。在又其他实施例中,引入的多核苷酸编码与SEQ ID NO:155具有约90%序列同一性的天然prsA蛋白。在某些其他实施例中,引入的编码天然prsA蛋白的多核苷酸整合到经修饰的地衣芽孢杆菌细胞的基因组中。在另一个实施例中,经修饰的细胞包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏。在另一个实施例中,经修饰的细胞包含与SEQ ID NO:121或SEQ IDNO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。在一个优选的实施例中,经修饰的细胞包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏和与SEQ ID NO:121或SEQ ID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。在某些其他实施例中,经修饰的细胞包含编码异源目的蛋白(POI)的引入的表达构建体。在其他实施例中,异源POI是蛋白酶或淀粉酶。因此,本公开的某些实施例涉及获得、分离、纯化以及类似地处理由本公开的经修饰的地衣芽孢杆菌细胞产生的目的蛋白。
因此,本公开的某些其他实施例涉及经修饰的地衣芽孢杆菌细胞,其相对于衍生它们的亲本地衣芽孢杆菌细胞产生增加量的目的蛋白(POI)。因此,在某些实施例中,本公开涉及相对于亲本地衣芽孢杆菌细胞产生增加量的目的蛋白(POI)的经修饰的地衣芽孢杆菌细胞,其中经修饰的细胞衍生自表达POI的亲本地衣芽孢杆菌细胞,其中经修饰的细胞包含引入的包含可操作地连接到天然prsA可读框(ORF)的天然prsA启动子的多核苷酸并且包含与SEQ ID NO:121或SEQ ID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏,其中当在相同条件下发酵时,经修饰的细胞相对于亲本菌株产生增加量的POI。在另一个实施例中,经修饰的地衣芽孢杆菌细胞包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏。在又另一个实施例中,天然prsA启动子与SEQ ID NO:100具有至少95%序列同一性。在某些其他实施例中,天然prsA ORF与SEQ ID NO:101具有至少90%序列同一性。在另一个实施例中,天然prsA蛋白与SEQ ID NO:155具有约90%序列同一性。在特定实施例中,目的蛋白(POI)是蛋白酶或淀粉酶。因此,本公开的某些其他实施例涉及获得、分离、纯化以及类似地处理由经修饰的地衣芽孢杆菌细胞产生的目的蛋白。
在其他实施例中,本公开涉及相对于亲本地衣芽孢杆菌细胞产生增加量的目的蛋白(POI)的经修饰的地衣芽孢杆菌细胞,其中经修饰的细胞衍生自表达POI的亲本地衣芽孢杆菌细胞,其中经修饰的细胞包含引入的包含可操作地连接到天然prsA可读框(ORF)的天然prsA启动子的多核苷酸并且包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏,其中当在相同条件下发酵时,经修饰的细胞相对于亲本菌株产生增加量的POI。在其他实施例中,经修饰的细胞进一步包含与SEQ ID NO:121或SEQ ID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。在又另一个实施例中,天然prsA启动子与SEQ ID NO:100具有至少95%序列同一性。在某些其他实施例中,天然prsA ORF与SEQ IDNO:101具有至少90%序列同一性。在另一个实施例中,天然prsA蛋白与SEQ ID NO:155具有约90%序列同一性。在特定实施例中,目的蛋白(POI)是蛋白酶或淀粉酶。因此,本公开的某些其他实施例涉及获得、分离、纯化以及类似地处理由经修饰的地衣芽孢杆菌细胞产生的目的蛋白。
生物序列说明
SEQ ID NO:1是编码天然化脓链球菌Cas9蛋白的氨基酸序列。
SEQ ID NO:2是编码SEQ ID NO:1的Cas9蛋白的核酸序列,该核酸序列已针对在芽孢杆菌属物种细胞中表达进行了密码子优化。
SEQ ID NO:3是合成的N-末端核定位信号(NLS)的氨基酸序列。
SEQ ID NO:4是合成的C-末端核定位信号(NLS)的氨基酸序列。
SEQ ID NO:5是合成的十组氨酸标签的氨基酸序列。
SEQ ID NO:6是枯草芽孢杆菌aprE启动子序列。
SEQ ID NO:7是合成的终止子核酸序列。
SEQ ID NO:8是正向引物核酸序列。
SEQ ID NO:9是反向引物核酸序列。
SEQ ID NO:10是合成的pKB320骨架核酸序列。
SEQ ID NO:11是合成的pKB320核酸序列。
SEQ ID NO:12是引物核酸序列。
SEQ ID NO:13是引物核酸序列。
SEQ ID NO:14是引物核酸序列。
SEQ ID NO:15是引物核酸序列。
SEQ ID NO:16是引物核酸序列。
SEQ ID NO:17是引物核酸序列。
SEQ ID NO:18是引物核酸序列。
SEQ ID NO:19是引物核酸序列。
SEQ ID NO:20是引物核酸序列。
SEQ ID NO:21是引物核酸序列。
SEQ ID NO:22是引物核酸序列。
SEQ ID NO:23是引物核酸序列。
SEQ ID NO:24是引物核酸序列。
SEQ ID NO:25是合成的pRF694核酸序列。
SEQ ID NO:26是合成的pRF801核酸序列。
SEQ ID NO:27是合成的pRF806核酸序列。
SEQ ID NO:28是地衣芽孢杆菌靶位点1(TS1)核酸序列。
SEQ ID NO:29是地衣芽孢杆菌靶位点2(TS2)核酸序列。
SEQ ID NO:30是地衣芽孢杆菌serA1可读框(ORF)序列。
SEQ ID NO:31是包含核苷酸“AGG”的靶位点1PAM序列。
SEQ ID NO:32是编码可变靶向(VT)位点1的核酸序列。
SEQ ID NO:33是编码CER结构域的合成的核酸序列。
SEQ ID NO:34是靶向位点1的合成的指导RNA(gRNA)序列。
SEQ ID NO:35是合成的spac启动子核酸序列。
SEQ ID NO:36是合成的t0终止子核酸序列。
SEQ ID NO:37是地衣芽孢杆菌serA1同源臂1核酸序列。
SEQ ID NO:38是合成的serA1同源臂1正向引物序列。
SEQ ID NO:39是合成的serA1同源臂1反向引物序列。
SEQ ID NO:40是地衣芽孢杆菌serA1同源臂2核酸序列。
SEQ ID NO:41是合成的serA1同源臂2正向引物序列。
SEQ ID NO:42是合成的serA1同源臂2正向引物序列。
SEQ ID NO:43是编码靶位点1(TS1)gRNA的表达盒。
SEQ ID NO:44是合成的serA1缺失编辑模板。
SEQ ID NO:45是地衣芽孢杆菌rghR1可读框(ORF)序列。
SEQ ID NO:46是包含核苷酸“CGG”的靶位点2PAM序列。
SEQ ID NO:47是靶向位点2的合成的指导RNA(gRNA)序列。
SEQ ID NO:48是地衣芽孢杆菌rghR1同源臂1核酸序列。
SEQ ID NO:49是合成的rghR1同源臂1正向引物序列。
SEQ ID NO:50是合成的rghR1同源臂1反向引物序列。
SEQ ID NO:51是地衣芽孢杆菌rghR1同源臂2核酸序列。
SEQ ID NO:52是合成的rghR1同源臂2正向引物序列。
SEQ ID NO:53是合成的rghR1同源臂2反向引物序列。
SEQ ID NO:54是编码靶位点2(TS2)gRNA的表达盒。
SEQ ID NO:55是合成的rghR1缺失编辑模板。
SEQ ID NO:56是编码Cas9(Y155H)变体蛋白的氨基酸序列。
SEQ ID NO:57是合成的Y155H正向引物序列。
SEQ ID NO:58是合成的Y155H反向引物序列。
SEQ ID NO:59是合成的pRF827核酸序列。
SEQ ID NO:60是编码SEQ ID NO:56的变体Cas9(Y155H)蛋白的表达盒。
SEQ ID NO:61是合成的pRF856核酸序列。
SEQ ID NO:62是合成的pRF862核酸序列。
SEQ ID NO:63是合成的Y155H片段序列。
SEQ ID NO:64是合成的Y155H片段正向引物序列。
SEQ ID NO:65是合成的Y155H片段反向引物序列。
SEQ ID NO:66是合成的pRF694片段序列。
SEQ ID NO:67是合成的pRF694片段正向引物序列。
SEQ ID NO:68是合成的pRF694片段反向引物序列。
SEQ ID NO:69是合成的pRF869核酸序列。
SEQ ID NO:70是地衣芽孢杆菌rghR2可读框(ORF)序列。
SEQ ID NO:71是合成的rghR2终止片段。
SEQ ID NO:72是合成的rghR2终止编辑模板。
SEQ ID NO:73是编码rghR2 gRNA的表达盒。
SEQ ID NO:74是合成的片段正向引物。
SEQ ID NO:75是合成的片段反向引物。
SEQ ID NO:76是合成的pRF862骨架正向引物。
SEQ ID NO:77是合成的pRF862骨架反向引物。
SEQ ID NO:78是合成的pRF879核酸序列。
SEQ ID NO:79是地衣芽孢杆菌pRF879靶位点和PAM核酸序列。
SEQ ID NO:80是合成的pRF879编辑模板序列。
SEQ ID NO:81是合成的pRF946核酸序列。
SEQ ID NO:82是地衣芽孢杆菌pR946靶位点和PAM核酸序列。
SEQ ID NO:83是合成的pR946编辑模板序列。
SEQ ID NO:84是合成的pZM221核酸序列。
SEQ ID NO:85是合成的pZM221靶位点和PAM核酸序列。
SEQ ID NO:86是合成的pZM221编辑模板序列。
SEQ ID NO:87是地衣芽孢杆菌lysA可读框(ORF)序列。
SEQ ID NO:88是合成的pBl.comK核酸序列。
SEQ ID NO:89是合成的壮观霉素标记核酸序列。
SEQ ID NO:90是枯草芽孢杆菌xylR核酸序列。
SEQ ID NO:91是枯草芽孢杆菌xylAp核酸序列。
SEQ ID NO:92是合成的comK核酸序列。
SEQ ID NO:93是合成的cat_prsA核酸序列。
SEQ ID NO:94是地衣芽孢杆菌cat上游核酸序列。
SEQ ID NO:95是地衣芽孢杆菌cat启动子核酸序列。
SEQ ID NO:96是地衣芽孢杆菌catH核酸序列。
SEQ ID NO:97是合成的双终止子核酸序列。
SEQ ID NO:98是地衣芽孢杆菌catH终止子核酸序列。
SEQ ID NO:99是枯草芽孢杆菌spoVG终止子核酸序列。
SEQ ID NO:100是地衣芽孢杆菌prsA启动子核酸序列。
SEQ ID NO:101是地衣芽孢杆菌prsA可读框(ORF)序列。
SEQ ID NO:102地衣芽孢杆菌amyL终止子核酸序列。
SEQ ID NO:103是地衣芽孢杆菌cat下游核酸序列。
SEQ ID NO:104是合成的正向引物核酸序列。
SEQ ID NO:105是合成的反向引物核酸序列。
SEQ ID NO:106是合成的prsA(第2拷贝)验证核酸序列。
SEQ ID NO:107是合成的引物序列。
SEQ ID NO:108是合成的引物序列。
SEQ ID NO:109是合成的引物序列。
SEQ ID NO:110是地衣芽孢杆菌缺失的catHP和catH编码核酸序列。
SEQ ID NO:111是cat catH缺失中的合成prsA(第2拷贝)表达盒
SEQ ID NO:112是合成的catH(第2拷贝)缺失验证PCR产物。
SEQ ID NO:113是合成的正向引物序列。
SEQ ID NO:114是合成的反向引物序列。
SEQ ID NO:115是合成的dltA-2验证PCR产物。
SEQ ID NO:116是合成的dltA-2亲本验证PCR产物。
SEQ ID NO:117是合成的正向引物序列。
SEQ ID NO:118是合成的反向引物序列。
SEQ ID NO:119是合成的rghR2缺失验证PCR产物。
SEQ ID NO:120是地衣芽孢杆菌亲本rghR2缺失验证PCR产物。
SEQ ID NO:121是地衣芽孢杆菌亲本rghR2基因座。
SEQ ID NO:122是地衣芽孢杆菌亲本dltA基因座。
SEQ ID NO:123是地衣芽孢杆菌亲本cat基因座。
SEQ ID NO:124是合成的cat 2x prsA基因座
SEQ ID NO:125是合成的dltA-2基因座。
SEQ ID NO:126是地衣芽孢杆菌淀粉酶1蛋白的氨基酸序列。
SEQ ID NO:127是合成的serA1淀粉酶1盒。
SEQ ID NO:128是合成的p3启动子序列。
SEQ ID NO:129是合成的经修饰的aprE 5'-UTR序列。
SEQ ID NO:130是编码amyL信号序列的地衣芽孢杆菌核酸序列。
SEQ ID NO:131是编码SEQ ID NO:126的淀粉酶1蛋白的地衣芽孢杆菌核酸序列。
SEQ ID NO:132是合成的lysA淀粉酶1盒。
SEQ ID NO:133是合成的lysA亲本基因座核酸序列。
SEQ ID NO:134是编码lysA的地衣芽孢杆菌核酸序列。
SEQ ID NO:135是合成的p2启动子序列。
SEQ ID NO:136是淀粉酶2蛋白的氨基酸序列。
SEQ ID NO:137是合成的serA1淀粉酶2盒。
SEQ ID NO:138是枯草芽孢杆菌rrnI启动子序列。
SEQ ID NO:139是枯草芽孢杆菌aprE 5'-UTR序列。
SEQ ID NO:140是编码SEQ ID NO:136的淀粉酶2蛋白的合成核酸序列。
SEQ ID NO:141是合成的amyL或lysA淀粉酶2盒。
SEQ ID NO:142是合成的amyL亲本基因座。
SEQ ID NO:143是淀粉酶3蛋白的氨基酸序列。
SEQ ID NO:144是合成的serA1淀粉酶3盒。
SEQ ID NO:145是编码SEQ ID NO:143的淀粉酶3蛋白的合成的核酸序列。
SEQ ID NO:146是合成的lysA淀粉酶3盒。
SEQ ID NO:147是淀粉酶4蛋白的氨基酸序列。
SEQ ID NO:148是合成的serA1淀粉酶4盒。
SEQ ID NO:149是编码SEQ ID NO:147的淀粉酶4蛋白的合成的核酸序列。
SEQ ID NO:150是合成的lysA淀粉酶4盒。
SEQ ID NO:151是淀粉酶5蛋白的氨基酸序列。
SEQ ID NO:152是合成的serA1淀粉酶5盒。
SEQ ID NO:153是编码SEQ ID NO:151的淀粉酶5蛋白的合成的核酸序列。
SEQ ID NO:154是合成的lysA淀粉酶5盒。
SEQ ID NO:155是天然地衣芽孢杆菌prsA蛋白的氨基酸序列。
SEQ ID NO:156是天然地衣芽孢杆菌RghR2蛋白的氨基酸序列。
SEQ ID NO:157是变体地衣芽孢杆菌RghR2蛋白的氨基酸序列。
SEQ ID NO:158是编码SEQ ID NO:157的变体RghR2蛋白的变体地衣芽孢杆菌rghR2基因的核酸序列。
具体实施方式
本公开总体上涉及用于获得包含增强的蛋白产生能力的地衣芽孢杆菌细胞(例如,蛋白产生型宿主)的组合物和方法。本公开的某些实施例涉及衍生自亲本地衣芽孢杆菌细胞/菌株的遗传修饰的地衣芽孢杆菌细胞/菌株。因此,本公开的某些其他实施例涉及用于构建此类经修饰的地衣芽孢杆菌细胞/菌株的方法,所述细胞/菌株产生增加量的一种或多种目的蛋白。
例如,本公开的某些实施例涉及在经修饰的地衣芽孢杆菌细胞中产生增加量的目的蛋白(POI)的方法,所述细胞包括(a)通过在表达POI的亲本地衣芽孢杆菌细胞中引入包含与天然prsA可读框(ORF)可操作地连接的天然prsA启动子的多核苷酸来修饰所述表达POI的亲本地衣芽孢杆菌细胞以及(b)在用于产生所述POI的合适条件下发酵所述经修饰的细胞,其中当在相同条件下发酵时,相对于所述亲本细胞,所述经修饰的细胞产生增加量的所述POI。在某些实施例中,经修饰的细胞进一步包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏和/或与SEQ ID NO:121具有至少90%序列同一性的rghR2基因的缺失或破坏。在某些实施例中,目的蛋白(POI)是酶。在某些实施例中,酶是蛋白酶或淀粉酶。
本公开的其他实施例涉及经修饰的地衣芽孢杆菌细胞/菌株,其衍生自包含编码天然prsA蛋白的内源prsA基因的亲本地衣芽孢杆菌细胞/菌株。因此,在某些实施例中,本公开的经修饰的地衣芽孢杆菌细胞包含引入的多核苷酸,所述多核苷酸包含可操作地连接至天然prsA可读框(ORF)的天然prsA启动子。在特定实施例中,引入的多核苷酸编码与SEQID NO:155具有约90%序列同一性的天然prsA蛋白。在其他实施例中,经修饰的细胞包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏和/或与SEQ ID NO:121或SEQ ID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。
因此,本公开的某些实施例涉及获得、分离、纯化以及类似地处理由本公开的经修饰的地衣芽孢杆菌细胞产生的目的蛋白。
I.定义
鉴于本公开的经修饰的地衣芽孢杆菌细胞及本文所述的其方法,定义了以下术语和短语。本文未定义的术语应当符合如本领域使用的其常规含义。
除非另外定义,否则本文使用的所有技术和科学术语都具有与本发明组合物和方法应用的领域的普通技术人员通常所理解的相同的含义。尽管在本发明组合物和方法的实践或测试中也可以使用与本文所述的那些方法和材料类似或等效的任何方法和材料,但现在描述代表性的说明性方法和材料。将本文引用的所有出版物和专利都通过引用以其全文而并入。
应进一步注意,可以撰写权利要求书以排除任何任选的要素。因此,此陈述旨在作为使用与权利要求要素的叙述有关的排他性术语如“单独”、“仅”、“排除”、“不包括”等或使用“否定型”限定的前提基础(或其条件)。
在阅读本公开后,如将对于本领域技术人员应显而易见的,本文描述和说明的单独实施例中的每一个具有离散的组分和特征,这些组分和特征可以在不偏离本文所述的本发明组合物和方法的范围或精神的情况下容易地与其他若干个实施例中的任何一个的特征分离或组合。可以按照所叙述的事件的顺序或按照逻辑上可行的任何其他顺序来进行任何叙述的方法。
如本文所用,“芽孢杆菌属”包括如本领域技术人员已知的“芽孢杆菌”属内的所有物种,包括但不限于:枯草芽孢杆菌、地衣芽孢杆菌、迟缓芽孢杆菌(B.lentus)、短芽孢杆菌(B.brevis)、嗜热脂肪芽孢杆菌(B.stearothermophilus)、嗜碱芽孢杆菌(B.alkalophilus)、解淀粉芽孢杆菌、克劳氏芽孢杆菌(B.clausii)、耐盐芽孢杆菌(B.halodurans)、巨大芽孢杆菌(B.megaterium)、凝结芽孢杆菌(B.coagulans)、环状芽孢杆菌(B.circulans)、灿烂芽孢杆菌(B.lautus)和苏云金芽孢杆菌(B.thuringiensis)。应认识到,芽孢杆菌属不断进行分类学重组。因此,该属旨在包括已重新分类的物种,包括但不限于诸如嗜热脂肪芽孢杆菌(其现在名为“嗜热脂肪土芽孢杆菌(Geobacillusstearothermophilus)”)等的生物体。
如本文所用,“亲本细胞”是指“未修饰的细胞”(例如,比如未修饰的地衣芽孢杆菌亲本细胞)。
如本文所用,“经修饰的细胞”或“子细胞”可互换地使用,并且是指包含至少一个遗传修饰的重组地衣芽孢杆菌细胞,该遗传修饰不存在于经修饰的(子)细胞所来源的“亲本细胞”中。
在某些实施例中,“未经修饰的”地衣芽孢杆菌(亲本)细胞可以被称为“对照细胞”,特别是当与“经修饰的”的地衣芽孢杆菌(子代)细胞进行比较或相对于“经修饰的”的地衣芽孢杆菌(子代)细胞时。
如本文所用,当将“未经修饰的”(亲本)细胞中目的蛋白(POI)的表达和/或产生与“经修饰的”(子代)细胞中相同POI的表达和/产生相比较时,应理解,在相同条件(例如,相同条件如培养基、温度、pH等)下生长/培养/发酵“经修饰的”和“未经修饰的”细胞。
如本文所用,“宿主细胞”是指具有作为新引入的DNA序列的宿主或表达媒介物的能力的细胞。在本公开的某些实施例中,宿主细胞是芽孢杆菌属物种或大肠杆菌(E.coli)细胞。
如本文所用,本公开的“天然地衣芽孢杆菌prsA启动子”与SEQ ID NO:100具有约95%序列同一性。在某些实施例中,天然地衣芽孢杆菌prsA启动子与SEQ ID NO:100具有约95%、96%、97%、98%、99%或100%序列同一性。
如本文所用,“天然地衣芽孢杆菌prsA可读框(ORF)”与SEQ ID NO:101具有约90%或更高序列同一性。在某些实施例中,天然地衣芽孢杆菌prsA ORF与SEQ ID NO:101具有约90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%序列同一性。
枯草芽孢杆菌的prsA基因已在Kontinen和Sarvas(1993)以及PCT公开号WO 1994/019471中进行了描述,这些公开表明prsA基因参与蛋白分泌(即,编码细胞分泌机构的组成部分),其中prsA基因产物是膜相关脂蛋白。
如本文所用,“天然地衣芽孢杆菌prsA蛋白”与SEQ ID NO:155具有约90%或更高序列同一性并包含肽基-脯氨酰顺-反异构酶活性(EC5.2.1.8)。在某些实施例中,天然地衣芽孢杆菌prsA蛋白与SEQ ID NO:155具有约90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%序列同一性。
如本文所用,“亲本地衣芽孢杆菌细胞包含编码天然prsA蛋白的内源(野生型)prsA基因”,因此,当编码与SEQ ID NO:155具有约90%序列同一性的prsA蛋白的多核苷酸被引入本公开的经修饰的地衣芽孢杆菌细胞中时,引入的多核苷酸在本文中可称为作为第二(第2)prsA拷贝。例如,包含编码与SEQ ID NO:155具有约90%序列同一性的prsA蛋白的引入的多核苷酸的本公开的经修饰的地衣芽孢杆菌细胞在本文中可称为二(2)拷贝prsA(经修饰的)地衣芽孢杆菌细胞,其包含编码天然prsA蛋白的第一(第1)内源(野生型)prsA基因,以及编码prsA蛋白的第二(第2)引入的多核苷酸。
在枯草芽孢杆菌中,dlt操纵子包含分别编码名为DltA、DltB、DltC、DltD和DltE的蛋白的五(5)个ORF(dltA、dltB、dltC、dltD和dltE)(May等人,2005)。例如,如May等人(2005)所述,DltA蛋白是D-丙氨酸:D-丙氨酰载体蛋白连接酶,其参与将D-Ala掺入细胞壁的脂磷壁酸中。
如本文所用,“dltA基因”与SEQ ID NO:122具有约90%序列同一性。在某些实施例中,dltA基因与SEQ ID NO:155具有约90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%序列同一性。
枯草芽孢杆菌rghR基因编码称为RghR的转录调节蛋白,其在本领域中已被描述为rapG、rapH(Hayashi等人,2006)和rapD(Ogura和Fujita,2007)的阻遏物。相比之下,如最近在PCT公开号WO 2018/156705中所述,地衣芽孢杆菌编码RghR转录调节蛋白的两(2)个同源物,称为RghR1和RghR2。如下文所述,本公开的某些实施例涉及地衣芽孢杆菌细胞,其包含经修饰的(例如,缺失或破坏)rghr2基因。
如本文所用,适用于本文所述遗传修饰的“地衣芽孢杆菌rghR2基因”可以是野生型地衣芽孢杆菌rghR2基因(SEQ ID NO:121),其编码与SEQ ID NO:156具有约90%的序列同一性的天然RhgR2蛋白(例如与SEQ ID NO:156具有约90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%序列同一性),或者它可以是变体地衣芽孢杆菌rghR2基因(SEQ ID NO:158),其编码与SEQ ID NO:157具有约90%序列同一性的变体RhgR2蛋白(例如与SEQ ID NO:156具有约90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%序列同一性)。例如,如SEQ ID NO:157所示,变体RhgR2蛋白在SEQ ID NO:157的氨基酸残基36-41处包含“Ala-Ala-Ala-Ile-Ser-Arg”的六(6)氨基酸残基重复,其中六(6)氨基酸重复不存在于天然RghR2蛋白中(即,SEQ ID NO:156的氨基酸残基1-134)。
因此,在某些其他实施例中,rghR2基因或其可读框与天然rghR2基因具有约90%的序列同一性(例如与SEQ ID NO:121具有约90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%序列同一性);或与变体rghR2基因具有约90%的序列同一性(例如,与SEQ ID NO:158具有约90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%序列同一性)。
如本文所用,名为“BF140”或“BF140(ΔserA_ΔlysA)”的亲本地衣芽孢杆菌菌株包含serA基因缺失(ΔserA)和lysA基因缺失(ΔlysA)。
如本文所用,名为“BF561”或“BF561(第2拷贝prsA)”的经修饰的地衣芽孢杆菌菌株衍生自亲本菌株BF140,其中经修饰的BF561菌株包含编码天然prsA蛋白的野生型地衣芽孢杆菌prsA基因的引入的第2拷贝。
如本文所用,经修饰的地衣芽孢杆菌菌株命名为“BF598”或“BF598(ΔdltA_第2拷贝prsA)”衍生自BF561菌株,其中经修饰的BF598进一步包含地衣芽孢杆菌dltA基因的缺失。
如本文所用,经修饰的地衣芽孢杆菌菌株命名为“BF602”或“BF602(ΔrghR2_第2拷贝prsA)”衍生自BF561菌株,其中经修饰的BF602进一步包含地衣芽孢杆菌rghR2基因的缺失。
如本文所用,命名为“BF613”或“BF613”的经修饰的地衣芽孢杆菌菌株(ΔrghR2_ΔdltA_第2拷贝prsA)”衍生自BF598(ΔdltA_第2拷贝prsA)菌株,其中经修饰的BF613进一步包含地衣芽孢杆菌rghR2基因的缺失。
如本文所用,“淀粉酶1”是本领域通常称为AmyL的天然地衣芽孢杆菌α-淀粉酶并且包含SEQ ID NO:126的氨基酸序列。
如本文所用,“淀粉酶2”是包含SEQ ID NO:136的变体芽孢杆菌属物种α-淀粉酶,如国际PCT公开号WO 2018/184004(通过引用以其整体并入本文)中一般描述的。
如本文所用,“淀粉酶3”是包含SEQ ID NO:143的噬细胞菌属物种α-淀粉酶,如国际PCT公开号WO 2014/164777;WO 2012/164800和WO 2014/164834(各自通过引用以其整体并入本文)中一般描述的。
如本文所用,“淀粉酶4”是包含SEQ ID NO:147的噬细胞菌属物种α-淀粉酶,如国际PCT公开号WO 2014/164777;WO 2012/164800和WO 2014/164834(各自通过引用以其整体并入本文)中一般描述的。
如本文所用,“淀粉酶5”是包含SEQ ID NO:151的芽孢杆菌属物种707碱性α-淀粉酶,如国际PCT公开号WO 2008/153805和美国专利公开号US 2014/0057324(自通过引用以其整体并入本文)中一般描述的。
如本文所用,本文名为“Cas9 Y155H”的变体Cas9蛋白已在PCT公开号WO 2019/118463(通过引用以其整体并入本文)中进行了描述。
如本文所用,术语“修饰”和“遗传修饰”可互换地使用,并且包括:(a)引入、取代、或去除基因(或其ORF)中的一个或多个核苷酸,或者引入、取代、或去除基因或其ORF的转录或翻译所需要的调控元件中的一个或多个核苷酸,(b)基因破坏,(c)基因转换,(d)基因缺失,(e)基因下调,(f)特异性诱变,和/或(g)随机诱变本文公开的任何一个或多个基因。
如本文所用,当在短语如“相对于(未经修饰的)亲本宿主细胞,经修饰的宿主细胞‘表达/产生增加量’的一种或多种目的蛋白”中使用时,“增加量”特别是指在经修饰的宿主细胞中表达/产生的任何“增加量”的目的蛋白(POI),所述“增加量”总是相对于表达/产生相同POI的(未经修饰的)亲本地衣芽孢杆菌细胞而言,其中所述经修饰的和未经修饰的细胞在相同的条件下(例如,相同的条件如培养基、温度、pH等)生长/培养/发酵。例如,增加量的POI可以是在本公开的经修饰的芽孢杆菌属物种细胞中表达的内源芽孢杆菌属物种POI或异源POI。
如本文所用,“增加”蛋白产生或“增加的”蛋白产生意指产生的蛋白(例如,目的蛋白)的量增加。蛋白可以在宿主细胞内产生,或分泌(或转运)到培养基中。在某些实施例中,目的蛋白被产生(被分泌)到培养基中。与亲本宿主细胞相比,增加的蛋白产生可以被检测为例如蛋白或酶活性(例如像蛋白酶活性、淀粉酶活性、纤维素酶活性、半纤维素酶活性等)或者所产生的总细胞外蛋白的更高的最大水平。
如本文所用,术语“表达”是指衍生自本公开的核酸分子的有义(mRNA)或反义RNA的转录和稳定积累。表达也可指将mRNA翻译成多肽。因此,术语“表达”包括涉及多肽的产生的任何步骤,包括但不限于转录、转录后修饰、翻译、翻译后修饰、分泌等。
如本文所用,“核酸”是指核苷酸或多核苷酸序列及其片段或部分,以及基因组或合成起点的DNA、cDNA和RNA,其可能是双链或单链,无论代表正义或反义链。应理解,由于遗传密码的简并性,许多核苷酸序列可以编码给定的蛋白。
应理解,本文所述的多核苷酸(或核酸分子)包括“基因”、“载体”和“质粒”。
因此,术语“基因”是指编码氨基酸的特定序列的多核苷酸,其包含所有或部分蛋白编码序列,并且可以包括调控(非转录的)DNA序列,如启动子序列,该启动子序列决定例如基因在其下表达的条件。基因的转录区可以包括非翻译区(UTR)(包括内含子、5'-非翻译区(UTR)和3'-UTR)以及编码序列。
如本文所用,术语“编码序列”是指直接指定其(所编码的)蛋白产物的氨基酸序列的核苷酸序列。编码序列的边界一般由通常以ATG起始密码子开始的可读框(在下文质,“ORF”)决定。编码序列典型地包括DNA、cDNA和重组核苷酸序列。
如本文使用的,术语“启动子”是指能够控制编码序列或功能性RNA表达的核酸序列。通常,编码序列位于启动子序列3'(下游)。启动子可以全部衍生自天然基因,或者由衍生自在自然界中发现的不同启动子的不同元件构成,或者甚至包含合成核酸片段。本领域技术人员应理解,不同启动子可以指导基因在不同细胞类型中、或在不同发育阶段、或响应于不同环境或生理条件而表达。使基因在大多数细胞类型中在大多数时间表达的启动子通常被称为“组成型启动子”。应进一步认识到,由于在大多数情况下,尚未完全界定调节序列的确切边界,因此不同长度的DNA片段可以具有相同启动子活性。
如本文使用的术语“可操作地连接”是指核酸序列在单个核酸片段上的缔合,使得一者的功能受到另一者影响。例如,当能够实现该编码序列的表达(即,编码序列在启动子的转录控制下)时,启动子与该编码序列(例如ORF)可操作地连接。编码序列可以在有义或反义方向上可操作地连接到调节序列。
当核酸被放置成与另一个核酸序列具有功能关系时,该核酸与另一个核酸序列“可操作地连接”。例如,如果编码分泌性前导序列(即,信号肽)的DNA被表达为参与多肽分泌的前蛋白,则该编码分泌性前导序列(即,信号肽)的DNA可操作地连接到该多肽的DNA;如果启动子或增强子影响编码序列的转录,则该启动子或增强子可操作地连接到该序列;或者如果核糖体结合位点被定位以便促进翻译,则该核糖体结合位点可操作地连接到编码序列。通常,“可操作地连接”意指被连接的DNA序列是连续的,并且在分泌性前导序列的情况下,是连续的并且处于阅读相中。然而,增强子不必是连续的。通过在方便的限制位点处连接来实现连接。如果此类位点不存在,则按照常规实践使用合成的寡核苷酸衔接子或接头。
如本文所用,“控制目的基因(或其可读框)的表达的与目的基因的蛋白编码序列连接的功能性启动子序列”是指控制编码序列在芽孢杆菌属中的转录和翻译的启动子序列。例如,在某些实施例中,本公开涉及包含5′启动子(或5′启动子区、或串联5′启动子等)的多核苷酸,其中启动子区可操作地连接到编码蛋白的核酸序列(例如,ORF)。
如本文所用,“合适的调节序列”是指位于编码序列上游(5'非编码序列)、内部或下游(3'非编码序列),并且影响相关编码序列的转录、RNA加工或稳定性或者翻译的核苷酸序列。调节序列可以包括启动子、翻译前导序列、RNA加工位点、效应子结合位点和茎环结构。
如本文所用,术语“引入”,如短语中所使用的,如“向细菌细胞中引入”或“向地衣芽孢杆菌中引入”至少一种多核苷酸可读框(ORF)、或其基因、或其载体,包括本领域已知用于将多核苷酸引入细胞的方法,这些方法包括但不限于原生质体融合、自然或人工转化(例如,氯化钙、电穿孔)、转导、转染、缀合等(例如,参见Ferrari等人,1989)。
如本文所用,“转化的”或“转化”意指通过使用重组DNA技术转化的细胞。转化典型地通过将一个或多个核苷酸序列(例如,多核苷酸、ORF或基因)***细胞中而发生。所***的核苷酸序列可以是异源核苷酸序列(即在待转化的细胞中不是天然存在的序列)。因此,转化通常是指将外源DNA引入宿主细胞中,使得DNA保持为染色体整合体或自我复制的染色体外载体。
如本文所用,“转化DNA”、“转化序列”和“DNA构建体”是指用于将序列引入宿主细胞或生物体中的DNA。转化DNA是用于将序列引入宿主细胞或生物体中的DNA。可以通过PCR或任何其他合适的技术在体外产生DNA。在一些实施例中,转化DNA包含输入序列,而在其他实施例中,它进一步包含侧翼为同源盒的输入序列。在又另一个实施例中,转化DNA包含其他非同源序列,添加到末端(即,填充序列或侧翼)。末端可以闭合,使得转化DNA形成闭环,例如像***载体中。
如本文所用,“基因的破坏”或“基因破坏”可互换使用,并且广泛地是指基本上防止宿主细胞产生功能性基因产物(例如,蛋白)的任何遗传修饰。因此,如本文所用,基因破坏包括但不限于移码突变、过早终止密码子(即使得不产生功能性蛋白)、消除或降低蛋白活性的取代、内部缺失(使得不产生功能性蛋白)、破坏编码序列的***、去除转录所需的天然启动子和可读框之间的可操作连接的突变等。
如本文使用的,“输入序列”是指引入芽孢杆菌属物种染色体中的DNA序列。在一些实施例中,输入序列是DNA构建体的一部分。在其他实施例中,输入序列编码一种或多种目的蛋白。在一些实施例中,输入序列包含可以已经或可以不存在于待转化的细胞的基因组中的序列(即,它可以是同源或异源序列)。在一些实施例中,输入序列编码一种或多种目的蛋白、基因和/或经突变的或经修饰的基因。在可替代的实施例中,输入序列编码功能性野生型基因或操纵子、功能性突变型基因或操纵子或者非功能性基因或操纵子。在一些实施例中,可以将非功能性序列***基因中以破坏基因的功能。在另一个实施例中,输入序列包括选择性标记。在进一步的实施例中,输入序列包括两个同源盒。
如本文所用,“同源盒”是指与芽孢杆菌属染色体中的序列同源的核酸序列。更具体地,根据本发明,同源盒是上游或下游区,该上游或下游区与被缺失、破坏、灭活的、下调等的基因或基因的一部分的直接侧翼编码区具有约80%与100%之间的序列同一性、约90%与100%之间的序列同一性、或约95%与100%之间的序列同一性。这些序列指导在芽孢杆菌属染色体中,DNA构建体被整合,并且指导哪一部分芽孢杆菌属染色体被输入序列替代。虽然不意在限制本公开,但同源盒可以包括约1个碱基对(bp)至200千碱基(kb)之间。优选地,同源盒包括约1bp与10.0kb之间;1bp与5.0kb之间;1bp与2.5kb之间;1bp与1.0kb之间;以及0.25kb与2.5kb之间。同源盒还可以包括约10.0kb、5.0kb、2.5kb、2.0kb、1.5kb、1.0kb、0.5kb、0.25kb和0.1kb。在一些实施例中,选择性标记的5'和3'端的侧翼为同源盒,其中同源盒包含紧密侧翼基因的编码区的核酸序列。
如本文所用,术语“编码可选择性标记的核苷酸序列”是指核苷酸序列,所述核苷酸序列能够在宿主细胞中表达并且其中可选择性标记的表达赋予含有表达的基因的细胞在不存在对应的选择性试剂或缺乏必需营养素的情况下生长的能力。
如本文所用,术语“可选择标记”和“选择性标记”是指能够在宿主细胞中表达的核酸(例如,基因),其允许容易地选择包含载体的那些宿主。此类可选择性标记的实例包括但不限于抗微生物剂。因此,术语“可选择标记”是指提供宿主细胞已经摄取了输入性目的DNA或者已经发生了一些其他反应的指示的基因。典型地,可选择性标记是赋予宿主细胞抗微生物抗性或代谢优势的基因,以允许在转化期间将含有外源DNA的细胞与未接受任何外源序列的细胞区分开来。
“驻留可选择性标记(residing selectable marker)”是位于待转化微生物的染色体上的标记。驻留可选择性标记编码与转化DNA构建体上的可选择性标记不同的基因。选择性标记是本领域技术人员熟知的。如上所述,标记可以是抗微生物抗性标记(例如,ampR、phleoR、specR、kanR、eryR、tetR、cmpR和neoR)(参见例如,Guerot-Fleury,1995;Palmeros等人,2000;和Trieu-Cuot等人,1983)。在一些实施例中,本发明提供氯霉素抗性基因(例如,存在于pC194上的基因,以及存在于地衣芽孢杆菌基因组中的抗性基因)。此抗性基因在本发明中以及涉及染色体整合的盒和整合质粒的染色体扩增的实施例中特别有用(参见例如,Albertini和Galizzi,1985;Stahl和Ferrari,1984)。根据本发明有用的其他标记包括但不限于营养缺陷型标记,如丝氨酸、赖氨酸、色氨酸;和检测标记,如β-半乳糖苷酶。
如本文定义的,宿主细胞“基因组”、细菌(宿主)细胞“基因组”、或芽孢杆菌属物种(宿主)细胞“基因组”包括染色体基因和染色体外基因。
如本文所用,术语“质粒”、“载体”和“盒”是指染色体外元件,其通常携带典型地不是细胞的中心代谢的一部分的基因,并且通常呈环状双链DNA分子的形式。此类元件可以是源自任何来源的单链或双链DNA或RNA的线性或环状自主复制序列、基因组整合序列、噬菌体或核苷酸序列,其中许多核苷酸序列已连接或重组到单一结构中,该单一结构能够将针对选定基因产物的启动子片段和DNA序列连同适当3'未翻译序列引入到细胞中。
如本文所用,术语“质粒”是指用作克隆载体,并且在许多细菌和一些真核生物中形成染色体外自我复制遗传元件的环状双链(ds)DNA构建体。在一些实施例中,将质粒掺入宿主细胞的基因组中。在一些实施例中,质粒存在于亲本细胞中并且在子代细胞中丢失。
如本文使用的,“转化盒”是指包含基因(或其ORF)并且除了外源基因之外还具有促进特定宿主细胞的转化的元件的特定载体。
如本文所用,术语“载体”是指可以在细胞中复制(传播)并且可以携带新基因或DNA区段到细胞中的任何核酸。因此,该术语是指设计用于在不同宿主细胞之间转移的核酸构建体。载体包括为“附加体”(即,其自主复制或可以整合到宿主生物体的染色体中)的病毒、噬菌体、前病毒、质粒、噬菌粒、转座子、和人工染色体如YAC(酵母人工染色体)、BAC(细菌人工染色体)、PLAC(植物人工染色体)等。
“表达载体”是指具有在细胞中掺入并且表达异源DNA能力的载体。许多原核和真核表达载体可商购获得并且是本领域技术人员已知的。适当的表达载体的选择在本领域技术人员的知识范围内。
如本文所用,术语“表达盒”和“表达载体”是指重组或合成生成的具有允许特定核酸在靶细胞中转录的一系列特定核酸元件(即,这些是载体或载体元件,如上所述)的核酸构建体。可以将重组表达盒掺入质粒、染色体、线粒体DNA、质体DNA、病毒或核酸片段中。典型地,表达载体的重组表达盒部分包括(除了其他序列之外)待转录的核酸序列和启动子。在一些实施例中,DNA构建体还包括允许特定核酸在靶细胞中转录的一系列指定核酸元件。在某些实施例中,本公开的DNA构建体包含如本文定义的选择性标记和失活染色体或基因或DNA区段。
如本文所用,“靶向载体”是这样的载体,该载体包括与该靶向载体转化至其中的宿主细胞的染色体中的区同源的多核苷酸序列,并且该载体可以驱动在该区处的同源重组。例如,靶向载体可用于通过同源重组将突变引入宿主细胞的染色体中。在一些实施例中,靶向载体包含其他非同源序列,例如添加到末端(即,填充序列或侧翼序列)。末端可以闭合,使得靶向载体形成闭环,例如像***载体中。例如,在某些实施例中,通过在其中引入一个或多个“靶向载体”来修饰(例如,转化)亲本地衣芽孢杆菌(宿主)细胞。
如本文所用,术语“目的蛋白”或“POI”是指期望在经修饰的地衣芽孢杆菌(子代)宿主细胞中表达的目的多肽,其中所述POI优选地以增加的水平表达(即,相对于“未经修饰的”(亲本)细胞)。因此,如本文所用,POI可以是酶、底物结合蛋白、表面活性蛋白、结构蛋白、受体蛋白等。在某些实施例中,相对于亲本细胞,本公开的经修饰的细胞产生增加量的异源目的蛋白或内源目的蛋白。在特别地实施例中,由本公开的经修饰的细胞产生的目的蛋白的增加量是相对于亲本细胞,至少0.5%增加、至少1.0%增加、至少5.0%增加、或超过5.0%增加。
类似地,如本文定义的,“目的基因”或“GOI”是指编码POI的核酸序列(例如,多核苷酸、基因、或ORF)。编码“目的蛋白”的“目的基因”可以是天然存在的基因、经突变的基因或合成基因。
如本文所用,术语“多肽”和“蛋白”可互换地使用,并且是指包含通过肽键连接的氨基酸残基的任何长度的聚合物。在本文中使用氨基酸残基的常规单(1)字母或三(3)字母代码。多肽可以是线性的或支化的,它可以包含经修饰的氨基酸,并且它可以被非氨基酸中断。术语多肽还涵盖已经天然地或通过干预修饰;例如,二硫键形成、糖基化、脂化、乙酰化、磷酸化或任何其他操作或修饰,诸如与标记组分缀合而修饰的氨基酸聚合物。这些定义内还包括例如含有一种或多种氨基酸类似物(包括例如非天然氨基酸等)以及本领域已知的其他修饰的多肽。
在某些实施例中,本公开的基因编码商业上相关的工业目的蛋白,例如酶(例如,乙酰酯酶、氨肽酶、淀粉酶、***糖酶、***呋喃糖苷酶、碳酸酐酶、羧肽酶、过氧化氢酶、纤维素酶、几丁质酶、凝乳酶、角质酶、脱氧核糖核酸酶、差向异构酶、酯酶、α-半乳糖苷酶、β-半乳糖苷酶、α-葡聚糖酶、葡聚糖裂解酶、内切-β-葡聚糖酶、葡糖淀粉酶、葡萄糖氧化酶、α-葡糖苷酶、β-葡糖苷酶、葡萄糖醛酸酶、糖基水解酶、半纤维素酶、己糖氧化酶、水解酶、转化酶、异构酶、漆酶、脂肪酶、裂解酶、甘露糖苷酶、氧化酶、氧化还原酶、果胶裂解酶、果胶乙酰酯酶、果胶解聚酶、果胶甲酯酶、果胶分解酶、过水解酶、多元醇氧化酶、过氧化物酶、酚氧化酶、植酸酶、聚半乳糖醛酸酶、蛋白酶、肽酶、鼠李糖-半乳糖醛酸酶、核糖核酸酶、转移酶、转运蛋白、转谷氨酰胺酶、木聚糖酶、己糖氧化酶、及其组合)。
如本文所用,“变体”多肽是指典型地通过重组DNA技术,通过取代、添加或缺失一个或多个氨基酸,衍生自亲本(或参考)多肽的多肽。变体多肽与亲本多肽可以相差小数量的氨基酸残基,并且可以通过它们与亲本(参考)多肽的一级氨基酸序列同源性/同一性的水平来定义。
优选地,变体多肽与亲本(参考)多肽序列具有至少70%、至少75%、至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或甚至至少99%氨基酸序列同一性。如本文所用,“变体”多核苷酸是指编码变体多肽的多核苷酸,其中“变体多核苷酸”与亲本多核苷酸具有指定程度的序列同源性/同一性,或者与亲本多核苷酸(或其互补序列)在严格杂交条件下杂交。优选地,变体多核苷酸与亲本(参考)多核苷酸序列具有至少70%、至少75%、至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或甚至至少99%核苷酸序列同一性。
如本文所用,“突变”是指核酸序列中的任何变化或改变。存在几种类型的突变,包括点突变、缺失突变、沉默突变、移码突变、剪接突变等。突变可以特异性地(例如,经由定点诱变)或随机地(例如,经由化学试剂、通过修复减去细菌菌株传代)进行。
如本文所用,在多肽或其序列的上下文中,术语“取代”意指一个氨基酸被另一个氨基酸替代(即,取代)。
如本文定义的,“内源基因”是指位于生物体基因组的其天然位置中的基因。
如本文定义的,“异源”基因、“非内源”基因或“外源”基因是指通常不在宿主生物体中被发现,但是通过基因转移引入宿主生物体中的基因(或ORF)。如本文所用,术语一个或多个“外源”基因包括***非天然生物体中的天然基因(或ORF)和/或***天然或非天然生物体中的嵌合基因。
如本文定义的,“异源控制序列”是指在自然界中不起调节(控制)目的基因表达的作用的基因表达控制序列(例如,启动子或增强子)。通常,异源核酸序列对于它们存在的细胞或基因组的一部分而言不是内源(天然)的,并且已通过感染、转染、转化、显微注射、电穿孔等添加到细胞中。“异源”核酸构建体可以含有与在天然宿主细胞中发现的控制序列/DNA编码序列组合相同或不同的控制序列/DNA编码(ORF)序列组合。
如本文所用,术语“信号序列”和“信号肽”是指可以参与成熟蛋白或蛋白的前体形式的分泌或定向转运的氨基酸残基的序列。典型地,信号序列位于前体或成熟蛋白序列的N-末端。信号序列可以是内源的或外源的。成熟蛋白中通常不存在信号序列。典型地,在蛋白被转运后,信号序列通过信号肽酶从蛋白上切割下来。
术语“衍生的”涵盖术语“起源的”、“获得的”“可获得的”和“产生的”,并且通常指示一种指定的材料或组合物在另一种指定的材料或组合物中找到它的起源,或者具有可以参考另一种指定的材料或组合物描述的特征。
如本文所用,术语“同源性”涉及同源多核苷酸或多肽。如果两个或更多个多核苷酸或者两个或更多个多肽是同源的,则这意味着同源多核苷酸或多肽具有至少60%、更优选至少70%、甚至更优选至少85%、仍更优选至少90%、更优选至少95%、最优选至少98%的“同一性程度”。两个多核苷酸或多肽序列是否具有如本文定义的同源程度足够高的同一性,可以通过使用本领域已知的计算机程序比对两个序列来适当地研究,该计算机程序如GCG程序包中提供的“GAP”(威斯康星程序包手册(Program Manual for the WisconsinPackage),第8版,1994年8月,遗传学计算机集团(Genetics Computer Group),科学大道575号(575Science Drive),麦迪逊,威斯康星州,美国53711)(Needleman和Wunsch,(1970))。使用具有以下设置的GAP进行DNA序列比较:GAP产生罚分5.0和GAP扩展罚分0.3。
如本文所用,术语“百分比(%)同一性”是指当使用序列比对程序进行比对时,编码多肽的核酸序列或多肽的氨基酸序列之间的核酸或氨基酸序列同一性的水平。
如本文所用,“比生产力”是在给定时间段内产生的蛋白的总量/细胞/时间。
如本文定义的,术语“纯化的”、“分离的”或“富集的”意指生物分子(例如,多肽或多核苷酸)通过将其与它在自然界中相关联的天然存在的成分中的一些或所有分离而从其天然状态改变。这样的分离或纯化可以通过本领域公认的分离技术如离子交换色谱法、亲和色谱法、疏水分离、透析、蛋白酶处理、硫酸铵沉淀或其他蛋白盐沉淀、离心、尺寸排阻色谱法、过滤、微量过滤、凝胶电泳或梯度分离完成,以去除最终组合物中所不希望的全细胞、细胞碎片、杂质、外来蛋白或酶。然后可以进一步向纯化的或分离的生物分子组合物中添加提供另外益处的成分,例如活化剂、抗抑制剂、期望的离子、控制pH的化合物或者其他酶或化学品。
如本文所用,术语“ComK多肽”定义为comK基因的产物;一种转录因子,在感受态发展之前充当最终的自动调节控制开关;参与激活参与DNA结合和摄取以及重组的晚期感受态基因的表达(Liu和Zuber,1998,Hamoen等人,1998)。示例性ComK核酸如SEQ ID NO:92所示。
如本文所用,“重组体”包括提及细胞或载体,其已通过引入异源核酸序列而被修饰,或者该细胞衍生自如此修饰的细胞。因此,例如,重组细胞表达未在细胞的天然(非重组)形式内发现的相同形式的基因或表达以其他方式异常表达的、低表达的或根本不表达的天然基因(由于故意的人为干预)。“重组(recombination、recombining)”或产生“重组的(recombined)”核酸通常是两个或更多个核酸片段的组装,其中所述组装产生嵌合基因。
如本文所用,“侧翼序列”是指正在讨论的序列的上游或下游的任何序列(例如,针对基因A-B-C,基因B以A和C基因序列为侧翼)。在某些实施例中,输入序列在每侧的侧翼为同源盒。在另一个实施例中,输入序列和同源盒包含在每侧的侧翼为填充序列的单元。在一些实施例中,侧翼序列仅存在于单侧(3'或5'),但是在优选的实施例中,它在被侧翼的序列的每侧。每个同源盒的序列与芽孢杆菌属染色体中的序列同源。这些序列指导在芽孢杆菌属染色体中新构建体的整合位置以及芽孢杆菌属染色体的哪部分将被输入序列替代。在其他实施例中,选择性标记的5'和3'端的侧翼为包含失活染色体区段的部分的多核苷酸序列。在一些实施例中,侧翼序列仅存在于单侧(3'或5'),而在其他实施例中,它存在于被侧翼的序列的每侧。
II.包含增强的蛋白产生表型的经修饰的地衣芽孢杆菌细胞
如以下实例部分中一般阐述的,申请人构建了一系列宿主修饰并将其引入亲本地衣芽孢杆菌菌株中。更具体地,如以下实例中所示(例如,参见表18),本实例中使用的亲本地衣芽孢杆菌菌株包含serA1基因(SEQ ID NO:30)和lysA基因(SEQ ID NO:87)的缺失,并命名为BF140(ΔserA_ΔlysA)。申请人随后将某些遗传修饰引入亲本地衣芽孢杆菌菌株(BF140),包括引入(1)编码天然prsA蛋白的野生型地衣芽孢杆菌prsA基因的第2拷贝(命名为BF561;第2拷贝prsA),(2)地衣芽孢杆菌dltA基因的缺失(命名为BF598;ΔdltA_第2拷贝prsA),(3)地衣芽孢杆菌rghR2基因的缺失(命名为BF602;ΔrghR2_第2拷贝prsA)和(4)地衣芽孢杆菌rghR2基因和dltA基因的组合缺失(命名为BF613;ΔrghR2_ΔdltA_第2拷贝prsA)。
在构建上述经修饰的菌株之后,将一系列α-淀粉酶表达盒引入经修饰的地衣芽孢杆菌菌株(BF561、BF598、BF602和BF613)和亲本地衣芽孢杆菌菌株(BF140)。更具体地,如下文实例4中所示,将五(5)个不同的α-淀粉酶表达盒(即,“淀粉酶1”、“淀粉酶2”、“淀粉酶3”、“淀粉酶4”和“淀粉酶5”)的两个(2)拷贝引入地衣芽孢杆菌菌株。
如下文实例5中进一步描述的,测定了亲本(BF140)和含有淀粉酶1-5表达盒的两(2)个拷贝的经修饰(BF561、BF598、BF602和BF613)的地衣芽孢杆菌菌株的淀粉酶产生(例如,参见表19)。例如,从不同的α-淀粉酶组中测试的所有五(5)种淀粉酶都证明了与未修饰的亲本宿主BF140相比在BF613修饰的背景(ΔrghR2_ΔdltA_第2拷贝prsA)(其包括dltA-2等位基因(SEQ ID NO:125)缺失(ΔdltA-2),rghR2等位基因(SEQ ID NO:80)缺失(ΔrghR2)和由天然prsA启动子(SEQ ID NO:124)控制的天然prsA基因的第二拷贝的***)中α-淀粉酶产生的改善。对于淀粉酶2和淀粉酶3,在BF602修饰的背景(ΔrghR2_第2拷贝prsA)(其包含rghR2(ΔrghR2)等位基因(SEQ ID NO:80)缺失和由天然prsA启动子(SEQ IDNO:124)控制的天然prsA基因的第二拷贝)中α-淀粉酶产生方面的改善几乎与在BF613修饰的宿主中观察到的生产力改善一样好。这一观察表明,对于某些淀粉酶,生产力的提高只需要存在这两个(2)等位基因(即,ΔrghR2_第2拷贝prsA),并且ΔdltA-2等位基因的存在对这种改善没有害处。
III.分子生物学
如上文一般阐述的,本公开的某些实施例涉及衍生自亲本地衣芽孢杆菌细胞的经修饰的地衣芽孢杆菌(子代)细胞。更特别地,本公开的某些实施例涉及经修饰的芽孢杆菌属(子代)细胞及其用于产生和构建此类具有增加的蛋白产生能力、增加的次级代谢产物产生能力等的经修饰的芽孢杆菌属(宿主)细胞(例如,蛋白产生宿主细胞、细胞工厂)的方法。
在某些实施例中,本公开的经修饰的地衣芽孢杆菌细胞包含编码天然prsA蛋白的基因或ORF的引入的第2拷贝。在其他实施例中,本公开的经修饰的地衣芽孢杆菌细胞包含dltA基因的缺失。在某些其他实施例中,本公开的经修饰的地衣芽孢杆菌细胞包含编码天然prsA蛋白的基因或ORF的引入的第2拷贝和dltA基因的缺失。在其他实施例中,本公开的经修饰的地衣芽孢杆菌细胞包含rghR2基因的缺失。在某些其他实施例中,本公开的经修饰的地衣芽孢杆菌细胞包含编码天然prsA蛋白的基因或ORF的引入的第2拷贝和rghR2基因的缺失。在其他实施例中,本公开的经修饰的地衣芽孢杆菌细胞包含dltA基因的缺失和rghR2基因的缺失。在某些其他实施例中,本公开的经修饰的地衣芽孢杆菌细胞包含编码天然prsA蛋白的基因或ORF的引入的第2拷贝、dltA基因的缺失和rghR2基因的缺失。
因此,本公开的某些实施例提供了用于遗传修饰(改变)本公开的亲本芽孢杆菌属细胞以产生其经修饰的芽孢杆菌属细胞的组合物和方法,并且更特别地,经修饰的芽孢杆菌属细胞相对于(未经修饰的)亲本地衣芽孢杆菌细胞,产生增加量的内源和/或异源目的蛋白。
因此,本公开的某些实施例涉及用于遗传修饰芽孢杆菌属细胞的方法,其中所述修饰包括(a)引入、取代、或去除基因(或其ORF)中的一个或多个核苷酸,或者引入、取代、或去除基因或其ORF的转录或翻译所需要的调控元件中的一个或多个核苷酸,(b)基因破坏,(c)基因转换,(d)基因缺失,(e)下调基因,(f)位点特异性诱变,和/或(g)随机诱变。
在某些实施例中,通过使用本领域熟知的方法(例如,***、破坏、替换、或缺失)减少或消除上述基因的表达来构建本公开的经修饰的芽孢杆菌属细胞。待修饰或失活的基因的部分可以是例如编码区或编码区表达所需的调节元件。
此类调控或控制序列的实例可以是启动子序列或其功能部分(即,足以影响核酸序列表达的部分)。用于修饰的其他控制序列包括但不限于前导序列、前肽序列、信号序列、转录终止子、转录激活子等。
在某些其他实施例中,通过基因缺失构建经修饰的芽孢杆菌属细胞以消除或减少本公开的至少一种前述基因的表达。基因缺失技术使得能够部分或完全去除一个或多个基因,从而消除它们的表达,或者表达非功能性(或活性降低的)蛋白产物。在此类方法中,一个或多个基因的缺失可以通过使用已构建为连续含有侧翼于该基因的5'和3'区的质粒进行同源重组来完成。可以将连续的5'和3'区引入芽孢杆菌属细胞中,例如,在温度敏感的质粒(如pE194)上,在允许温度下与第二可选择标记缔和以允许质粒在细胞中建立。然后将细胞移至非允许温度,以选择在同源侧翼区之一处具有整合到染色体中的质粒的细胞。通过选择第二可选择性标记来实现质粒整合的选择。在整合后,通过将细胞移至允许温度持续几代而不进行选择来刺激第二同源侧翼区处的重组事件。将细胞铺板以获得单菌落,并检查菌落是否损失两种可选择标记(参见例如,Perego,1993)。因此,本领域的技术人员可容易地鉴定在基因的编码序列和/或基因的非编码序列中的核苷酸区(适于完全或部分缺失)。
在其他实施例中,通过在基因或其转录或翻译所需的调控元件中引入、取代、或去除一个或多个核苷酸来构建本公开的经修饰的芽孢杆菌属细胞。例如,可以***或去除核苷酸,以便导致终止密码子的引入、起始密码子的去除或可读框的移码。此种修饰可以根据本领域已知的方法通过定点诱变或PCR产生的诱变来完成(例如,参见Botstein和Shortle,1985;Lo等人,1985;Higuchi等人,1988;Shimada,1996;Ho等人,1989;Horton等人,1989;以及Sarkar和Sommer,1990)。因此,在某些实施例中,通过完全或部分缺失使本公开的基因失活。
在另一个实施例中,通过基因转换的过程构建经修饰的芽孢杆菌属细胞(例如,参见Iglesias和Trautner,1983)。例如,在基因转换方法中,对应于一个或多个基因的核酸序列在体外诱变以产生缺陷核酸序列,然后将所述核酸序列转化到亲本芽孢杆菌属细胞中以产生缺陷基因。通过同源重组,缺陷核酸序列替代内源基因。可能期望的是,缺陷基因或基因片段也编码可以用于选择含有缺陷基因的转化体的标记。例如,可以将缺陷基因与可选择标记缔和引入非复制或温度敏感的质粒上。通过在不允许质粒复制的条件下选择标记来实现质粒整合的选择。通过检查菌落是否丢失可选择性标记和是否获得经突变的基因来实现导致基因替代的第二重组事件的选择(Perego,1993)。可替代地,缺陷核酸序列可以含有基因的一个或多个核苷酸的***、取代或缺失,如下文所述。
在其他实施例中,通过确立已久的反义技术使用与基因的核酸序列互补的核苷酸序列来构建经修饰的芽孢杆菌属细胞(Parish和Stoker,1997)。更具体地,可以通过引入与基因的核酸序列互补的核苷酸序列来减少(下调)或消除芽孢杆菌属细胞的基因的表达,该核苷酸序列可以在细胞中转录并且能够与细胞中产生的mRNA杂交。在允许互补的反义核苷酸序列与mRNA杂交的条件下,由此降低或消除所翻译的蛋白的量。此类反义方法包括但不限于RNA干扰(RNAi)、小干扰RNA(siRNA)、微小RNA(miRNA)、反义寡核苷酸等,所有这些都是熟练技术人员熟知的。
在其他实施例中,经由CRISPR-Cas9编辑产生/构建经修饰的芽孢杆菌属细胞。例如,编码目的蛋白的基因可以通过核酸指导的内切核酸酶的方式编辑或破坏(或缺失或下调),所述核酸指导的内切核酸酶通过结合指导RNA(例如,Cas9)和Cpf1或指导DNA(例如,NgAgo)发现其靶DNA,这将内切核酸酶募集到DNA上的靶序列上,其中所述内切核酸酶可以在DNA中产生单链或双链断裂。此靶向DNA断裂成为DNA修复的底物,并且可以与所提供的编辑模板重组以使基因破坏或缺失。例如,编码核酸指导的内切核酸酶(出于此目的,来自化脓链球菌的Cas9)的基因或编码Cas9核酸酶的密码子优化的基因可操作地连接到在芽孢杆菌属细胞中有活性的启动子和在芽孢杆菌属细胞中有活性的终止子,从而产生芽孢杆菌Cas9表达盒。同样,本领域技术人员容易地鉴定目的基因特有的一个或多个靶位点。例如,为了构造编码gRNA的DNA构建体-指向目的基因内的靶位点,可变的靶向结构域(VT)将包含靶位点的核苷酸,所述核苷酸为(PAM)前间区序列邻近基序(TGG)的5′,所述核苷酸与编码化脓链球菌Cas9的Cas9核酸内切酶识别结构域(CER)的DNA融合。将编码VT结构域的DNA和编码CER结构域的DNA组合,从而产生编码gRNA的DNA。因此,通过将编码gRNA的DNA可操作地连接到在芽孢杆菌属细胞中有活性的启动子和在芽孢杆菌属细胞中有活性的终止子来产生gRNA的芽孢杆菌属表达盒。
在某些实施例中,将由核酸内切酶诱导的DNA断裂用输入序列修复/替代。例如,为了精确修复由上述Cas9表达盒和gRNA表达盒产生的DNA断裂,提供了核苷酸编辑模板,使得细胞的DNA修复机器可以利用编辑模板。例如,靶基因的约500bp 5'可以与靶基因的约500bp 3'融合以产生编辑模板,所述模板由芽孢杆菌宿主的机器用于修复由RGEN产生的DNA断裂。
可以使用许多不同的方法(例如,原生质体融合、电穿孔、自然感受态或诱导感受态)将Cas9表达盒、gRNA表达盒、和编辑模板共同递送至丝状真菌细胞。通过用正向和反向引物扩增基因座,通过PCR扩增靶基因座来筛选经转化的细胞。这些引物可以扩增野生型基因座或已由RGEN编辑的经修饰的基因座。然后使用测序引物对这些片段进行测序以鉴定经编辑的菌落。
在又其他实施例中,使用本领域已知的方法(包括但不限于,化学诱变(参见例如,Hopwood,1970)和转座(参见例如,Youngman等人,1983)),通过随机或特异性诱变构建经修饰的芽孢杆菌属细胞。可以通过使亲本细胞经受诱变并且筛选基因表达已减少或消除的突变型细胞来进行基因的修饰。可以例如通过使用合适的物理或化学诱变剂、使用合适的寡核苷酸或使DNA序列经受PCR产生的诱变来进行可以是特异性的或随机的诱变。此外,诱变可以通过使用这些诱变方法的任何组合来进行。
适用于本发明目的的物理或化学诱变剂的实例包括紫外线(UV)照射、羟胺、N-甲基-N'-硝基-N-亚硝基胍(MNNG)、N-甲基-N'-亚硝基胍(NTG)、邻甲基羟胺、亚硝酸、甲烷磺酸乙酯(EMS)、亚硫酸氢钠、甲酸、和核苷酸类似物。当使用此类试剂时,典型地通过如下方式来进行诱变:在合适的条件下在选择的诱变剂的存在下孵育待诱变的亲本细胞,并且选择展现出基因的表达降低或无表达的突变型细胞。
在某些其他实施例中,经修饰的芽孢杆菌属细胞包含内源基因的缺失。在其他实施例中,经修饰的芽孢杆菌属细胞包含内源基因的破坏。在某些实施例中,本公开的多核苷酸破坏盒包含标记基因。
在其他实施例中,经修饰的芽孢杆菌属细胞包含内源基因的下调。例如,在某些实施例中,下调上述一个或多个基因包含缺失或破坏基因的上游或下游调控元件。
PCT公开号WO 2003/083125公开了用于修饰芽孢杆菌属细胞的方法,如使用PCR融合来产生芽孢杆菌属缺失菌株和DNA构建体以绕过大肠杆菌。
PCT公开号WO 2002/14490公开了用于修饰芽孢杆菌属细胞的方法,这些方法包括(1)构建和转化整合的质粒(pComK),(2)随机诱变编码序列、信号序列和前肽序列,(3)同源重组,(4)通过向转化DNA添加非同源侧翼提高转化效率,(5)优化双交叉整合,(6)定向诱变和(7)无标记缺失。
本领域技术人员熟知用于将多核苷酸序列引入细菌细胞(例如,大肠杆菌和芽孢杆菌属物种)中的合适方法(例如Ferrari等人,1989;Saunders等人,1984;Hoch等人,1967;Mann等人,1986;Holubova,1985;Chang等人,1979;Vorobjeva等人,1980;Smith等人,1986;Fisher等人,1981;以及McDonald,1984)。实际上,包括原生质体转化和中板集合、转导和原生质体融合在内的诸如转化等的方法是已知的并且适合用于本公开。转化方法特别优选地用于将本公开的DNA构建体引入宿主细胞中。
除了常用方法之外,在一些实施例中,直接转化宿主细胞(即,在引入宿主细胞之前,中间细胞不用于扩增DNA构建体或以其他方式处理DNA构建体)。将DNA构建体引入宿主细胞中包括本领域已知的将DNA引入宿主细胞中而不***质粒或载体中的那些物理和化学方法。此类方法包括但不限于氯化钙沉淀、电穿孔、裸DNA、脂质体等。在另外的实施例中,将DNA构建体与质粒一起共转化而不***该质粒中。在另外的实施例中,通过本领域已知的方法,将选择性标记从经修饰的芽孢杆菌属菌株中缺失或基本上切除(例如,Stahl等人,1984和Palmeros等人,2000)。在一些实施例中,载体从宿主染色体上分解下来,将侧翼区留在染色体上,而将固有的染色体区去除。
用于在芽孢杆菌属细胞中表达基因、其可读框(ORF)和/或其变体序列的启动子和启动子序列区通常是本领域技术人员已知的。通常选择本公开的启动子序列,使得它们在芽孢杆菌属细胞(例如,地衣芽孢杆菌细胞、枯草芽孢杆菌属细胞等)中起作用。某些示例性芽孢杆菌属启动子序列呈现于表6中。同样地,用于驱动芽孢杆菌属细胞中基因表达的启动子包括但不限于枯草芽孢杆菌碱性蛋白酶(aprE)启动子(Stahl等人,1984)、枯草芽孢杆菌的α-淀粉酶启动子(Yang等人,1983)、解淀粉芽孢杆菌的α-淀粉酶启动子(Tarkinen等人,1983)、来自枯草芽孢杆菌的中性蛋白酶(nprE)启动子(Yang等人,1984)、突变体aprE启动子(PCT公开号WO 2001/51643)或来自地衣芽孢杆菌或其他相关的芽孢杆菌属的任何其他启动子。在某些其他实施例中,启动子是美国专利公开号2014/0329309中公开的核糖体蛋白启动子或核糖体RNA启动子(例如,rrnI启动子)。在PCT公开号WO 2003/089604中描述了用于在芽孢杆菌属细胞中筛选和产生具有一系列活性(启动子强度)的启动子文库的方法。
IV.培养用于产生目的蛋白的芽孢杆菌属细胞
在其他实施例中,本公开提供了与未经修饰的(亲本)细胞相比(即,相对于),用于提高经修饰的细菌细胞的蛋白生产力的方法。在某些实施例中,本公开内容针对产生目的蛋白(POI)的方法,这些方法包括发酵培养经修饰的细菌细胞,其中经修饰的细胞将POI分泌到培养基中。本领域熟知的发酵方法可用于发酵本公开的经修饰的和未经修饰的芽孢杆菌属细胞。
在一些实施例中,将细胞在分批或连续发酵条件下培养。经典的分批发酵是封闭的***,其中在发酵开始时设定培养基的组成,并且该组成在发酵期间不改变。在发酵开始时,将培养基用一种或多种所希望的生物体接种。在这种方法中,允许发酵发生而不向***中添加任何组分。典型地,分批发酵符合关于添加碳源的“分批”的资格,并且经常对控制因素(如pH和氧浓度)进行尝试。分批***的代谢物和生物质组成不断变化直到发酵停止时。在典型分批培养中,细胞可以通过静态停滞期进展到高生长对数期,并且最后进入生长速率减小或停止的静止期。如果不经处理,处于静止期的细胞最终死亡。通常,处于对数期的细胞负责产物的大量产生。
标准分批***的合适的变体是“补料分批发酵”***。在典型分批***的这种变体中,随着发酵的进展,以增量添加底物。当分解代谢物阻遏可能抑制细胞的代谢时并且在培养基中希望具有有限量的底物的情况下,补料分批***是有用的。在补料分批***中实际底物浓度的测量是困难的并且因此基于可测量因素(如pH、溶解氧和废气(如CO2)的分压)的变化对其进行估计。分批和补料分批发酵在本领域中是常用并且已知的。
连续发酵是开放的***,其中将确定的发酵培养基连续添加到生物反应器中,同时去除等量的条件培养基以用于处理。连续发酵通常将培养物保持在恒定的高密度,其中细胞主要处于对数期生长。连续发酵允许对影响细胞生长和/或产物浓度的一种或多种因素进行调节。例如,在一个实施例中,将限制营养素(如碳源或氮源)保持在固定的速率,并且允许调控所有其他参数。在其他***中,影响生长的许多因素可以不断改变,而通过培养基浊度测量的细胞浓度保持不变。连续***努力保持稳态生长条件。因此,由于转移培养基而引起的细胞损失应当与发酵中的细胞生长速率相平衡。调节用于连续发酵过程的营养素和生长因子的方法以及用于最大化产物形成速率的技术是工业微生物学领域中熟知的。
因此,在某些实施例中,可以通过常规程序从培养基中回收由转化的(经修饰的)宿主细胞产生的POI,所述常规程序包括通过离心或过滤从培养基中分离宿主细胞,或者如果需要,破坏细胞并从细胞部分和碎片中去除上清液。典型地,在澄清后,将上清液或滤液的蛋白组分借助盐(例如,硫酸铵)沉淀。然后将沉淀出的蛋白溶解,并且可以通过各种色谱程序(例如,离子交换色谱法、凝胶过滤)进行纯化。
V.经修饰的(宿主)细胞产生的目的蛋白
本公开的目的蛋白(POI)可以是任何内源或异源蛋白,并且它可以是此种POI的变体。蛋白可以包含一个或多个二硫桥,或者是其功能形式是单体或多聚体的蛋白,即蛋白具有四级结构并且由多个相同(同源的)或不相同的(异源的)亚基构成,其中POI或其变体POI优选是具有目的特性的POI。
例如,如在以下实例中所述,本公开的经修饰的芽孢杆菌属细胞产生增加量的内源和/或异源目的蛋白。因此,在某些实施例中,本公开的经修饰的细胞表达内源POI、异源POI或一种或多种此类POI的组合。例如,在某些实施例中,本公开的经修饰的芽孢杆菌属(子代)细胞相对于亲本芽孢杆菌属细胞产生增加量的内源POI。在其他实施例中,本公开的经修饰的芽孢杆菌属(子代)细胞相对于亲本芽孢杆菌属细胞产生增加量的异源POI。
因此,在某些实施例中,本公开的经修饰的芽孢杆菌属(子代)细胞相对于亲本芽孢杆菌(对照)细胞产生增加量的POI,其中所述增加量的POI是至少约0.01%增加、至少约0.10%增加、至少约0.50%增加、至少约1.0%增加、至少约2.0%增加、至少约3.0%增加、至少约4.0%增加、至少约5.0%增加、或超过5.0%增加。在某些实施例中,增加量的POI通过测定其酶活性和/或通过测定/定量其比生产力(Qp)来确定。同样地,本领域技术人员可以利用本领域已知的其它规方法和技术来检测、测定、测量等一种或多种目的蛋白的表达或产生。
在某些实施例中,相对于(未经修饰的)亲本芽孢杆菌属细胞,本公开的经修饰的芽孢杆菌属细胞展示POI的增加的比生产力(Qp)。例如,比生产力(Qp)的检测是用于评价蛋白产生的合适方法。比生产力(Qp)可以使用以下等式确定:
“Qp=gP/gDCW·hr”
其中,“gP”是罐中产生的蛋白的克数;“gDCW”是罐中的干细胞重量(DCW)的克数;并且“hr”是从接种时间开始的以小时计的发酵时间,其包括生产时间以及生长时间。
因此,在某些其他实施例中,相比于未经修饰的(亲本)细胞,本公开的经修饰的芽孢杆菌属细胞包含至少约0.1%、至少约1%、至少约5%、至少约6%、至少约7%、至少约8%、至少约9%、或至少约10%或更多的比生产力(Qp)增加。
在某些实施例中,POI或其变体POI选自由以下组成的组:乙酰酯酶、氨肽酶、淀粉酶、***糖酶、***呋喃糖苷酶、碳酸酐酶、羧肽酶、过氧化氢酶、纤维素酶、几丁质酶、凝乳酶、角质酶、脱氧核糖核酸酶、差向异构酶、酯酶、α-半乳糖苷酶、β-半乳糖苷酶、α-葡聚糖酶、葡聚糖裂解酶、内切-β-葡聚糖酶、葡糖淀粉酶、葡萄糖氧化酶、α-葡糖苷酶、β-葡糖苷酶、葡萄糖醛酸酶、糖基水解酶、半纤维素酶、己糖氧化酶、水解酶、转化酶、异构酶、漆酶、连接酶、脂肪酶、裂解酶、甘露糖苷酶、氧化酶、氧化还原酶、果胶裂解酶、果胶乙酰酯酶、果胶解聚酶、果胶甲酯酶、果胶分解酶、过水解酶、多元醇氧化酶、过氧化物酶、酚氧化酶、植酸酶、聚半乳糖醛酸酶、蛋白酶、肽酶、鼠李糖-半乳糖醛酸酶、核糖核酸酶、转移酶、转运蛋白、转谷氨酰胺酶、木聚糖酶、己糖氧化酶、及其组合。
因此,在某些实施例中,POI或其变体POI是选自酶学委员会(EC)编号EC 1、EC 2、EC 3、EC 4、EC 5或EC 6的酶。
例如,在某些实施例中,POI是包括但不限于选自以下的EC 1(氧化还原酶)酶的氧化还原酶:EC 1.10.3.2(例如,漆酶)、EC 1.10.3.3(例如,L-抗坏血酸氧化酶)、EC 1.1.1.1(例如,醇脱氢酶)、EC 1.11.1.10(例如,氯化物过氧化物酶)、EC 1.11.1.17(例如,过氧化物酶)、EC 1.1.1.27(例如,L-乳酸脱氢酶)、EC 1.1.1.47(例如,葡萄糖1-脱氢酶)、EC1.1.3.X(例如,葡萄糖氧化酶)、EC 1.1.3.10(例如,吡喃糖氧化酶)、EC 1.13.11.X(例如,加双氧酶)、EC 1.13.11.12(例如,亚油酸13S-脂氧合酶(lineolate 13S-lipozygenase))、EC 1.1.3.13(例如,醇氧化酶)、EC 1.14.14.1(例如,单加氧酶)、EC 1.14.18.1(例如,单酚单加氧酶(monophenol monooxigenase))、EC 1.15.1.1(例如,超氧化物歧化酶)、EC1.1.5.9(先前,EC 1.1.99.10,例如,葡萄糖脱氢酶)、EC 1.1.99.18(例如,纤维二糖脱氢酶)、EC 1.1.99.29(例如,吡喃糖脱氢酶)、EC 1.2.1.X(例如,脂肪酸还原酶)、EC 1.2.1.10(例如,乙醛脱氢酶)、EC 1.5.3.X(例如,果糖基胺还原酶)、EC 1.8.1.X(例如,二硫还原酶)和EC 1.8.3.2(例如,硫醇氧化酶)。
在某些实施例中,POI是包括但不限于EC 2(转移酶)酶的转移酶,所述EC 2(转移酶)酶选自EC 2.3.2.13(例如,转谷氨酰胺酶)、EC 2.4.1.X(例如,己糖基转移酶)、EC2.4.1.40(例如,交替蔗糖酶(alternasucrase))、EC 2.4.1.18(例如,1,4α-葡聚糖分支酶)、EC 2.4.1.19(例如,环麦芽糖糊精葡聚糖转移酶)、EC 2.4.1.2(例如,糊精葡聚糖酶)、EC 2.4.1.20(例如,纤维二糖磷酸化酶)、EC 2.4.1.25(例如,4-α-葡聚糖转移酶)、EC2.4.1.333(例如,1,2-β-低聚葡糖磷酸转移酶)、EC 2.4.1.4(例如,淀粉蔗糖酶)、EC2.4.1.5(例如,葡聚糖蔗糖酶)、EC 2.4.1.69(例如,半乳糖苷2-α-L-岩藻糖基转移酶)、EC2.4.1.9(例如,菊粉蔗糖酶(inulosucrase))、EC 2.7.1.17(例如,木酮糖激酶)、EC2.7.7.89(原名EC 3.1.4.15,例如,[谷氨酰氨合成酶]-腺苷-L-酪氨酸磷酸化酶)、EC2.7.9.4(例如,α葡聚糖激酶)和EC 2.7.9.5(例如,磷酸葡聚糖激酶)。
在其他实施例中,POI是水解酶,包括但不限于选自以下的EC 3(水解酶)酶:EC3.1.X.X(例如,酯酶)、EC 3.1.1.1(例如,果胶酶)、EC 3.1.1.14(例如,叶绿素酶)、EC3.1.1.20(例如,鞣酸酶)、EC 3.1.1.23(例如,甘油酯酰基水解酶)、EC 3.1.1.26(例如,半乳糖脂酶)、EC 3.1.1.32(例如,磷脂酶A1)、EC 3.1.1.4(例如,磷脂酶A2)、EC 3.1.1.6(例如,乙酰酯酶)、EC 3.1.1.72(例如,乙酰木聚糖酯酶)、EC 3.1.1.73(例如,阿魏酸酯酶)、EC3.1.1.74(例如,角质酶)、EC 3.1.1.86(例如,鼠李糖半乳糖醛酸乙酰酯酶)、EC 3.1.1.87(例如,伏马菌素B1酯酶)、EC 3.1.26.5(例如,核糖核酸酶P)、EC 3.1.3.X(例如,磷酸单酯水解酶)、EC 3.1.30.1(例如,曲霉属核酸酶S1)、EC 3.1.30.2(例如,粘质沙雷氏菌(Serratia marcescens)核酸酶)、EC 3.1.3.1(例如,碱性磷酸酶)、EC 3.1.3.2(例如,酸性磷酸酶)、EC 3.1.3.8(例如,3-植酸酶)、EC 3.1.4.1(例如,磷酸二酯酶I)、EC 3.1.4.11(例如,磷酸肌醇磷脂酶C)、EC 3.1.4.3(例如,磷脂酶C)、EC 3.1.4.4(例如,磷脂酶D)、EC3.1.6.1(例如,芳基硫酸酯酶)、EC 3.1.8.2(例如,二异丙基-氟磷酸酶)、EC 3.2.1.10(例如,寡-1,6-葡糖苷酶)、EC 3.2.1.101(例如,甘露聚糖内切-1,6-α-甘露糖苷酶)、EC3.2.1.11(例如,α-1,6-葡聚糖-6-葡聚糖水解酶)、EC 3.2.1.131(例如,木聚糖α-1,2-葡萄糖醛酸苷酶(glucuronosidase))、EC 3.2.1.132(例如,壳聚糖N-乙酰葡糖氨基水解酶)、EC3.2.1.139(例如,α-葡萄糖醛酸酶)、EC 3.2.1.14(例如,几丁质酶)、EC 3.2.1.151(例如,木葡聚糖特异性内切-β-1,4-葡聚糖酶)、EC 3.2.1.155(例如,木葡聚糖特异性外切-β-1,4-葡聚糖酶)、EC 3.2.1.164(例如,半乳聚糖内切-1,6-β-半乳糖苷酶)、EC 3.2.1.17(例如,溶菌酶)、EC 3.2.1.171(例如,鼠李半乳糖醛酸聚糖水解酶)、EC3.2.1.174(例如,鼠李半乳糖醛酸聚糖鼠李糖水解酶)、EC 3.2.1.2(例如,β-淀粉酶)、EC 3.2.1.20(例如,α-葡糖苷酶)、EC 3.2.1.22(例如,α-半乳糖苷酶)、EC 3.2.1.25(例如,β-甘露糖苷酶)、EC3.2.1.26(例如,β-果糖呋喃糖苷酶)、EC 3.2.1.37(例如,木聚糖1,4-β-木糖苷酶)、EC3.2.1.39(例如,葡聚糖内切-1,3-β-D-葡糖苷酶)、EC 3.2.1.40(例如,α-L-鼠李糖苷酶)、EC 3.2.1.51(例如,α-L-岩藻糖苷酶)、EC 3.2.1.52(例如,β-N-乙酰己糖胺酶)、EC3.2.1.55(例如,α-N-***呋喃糖苷酶)、EC 3.2.1.58(例如,葡聚糖1,3-β-葡糖苷酶)、EC3.2.1.59(例如,葡聚糖内切-1,3-α-葡糖苷酶)、EC 3.2.1.67(例如,半乳糖醛酸1,4-α-半乳糖醛酸酶)、EC 3.2.1.68(例如,异淀粉酶)、EC 3.2.1.7(例如,1-β-D-果聚糖果糖水解酶)、EC 3.2.1.74(例如,葡聚糖1,4-β-葡糖苷酶)、EC 3.2.1.75(例如,葡聚糖内切-1,6-β-葡糖苷酶)、EC 3.2.1.77(例如,甘露聚糖1,2-(1,3)-α-甘露糖苷酶)、EC 3.2.1.80(例如,果聚糖β-果糖苷酶)、EC 3.2.1.82(例如,外切-聚-α-半乳糖醛酸苷酶)、EC 3.2.1.83(例如,κ-角叉菜胶酶)、EC 3.2.1.89(例如,***半乳聚糖内切-1,4-β-半乳糖苷酶)、EC3.2.1.91(例如,纤维素1,4-β-纤维二糖苷酶)、EC 3.2.1.96(例如,甘露糖基-糖蛋白内切-β-N-乙酰葡糖胺糖苷酶)、EC 3.2.1.99(例如,***聚糖内切-1,5-α-L-***糖苷酶)、EC 3.4.X.X(例如,肽酶)、EC 3.4.11.X(例如,氨肽酶)、EC 3.4.11.1(例如,亮氨酰氨肽酶)、EC 3.4.11.18(例如,甲硫氨酰氨肽酶)、EC 3.4.13.9(例如,Xaa-Pro二肽酶)、EC3.4.14.5(例如,二肽基-肽酶IV)、EC 3.4.16.X(例如,丝氨酸型羧肽酶)、EC 3.4.16.5(例如,羧肽酶C)、EC 3.4.19.3(例如,焦谷氨酰肽酶I)、EC 3.4.21.X(例如,丝氨酸内肽酶)、EC3.4.21.1(例如,胰凝乳蛋白酶)、EC 3.4.21.19(例如,谷氨酰内肽酶)、EC 3.4.21.26(例如,脯氨酰寡肽酶)、EC 3.4.21.4(例如,胰蛋白酶)、EC 3.4.21.5(例如,凝血酶)、EC3.4.21.63(例如,蜂蜜曲霉蛋白酶)、EC 3.4.21.65(例如,热霉菌素)、EC 3.4.21.80(例如,链霉菌素A)、EC 3.4.22.X(例如,半胱氨酸内肽酶)、EC 3.4.22.14(例如,猕猴桃蛋白酶(actinidain))、EC 3.4.22.2(例如,木瓜蛋白酶)、EC 3.4.22.3(例如,无花果蛋白酶)、EC3.4.22.32(例如,茎菠萝蛋白酶)、EC 3.4.22.33(例如,水果菠萝蛋白酶)、EC 3.4.22.6(例如,木瓜凝乳蛋白酶)、EC 3.4.23.1(例如,胃蛋白酶A)、EC 3.4.23.2(例如,胃蛋白酶B)、EC3.4.23.22(例如,栗疫霉胃蛋白酶)、EC 3.4.23.23(例如,毛霉胃蛋白酶)、EC 3.4.23.3(例如,胃亚蛋白酶)、EC 3.4.24.X(例如,金属内肽酶)、EC 3.4.24.39(例如,氘代溶素(deuterolysin))、EC 3.4.24.40(例如,舍雷肽酶)、EC 3.5.1.1(例如,天冬酰胺酶)、EC3.5.1.11(例如,青霉素酰胺酶)、EC 3.5.1.14(例如,N-酰基-脂族-L-氨基酸酰胺水解酶)、EC 3.5.1.2(例如,L-谷氨酰胺酰胺水解酶)、EC 3.5.1.28(例如,N-乙酰胞壁酰-L-丙氨酸酰胺酶)、EC 3.5.1.4(例如,酰胺酶)、EC 3.5.1.44(例如,蛋白-L-谷氨酰胺酰胺水解酶)、EC 3.5.1.5(例如,脲酶)、EC 3.5.1.52(例如,肽-N(4)-(N-乙酰基-β-葡糖胺基)天冬酰胺酰胺酶)、EC 3.5.1.81(例如,N-酰基-D-氨基酸脱酰基酶)、EC 3.5.4.6(例如,AMP脱氨酶)、和EC 3.5.5.1(例如,腈水解酶)。
在其他实施例中,POI是包括但不限于选自以下的EC 4(裂解酶)酶的裂解酶:EC4.1.2.10(例如,苯乙醇腈裂解酶)、EC 4.1.3.3(例如,N-乙酰神经氨酸裂解酶)、EC4.2.1.1(例如,碳酸脱水酶)、EC 4.2.2.-(例如,鼠李半乳糖醛酸聚糖裂解酶)、EC4.2.2.10(例如,果裂解酶)、EC 4.2.2.22(例如,果胶三糖-裂解酶)、EC 4.2.2.23(例如,鼠李半乳糖醛酸聚糖内切裂解酶)和EC 4.2.2.3(例如,甘露糖醛特异性海藻酸裂解酶)。
在某些其他实施例中,POI是包括但不限于选自以下的EC 5(异构酶)酶的异构酶:EC 5.1.3.3(例如,醛糖1-差向异构酶)、EC 5.1.3.30(例如,D-阿洛酮糖3-差向异构酶)、EC5.4.99.11(例如,异麦芽酮糖合酶)和EC 5.4.99.15(例如,(1→4)-α-D-葡聚糖1-α-D-葡萄糖基变位酶((1→4)-α-D-glucan 1-α-D-glucosylmutase))。
在又其他实施例中,POI是连接酶,该连接酶包括但不限于选自以下的EC 6(连接酶)酶:EC 6.2.1.12(例如,4-香豆酸:辅酶A连接酶)和EC 6.3.2.28(例如,L-氨基酸α-连接酶)9
因此,在某些实施例中,产生工业蛋白酶的芽孢杆菌属宿主细胞提供特别优选的表达宿主。同样地,在某些其他实施例中,产生工业淀粉酶的芽孢杆菌属宿主细胞提供特别优选的表达宿主。
例如,存在两种一般类型的蛋白酶通常由芽孢杆菌属物种分泌,即中性(或“金属蛋白酶”)和碱性(或“丝氨酸”)蛋白酶。例如,芽孢杆菌枯草杆菌蛋白酶蛋白(酶)是可用于在本公开中使用的示例性丝氨酸蛋白酶。已经鉴定并测序了多种枯草芽孢杆菌蛋白酶,例如枯草杆菌蛋白酶168、枯草杆菌蛋白酶BPN’、枯草杆菌蛋白酶Carlsberg、枯草杆菌蛋白酶DY、枯草杆菌蛋白酶147和枯草杆菌蛋白酶309(例如,WO 1989/06279和Stahl等人,1984)。在本公开的一些实施例中,经修饰的芽孢杆菌属细胞产生突变体(即,变体)蛋白酶。许多参考文献提供变体蛋白酶的实例,如PCT公开号WO 1999/20770;WO 1999/20726;WO 1999/20769;WO 1989/06279;US RE34,606;US专利号4,914,031;4,980,288;5,208,158;5,310,675;5,336,611;5,399,283;5,441,882;5,482,849;5,631,217;5,665,587;5,700,676;5,741,694;5,858,757;5,880,080;6,197,567和6,218,165。因此,在某些实施例中,本公开的经修饰的芽孢杆菌属细胞包含编码蛋白酶的表达构建体。
在某些其他实施例中,本公开的经修饰的芽孢杆菌属细胞包含编码淀粉酶的表达构建体。多种淀粉酶及其变体是本领域技术人员已知的。例如,国际PCT公开号WO 2006/037484和WO 2006/037483描述了具有改善的溶剂稳定性的变体α-淀粉酶,公开号WO 1994/18314公开了氧化稳定的α-淀粉酶变体,公开号WO 1999/19467、WO 2000/29560和WO 2000/60059公开了Termamyl-样α-淀粉酶变体,公开号WO 2008/112459公开了衍生自芽孢杆菌属物种编号707的α-淀粉酶变体,公开号WO 1999/43794公开了麦芽生成的α-淀粉酶变体,公开号WO 1990/11352公开了超热稳定的(hyper-thermostable)α-淀粉酶变体,公开号WO2006/089107公开了具有颗粒淀粉水解活性的α-淀粉酶变体。
在其他实施例中,在本公开的经修饰细胞中表达和产生的POI或变体POI是肽、肽激素、生长因子、凝血因子、趋化因子、细胞因子、淋巴因子、抗体、受体、粘附分子、微生物抗原(例如,HBV表面抗原、HPV E7等)、其变体、其片段等。其他类型的目的蛋白(或其变体)可以是能够为食品或作物提供营养价值的蛋白或变体。非限制性实例包括可以抑制抗营养因子形成的植物蛋白和具有更理想的氨基酸组成的植物蛋白(例如比非转基因植物具有更高的赖氨酸含量)。
存在本领域普通技术人员已知的用于检测和测量细胞内和细胞外表达的蛋白的活性的各种测定。特别地,对于蛋白酶,存在基于来自酪蛋白或血红蛋白的酸溶性肽的释放的测定,其作为280nm下的吸光度测量或使用Folin方法进行比色测定(例如Bergmeyer等人,1984)。其他测定涉及生色底物的溶解(参见例如,Ward,1983)。其他示例性测定包括琥珀酰-Ala-Ala-Pro-Phe-对-硝基苯胺测定(SAAPFpNA)和2,4,6-三硝基苯磺酸钠盐测定(TNBS测定)。本领域技术人员已知的许多另外的参考文献提供了适合的方法(参见例如,Wells等人,1983;Christianson等人,1994和Hsia等人,1999)。
国际PCT公开号WO 2014/164777公开了可用于本文所述淀粉酶活性的Ceralphaα-淀粉酶活性测定。
用于确定宿主细胞中目的蛋白的分泌水平并且检测所表达的蛋白的手段包括使用对蛋白具有特异性的多克隆或单克隆抗体的免疫测定。实例包括酶联免疫吸附测定(ELISA)、放射免疫测定(RIA)、荧光免疫测定(FIA)以及荧光激活细胞分选术(FACS)。
VI.示例性实施例
本公开的非限制性实施例包括但不限于:
1.一种用于在经修饰的地衣芽孢杆菌细胞中产生增加量的目的蛋白(POI)的方法,所述方法包括:(a)通过在表达POI的亲本地衣芽孢杆菌细胞中引入包含与天然prsA可读框(ORF)序列可操作地连接的天然prsA启动子序列的多核苷酸来修饰所述表达POI的亲本地衣芽孢杆菌细胞,以及(b)在用于产生所述POI的合适条件下发酵所述经修饰的细胞,其中当在相同条件下发酵时,相对于所述亲本细胞,所述经修饰的细胞产生增加量的所述POI。
2.一种用于在经修饰的地衣芽孢杆菌细胞中产生增加量的目的蛋白(POI)的方法,所述方法包括:(a)通过向亲本地衣芽孢杆菌细胞中引入以下来修饰所述亲本地衣芽孢杆菌细胞(i)编码POI的表达盒和(ii)包含可操作地连接到天然prsA可读框(ORF)序列的天然prsA启动子序列的多核苷酸,以及(b)在用于产生所述POI的合适条件下发酵所述经修饰的细胞,其中当在相同条件下发酵时,相对于所述亲本细胞,所述经修饰的细胞产生增加量的所述POI。
3.如实施例1或实施例2所述的方法,其中所述引入的多核苷酸包含与SEQ ID NO:100具有至少95%序列同一性的天然prsA启动子序列。
4.如实施例1或实施例2所述的方法,其中所述引入的多核苷酸包含与SEQ ID NO:101具有至少90%序列同一性的天然prsA ORF。
5.如实施例1或实施例2所述的方法,其中所述亲本细胞包含编码天然prsA蛋白的内源prsA基因。
6.如实施例5所述的方法,其中所述内源prsA基因编码与SEQ ID NO:155具有约90%序列同一性的天然prsA蛋白。
7.如实施例1或实施例2所述的方法,其中将所述引入的多核苷酸整合到所述经修饰的地衣芽孢杆菌细胞的基因组中。
8.如实施例1或实施例2所述的方法,其中所述经修饰的细胞进一步包含与SEQ IDNO:122具有至少90%序列同一性的dltA基因的缺失或破坏。
9.如实施例1或实施例2所述的方法,其中所述经修饰的细胞进一步包含与SEQ IDNO:121或SEQ ID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。
10.如实施例1或实施例2所述的方法,其中所述经修饰的细胞进一步包含与SEQID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏和与SEQ ID NO:121或SEQID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。
11.如实施例1或实施例2所述的方法,其中所述POI是酶。
12.如实施例11所述的方法,其中所述酶是蛋白酶或淀粉酶。
13.一种衍生自亲本地衣芽孢杆菌细胞的经修饰的地衣芽孢杆菌细胞,其中所述经修饰的细胞包含含有可操作地连接到天然prsA可读框(ORF)序列的天然prsA启动子序列的引入的多核苷酸。
14.一种衍生自包含编码天然prsA蛋白的内源prsA基因的亲本地衣芽孢杆菌的经修饰的地衣芽孢杆菌细胞,其中所述经修饰的细胞包含含有可操作地连接到天然prsA可读框(ORF)序列的天然prsA启动子序列的引入的多核苷酸。
15.如实施例13或实施例14所述的经修饰的细胞,其中所述引入的多核苷酸包含与SEQ ID NO:100具有至少95%序列同一性的天然prsA启动子。
16.如实施例13或实施例14所述的经修饰的细胞,其中所述引入的多核苷酸包含与SEQ ID NO:101具有至少90%序列同一性的天然prsA ORF。
17.如实施例13或实施例14所述的经修饰的细胞,其中所述引入的多核苷酸编码与SEQ ID NO:155具有约90%序列同一性的天然prsA蛋白。
18.如实施例13或实施例14所述的经修饰的细胞,其中将所述引入的多核苷酸整合到所述经修饰的地衣芽孢杆菌细胞的基因组中。
19.如实施例13或实施例14所述的经修饰的细胞,其包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏。
20.如实施例13或实施例14所述的经修饰的细胞,其包含与SEQ ID NO:121或SEQID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。
21.如实施例13或实施例14所述的经修饰的细胞,其包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏和与SEQ ID NO:121或SEQ ID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。
22.如实施例13或实施例14所述的经修饰的细胞,其包含编码异源目的蛋白(POI)的引入的表达盒。
23.如实施例22所述的经修饰的细胞,其中所述POI是酶。
24.如实施例13或实施例14所述的经修饰的细胞,其中所述亲本细胞表达内源POI。
25.一种目的蛋白,所述目的蛋白由如实施例22或实施例24所述的经修饰的细胞产生。
26.一种相对于亲本地衣芽孢杆菌细胞产生增加量的目的蛋白(POI)的经修饰的地衣芽孢杆菌细胞,其中所述经修饰的细胞衍生自表达POI的亲本地衣芽孢杆菌细胞,其中所述经修饰的细胞包含含有可操作地连接到天然prsA可读框(ORF)序列的天然prsA启动子序列的引入的多核苷酸并且包含与SEQ ID NO:121或SEQ ID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏,其中当在相同条件下发酵时,经修饰的细胞相对于亲本菌株产生增加量的POI。
27.如实施例26所述的经修饰的细胞,其进一步包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏。
28.一种相对于亲本地衣芽孢杆菌细胞产生增加量的目的蛋白(POI)的经修饰的地衣芽孢杆菌细胞,其中经修饰的细胞衍生自表达POI的亲本地衣芽孢杆菌细胞,其中所述经修饰的细胞包含含有可操作地连接到天然prsA可读框(ORF)的天然prsA启动子的引入的多核苷酸并且包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏,其中当在相同条件下发酵时,所述经修饰的细胞相对于所述亲本菌株产生增加量的所述POI。
29.如实施例28所述的经修饰的细胞,其进一步包含与SEQ ID NO:121或SEQ IDNO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。
30.如实施例26或实施例28所述的经修饰的细胞,其中所述天然prsA启动子与SEQID NO:100具有至少95%序列同一性。
31.如实施例26或实施例28所述的经修饰的细胞,其中所述天然prsA ORF与SEQID NO:101具有至少90%序列同一性。
32.如实施例26或实施例28所述的经修饰的细胞,其中所述天然prsA蛋白与SEQID NO:155具有约90%序列同一性。
33.如实施例26或实施例28所述的经修饰的细胞,其中所述POI是酶。
34.如实施例33所述的经修饰的细胞,其中所述酶是蛋白酶或淀粉酶。
35.一种目的蛋白,所述目的蛋白由如实施例26或实施例28所述的经修饰的细胞产生。
实例
根据以下实例可以进一步理解本发明的某些方面,这些实例不应被解释为限制性的。对材料和方法的修改对于本领域技术人员而言应是显而易见的。
实例1
靶向RGHR2通路基因的CAS9载体的构建
对来自化脓链球菌的Cas9蛋白(SEQ ID NO:1)进行密码子优化用于芽孢杆菌(SEQID NO:2),其中添加N-末端核定位序列(NLS;“APKKKRKV”;SEQ ID NO:3)、C-末端NLS(“KKKKLK”;SEQ ID NO:4)、十组氨酸标签(“HHHHHHHHHH”;SEQ ID NO:5)、来自枯草芽孢杆菌的aprE启动子(SEQ ID NO:6)和终止子序列(SEQ ID NO:7),并使用Q5 DNA聚合酶(NEB)(按照制造商的说明书)用下面表1中列出的正向(SEQ ID NO:8)和反向(SEQ ID NO:9)引物对扩增。
正向 ATATATGAGTAAACTTGGTCTGACAGAATTCCTCCATTTTCTTCTGCTAT SEQ ID NO:8
反向 TGCGGCCGCGAATTCGATTACGAATGCCGTCTCCC SEQ ID NO:9
表1
正向和反向引物对
使用Q5 DNA聚合酶(NEB)(按照制造商的说明书)用下表2中列出的正向(SEQ IDNO:12)和反向(SEQ ID NO:13)引物对扩增质粒pKB320(SEQ ID NO:11)的主链(SEQ ID NO:10)。
正向 GGGAGACGGCATTCGTAATCGAATTCGCGGCCGCA SEQ ID NO:12
反向 ATAGCAGAAGAAAATGGAGGAATTCTGTCAGACCAAGTTTACTCATATAT SEQ ID NO:13
表2
正向和反向引物对
按照制造商的说明书用Zymo清洁和浓缩(Zymo clean and concentrate)5柱纯化PCR产物。随后,用Q5聚合酶(NEB)与等摩尔比的两个片段混合,使用延长重叠延伸PCR(POE-PCR)组装PCR产物。POE-PCR反应循环进行:98℃持续五(5)秒,64℃持续十(10)秒,72℃持续四(4)分钟十五(15)秒,持续30个循环。将五(5)μlPOE-PCR(DNA)按照制造商的说明书转化进入Top10大肠杆菌(英杰公司(Invitrogen)),并在含有五十(50)μg/ml硫酸卡那霉素并用1.5%琼脂固化的溶源性(L)培养液(Miller配方;1%(w/v)胰蛋白胨、0.5%酵母提取物(w/v)、1%NaCl(w/v))中选择。允许菌落在37℃生长十八(18)小时。挑取菌落,并使用QiaprepDNA迷你制备试剂盒(按照制造商的说明书)制备质粒DNA,并用五十五(55)μl ddH2O进行洗脱。使用下面表3中列出的测序引物对质粒DNA进行桑格(Sanger)测序以验证正确组装。
表3
测序引物
反向 CCGACTGGAGCTCCTATATTACC SEQ ID NO:14
反向 GCTGTGGCGATCTGTATTCC SEQ ID NO:15
正向 GTCTTTTAAGTAAGTCTACTCT SEQ ID NO:16
正向 CCAAAGCGATTTTAAGCGCG SEQ ID NO:17
正向 CCTGGCACGTGGTAATTCTC SEQ ID NO:18
正向 GGATTTCCTCAAATCTGACG SEQ ID NO:19
正向 GTAGAAACGCGCCAAATTACG SEQ ID NO:20
正向 GCTGGTGGTTGCTAAAGTCG SEQ ID NO:21
正向 GGACGCAACCCTCATTCATC SEQ ID NO:22
反向 CAGGCATCCGATTTGCAAGG SEQ ID NO:23
正向 GCAAGCAGCAGATTACGCG SEQ ID NO:24
将正确组装的质粒pRF694(SEQ ID NO:25)用于构建质粒pRF801(SEQ ID NO:26)和pRF806(SEQ ID NO:27),用于编辑地衣芽孢杆菌基因组的靶位点1(TS1;SEQ ID NO:28)和靶位点2(TS2;SEQ ID NO:29),如下所述。
地衣芽孢杆菌的serA1可读框(SEQ ID NO:30)含有独特的靶位点,即反向定向的靶位点1(TS1;SEQ ID NO:28)。靶位点与反向定向的原间隔子邻近基序(SEQ ID NO:31)相邻。靶位点可转换为编码可变靶向结构域(SEQ ID NO:32)的DNA。
编码VT结构域的DNA序列(SEQ ID NO:32)可操作地融合到编码Cas9核酸内切酶识别结构域(CER;SEQ ID NO:33)的DNA序列,使得当被细菌细胞的RNA聚合酶转录时,它产生靶向靶位点1的功能性gRNA(SEQ ID NO:34)。编码gRNA的DNA可操作地连接到在芽孢杆菌属物种细胞中有效的启动子(例如spac启动子;SEQ ID NO:35)和在芽孢杆菌属物种细胞中有效的终止子(例如噬菌体λ的t0终止子;SEQ ID NO:36),使得所述启动子位于编码gRNA的DNA(SEQ ID NO:33)的上游(5'),并且所述终止子位于编码gRNA的DNA(SEQ ID NO:33)的下游(3')。
通过扩增地衣芽孢杆菌基因组DNA(gDNA)的两个同源臂,创建了编辑模板,用以响应Cas9/gRNA切割而缺失serA1基因。第一片段对应于serA1可读框(SEQ ID NO:37)直接上游500bp。使用Q5 DNA聚合酶(按照制造商的说明书),以及下表4中列出的正向(SEQ ID NO:38)和反向(SEQ ID NO:39)引物扩增此片段。所述引物在第一片段的3'末端并入了与第二片段5'末端同源的18bp,在第一片段的5'末端并入了与pRF694同源的20bp。
表4
正向和反向引物对
正向 TGAGTAAACTTGGTCTGACAAATGGTTCTTTCCCCTGTCC SEQ ID NO:38
反向 AGGTTCCGCAGCTTCTGTGTAAGATTTCCTCCTAAATAAGCGTCAT SEQ ID NO:39
第二片段对应于serA1可读框(SEQ ID NO:40)的3'末端直接下游的500bp。使用Q5DNA聚合酶(按照制造商的说明书),以及下表5中列出的正向(SEQ ID NO:41)和反向(SEQID NO:42)引物扩增此片段。所述引物在第二片段的5'末端并入了与第一片段3'末端同源的28bp,在第二片段的3'末端并入了与pRF694同源的21bp。
表5
正向和反向引物对
正向 ATGACGCTTATTTAGGAGGAAATCTTACACAGAAGCTGCGGAACCT SEQ ID NO:41
反向 CAGAAGAAAATGGAGGAATTCGAATATCGACCGGAACCCAC SEQ ID NO:42
使用标准分子生物学技术将编码靶位点1gRNA表达盒(SEQ ID NO:43)、第一(SEQID NO:37)和第二同源臂(SEQ ID NO:40)的DNA组装到pRF694(SEQ ID NO:25)中,产生pRF801(SEQ ID NO:26),包含以下的大肠杆菌-地衣芽孢杆菌穿梭质粒:Cas9表达盒(SEQID NO:2)、编码靶向serA1可读框内的靶位点1的gRNA的gRNA表达盒(SEQ ID NO:43)和由第一(SEQ ID NO:37)和第二(SEQ ID NO:40)同源臂构成的编辑模板(SEQ ID NO:44)。用表3中列出的寡核苷酸通过桑格测序验证质粒。
地衣芽孢杆菌的rghR1可读框(SEQ ID NO:45)在反向链上含有唯一的靶位点,即靶位点2(TS2;SEQ ID NO:29)。靶位点与反向链上的原间隔子邻近基序(SEQ ID NO:46)相邻。编码靶位点的DNA序列(SEQ ID NO:29)可操作地融合到编码Cas9核酸内切酶识别结构域(CER,SEQ ID NO:33)的DNA序列,使得当被细菌细胞的RNA聚合酶转录时,它产生靶向靶位点2的功能性gRNA(SEQ ID NO:47)。编码gRNA的DNA可操作地连接到在芽孢杆菌属物种细胞中有效的启动子(例如来自枯草芽孢杆菌的spac启动子;SEQ ID NO:35)和在芽孢杆菌属物种细胞中有效的终止子(例如噬菌体λ的t0终止子;SEQ ID NO:36),使得该启动子位于编码gRNA的DNA(SEQ ID NO:47)的5',并且该终止子位于编码gRNA的DNA(SEQ ID NO:47)的3'。
通过扩增地衣芽孢杆菌基因组DNA(gDNA)的两个同源臂,构建了编辑模板,用以响应Cas9/gRNA切割而修饰rghR1基因。第一片段对应于rghR1可读框(SEQ ID NO:48)直接上游500bp。使用Q5 DNA聚合酶(根据制造商的说明书)和下面表6中列出的引物扩增该片段。所述引物在第一片段的3'末端并入了与第二片段5'末端同源的23bp,在第一片段的5'末端并入了与pRF694同源的20bp。
表6
正向和反向引物对
正向 TGAGTAAACTTGGTCTGACATTGATATTCAGCACCCTGCG SEQ ID NO:49
反向 TGTGCCGCGGAGAAGTATGGCCAAAACCTCGCAATCTC SEQ ID NO:50
第二片段对应于rghR1可读框(SEQ ID NO:51)的3'末端直接下游的500bp。使用Q5DNA聚合酶(根据制造商的说明书)和下面表7中列出的引物扩增该片段。所述引物在第二片段的5'末端并入了与第一片段3'末端同源的20bp,在第二片段的3'末端并入了与pRF694同源的21bp。
表7
正向和反向引物对
正向 GAGATTGCGAGGTTTTGGCCATACTTCTCCGCGGCACA SEQ ID NO:52
反向 CAGAAGAAAATGGAGGAATTCATTTCTCGGGTTTAAACAGCCAC SEQ ID NO:53
使用标准分子生物学技术将编码靶位点2gRNA表达盒(SEQ ID NO:54)、第一(SEQID NO:48)和第二同源臂(SEQ ID NO:51)的DNA组装到pRF694(SEQ ID NO:25)中,产生pRF806(SEQ ID NO:27),包含以下的大肠杆菌-地衣芽孢杆菌穿梭质粒:Cas9表达盒(SEQID NO:2)、编码靶向rghR1可读框内的靶位点2的gRNA的gRNA表达盒(SEQ ID NO:54)和由第一(SEQ ID NO:48)和第二(SEQ ID NO:51)同源臂构成的编辑模板(SEQ ID NO:55)。用表3列出的寡核苷酸通过桑格序列验证质粒。
实例2
CAS9 Y155H变体和相关靶向质粒的构建
在本例中,在pRF801(SEQ ID NO:26)和pRF806质粒(SEQ ID NO:27)中构建化脓链球菌Cas9的Y155H变体(SEQ ID NO:56)。为了在pRF801质粒(SEQ ID NO:26)或pRF806质粒(SEQ ID NO:27)中引入Y155H变体,使用Quikchange诱变试剂盒(按照制造商说明书)和下表8中的寡核苷酸,使用pRF801(SEQ ID NO:26)或pRF806(SEQ ID NO:27)作为模板DNA进行定点诱变。
表8
正向和反向引物对
正向 GATCTGCGTTTAATCCATCTTGCGTTAGCGCAC SEQ ID NO:57
反向 GTGCGCTAACGCAAGATGGATTAAACGCAGATC SEQ ID NO:58
所产生的反应产物,pRF827(SEQ ID NO:59)包含Cas9 Y155H变体表达盒(SEQ IDNO:60)、gRNA表达盒(SEQ ID NO:43)(其编码靶向serA1可读框内的靶位点1的gRNA)和由第一(SEQ ID NO:37)和第二(SEQ ID NO:40)同源臂构成的编辑模板(SEQ ID NO:44),或者pRF856(SEQ ID NO:61),其含有Cas9 Y155H变体表达盒(SEQ ID NO:60)、靶向rghR1可读框内的靶位点2的gRNA表达盒(SEQ ID NO:54)和由第一(SEQ ID NO:48)和第二(SEQ ID NO:51)同源臂构成的编辑模板(SEQ ID NO:55)。使用表3中列出的测序引物对质粒DNA进行桑格测序以验证正确组装。
质粒pRF862的构建
质粒pRF862(SEQ ID NO:62)通过以下来构建:从pRF827(SEQ ID NO:59)移出Cas9可读框的含有Y155H取代的片段(SEQ ID NO:63),使用表9中所列引物扩增。
表9
正向和反向引物对
正向 CACGTCGTAAAAATCGTATT SEQ ID NO:64
反向 CAAACAGACCATTTTTCTTT SEQ ID NO:65
从pRF694(SEQ ID NO:25)扩增第二片段(SEQ ID NO:66),这样使得它包含整个质粒,除了以上pRF827片段(SEQ ID NO:63)上含有的片段。该片段与pRF827片段(SEQ ID NO:60)的5'和3'末端具有同源性用于组装,并使用表10中列出的引物进行扩增。
表10
正向和反向引物对
正向 AAAGAAAAATGGTCTGTTTG SEQ ID NO:67
反向 AATACGATTTTTACGACGTG SEQ ID NO:68
根据制造商的说明书,使用NEBuilder组装两个片段,并且转化到大肠杆菌感受态细胞中。如表3中列出通过桑格的方法验证质粒序列。经序列验证的分离物以质粒pRF862存储(SEQ ID NO:62)。
使用两个部分构建pRF869(SEQ ID NO:69),它是靶向rghR2 ORF(SEQ ID NO:70)并且***三个(3)框内终止密码子的质粒。通过IDT合成第一部分(SEQ ID NO:71),其包含用来修饰rghR2 ORF(SEQ ID NO:70)的编辑模板(SEQ ID NO:72),和靶向rghR2 ORF(SEQID NO:70)的gRNA表达盒(SEQ ID NO:73),并且使用表11中列出的引物扩增该第一部分。
表11
正向和反向引物对
正向 CGTGCGGCCGCGAATTC SEQ ID NO:74
反向 CCTGATACCGGGAGACGGCATTCGTAATC SEQ ID NO:75
通过使用表12中列出的引物扩增pRF862,将合成的片段***pRF862(SEQ ID NO:62)。
表12
正向和反向引物对
正向 GAATTCGCGGCCGCACG SEQ ID NO:76
反向 GATTACGAATGCCGTCTCCCGGTATCAGG SEQ ID NO:77
根据制造商的说明书,使用NEBuilder组装这两个部分并且转化到大肠杆菌中。如表3中列出通过桑格的方法验证质粒序列。经序列验证的分离物以pRF869(SEQ ID NO:69)存储。
如以上实例1和2中所述组装几个另外的Cas9质粒。下表13中列出了那些质粒,连同靶位点序列和编辑模板效应。
表13
用于编辑地衣芽孢杆菌细胞的另外的CAS9质粒
Figure BDA0003748329410000561
对于所有质粒,使用滚环扩增(RCA)扩增质粒并使质粒适合使用TruPrime RCA试剂盒(Sygnis)进行转化。
实例3
经修饰的宿主菌株的构建
在本实例中,将一系列宿主修饰引入亲本地衣芽孢杆菌菌株。亲本地衣芽孢杆菌菌株含有serA1(SEQ ID NO:30)和lysA基因(SEQ ID NO:87)的缺失,并命名为BF140。
将含有pBl.comK质粒(SEQ ID NO:88)的BF140版本(Liu和Zuber,1998,Hamoen等人,1998)(该质粒含有壮观霉素标志物(SEQ ID NO:89)、编码枯草芽孢杆菌的XylR阻遏物(SEQ ID NO:90)和xylA启动子(SEQ ID NO:91)的DNA(其可操作地连接到编码地衣芽孢杆菌ComK蛋白(SEQ ID NO:92)的DNA上)用靶向catH基因座的线性PCR产物转化,该基因座用于整合地衣芽孢杆菌的prsA基因(SEQ ID NO:93)的第二拷贝。该构建体含有与catH启动子(SEQ ID NO:95)可操作地连接的catH基因座(SEQ ID NO:94)的上游同源臂、编码与双终止子(SEQ ID NO:97)可操作地连接的CatH蛋白的DNA(SEQ ID NO:96)(双终止子由与枯草芽孢杆菌的spoVG终止子(SEQ ID NO:99)可操作地连接的catH终止子(SEQ ID NO:98)构成)。
然后该构建体包含以下:地衣芽孢杆菌的prsA启动子(SEQ ID NO:100),其可操作地连接至prsA编码序列(SEQ ID NO:101),其可操作地连接至地衣芽孢杆菌的amyL基因的终止子(SEQ ID NO:102),其可操作地连接至catH基因座的下游同源臂(SEQ ID NO:103)。简而言之,产生了BF140/pBl.comK感受态细胞。在37℃和250RPM振荡下,在含有一百(100)ppm壮观霉素的L培养液中将BF140/pBl.comK菌株生长过夜。第二天在含有一百(100)ppm壮观霉素的新鲜L培养液中将培养物稀释至0.7的OD600。这种新的培养物在37℃、250RPM振荡下生长一(1)小时。将D-木糖添加至0.1%w·v-1。将培养物在37℃和250RPM振荡下再生长四(4)小时。将细胞以1700·g收获七(7)分钟。将细胞重新悬浮在1/4体积的含有10%v·v- 1DMSO的用过的培养基中。一百(100)μl细胞与十(10)μl catH::[catH prsAp-prsA]整合片段(SEQ ID NO:94)混合。将细胞/DNA混合物在1400RPM、37℃孵育一个半(1.5)小时。然后将混合物铺在含有十(10)ppm氯霉素的L琼脂板上。将接种的板在37℃孵育四十八(48)小时。
使用菌落PCR筛选在含有十(10)ppm氯霉素的L琼脂上形成的菌落,以使用表14中列出的引物和标准PCR技术确认catH基因座的修饰。
表14
正向和反向引物对
正向 TCGTTCTGAATGAGCAAGCA SEQ ID NO:104
反向 TGTTAATCAGGCCGACGATC SEQ ID NO:105
将该PCR产物,2676bp片段(SEQ ID NO:106),使用桑格的方法和表15中列出的引物进行测序。
表15
桑格测序引物
1915 AACCTATATGACAGCCGGAG SEQ ID NO:107
1916 GGCAAAATCCACTTAAGCCAC SEQ ID NO:108
1054 AACGAGTTGGAACGGCTTGC SEQ ID NO:109
具有正确catH::[catH prsAp-prsA]整合(SEQ ID NO:93)的分离株作为菌株BF547储存。
如上所述,使含有pBl.comK质粒(SEQ ID NO:88)的BF547版本成为感受态。将一百(100)μl感受态细胞与五(5)μl pRF946(SEQ ID NO:81)RCA混合,并在1400RPM、37℃孵育一个半(1.5)小时。将混合物铺在含有二十(20)ppm卡那霉素的L琼脂平板上,以选择质粒转化。将板在37℃孵育四十八(48)小时。
筛选在含有二十(20)ppm卡那霉素的L琼脂上形成的菌落进行菌落PCR,以使用标准PCR技术和上表14中列出的引物确认编码catH启动子3’末端的DNA和编码CatH蛋白的DNA(SEQ ID NO:110)的缺失,同时保留catH::[prsAp-prsA]盒(SEQ ID NO:111)。
含有catH::[prsAp-prsA]盒(SEQ ID NO:111)的正确菌落产生1990bp(SEQ IDNO:112)的PCR产物,而含有catH::[catH prsAp-prsA]盒(SEQ ID NO:93)的亲本菌落产生PCR产物长度为2676bp(SEQ ID NO:106)。使用标准凝胶电泳技术目测评估差异。使用上面表15中的引物1915(SEQ ID NO:107)和引物1916(SEQ ID NO:108)对具有正确大小的PCR产物的分离株进行测序。
含有catH::[prsAp-prsA]盒(SEQ ID NO:111)并且对氯霉素(10ppm)表型敏感的经序列验证的分离株被储存为BF561。
如上所述,使含有pBl.comK质粒(SEQ ID NO:88)的BF561版本成为感受态。将一百(100)μl感受态细胞与五(5)μl pZM221(SEQ ID NO:84)或pRF879(SEQ ID NO:78)RCA混合,并在1400RPM和37℃孵育一个半(1.5)小时。将混合物铺在含有二十(20)ppm卡那霉素的L琼脂平板上以选择用质粒转化的细胞。
对于用pZM221(SEQ ID NO:84)转化并在含有二十(20)ppm卡那霉素的L琼脂平板上形成菌落的细胞,针对ΔdltA-2等位基因(SEQ ID NO:86),dltA编码序列的700bp缺失,使用标准PCR技术和表16中的引物筛选菌落。
表16
正向和反向引物对
正向 GGGTACCTCCATGGTAAAGT SEQ ID NO:113
反向 ACGTATTAATGCAGTAGCCG SEQ ID NO:114
具有ΔdltA-2等位基因的菌落在使用表16中的引物时产生2067bp的PCR产物(SEQID NO:115),而含有完整dltA基因的亲本细胞产生2767bp的PCR产物(SEQ ID NO:116)。这可以使用标准电泳技术来区分。含有dltA(SEQ ID NO:86)的700bp内部缺失的菌落作为BF598储存。
对于用pRF879(SEQ ID NO:78)转化并在含有二十(20)ppm卡那霉素的L琼脂平板上形成菌落的细胞,针对ΔrghR2等位基因(SEQ ID NO:80)、rghR2编码序列的缺失(除了前九个(9)和后九个(9)bp)使用标准PCR技术和下表17中的引物筛选菌落。
表17
正向和反向引物对
正向 GGATACGCCGATTTCAATGGC SEQ ID NO:117
反向 GGCTATGTGCTGGGGGAATT SEQ ID NO:118
具有ΔrghR2等位基因(SEQ ID NO:80)的菌落在使用表17中的引物时产生1523bp的PCR产物(SEQ ID NO:119),而含有完整rghR2基因的亲本细胞产生1922bp的PCR产物(SEQID NO:120)。使用标准电泳技术可以区分这两种产物之间的差异。将含有rghR2基因(SEQID NO:84)缺失的菌落作为BF602储存。
如上所述,使含有pBl.comK质粒(SEQ ID NO:88)的BF598版本成为感受态。将一百(100)μl感受态细胞与五(5)μl pRF879(SEQ ID NO:78)RCA混合,并在1400RPM且37℃孵育一个半(1.5)小时。将混合物铺在含有二十(20)ppm卡那霉素的L琼脂平板上以选择用质粒转化的细胞。
对于用pRF879(SEQ ID NO:78)转化并在含有二十(20)ppm卡那霉素的L琼脂平板上形成菌落的细胞,针对ΔrghR2等位基因(SEQ ID NO:80)、rghR2编码序列的缺失(除了前九个(9)和后九个(9)bp)使用标准PCR技术和上表17中的引物筛选菌落。
具有ΔrghR2等位基因(SEQ ID NO:80)的菌落在使用表17中的引物时产生1523bp的PCR产物(SEQ ID NO:119),而含有完整rghR2基因的亲本细胞产生1922bp的PCR产物(SEQID NO:120)。使用标准电泳技术可以区分这两种产物之间的差异。将含有rghR2基因(SEQID NO:80)缺失的菌落作为BF613储存。下表18显示了在本实例中产生的经修饰的宿主菌株,其具有在该实例中的三(3)个经修饰的基因座的SEQ ID号。
表18
经修饰的宿主菌株
Figure BDA0003748329410000601
实例4
在经修饰的宿主菌株中构建表达淀粉酶的菌株
在本实例中,将一系列淀粉酶和淀粉酶变体表达盒引入以上实例2表18中列出的菌株谱系中。
淀粉酶1
淀粉酶1(SEQ ID NO:126)是地衣芽孢杆菌的天然α淀粉酶,通常称为AmyL。淀粉酶1(SEQ ID NO:127)的第一盒被整合到serA1基因座(SEQ ID NO:44)中并包含以下:serA1ORF(SEQ ID NO:30)和合成的p3启动子(SEQ ID NO:128),其可操作地连接至编码经修饰的枯草芽孢杆菌aprE 5’UTR(SEQ ID NO:129)的DNA,其可操作地连接至编码地衣芽孢杆菌AmyL信号序列(SEQ ID NO:130)的DNA,其可操作地连接至编码淀粉酶1(SEQ ID NO:131)的DNA,其可操作地连接至地衣芽孢杆菌amyL转录终止子(SEQ ID NO:102)。整合在lysA基因座(SEQ ID NO:133)中的淀粉酶1的第二盒(SEQ ID NO:132)包含编码LysA的DNA(SEQ IDNO:134)和合成的p2启动子(SEQ ID NO:135),其可操作地连接至编码经修饰的枯草芽孢杆菌aprE 5’UTR(SEQ ID NO:129)的DNA,其可操作地连接至编码地衣芽孢杆菌AmyL信号序列(SEQ ID NO:130)的DNA,其可操作地连接至编码淀粉酶1(SEQ ID NO:131)的DNA,其可操作地连接至地衣芽孢杆菌amyL转录终止子(SEQ ID NO:102)。
淀粉酶2
淀粉酶2(SEQ ID NO:136)是在PCT公开号WO 2018/184004(通过引用以其整体并入本文)中描述的变体芽孢杆菌属物种α-淀粉酶。淀粉酶2(SEQ ID NO:137)的第一盒被整合到serA1基因座(SEQ ID NO:44)中并包含以下:serA1 ORF(SEQ ID NO:30)和枯草芽孢杆菌rrnI启动子(SEQ ID NO:138),其可操作地连接至编码枯草芽孢杆菌aprE5’UTR(SEQ IDNO:139)的DNA,其可操作地连接至编码地衣芽孢杆菌AmyL信号序列(SEQ ID NO:130)的DNA,其可操作地连接至编码淀粉酶2(SEQ ID NO:140)的DNA,其可操作地连接至地衣芽孢杆菌amyL转录终止子(SEQ ID NO:102)。整合在lysA基因座(SEQ ID NO:133)或amyL基因座(SEQ ID NO:142)中的淀粉酶2(SEQ ID NO:141)的第二盒包含编码LysA的DNA(SEQ ID NO:134)和合成的p3启动子(SEQ ID NO:128),其可操作地连接至编码枯草芽孢杆菌aprE 5’UTR(SEQ ID NO:139)的DNA,其可操作地连接至编码地衣芽孢杆菌AmyL信号序列(SEQ IDNO:130)的DNA,其可操作地连接至编码淀粉酶2(SEQ ID NO:140)的DNA,其可操作地连接至地衣芽孢杆菌amyL转录终止子(SEQ ID NO:102)。
淀粉酶3
淀粉酶3(SEQ ID NO:143)是变体噬细胞菌属物种α-淀粉酶(例如,参见PCT公开号WO 2014/164777;WO 2012/164800和WO2014/16483,各自通过引用以其整体并入本文)。淀粉酶3(SEQ ID NO:144)的第一盒被整合到serA1基因座(SEQ ID NO:44)中并包含以下:serA1 ORF(SEQ ID NO:30)和合成的p3启动子(SEQ ID NO:128),其可操作地连接至编码经修饰的枯草芽孢杆菌aprE 5’UTR(SEQ ID NO:129)的DNA,其可操作地连接至编码地衣芽孢杆菌AmyL信号序列(SEQ ID NO:130)的DNA,其可操作地连接至编码淀粉酶3(SEQ ID NO:145)的DNA,其可操作地连接至地衣芽孢杆菌amyL转录终止子(SEQ ID NO:102)。整合在lysA基因座(SEQ ID NO:133)中的第二淀粉酶3盒(SEQ ID NO:146)包含编码LysA的DNA(SEQ ID NO:134)和合成的p2启动子(SEQ ID NO:135),其可操作地连接至编码经修饰的枯草芽孢杆菌aprE 5’UTR(SEQ ID NO:129)的DNA,其可操作地连接至编码地衣芽孢杆菌AmyL信号序列(SEQ ID NO:130)的DNA,其可操作地连接至编码淀粉酶3(SEQ ID NO:145)的DNA,其可操作地连接至地衣芽孢杆菌amyL转录终止子(SEQ ID NO:102)。
淀粉酶4
淀粉酶4(SEQ ID NO:147)是变体噬细胞菌属物种α-淀粉酶(例如,参见PCT公开号WO 2014/164777;WO 2012/164800和WO2014/16483,各自通过引用以其整体并入本文)。淀粉酶4(SEQ ID NO:148)的第一盒被整合到serA1基因座(SEQ ID NO:44)中并包含以下:serA1 ORF(SEQ ID NO:30)和合成的p3启动子(SEQ ID NO:128),其可操作地连接至编码枯草芽孢杆菌aprE 5’UTR(SEQ ID NO:139)的DNA,其可操作地连接至编码地衣芽孢杆菌AmyL信号序列(SEQ ID NO:130)的DNA,其可操作地连接至编码淀粉酶4(SEQ ID NO:149)的DNA,其可操作地连接至地衣芽孢杆菌amyL转录终止子(SEQ ID NO:129)。整合在lysA基因座(SEQ ID NO:133)中的第二淀粉酶4盒(SEQ ID NO:150)包含编码LysA的DNA(SEQ ID NO:134)和合成的p2启动子(SEQ ID NO:135),其可操作地连接至编码枯草芽孢杆菌aprE 5’UTR(SEQ ID NO:139)的DNA,其可操作地连接至编码地衣芽孢杆菌AmyL信号序列(SEQ IDNO:130)的DNA,其可操作地连接至编码淀粉酶4(SEQ ID NO:149)的DNA,其可操作地连接至地衣芽孢杆菌amyL转录终止子(SEQ ID NO:102)。
淀粉酶5
淀粉酶5(SEQ ID NO:151)是变体芽孢杆菌属物种707α-淀粉酶(参见PCT公开号WO2008/153805和美国专利公开号US 2014/0057324)。淀粉酶5(SEQ ID NO:152)的第一盒被整合到serA1基因座(SEQ ID NO:44)中并包含以下:serA1 ORF(SEQ ID NO:30)和合成的p3启动子(SEQ ID NO:128),其可操作地连接至编码枯草芽孢杆菌aprE 5’UTR(SEQ ID NO:139)的DNA,其可操作地连接至编码地衣芽孢杆菌AmyL信号序列(SEQ ID NO:130)的DNA,其可操作地连接至编码淀粉酶5(SEQ ID NO:153)的DNA,其可操作地连接至地衣芽孢杆菌amyL转录终止子(SEQ ID NO:102)。整合在lysA基因座(SEQ ID NO:133)中的第二淀粉酶5盒(SEQ ID NO:154)包含编码LysA的DNA(SEQ ID NO:134)和合成的p2启动子(SEQ ID NO:135),其可操作地连接至编码枯草芽孢杆菌aprE 5’UTR(SEQ ID NO:139)的DNA,其可操作地连接至编码地衣芽孢杆菌AmyL信号序列(SEQ ID NO:130)的DNA,其可操作地连接至编码淀粉酶5(SEQ ID NO:153)的DNA,其可操作地连接至地衣芽孢杆菌amyL转录终止子(SEQ IDNO:102)。
使用PCT公开号WO 2019/040412(通过引用以其整体并入本文)所述的方法将所有淀粉酶表达盒转化到经修饰的宿主菌株中。
实例5
经修饰的宿主背景对淀粉酶产生的影响
在本实例中,使用标准的小规模或实验室规模发酵条件测定含有淀粉酶1-5表达盒(实例4)的两个拷贝的经修饰宿主菌株(即,表19;BF140、BF561、BF598、BF602和BF613)的α-淀粉酶产生(如PCT公开号WO 2018/156705和WO 2019/055261中描述,各自通过引用并入本文)。使用Bradford方法或Ceralpha测定法对α-淀粉酶产生进行量化。将淀粉酶产生方面的相对改善与包含下表19中呈现的相同α-淀粉酶表达盒的未修饰的宿主进行比较。
表19
不同的经修饰的宿主菌株的关于淀粉酶生产的相对性能
Figure BDA0003748329410000641
因此,从不同的α-淀粉酶组中测试的所有五(5)种淀粉酶都证明了与未修饰的宿主BF140相比在BF613修饰的背景(其包括dltA-2等位基因(SEQ ID NO:125)缺失(ΔdltA-2),rghR2等位基因(SEQ ID NO:80)缺失(ΔrghR2)和由天然prsA启动子(SEQ ID NO:124)控制的天然prsA基因的第二拷贝的***)中α-淀粉酶产生的改善。
对于淀粉酶2和淀粉酶3,在BF602修饰的背景中α-淀粉酶产生方面的改善(包括rghR2(ΔrghR2)等位基因(SEQ ID NO:80)缺失和由天然prsA启动子(SEQ ID NO:124)控制的天然prsA基因的第二拷贝)几乎与BF613修饰的宿主中看到的改善一样好,这表明对于某些淀粉酶,改善只需要这两个等位基因的存在,而且ΔdltA-2等位基因的存在对这种改善没有害处。
参考文献
PCT Publication No.WO 1989/06279
PCT Publication No.WO 1990/11352
PCT Publication No.WO 1994/18314
PCT Publication No.WO 1999/19467
PCT Publication No.WO 1999/20726
PCT Publication No.WO 1999/20769
PCT Publication No.WO 1999/20770
PCT Publication No.WO 1999/43794
PCT Publication No.WO 2000/29560
PCT Publication No.WO 2000/60059
PCT Publication No.WO 2001/51643
PCT Publication No.WO 2002/14490
PCT Publication No.WO 2003/083125
PCT Publication No.WO 2003/089604
PCT Publication No.WO 2006/037483
PCT Publication No.WO 2006/037484
PCT Publication No.WO 2006/089107
PCT Publication No.WO 2008/112459
PCT Publication No.WO 2014/164777
PCT Publication No.WO 2019/040412
PCT Publication No.WO 2018/156705
PCT Publication No.WO 2019/055261
U.S.Publication No.US 2014/0329309
US Patent No.4,914,031
US Patent No.4,980,288
US Patent No.5,208,158
US Patent No.5,310,675
US Patent No.5,336,611
US Patent No.5,399,283
US Patent No.5,441,882
US Patent No.5,482,849
US Patent No.5,665,587
US Patent No.5,700,676
US Patent No.5,741,694
US Patent No.5,858,757
US Patent No.5,880,080
US Patent No.6,197,567
US Patent No.6,218,165.
US RE34,606
Albertini and Galizzi,Bacteriol.,162:1203-1211,1985.
Bergmeyer et al.,"Methods of Enzymatic Analysis"vol.5,Peptidases,Proteinases and their Inhibitors,Verlag Chemie,Weinheim,1984.
Botstein and Shortle,Science 229:4719,1985.
Brode et al.,“Subtilisin BPN'variants:increased hydrolytic activityonsurface-bound substrates via decreased surface activity”,Biochemistry,35(10):3162-3169,1996.
Caspers et al.,“Improvement of Sec-dependent secretion ofaheterologous model protein in Bacillus subtilis by saturation mutagenesisofthe N-domain of the AmyE signal peptide”,Appl.Microbiol.Biotechnol.,86(6):1877-1885,2010.
Chang et al.,Mol.Gen.Genet.,168:11-115,1979.
Christianson et al.,Anal.Biochem.,223:119-129,1994.
Devereux et a/.,Nucl.Acid Res.,12:387-395,1984.
Earl et al.,“Ecology and genomics of Bacillus subtilis”,Trends inMicrobiology.,16(6):269-275,2008.
Ferrari et al.,"Genetics,"in Harwood et al.(ed.),Bacillus,PlenumPublishing Corp.,1989.
Fisher et.al.,Arch.Microbiol.,139:213-217,1981.
Guerot-Fleury,Gene,167:335-337,1995.
Hamoen et al.,“Controlling competence in Bacillus subtilis:sharedused of regulators”,Microbiology,149:9-17,2003.
Hamoen et al.,Genes Dev.12:1539-1550,1998.
Higuchi et al.,Nucleic Acids Research 16:7351,1988.
Ho et al.,Gene 77:61,1989.
Hoch et al.,J.Bacteriol.,93:1925-1937,1967.
Holubova,Folia Microbiol.,30:97,1985.
Hopwood,The Isolation of Mutants in Methods in Microbiology(J.R.Norris and D.W.Ribbons,eds.)pp 363-433,Academic Press,New York,1970.
Horton et al.,Gene 77:61,1989.
Hsia et al.,Anal Biochem.,242:221-227,1999.
Iglesias and Trautner,Molecular General Genetics 189:73-76,1983.
Jensen et al.,“Cell-associated degradation affects the yield ofsecreted engineered and heterologous proteins in the Bacillus subtilisexpression system”Microbiology,146(Pt 10:2583-2594,2000.
Kontinen and Sarvas,“The PrsA lipoprotein is essential for proteinsecretion in Bacillus subtilis and sets a limit for high-level secretion”,Mol.Microbiol.May;8(4):727-737,1993.
Liu and Zuber,1998,
Lo et al.,Proceedings of the National Academy of Sciences USA 81:2285,1985.
May et al.“Inhibition of the D-alanine:D-alanyl carrier proteinligase in Bacillus subtilis increases the bacterium’s susceptibility toantibiotics that target the cell wall”,FEBS Journal,272:2993-3003,2005.
McDonald,J.Gen.Microbiol.,130:203,1984.
Needleman and Wunsch,J.Mol.Biol.,48:443,1970.
Ogura&Fujita,FEMS Microbiol Lett.,268(1):73-80.2007.
Olempska-Beer et al.,“Food-processing enzymes from recombinantmicroorganisms--a review”’Regul.Toxicol.Pharmacol.,45(2):144-158,2006.
Palmeros et al.,Gene 247:255-264,2000.
Raul et al.,“Production and partial purification of alpha amylasefrom Bacillus subtilis(MTCC 121)using solid state fermentation”,BiochemistryResearch International,2014.
Sarkar and Sommer,BioTechniques 8:404,1990.
Saunders et al.,J.Bacteriol.,157:718-726,1984.
Shimada,Meth.Mol.Biol.57:157;1996
Smith and Waterman,Adv.Appl.Math.,2:482,1981.
Smith et al.,Appl.Env.Microbiol.,51:634 1986.
Stahl and Ferrari,J.Bacteriol.,158:411-418,1984.
Stahl et al,J.Bacteriol.,158:411-418,1984.
Trieu-Cuot et al.,Gene,23:331-341,1983.
Van Dijl and Hecker,“Bacillus subtilis:from soil bacterium to super-secreting cell factory”,Microbial Cell Factories,12(3).2013.
Vorobjeva et al.,FEMS Microbiol.Lett.,7:261-263,1980.
Ward,"Proteinases,"in Fogarty(ed.).,Microbial Enzymes andBiotechnology.Applied Science,London,pp 251-317,1983.
Wells et al.,Nucleic Acids Res.11:7911-7925,1983.
Westers et al.,“Bacillus subtilis as cell factory for pharmaceuticalproteins:a biotechnological approach to optimize the host organism”,Biochimica et Biophysica Acta.,1694:299-310,2004.
序列表
<110> 美国丹尼斯科公司(DANISCO US INC. )
<120> 用于增强地衣芽孢杆菌中蛋白产生的组合物和方法
<130> NB41684-WO-PCT
<160> 158
<170> PatentIn 3.5版
<210> 1
<211> 1368
<212> PRT
<213> 化脓链球菌(Streptococcus pyogenes)
<400> 1
Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val
1 5 10 15
Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe
20 25 30
Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile
35 40 45
Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu
50 55 60
Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys
65 70 75 80
Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser
85 90 95
Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys
100 105 110
His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr
115 120 125
His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp
130 135 140
Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His
145 150 155 160
Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro
165 170 175
Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr
180 185 190
Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala
195 200 205
Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn
210 215 220
Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn
225 230 235 240
Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe
245 250 255
Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp
260 265 270
Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp
275 280 285
Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp
290 295 300
Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser
305 310 315 320
Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys
325 330 335
Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe
340 345 350
Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser
355 360 365
Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp
370 375 380
Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg
385 390 395 400
Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu
405 410 415
Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe
420 425 430
Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile
435 440 445
Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp
450 455 460
Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu
465 470 475 480
Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr
485 490 495
Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser
500 505 510
Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys
515 520 525
Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln
530 535 540
Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr
545 550 555 560
Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp
565 570 575
Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly
580 585 590
Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp
595 600 605
Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr
610 615 620
Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala
625 630 635 640
His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr
645 650 655
Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp
660 665 670
Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe
675 680 685
Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe
690 695 700
Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu
705 710 715 720
His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly
725 730 735
Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly
740 745 750
Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln
755 760 765
Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile
770 775 780
Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro
785 790 795 800
Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu
805 810 815
Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg
820 825 830
Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys
835 840 845
Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg
850 855 860
Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys
865 870 875 880
Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys
885 890 895
Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp
900 905 910
Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr
915 920 925
Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp
930 935 940
Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser
945 950 955 960
Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg
965 970 975
Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val
980 985 990
Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe
995 1000 1005
Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala
1010 1015 1020
Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe
1025 1030 1035
Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala
1040 1045 1050
Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu
1055 1060 1065
Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val
1070 1075 1080
Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr
1085 1090 1095
Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys
1100 1105 1110
Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro
1115 1120 1125
Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val
1130 1135 1140
Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys
1145 1150 1155
Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser
1160 1165 1170
Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys
1175 1180 1185
Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu
1190 1195 1200
Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly
1205 1210 1215
Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val
1220 1225 1230
Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser
1235 1240 1245
Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys
1250 1255 1260
His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys
1265 1270 1275
Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala
1280 1285 1290
Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn
1295 1300 1305
Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala
1310 1315 1320
Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser
1325 1330 1335
Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr
1340 1345 1350
Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp
1355 1360 1365
<210> 2
<211> 4188
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 2
gtggccccaa aaaagaaacg caaggttatg gataaaaaat acagcattgg tctggatatc 60
ggaaccaaca gcgttgggtg ggcagtaata acagatgaat acaaagtgcc gtcaaaaaaa 120
tttaaggttc tggggaatac agatcgccac agcataaaaa agaatctgat tggggcattg 180
ctgtttgatt cgggtgagac agctgaggcc acgcgtctga aacgtacagc aagaagacgt 240
tacacacgtc gtaaaaatcg tatttgctac ttacaggaaa ttttttctaa cgaaatggcc 300
aaggtagatg atagtttctt ccatcgtctc gaagaatctt ttctggttga ggaagataaa 360
aaacacgaac gtcaccctat ctttggcaat atcgtggatg aagtggccta tcatgaaaaa 420
taccctacga tttatcatct tcgcaagaag ttggttgata gtacggacaa agcggatctg 480
cgtttaatct atcttgcgtt agcgcacatg atcaaatttc gtggtcattt cttaattgaa 540
ggtgatctga atcctgataa ctctgatgtg gacaaattgt ttatacaatt agtgcaaacc 600
tataatcagc tgttcgagga aaaccccatt aatgcctctg gagttgatgc caaagcgatt 660
ttaagcgcga gactttctaa gtcccggcgt ctggagaatc tgatcgccca gttaccaggg 720
gaaaagaaaa atggtctgtt tggtaatctg attgccctca gtctggggct taccccgaac 780
ttcaaatcca attttgacct ggctgaggac gcaaagctgc agctgagcaa agatacttat 840
gatgatgacc tcgacaatct gctcgcccag attggtgacc aatatgcgga tctgtttctg 900
gcagcgaaga atctttcgga tgctatcttg ctgtcggata ttctgcgtgt taataccgaa 960
atcaccaaag cgcctctgtc tgcaagtatg atcaagagat acgacgagca ccaccaggac 1020
ctgactcttc ttaaggcact ggtacgccaa cagcttccgg agaaatacaa agaaatattc 1080
ttcgaccagt ccaagaatgg ttacgcgggc tacatcgatg gtggtgcatc acaggaagag 1140
ttctataaat ttattaaacc aatccttgag aaaatggatg gcacggaaga gttacttgtt 1200
aaacttaacc gcgaagactt gcttagaaag caacgtacat tcgacaacgg ctccatccca 1260
caccagattc atttaggtga acttcacgcc atcttgcgca gacaagaaga tttctatccc 1320
ttcttaaaag acaatcggga gaaaatcgag aagatcctga cgttccgcat tccctattat 1380
gtcggtcccc tggcacgtgg taattctcgg tttgcctgga tgacgcgcaa aagtgaggaa 1440
accatcaccc cttggaactt tgaagaagtc gtggataaag gtgctagcgc gcagtctttt 1500
atagaaagaa tgacgaactt cgataaaaac ttgcccaacg aaaaagtcct gcccaagcac 1560
tctcttttat atgagtactt tactgtgtac aacgaactga ctaaagtgaa atacgttacg 1620
gaaggtatgc gcaaacctgc ctttcttagt ggcgagcaga aaaaagcaat tgtcgatctt 1680
ctctttaaaa cgaatcgcaa ggtaactgta aaacagctga aggaagatta tttcaaaaag 1740
atcgaatgct ttgattctgt cgagatctcg ggtgtcgaag atcgtttcaa cgcttcctta 1800
gggacctatc atgatttgct gaagataata aaagacaaag actttctcga caatgaagaa 1860
aatgaagata ttctggagga tattgttttg accttgacct tattcgaaga tagagagatg 1920
atcgaggagc gcttaaaaac ctatgcccac ctgtttgatg acaaagtcat gaagcaatta 1980
aagcgccgca gatatacggg gtggggccgc ttgagccgca agttgattaa cggtattaga 2040
gacaagcaga gcggaaaaac tatcctggat ttcctcaaat ctgacggatt tgcgaaccgc 2100
aattttatgc agcttataca tgatgattcg cttacattca aagaggatat tcagaaggct 2160
caggtgtctg ggcaaggtga ttcactccac gaacatatag caaatttggc cggctctcct 2220
gcgattaaga aggggatcct gcaaacagtt aaagttgtgg atgaacttgt aaaagtaatg 2280
ggccgccaca agccggagaa tatcgtgata gaaatggcgc gcgagaatca aacgacacaa 2340
aaaggtcaaa agaactcaag agagagaatg aagcgcattg aggaggggat aaaggaactt 2400
ggatctcaaa ttctgaaaga acatccagtt gaaaacactc agctgcaaaa tgaaaaattg 2460
tacctgtact acctgcagaa tggaagagac atgtacgtgg atcaggaatt ggatatcaat 2520
agactctcgg actatgacgt agatcacatt gtccctcaga gcttcctcaa ggatgattct 2580
atagataata aagtacttac gagatcggac aaaaatcgcg gtaaatcgga taacgtccca 2640
tcggaggaag tcgttaaaaa gatgaaaaac tattggcgtc aactgctgaa cgccaagctg 2700
atcacacagc gtaagtttga taatctgact aaagccgaac gcggtggtct tagtgaactc 2760
gataaagcag gatttataaa acggcagtta gtagaaacgc gccaaattac gaaacacgtg 2820
gctcagatcc tcgattctag aatgaataca aagtacgatg aaaacgataa actgatccgt 2880
gaagtaaaag tcattacctt aaaatctaaa cttgtgtccg atttccgcaa agattttcag 2940
ttttacaagg tccgggaaat caataactat caccatgcac atgatgcata tttaaatgcg 3000
gttgtaggca cggcccttat taagaaatac cctaaactcg aaagtgagtt tgtttatggg 3060
gattataaag tgtatgacgt tcgcaaaatg atcgcgaaat cagaacagga aatcggtaag 3120
gctaccgcta aatacttttt ttattccaac attatgaatt tttttaagac cgaaataact 3180
ctcgcgaatg gtgaaatccg taaacggcct cttatagaaa ccaatggtga aacgggagaa 3240
atcgtttggg ataaaggtcg tgactttgcc accgttcgta aagtcctctc aatgccgcaa 3300
gttaacattg tcaagaagac ggaagttcaa acagggggat tctccaaaga atctatcctg 3360
ccgaagcgta acagtgataa acttattgcc agaaaaaaag attgggatcc aaaaaaatac 3420
ggaggctttg attcccctac cgtcgcgtat agtgtgctgg tggttgctaa agtcgagaaa 3480
gggaaaagca agaaattgaa atcagttaaa gaactgctgg gtattacaat tatggaaaga 3540
tcgtcctttg agaaaaatcc gatcgacttt ttagaggcca aggggtataa ggaagtgaaa 3600
aaagatctca tcatcaaatt accgaagtat agtctttttg agctggaaaa cggcagaaaa 3660
agaatgctgg cctccgcggg cgagttacag aagggaaatg agctggcgct gccttccaaa 3720
tatgttaatt ttctgtacct tgccagtcat tatgagaaac tgaagggcag ccccgaagat 3780
aacgaacaga aacaattatt cgtggaacag cataagcact atttagatga aattatagag 3840
caaattagtg aattttctaa gcgcgttatc ctcgcggatg ctaatttaga caaagtactg 3900
tcagcttata ataaacatcg ggataagccg attagagaac aggccgaaaa tatcattcat 3960
ttgtttacct taaccaacct tggagcacca gctgccttca aatatttcga taccacaatt 4020
gatcgtaaac ggtatacaag tacaaaagaa gtcttggacg caaccctcat tcatcaatct 4080
attactggat tatatgagac acgcattgat ctttcacagc tgggcggaga caagaagaaa 4140
aaactgaaac tgcaccatca tcaccatcat catcaccatc attgataa 4188
<210> 3
<211> 8
<212> PRT
<213> 人工序列
<220>
<223> 合成的
<400> 3
Ala Pro Lys Lys Lys Arg Lys Val
1 5
<210> 4
<211> 6
<212> PRT
<213> 人工序列
<220>
<223> 合成的
<400> 4
Lys Lys Lys Lys Leu Lys
1 5
<210> 5
<211> 10
<212> PRT
<213> 人工序列
<220>
<223> 合成的
<400> 5
His His His His His His His His His His
1 5 10
<210> 6
<211> 607
<212> DNA
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 6
attcctccat tttcttctgc tatcaaaata acagactcgt gattttccaa acgagctttc 60
aaaaaagcct ctgccccttg caaatcggat gcctgtctat aaaattcccg atattggtta 120
aacagcggcg caatggcggc cgcatctgat gtctttgctt ggcgaatgtt catcttattt 180
cttcctccct ctcaataatt ttttcattct atcccttttc tgtaaagttt atttttcaga 240
atacttttat catcatgctt tgaaaaaata tcacgataat atccattgtt ctcacggaag 300
cacacgcagg tcatttgaac gaattttttc gacaggaatt tgccgggact caggagcatt 360
taacctaaaa aagcatgaca tttcagcata atgaacattt actcatgtct attttcgttc 420
ttttctgtat gaaaatagtt atttcgagtc tctacggaaa tagcgagaga tgatatacct 480
aaatagagat aaaatcatct caaaaaaatg ggtctactaa aatattattc catctattac 540
aataaattca cagaatagtc ttttaagtaa gtctactctg aattttttta aaaggagagg 600
gtaacta 607
<210> 7
<211> 247
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 7
acataaaaaa ccggccttgg ccccgccggt tttttattat ttttcttcct ccgcatgttc 60
aatccgctcc ataatcgacg gatggctccc tctgaaaatt ttaacgagaa acggcgggtt 120
gacccggctc agtcccgtaa cggccaagtc ctgaaacgtc tcaatcgccg cttcccggtt 180
tccggtcagc tcaatgccgt aacggtcggc ggcgttttcc tgataccggg agacggcatt 240
cgtaatc 247
<210> 8
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 8
atatatgagt aaacttggtc tgacagaatt cctccatttt cttctgctat 50
<210> 9
<211> 35
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 9
tgcggccgcg aattcgatta cgaatgccgt ctccc 35
<210> 10
<211> 3290
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 10
gaattcgcgg ccgcacgcgt ccatggggat ccccgcgggt cgacctcgag agttacgcta 60
gggataacag ggtaatatag gagctccagt cggcttaaac cagttttcgc tggtgcgaaa 120
aaagagtgtc ttgtgacacc taaattcaaa atctatcggt cagatttata ccgatttgat 180
tttatatatt cttgaataac atacgccgag ttatcacata aaagcgggaa ccaatcataa 240
aatttaaact tcattgcata atccattaaa ctcttaaatt ctacgattcc ttgttcatca 300
ataaactcaa tcatttcttt aattaattta tatctatctg ttgttgtttt ctttaataat 360
tcattaacat ctacaccgcc ataaactatc atatcttctt tttgatattt aaatttatta 420
ggatcgtcca tgtgaagcat atatctcaca agacctttca cacttcctgc aatctgcgga 480
atagtcgcat tcaattcttc tgttaattat ttttatctgt tcataagatt tattaccctc 540
atacatcact agaatatgat aatgctcttt tttcatccta ccttctgtat cagtatccct 600
atcatgtaat ggagacacta caaattgaat gtgtaactct tttaaatact ctaaccactc 660
ggcttttgct gattctggat ataaaacaaa tgtccaatta cgtcctcttg aatttttctt 720
gttttcagtt tcttttatta cattttcgct catgatataa taacggtgct aatacactta 780
acaaaattta gtcatagata ggcagcatgc cagtgctgtc tatctttttt tgtttaaaat 840
gcaccgtatt cctcctttgc atattttttt attagaatac cggttgcatc tgatttgcta 900
atattatatt tttctttgat tctatttaat atctcatttt cttctgttgt aagtcttaaa 960
gtaacagcaa cttttttctc ttcttttcta tctacaacta tcactgtacc tcccaacatc 1020
tgtttttttc actttaacat aaaaaacaac cttttaacat taaaaaccca atatttattt 1080
atttgtttgg acaatggaca ctggacacct aggggggagg tcgtagtacc cccctatgtt 1140
ttctccccta aataacccca aaaatctaag aaaaaaagac ctcaaaaagg tctttaatta 1200
acatctcaaa tttcgcattt attccaattt cctttttgcg tgtgatgcga gctcatcggc 1260
tccgtcgata ctatgttata cgccaacttt caaaacaact ttgaaaaagc tgttttctgg 1320
tatttaaggt tttagaatgc aaggaacagt gaattggagt tcgtcttgtt ataattagct 1380
tcttggggta tctttaaata ctgtagaaaa gaggaaggaa ataataaatg gctaaaatga 1440
gaatatcacc ggaattgaaa aaactgatcg aaaaataccg ctgcgtaaaa gatacggaag 1500
gaatgtctcc tgctaaggta tataagctgg tgggagaaaa tgaaaaccta tatttaaaaa 1560
tgacggacag ccggtataaa gggaccacct atgatgtgga acgggaaaag gacatgatgc 1620
tatggctgga aggaaagctg cctgttccaa aggtcctgca ctttgaacgg catgatggct 1680
ggagcaatct gctcatgagt gaggccgatg gcgtcctttg ctcggaagag tatgaagatg 1740
aacaaagccc tgaaaagatt atcgagctgt atgcggagtg catcaggctc tttcactcca 1800
tcgacatatc ggattgtccc tatacgaata gcttagacag ccgcttagcc gaattggatt 1860
acttactgaa taacgatctg gccgatgtgg attgcgaaaa ctgggaagaa gacactccat 1920
ttaaagatcc gcgcgagctg tatgattttt taaagacgga aaagcccgaa gaggaacttg 1980
tcttttccca cggcgacctg ggagacagca acatctttgt gaaagatggc aaagtaagtg 2040
gctttattga tcttgggaga agcggcaggg cggacaagtg gtatgacatt gccttctgcg 2100
tccggtcgat cagggaggat atcggggaag aacagtatgt cgagctattt tttgacttac 2160
tggggatcaa gcctgattgg gagaaaataa aatattatat tttactggat gaattgtttt 2220
agtgactgca gtgagatctg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag 2280
gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg 2340
agtaggacaa atccgccgct ctagctaagc agaaggccat cctgacggat ggcctttttg 2400
cgtttctaca aactcttgtt aactctagag ctgcctgccg cgtttcggtg atgaagatct 2460
tcccgatgat taattaattc agaacgctcg gttgccgccg ggcgtttttt atgaagcttc 2520
gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 2580
aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 2640
ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 2700
cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 2760
ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 2820
cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 2880
agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 2940
gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 3000
gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 3060
tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 3120
agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 3180
agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 3240
atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 3290
<210> 11
<211> 4204
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 11
gcggccgcac gcgtccatgg ggatccccgc gggtcgacct cgagagttac gctagggata 60
acagggtaat ataggagctc cagtcggctt aaaccagttt tcgctggtgc gaaaaaagag 120
tgtcttgtga cacctaaatt caaaatctat cggtcagatt tataccgatt tgattttata 180
tattcttgaa taacatacgc cgagttatca cataaaagcg ggaaccaatc ataaaattta 240
aacttcattg cataatccat taaactctta aattctacga ttccttgttc atcaataaac 300
tcaatcattt ctttaattaa tttatatcta tctgttgttg ttttctttaa taattcatta 360
acatctacac cgccataaac tatcatatct tctttttgat atttaaattt attaggatcg 420
tccatgtgaa gcatatatct cacaagacct ttcacacttc ctgcaatctg cggaatagtc 480
gcattcaatt cttctgttaa ttatttttat ctgttcataa gatttattac cctcatacat 540
cactagaata tgataatgct cttttttcat cctaccttct gtatcagtat ccctatcatg 600
taatggagac actacaaatt gaatgtgtaa ctcttttaaa tactctaacc actcggcttt 660
tgctgattct ggatataaaa caaatgtcca attacgtcct cttgaatttt tcttgttttc 720
agtttctttt attacatttt cgctcatgat ataataacgg tgctaataca cttaacaaaa 780
tttagtcata gataggcagc atgccagtgc tgtctatctt tttttgttta aaatgcaccg 840
tattcctcct ttgcatattt ttttattaga ataccggttg catctgattt gctaatatta 900
tatttttctt tgattctatt taatatctca ttttcttctg ttgtaagtct taaagtaaca 960
gcaacttttt tctcttcttt tctatctaca actatcactg tacctcccaa catctgtttt 1020
tttcacttta acataaaaaa caacctttta acattaaaaa cccaatattt atttatttgt 1080
ttggacaatg gacactggac acctaggggg gaggtcgtag taccccccta tgttttctcc 1140
cctaaataac cccaaaaatc taagaaaaaa agacctcaaa aaggtcttta attaacatct 1200
caaatttcgc atttattcca atttcctttt tgcgtgtgat gcgagctcat cggctccgtc 1260
gatactatgt tatacgccaa ctttcaaaac aactttgaaa aagctgtttt ctggtattta 1320
aggttttaga atgcaaggaa cagtgaattg gagttcgtct tgttataatt agcttcttgg 1380
ggtatcttta aatactgtag aaaagaggaa ggaaataata aatggctaaa atgagaatat 1440
caccggaatt gaaaaaactg atcgaaaaat accgctgcgt aaaagatacg gaaggaatgt 1500
ctcctgctaa ggtatataag ctggtgggag aaaatgaaaa cctatattta aaaatgacgg 1560
acagccggta taaagggacc acctatgatg tggaacggga aaaggacatg atgctatggc 1620
tggaaggaaa gctgcctgtt ccaaaggtcc tgcactttga acggcatgat ggctggagca 1680
atctgctcat gagtgaggcc gatggcgtcc tttgctcgga agagtatgaa gatgaacaaa 1740
gccctgaaaa gattatcgag ctgtatgcgg agtgcatcag gctctttcac tccatcgaca 1800
tatcggattg tccctatacg aatagcttag acagccgctt agccgaattg gattacttac 1860
tgaataacga tctggccgat gtggattgcg aaaactggga agaagacact ccatttaaag 1920
atccgcgcga gctgtatgat tttttaaaga cggaaaagcc cgaagaggaa cttgtctttt 1980
cccacggcga cctgggagac agcaacatct ttgtgaaaga tggcaaagta agtggcttta 2040
ttgatcttgg gagaagcggc agggcggaca agtggtatga cattgccttc tgcgtccggt 2100
cgatcaggga ggatatcggg gaagaacagt atgtcgagct attttttgac ttactgggga 2160
tcaagcctga ttgggagaaa ataaaatatt atattttact ggatgaattg ttttagtgac 2220
tgcagtgaga tctggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag 2280
tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg 2340
acaaatccgc cgctctagct aagcagaagg ccatcctgac ggatggcctt tttgcgtttc 2400
tacaaactct tgttaactct agagctgcct gccgcgtttc ggtgatgaag atcttcccga 2460
tgattaatta attcagaacg ctcggttgcc gccgggcgtt ttttatgaag cttcgttgct 2520
ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 2580
gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 2640
cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 2700
gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 2760
tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 2820
cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 2880
cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 2940
gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 3000
agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 3060
cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 3120
tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 3180
tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 3240
ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 3300
cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 3360
cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 3420
accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 3480
ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 3540
ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 3600
tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 3660
acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 3720
tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 3780
actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 3840
ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 3900
aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 3960
ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 4020
cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 4080
aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 4140
actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgga 4200
attc 4204
<210> 12
<211> 35
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 12
gggagacggc attcgtaatc gaattcgcgg ccgca 35
<210> 13
<211> 50
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 13
atagcagaag aaaatggagg aattctgtca gaccaagttt actcatatat 50
<210> 14
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 14
ccgactggag ctcctatatt acc 23
<210> 15
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 15
gctgtggcga tctgtattcc 20
<210> 16
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 16
gtcttttaag taagtctact ct 22
<210> 17
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 17
ccaaagcgat tttaagcgcg 20
<210> 18
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 18
cctggcacgt ggtaattctc 20
<210> 19
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 19
ggatttcctc aaatctgacg 20
<210> 20
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 20
gtagaaacgc gccaaattac g 21
<210> 21
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 21
gctggtggtt gctaaagtcg 20
<210> 22
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 22
ggacgcaacc ctcattcatc 20
<210> 23
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 23
caggcatccg atttgcaagg 20
<210> 24
<211> 19
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 24
gcaagcagca gattacgcg 19
<210> 25
<211> 8347
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 25
gaattcctcc attttcttct gctatcaaaa taacagactc gtgattttcc aaacgagctt 60
tcaaaaaagc ctctgcccct tgcaaatcgg atgcctgtct ataaaattcc cgatattggt 120
taaacagcgg cgcaatggcg gccgcatctg atgtctttgc ttggcgaatg ttcatcttat 180
ttcttcctcc ctctcaataa ttttttcatt ctatcccttt tctgtaaagt ttatttttca 240
gaatactttt atcatcatgc tttgaaaaaa tatcacgata atatccattg ttctcacgga 300
agcacacgca ggtcatttga acgaattttt tcgacaggaa tttgccggga ctcaggagca 360
tttaacctaa aaaagcatga catttcagca taatgaacat ttactcatgt ctattttcgt 420
tcttttctgt atgaaaatag ttatttcgag tctctacgga aatagcgaga gatgatatac 480
ctaaatagag ataaaatcat ctcaaaaaaa tgggtctact aaaatattat tccatctatt 540
acaataaatt cacagaatag tcttttaagt aagtctactc tgaatttttt taaaaggaga 600
gggtaactag tggccccaaa aaagaaacgc aaggttatgg ataaaaaata cagcattggt 660
ctggatatcg gaaccaacag cgttgggtgg gcagtaataa cagatgaata caaagtgccg 720
tcaaaaaaat ttaaggttct ggggaataca gatcgccaca gcataaaaaa gaatctgatt 780
ggggcattgc tgtttgattc gggtgagaca gctgaggcca cgcgtctgaa acgtacagca 840
agaagacgtt acacacgtcg taaaaatcgt atttgctact tacaggaaat tttttctaac 900
gaaatggcca aggtagatga tagtttcttc catcgtctcg aagaatcttt tctggttgag 960
gaagataaaa aacacgaacg tcaccctatc tttggcaata tcgtggatga agtggcctat 1020
catgaaaaat accctacgat ttatcatctt cgcaagaagt tggttgatag tacggacaaa 1080
gcggatctgc gtttaatcta tcttgcgtta gcgcacatga tcaaatttcg tggtcatttc 1140
ttaattgaag gtgatctgaa tcctgataac tctgatgtgg acaaattgtt tatacaatta 1200
gtgcaaacct ataatcagct gttcgaggaa aaccccatta atgcctctgg agttgatgcc 1260
aaagcgattt taagcgcgag actttctaag tcccggcgtc tggagaatct gatcgcccag 1320
ttaccagggg aaaagaaaaa tggtctgttt ggtaatctga ttgccctcag tctggggctt 1380
accccgaact tcaaatccaa ttttgacctg gctgaggacg caaagctgca gctgagcaaa 1440
gatacttatg atgatgacct cgacaatctg ctcgcccaga ttggtgacca atatgcggat 1500
ctgtttctgg cagcgaagaa tctttcggat gctatcttgc tgtcggatat tctgcgtgtt 1560
aataccgaaa tcaccaaagc gcctctgtct gcaagtatga tcaagagata cgacgagcac 1620
caccaggacc tgactcttct taaggcactg gtacgccaac agcttccgga gaaatacaaa 1680
gaaatattct tcgaccagtc caagaatggt tacgcgggct acatcgatgg tggtgcatca 1740
caggaagagt tctataaatt tattaaacca atccttgaga aaatggatgg cacggaagag 1800
ttacttgtta aacttaaccg cgaagacttg cttagaaagc aacgtacatt cgacaacggc 1860
tccatcccac accagattca tttaggtgaa cttcacgcca tcttgcgcag acaagaagat 1920
ttctatccct tcttaaaaga caatcgggag aaaatcgaga agatcctgac gttccgcatt 1980
ccctattatg tcggtcccct ggcacgtggt aattctcggt ttgcctggat gacgcgcaaa 2040
agtgaggaaa ccatcacccc ttggaacttt gaagaagtcg tggataaagg tgctagcgcg 2100
cagtctttta tagaaagaat gacgaacttc gataaaaact tgcccaacga aaaagtcctg 2160
cccaagcact ctcttttata tgagtacttt actgtgtaca acgaactgac taaagtgaaa 2220
tacgttacgg aaggtatgcg caaacctgcc tttcttagtg gcgagcagaa aaaagcaatt 2280
gtcgatcttc tctttaaaac gaatcgcaag gtaactgtaa aacagctgaa ggaagattat 2340
ttcaaaaaga tcgaatgctt tgattctgtc gagatctcgg gtgtcgaaga tcgtttcaac 2400
gcttccttag ggacctatca tgatttgctg aagataataa aagacaaaga ctttctcgac 2460
aatgaagaaa atgaagatat tctggaggat attgttttga ccttgacctt attcgaagat 2520
agagagatga tcgaggagcg cttaaaaacc tatgcccacc tgtttgatga caaagtcatg 2580
aagcaattaa agcgccgcag atatacgggg tggggccgct tgagccgcaa gttgattaac 2640
ggtattagag acaagcagag cggaaaaact atcctggatt tcctcaaatc tgacggattt 2700
gcgaaccgca attttatgca gcttatacat gatgattcgc ttacattcaa agaggatatt 2760
cagaaggctc aggtgtctgg gcaaggtgat tcactccacg aacatatagc aaatttggcc 2820
ggctctcctg cgattaagaa ggggatcctg caaacagtta aagttgtgga tgaacttgta 2880
aaagtaatgg gccgccacaa gccggagaat atcgtgatag aaatggcgcg cgagaatcaa 2940
acgacacaaa aaggtcaaaa gaactcaaga gagagaatga agcgcattga ggaggggata 3000
aaggaacttg gatctcaaat tctgaaagaa catccagttg aaaacactca gctgcaaaat 3060
gaaaaattgt acctgtacta cctgcagaat ggaagagaca tgtacgtgga tcaggaattg 3120
gatatcaata gactctcgga ctatgacgta gatcacattg tccctcagag cttcctcaag 3180
gatgattcta tagataataa agtacttacg agatcggaca aaaatcgcgg taaatcggat 3240
aacgtcccat cggaggaagt cgttaaaaag atgaaaaact attggcgtca actgctgaac 3300
gccaagctga tcacacagcg taagtttgat aatctgacta aagccgaacg cggtggtctt 3360
agtgaactcg ataaagcagg atttataaaa cggcagttag tagaaacgcg ccaaattacg 3420
aaacacgtgg ctcagatcct cgattctaga atgaatacaa agtacgatga aaacgataaa 3480
ctgatccgtg aagtaaaagt cattacctta aaatctaaac ttgtgtccga tttccgcaaa 3540
gattttcagt tttacaaggt ccgggaaatc aataactatc accatgcaca tgatgcatat 3600
ttaaatgcgg ttgtaggcac ggcccttatt aagaaatacc ctaaactcga aagtgagttt 3660
gtttatgggg attataaagt gtatgacgtt cgcaaaatga tcgcgaaatc agaacaggaa 3720
atcggtaagg ctaccgctaa atactttttt tattccaaca ttatgaattt ttttaagacc 3780
gaaataactc tcgcgaatgg tgaaatccgt aaacggcctc ttatagaaac caatggtgaa 3840
acgggagaaa tcgtttggga taaaggtcgt gactttgcca ccgttcgtaa agtcctctca 3900
atgccgcaag ttaacattgt caagaagacg gaagttcaaa cagggggatt ctccaaagaa 3960
tctatcctgc cgaagcgtaa cagtgataaa cttattgcca gaaaaaaaga ttgggatcca 4020
aaaaaatacg gaggctttga ttcccctacc gtcgcgtata gtgtgctggt ggttgctaaa 4080
gtcgagaaag ggaaaagcaa gaaattgaaa tcagttaaag aactgctggg tattacaatt 4140
atggaaagat cgtcctttga gaaaaatccg atcgactttt tagaggccaa ggggtataag 4200
gaagtgaaaa aagatctcat catcaaatta ccgaagtata gtctttttga gctggaaaac 4260
ggcagaaaaa gaatgctggc ctccgcgggc gagttacaga agggaaatga gctggcgctg 4320
ccttccaaat atgttaattt tctgtacctt gccagtcatt atgagaaact gaagggcagc 4380
cccgaagata acgaacagaa acaattattc gtggaacagc ataagcacta tttagatgaa 4440
attatagagc aaattagtga attttctaag cgcgttatcc tcgcggatgc taatttagac 4500
aaagtactgt cagcttataa taaacatcgg gataagccga ttagagaaca ggccgaaaat 4560
atcattcatt tgtttacctt aaccaacctt ggagcaccag ctgccttcaa atatttcgat 4620
accacaattg atcgtaaacg gtatacaagt acaaaagaag tcttggacgc aaccctcatt 4680
catcaatcta ttactggatt atatgagaca cgcattgatc tttcacagct gggcggagac 4740
aagaagaaaa aactgaaact gcaccatcat caccatcatc atcaccatca ttgataactc 4800
gagaaagctt acataaaaaa ccggccttgg ccccgccggt tttttattat ttttcttcct 4860
ccgcatgttc aatccgctcc ataatcgacg gatggctccc tctgaaaatt ttaacgagaa 4920
acggcgggtt gacccggctc agtcccgtaa cggccaagtc ctgaaacgtc tcaatcgccg 4980
cttcccggtt tccggtcagc tcaatgccgt aacggtcggc ggcgttttcc tgataccggg 5040
agacggcatt cgtaatcgaa ttcgcggccg cacgcgtcca tggggatccc cgcgggtcga 5100
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag 5160
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag 5220
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa 5280
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta 5340
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg 5400
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt 5460
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac 5520
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca 5580
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct 5640
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt 5700
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt 5760
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa 5820
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat 5880
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg 5940
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt 6000
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca 6060
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa 6120
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg 6180
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc 6240
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt 6300
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg 6360
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg 6420
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata 6480
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg 6540
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga 6600
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg 6660
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt 6720
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc 6780
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat 6840
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg 6900
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg 6960
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa 7020
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa 7080
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta 7140
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga 7200
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt 7260
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc 7320
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt 7380
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct 7440
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt 7500
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc 7560
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 7620
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 7680
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 7740
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 7800
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 7860
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 7920
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 7980
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 8040
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 8100
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 8160
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 8220
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 8280
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 8340
tctgaca 8347
<210> 26
<211> 9724
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 26
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt 60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg 120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca 180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggactcgac ttcgaataca 240
tccagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa 300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca 360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatgtcga 420
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag 480
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag 540
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa 600
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta 660
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg 720
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt 780
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac 840
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca 900
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct 960
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt 1020
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt 1080
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa 1140
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat 1200
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg 1260
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt 1320
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca 1380
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa 1440
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg 1500
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc 1560
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt 1620
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg 1680
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg 1740
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata 1800
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg 1860
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga 1920
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg 1980
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt 2040
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc 2100
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat 2160
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg 2220
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg 2280
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa 2340
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa 2400
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta 2460
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga 2520
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt 2580
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc 2640
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt 2700
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct 2760
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt 2820
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc 2880
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 2940
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 3000
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 3060
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 3120
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 3180
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 3240
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 3300
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 3360
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 3420
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 3480
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 3540
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 3600
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 3660
tctgacaaat ggttctttcc cctgtcctaa acaaaaaacc cgctttattg aaaaagcggg 3720
gctgttttac agacaggtca aataaacgtt tgaaaatgtt catttcaaaa cgcgcggaac 3780
ctccatcttc tcccatccag actatactgt cggcttcgga atcgcaccga atcctgccca 3840
taaaaaggct cgcgggctta gagcgcttgc tcatcaccgc cggtagggaa tttcaccctg 3900
ccccgaagat tgatcttatt tatttttaat actgatatta ttataaatta attgtgaaaa 3960
aatgtacagg tgcaaagctt attgcgctgt tttgggacat cctgcacgat atttcggtaa 4020
actcactttt tccgcatact aaaaaccgca cattcacagt tatttcattt ttaattttcg 4080
tctttccgcg tgaaactcat tgacactctt tatggaatat ggtaaattat cagatattta 4140
tgacgcttat ttaggaggaa atcttacaca gaagctgcgg aacctgaaaa gaattccttt 4200
caggttccgt tttttttagg aattctccct gatctcaagc atctggcggg gataaatccg 4260
ctctcctttc aaatcgttcc attctttgag gcgctgtaca gttacgccca ttttttcggc 4320
gatatgatga agcgtatccc ctttccgcac tacatatgta ccggtcttcg attcatcgtc 4380
atgaaggcgg agtgtttggc cggccttgag atttgaatgt ttcaacccgt ttattctcat 4440
gatctcctcg atggatatac cgctatcctt gctgattctc cagagcgtgt cccctttttg 4500
aacggtcacc gcaccgctca ttgtcccggc gttttgataa acgtggatag aattttgccg 4560
gaacgcctcc tcacgaagca ccgtcagcgg attgattgca tatcttttat cttcagtcca 4620
tgaaccgtga tgcatttcaa aatgcaggtg ggttccggtc gatattcgaa ttcctccatt 4680
ttcttctgct atcaaaataa cagactcgtg attttccaaa cgagctttca aaaaagcctc 4740
tgccccttgc aaatcggatg cctgtctata aaattcccga tattggttaa acagcggcgc 4800
aatggcggcc gcatctgatg tctttgcttg gcgaatgttc atcttatttc ttcctccctc 4860
tcaataattt tttcattcta tcccttttct gtaaagttta tttttcagaa tacttttatc 4920
atcatgcttt gaaaaaatat cacgataata tccattgttc tcacggaagc acacgcaggt 4980
catttgaacg aattttttcg acaggaattt gccgggactc aggagcattt aacctaaaaa 5040
agcatgacat ttcagcataa tgaacattta ctcatgtcta ttttcgttct tttctgtatg 5100
aaaatagtta tttcgagtct ctacggaaat agcgagagat gatataccta aatagagata 5160
aaatcatctc aaaaaaatgg gtctactaaa atattattcc atctattaca ataaattcac 5220
agaatagtct tttaagtaag tctactctga atttttttaa aaggagaggg taactagtgg 5280
ccccaaaaaa gaaacgcaag gttatggata aaaaatacag cattggtctg gatatcggaa 5340
ccaacagcgt tgggtgggca gtaataacag atgaatacaa agtgccgtca aaaaaattta 5400
aggttctggg gaatacagat cgccacagca taaaaaagaa tctgattggg gcattgctgt 5460
ttgattcggg tgagacagct gaggccacgc gtctgaaacg tacagcaaga agacgttaca 5520
cacgtcgtaa aaatcgtatt tgctacttac aggaaatttt ttctaacgaa atggccaagg 5580
tagatgatag tttcttccat cgtctcgaag aatcttttct ggttgaggaa gataaaaaac 5640
acgaacgtca ccctatcttt ggcaatatcg tggatgaagt ggcctatcat gaaaaatacc 5700
ctacgattta tcatcttcgc aagaagttgg ttgatagtac ggacaaagcg gatctgcgtt 5760
taatctatct tgcgttagcg cacatgatca aatttcgtgg tcatttctta attgaaggtg 5820
atctgaatcc tgataactct gatgtggaca aattgtttat acaattagtg caaacctata 5880
atcagctgtt cgaggaaaac cccattaatg cctctggagt tgatgccaaa gcgattttaa 5940
gcgcgagact ttctaagtcc cggcgtctgg agaatctgat cgcccagtta ccaggggaaa 6000
agaaaaatgg tctgtttggt aatctgattg ccctcagtct ggggcttacc ccgaacttca 6060
aatccaattt tgacctggct gaggacgcaa agctgcagct gagcaaagat acttatgatg 6120
atgacctcga caatctgctc gcccagattg gtgaccaata tgcggatctg tttctggcag 6180
cgaagaatct ttcggatgct atcttgctgt cggatattct gcgtgttaat accgaaatca 6240
ccaaagcgcc tctgtctgca agtatgatca agagatacga cgagcaccac caggacctga 6300
ctcttcttaa ggcactggta cgccaacagc ttccggagaa atacaaagaa atattcttcg 6360
accagtccaa gaatggttac gcgggctaca tcgatggtgg tgcatcacag gaagagttct 6420
ataaatttat taaaccaatc cttgagaaaa tggatggcac ggaagagtta cttgttaaac 6480
ttaaccgcga agacttgctt agaaagcaac gtacattcga caacggctcc atcccacacc 6540
agattcattt aggtgaactt cacgccatct tgcgcagaca agaagatttc tatcccttct 6600
taaaagacaa tcgggagaaa atcgagaaga tcctgacgtt ccgcattccc tattatgtcg 6660
gtcccctggc acgtggtaat tctcggtttg cctggatgac gcgcaaaagt gaggaaacca 6720
tcaccccttg gaactttgaa gaagtcgtgg ataaaggtgc tagcgcgcag tcttttatag 6780
aaagaatgac gaacttcgat aaaaacttgc ccaacgaaaa agtcctgccc aagcactctc 6840
ttttatatga gtactttact gtgtacaacg aactgactaa agtgaaatac gttacggaag 6900
gtatgcgcaa acctgccttt cttagtggcg agcagaaaaa agcaattgtc gatcttctct 6960
ttaaaacgaa tcgcaaggta actgtaaaac agctgaagga agattatttc aaaaagatcg 7020
aatgctttga ttctgtcgag atctcgggtg tcgaagatcg tttcaacgct tccttaggga 7080
cctatcatga tttgctgaag ataataaaag acaaagactt tctcgacaat gaagaaaatg 7140
aagatattct ggaggatatt gttttgacct tgaccttatt cgaagataga gagatgatcg 7200
aggagcgctt aaaaacctat gcccacctgt ttgatgacaa agtcatgaag caattaaagc 7260
gccgcagata tacggggtgg ggccgcttga gccgcaagtt gattaacggt attagagaca 7320
agcagagcgg aaaaactatc ctggatttcc tcaaatctga cggatttgcg aaccgcaatt 7380
ttatgcagct tatacatgat gattcgctta cattcaaaga ggatattcag aaggctcagg 7440
tgtctgggca aggtgattca ctccacgaac atatagcaaa tttggccggc tctcctgcga 7500
ttaagaaggg gatcctgcaa acagttaaag ttgtggatga acttgtaaaa gtaatgggcc 7560
gccacaagcc ggagaatatc gtgatagaaa tggcgcgcga gaatcaaacg acacaaaaag 7620
gtcaaaagaa ctcaagagag agaatgaagc gcattgagga ggggataaag gaacttggat 7680
ctcaaattct gaaagaacat ccagttgaaa acactcagct gcaaaatgaa aaattgtacc 7740
tgtactacct gcagaatgga agagacatgt acgtggatca ggaattggat atcaatagac 7800
tctcggacta tgacgtagat cacattgtcc ctcagagctt cctcaaggat gattctatag 7860
ataataaagt acttacgaga tcggacaaaa atcgcggtaa atcggataac gtcccatcgg 7920
aggaagtcgt taaaaagatg aaaaactatt ggcgtcaact gctgaacgcc aagctgatca 7980
cacagcgtaa gtttgataat ctgactaaag ccgaacgcgg tggtcttagt gaactcgata 8040
aagcaggatt tataaaacgg cagttagtag aaacgcgcca aattacgaaa cacgtggctc 8100
agatcctcga ttctagaatg aatacaaagt acgatgaaaa cgataaactg atccgtgaag 8160
taaaagtcat taccttaaaa tctaaacttg tgtccgattt ccgcaaagat tttcagtttt 8220
acaaggtccg ggaaatcaat aactatcacc atgcacatga tgcatattta aatgcggttg 8280
taggcacggc ccttattaag aaatacccta aactcgaaag tgagtttgtt tatggggatt 8340
ataaagtgta tgacgttcgc aaaatgatcg cgaaatcaga acaggaaatc ggtaaggcta 8400
ccgctaaata ctttttttat tccaacatta tgaatttttt taagaccgaa ataactctcg 8460
cgaatggtga aatccgtaaa cggcctctta tagaaaccaa tggtgaaacg ggagaaatcg 8520
tttgggataa aggtcgtgac tttgccaccg ttcgtaaagt cctctcaatg ccgcaagtta 8580
acattgtcaa gaagacggaa gttcaaacag ggggattctc caaagaatct atcctgccga 8640
agcgtaacag tgataaactt attgccagaa aaaaagattg ggatccaaaa aaatacggag 8700
gctttgattc ccctaccgtc gcgtatagtg tgctggtggt tgctaaagtc gagaaaggga 8760
aaagcaagaa attgaaatca gttaaagaac tgctgggtat tacaattatg gaaagatcgt 8820
cctttgagaa aaatccgatc gactttttag aggccaaggg gtataaggaa gtgaaaaaag 8880
atctcatcat caaattaccg aagtatagtc tttttgagct ggaaaacggc agaaaaagaa 8940
tgctggcctc cgcgggcgag ttacagaagg gaaatgagct ggcgctgcct tccaaatatg 9000
ttaattttct gtaccttgcc agtcattatg agaaactgaa gggcagcccc gaagataacg 9060
aacagaaaca attattcgtg gaacagcata agcactattt agatgaaatt atagagcaaa 9120
ttagtgaatt ttctaagcgc gttatcctcg cggatgctaa tttagacaaa gtactgtcag 9180
cttataataa acatcgggat aagccgatta gagaacaggc cgaaaatatc attcatttgt 9240
ttaccttaac caaccttgga gcaccagctg ccttcaaata tttcgatacc acaattgatc 9300
gtaaacggta tacaagtaca aaagaagtct tggacgcaac cctcattcat caatctatta 9360
ctggattata tgagacacgc attgatcttt cacagctggg cggagacaag aagaaaaaac 9420
tgaaactgca ccatcatcac catcatcatc accatcattg ataactcgag aaagcttaca 9480
taaaaaaccg gccttggccc cgccggtttt ttattatttt tcttcctccg catgttcaat 9540
ccgctccata atcgacggat ggctccctct gaaaatttta acgagaaacg gcgggttgac 9600
ccggctcagt cccgtaacgg ccaagtcctg aaacgtctca atcgccgctt cccggtttcc 9660
ggtcagctca atgccgtaac ggtcggcggc gttttcctga taccgggaga cggcattcgt 9720
aatc 9724
<210> 27
<211> 9724
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 27
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt 60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg 120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca 180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggagatgcc atcagttcct 240
catagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa 300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca 360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatgtcga 420
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag 480
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag 540
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa 600
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta 660
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg 720
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt 780
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac 840
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca 900
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct 960
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt 1020
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt 1080
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa 1140
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat 1200
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg 1260
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt 1320
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca 1380
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa 1440
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg 1500
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc 1560
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt 1620
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg 1680
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg 1740
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata 1800
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg 1860
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga 1920
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg 1980
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt 2040
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc 2100
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat 2160
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg 2220
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg 2280
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa 2340
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa 2400
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta 2460
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga 2520
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt 2580
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc 2640
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt 2700
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct 2760
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt 2820
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc 2880
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 2940
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 3000
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 3060
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 3120
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 3180
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 3240
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 3300
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 3360
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 3420
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 3480
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 3540
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 3600
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 3660
tctgacattg atattcagca ccctgcgcat ttcgaccggg agaacgactc tgccgagctc 3720
atcgattctc cggacaatcc cggtattttt cacgtttgaa aagcctcctt ttctcctttc 3780
tttattgact tttgtcaaca tctttataat aaaagagatc ttcaaatttt ttgttgaaat 3840
actgaatcat ctttccgatc acaagttgtc cgggcctcct ttcgccattt aaaactctgc 3900
tgagtgtcgc cggggatacg ccgatttcaa tggcaagctg atttaaggag agattgtgtt 3960
caatcatgta ctggagaaca aaatctcttt tgatatgaat cttttttacc atgattactc 4020
ccctttctaa tctcttatgt ttctttttat ctacattgaa catatacgat ttgttaactt 4080
ttgtcaatac ttttaccatc catatgtttc ctataggcaa tattcgtact aaaatatttt 4140
ataataagag attgcgaggt tttggccata cttctccgcg gcacactctc ctctctatca 4200
ttttcgtctg tttacgatcc tgctgttatt ttatccctta tgttaacttt tgtcaatatt 4260
tttcctgtct aagtatttcc tatagtcaac atttgtatta aaatgttcat atcatgaatt 4320
tgcggggggg atggcgatga caaggttcgg cgagcggctc aaagagctga gggaacaaag 4380
aagcctgtcg gttaatcagc ttgccatgta tgccggtgtg agcgccgcag ccatttccag 4440
agccgcagcc atttccagaa tcgaaaacgg ccaccgcggc gttcccaagc ccgcgacgat 4500
cagaaaattg gccgaggctc tgaaaatgcc gtacgagcag ctcatggata ttgccggtta 4560
tatgagagct gacgagattc gcgaacagcc gcgcggctat gtcacgatgc aggagatcgc 4620
ggccaagcac ggcgtcgaag acctgtggct gtttaaaccc gagaaatgaa ttcctccatt 4680
ttcttctgct atcaaaataa cagactcgtg attttccaaa cgagctttca aaaaagcctc 4740
tgccccttgc aaatcggatg cctgtctata aaattcccga tattggttaa acagcggcgc 4800
aatggcggcc gcatctgatg tctttgcttg gcgaatgttc atcttatttc ttcctccctc 4860
tcaataattt tttcattcta tcccttttct gtaaagttta tttttcagaa tacttttatc 4920
atcatgcttt gaaaaaatat cacgataata tccattgttc tcacggaagc acacgcaggt 4980
catttgaacg aattttttcg acaggaattt gccgggactc aggagcattt aacctaaaaa 5040
agcatgacat ttcagcataa tgaacattta ctcatgtcta ttttcgttct tttctgtatg 5100
aaaatagtta tttcgagtct ctacggaaat agcgagagat gatataccta aatagagata 5160
aaatcatctc aaaaaaatgg gtctactaaa atattattcc atctattaca ataaattcac 5220
agaatagtct tttaagtaag tctactctga atttttttaa aaggagaggg taactagtgg 5280
ccccaaaaaa gaaacgcaag gttatggata aaaaatacag cattggtctg gatatcggaa 5340
ccaacagcgt tgggtgggca gtaataacag atgaatacaa agtgccgtca aaaaaattta 5400
aggttctggg gaatacagat cgccacagca taaaaaagaa tctgattggg gcattgctgt 5460
ttgattcggg tgagacagct gaggccacgc gtctgaaacg tacagcaaga agacgttaca 5520
cacgtcgtaa aaatcgtatt tgctacttac aggaaatttt ttctaacgaa atggccaagg 5580
tagatgatag tttcttccat cgtctcgaag aatcttttct ggttgaggaa gataaaaaac 5640
acgaacgtca ccctatcttt ggcaatatcg tggatgaagt ggcctatcat gaaaaatacc 5700
ctacgattta tcatcttcgc aagaagttgg ttgatagtac ggacaaagcg gatctgcgtt 5760
taatctatct tgcgttagcg cacatgatca aatttcgtgg tcatttctta attgaaggtg 5820
atctgaatcc tgataactct gatgtggaca aattgtttat acaattagtg caaacctata 5880
atcagctgtt cgaggaaaac cccattaatg cctctggagt tgatgccaaa gcgattttaa 5940
gcgcgagact ttctaagtcc cggcgtctgg agaatctgat cgcccagtta ccaggggaaa 6000
agaaaaatgg tctgtttggt aatctgattg ccctcagtct ggggcttacc ccgaacttca 6060
aatccaattt tgacctggct gaggacgcaa agctgcagct gagcaaagat acttatgatg 6120
atgacctcga caatctgctc gcccagattg gtgaccaata tgcggatctg tttctggcag 6180
cgaagaatct ttcggatgct atcttgctgt cggatattct gcgtgttaat accgaaatca 6240
ccaaagcgcc tctgtctgca agtatgatca agagatacga cgagcaccac caggacctga 6300
ctcttcttaa ggcactggta cgccaacagc ttccggagaa atacaaagaa atattcttcg 6360
accagtccaa gaatggttac gcgggctaca tcgatggtgg tgcatcacag gaagagttct 6420
ataaatttat taaaccaatc cttgagaaaa tggatggcac ggaagagtta cttgttaaac 6480
ttaaccgcga agacttgctt agaaagcaac gtacattcga caacggctcc atcccacacc 6540
agattcattt aggtgaactt cacgccatct tgcgcagaca agaagatttc tatcccttct 6600
taaaagacaa tcgggagaaa atcgagaaga tcctgacgtt ccgcattccc tattatgtcg 6660
gtcccctggc acgtggtaat tctcggtttg cctggatgac gcgcaaaagt gaggaaacca 6720
tcaccccttg gaactttgaa gaagtcgtgg ataaaggtgc tagcgcgcag tcttttatag 6780
aaagaatgac gaacttcgat aaaaacttgc ccaacgaaaa agtcctgccc aagcactctc 6840
ttttatatga gtactttact gtgtacaacg aactgactaa agtgaaatac gttacggaag 6900
gtatgcgcaa acctgccttt cttagtggcg agcagaaaaa agcaattgtc gatcttctct 6960
ttaaaacgaa tcgcaaggta actgtaaaac agctgaagga agattatttc aaaaagatcg 7020
aatgctttga ttctgtcgag atctcgggtg tcgaagatcg tttcaacgct tccttaggga 7080
cctatcatga tttgctgaag ataataaaag acaaagactt tctcgacaat gaagaaaatg 7140
aagatattct ggaggatatt gttttgacct tgaccttatt cgaagataga gagatgatcg 7200
aggagcgctt aaaaacctat gcccacctgt ttgatgacaa agtcatgaag caattaaagc 7260
gccgcagata tacggggtgg ggccgcttga gccgcaagtt gattaacggt attagagaca 7320
agcagagcgg aaaaactatc ctggatttcc tcaaatctga cggatttgcg aaccgcaatt 7380
ttatgcagct tatacatgat gattcgctta cattcaaaga ggatattcag aaggctcagg 7440
tgtctgggca aggtgattca ctccacgaac atatagcaaa tttggccggc tctcctgcga 7500
ttaagaaggg gatcctgcaa acagttaaag ttgtggatga acttgtaaaa gtaatgggcc 7560
gccacaagcc ggagaatatc gtgatagaaa tggcgcgcga gaatcaaacg acacaaaaag 7620
gtcaaaagaa ctcaagagag agaatgaagc gcattgagga ggggataaag gaacttggat 7680
ctcaaattct gaaagaacat ccagttgaaa acactcagct gcaaaatgaa aaattgtacc 7740
tgtactacct gcagaatgga agagacatgt acgtggatca ggaattggat atcaatagac 7800
tctcggacta tgacgtagat cacattgtcc ctcagagctt cctcaaggat gattctatag 7860
ataataaagt acttacgaga tcggacaaaa atcgcggtaa atcggataac gtcccatcgg 7920
aggaagtcgt taaaaagatg aaaaactatt ggcgtcaact gctgaacgcc aagctgatca 7980
cacagcgtaa gtttgataat ctgactaaag ccgaacgcgg tggtcttagt gaactcgata 8040
aagcaggatt tataaaacgg cagttagtag aaacgcgcca aattacgaaa cacgtggctc 8100
agatcctcga ttctagaatg aatacaaagt acgatgaaaa cgataaactg atccgtgaag 8160
taaaagtcat taccttaaaa tctaaacttg tgtccgattt ccgcaaagat tttcagtttt 8220
acaaggtccg ggaaatcaat aactatcacc atgcacatga tgcatattta aatgcggttg 8280
taggcacggc ccttattaag aaatacccta aactcgaaag tgagtttgtt tatggggatt 8340
ataaagtgta tgacgttcgc aaaatgatcg cgaaatcaga acaggaaatc ggtaaggcta 8400
ccgctaaata ctttttttat tccaacatta tgaatttttt taagaccgaa ataactctcg 8460
cgaatggtga aatccgtaaa cggcctctta tagaaaccaa tggtgaaacg ggagaaatcg 8520
tttgggataa aggtcgtgac tttgccaccg ttcgtaaagt cctctcaatg ccgcaagtta 8580
acattgtcaa gaagacggaa gttcaaacag ggggattctc caaagaatct atcctgccga 8640
agcgtaacag tgataaactt attgccagaa aaaaagattg ggatccaaaa aaatacggag 8700
gctttgattc ccctaccgtc gcgtatagtg tgctggtggt tgctaaagtc gagaaaggga 8760
aaagcaagaa attgaaatca gttaaagaac tgctgggtat tacaattatg gaaagatcgt 8820
cctttgagaa aaatccgatc gactttttag aggccaaggg gtataaggaa gtgaaaaaag 8880
atctcatcat caaattaccg aagtatagtc tttttgagct ggaaaacggc agaaaaagaa 8940
tgctggcctc cgcgggcgag ttacagaagg gaaatgagct ggcgctgcct tccaaatatg 9000
ttaattttct gtaccttgcc agtcattatg agaaactgaa gggcagcccc gaagataacg 9060
aacagaaaca attattcgtg gaacagcata agcactattt agatgaaatt atagagcaaa 9120
ttagtgaatt ttctaagcgc gttatcctcg cggatgctaa tttagacaaa gtactgtcag 9180
cttataataa acatcgggat aagccgatta gagaacaggc cgaaaatatc attcatttgt 9240
ttaccttaac caaccttgga gcaccagctg ccttcaaata tttcgatacc acaattgatc 9300
gtaaacggta tacaagtaca aaagaagtct tggacgcaac cctcattcat caatctatta 9360
ctggattata tgagacacgc attgatcttt cacagctggg cggagacaag aagaaaaaac 9420
tgaaactgca ccatcatcac catcatcatc accatcattg ataactcgag aaagcttaca 9480
taaaaaaccg gccttggccc cgccggtttt ttattatttt tcttcctccg catgttcaat 9540
ccgctccata atcgacggat ggctccctct gaaaatttta acgagaaacg gcgggttgac 9600
ccggctcagt cccgtaacgg ccaagtcctg aaacgtctca atcgccgctt cccggtttcc 9660
ggtcagctca atgccgtaac ggtcggcggc gttttcctga taccgggaga cggcattcgt 9720
aatc 9724
<210> 28
<211> 20
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 28
ctcgacttcg aatacatcca 20
<210> 29
<211> 20
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 29
gatgccatca gttcctcata 20
<210> 30
<211> 1578
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 30
atgtttcgag tattggtctc agataaaatg tccagcgacg gcctcaaacc attaatggaa 60
gcagatttta ttgaaattgt agaaaagaat gttgcggaag cggaagacga gcttcatacg 120
tttgacgcgc tcttggtgcg gagcgccacg aaggtaaccg aagagctgtt taaaaagatg 180
acttcgctga aaatcgtcgc cagagcaggt gtcggcgtcg acaatatcga tattgacgag 240
gcgacaaaac acggtgttat cgtcgtaaac gcgccaaacg ggaatacaat ttcaaccgct 300
gaacatacct ttgcaatgtt ttcagcgtta atgagacata ttccgcaggc aaacatctcc 360
gtgaaatcaa gggagtggaa tcgttcggct tacgtcggtt cagagcttta cggaaaaacg 420
ctcggcatca tcggaatggg ccgcatcgga agcgaaatcg cgagccgcgc aaaagcattc 480
ggtatgaccg ttcatgtatt tgacccgttc ctgacccaag aaagggcaag caagctcggc 540
gttaacgcga acagctttga agaagttctg gcatgcgccg acatcattac ggttcatacc 600
ccgctcacga aagaaacgaa gggacttttg aacaaagaaa ccatcgcaaa aacgaaaaaa 660
ggcgttcgtc tcgttaactg tgcaagaggc ggcatcatcg atgaagcagc gcttttggaa 720
gctctggaaa gcggacatgt cgctggcgct gccttggatg tattcgaagt cgagcctccg 780
gtcgattcaa aactgatcga tcatccgctt gtagtcgcga ctcctcactt gggcgcctca 840
acaaaagaag cccagctgaa tgtcgctgca caagtgtccg aagaagtcct tcagtatgcg 900
caaggaaacc ctgtgatgtc cgcgatcaac cttccggcca tgacaaagga ttcattcgaa 960
aaaatccagc cttatcatca gtttgccaat acgatcggaa accttgtgtc tcagtgcatg 1020
aatgagcctg ttcaagatgt agccatccaa tatgaaggct ccatcgccaa acttgaaacg 1080
tcatttatta cgaaaagcct tttggccgga tttctgaagc cgagggtcgc ggctaccgtt 1140
aacgaagtga atgccggcac cgttgcgaaa gagcgcggca tcagcttcag cgaaaaaatt 1200
tcttccaatg agtcaggcta tgaaaactgc atctctgtga ctgtcacggg agatgtaaca 1260
acattctctt taagagcgac gtacattccg cacttcggcg gacgcatcgt tgccttaaac 1320
ggctttgata ttgattttta tccggctgga caccttgtct acattcacca ccaggataaa 1380
ccaggggcta tcggccatgt cggacgaatt ttaggagacc atgacatcaa tatcgccact 1440
atgcaggtag gccgaaaaga aaaaggcgga gaagcgatca tgatgctttc ctttgaccgc 1500
caccttgagg acgatatttt agctgagctg aaaaacatcc cggatatcgt gtctgttaaa 1560
gccatcgacc ttccttaa 1578
<210> 31
<211> 3
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 31
agg 3
<210> 32
<211> 20
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 32
ctcgacttcg aatacatcca 20
<210> 33
<211> 76
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 33
gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt 60
ggcaccgagt cggtgc 76
<210> 34
<211> 96
<212> RNA
<213> 人工序列
<220>
<223> 合成的
<400> 34
cucgacuucg aauacaucca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc 60
cguuaucaac uugaaaaagu ggcaccgagu cggugc 96
<210> 35
<211> 224
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 35
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt 60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg 120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca 180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tgga 224
<210> 36
<211> 95
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 36
gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc 60
tcggttgccg ccgggcgttt tttattggtg agaat 95
<210> 37
<211> 500
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 37
aatggttctt tcccctgtcc taaacaaaaa acccgcttta ttgaaaaagc ggggctgttt 60
tacagacagg tcaaataaac gtttgaaaat gttcatttca aaacgcgcgg aacctccatc 120
ttctcccatc cagactatac tgtcggcttc ggaatcgcac cgaatcctgc ccataaaaag 180
gctcgcgggc ttagagcgct tgctcatcac cgccggtagg gaatttcacc ctgccccgaa 240
gattgatctt atttattttt aatactgata ttattataaa ttaattgtga aaaaatgtac 300
aggtgcaaag cttattgcgc tgttttggga catcctgcac gatatttcgg taaactcact 360
ttttccgcat actaaaaacc gcacattcac agttatttca tttttaattt tcgtctttcc 420
gcgtgaaact cattgacact ctttatggaa tatggtaaat tatcagatat ttatgacgct 480
tatttaggag gaaatcttac 500
<210> 38
<211> 40
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 38
tgagtaaact tggtctgaca aatggttctt tcccctgtcc 40
<210> 39
<211> 46
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 39
aggttccgca gcttctgtgt aagatttcct cctaaataag cgtcat 46
<210> 40
<211> 500
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 40
acagaagctg cggaacctga aaagaattcc tttcaggttc cgtttttttt aggaattctc 60
cctgatctca agcatctggc ggggataaat ccgctctcct ttcaaatcgt tccattcttt 120
gaggcgctgt acagttacgc ccattttttc ggcgatatga tgaagcgtat cccctttccg 180
cactacatat gtaccggtct tcgattcatc gtcatgaagg cggagtgttt ggccggcctt 240
gagatttgaa tgtttcaacc cgtttattct catgatctcc tcgatggata taccgctatc 300
cttgctgatt ctccagagcg tgtccccttt ttgaacggtc accgcaccgc tcattgtccc 360
ggcgttttga taaacgtgga tagaattttg ccggaacgcc tcctcacgaa gcaccgtcag 420
cggattgatt gcatatcttt tatcttcagt ccatgaaccg tgatgcattt caaaatgcag 480
gtgggttccg gtcgatattc 500
<210> 41
<211> 46
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 41
atgacgctta tttaggagga aatcttacac agaagctgcg gaacct 46
<210> 42
<211> 41
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 42
cagaagaaaa tggaggaatt cgaatatcga ccggaaccca c 41
<210> 43
<211> 415
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 43
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt 60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg 120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca 180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggactcgac ttcgaataca 240
tccagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa 300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca 360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaat 415
<210> 44
<211> 1000
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 44
aatggttctt tcccctgtcc taaacaaaaa acccgcttta ttgaaaaagc ggggctgttt 60
tacagacagg tcaaataaac gtttgaaaat gttcatttca aaacgcgcgg aacctccatc 120
ttctcccatc cagactatac tgtcggcttc ggaatcgcac cgaatcctgc ccataaaaag 180
gctcgcgggc ttagagcgct tgctcatcac cgccggtagg gaatttcacc ctgccccgaa 240
gattgatctt atttattttt aatactgata ttattataaa ttaattgtga aaaaatgtac 300
aggtgcaaag cttattgcgc tgttttggga catcctgcac gatatttcgg taaactcact 360
ttttccgcat actaaaaacc gcacattcac agttatttca tttttaattt tcgtctttcc 420
gcgtgaaact cattgacact ctttatggaa tatggtaaat tatcagatat ttatgacgct 480
tatttaggag gaaatcttac acagaagctg cggaacctga aaagaattcc tttcaggttc 540
cgtttttttt aggaattctc cctgatctca agcatctggc ggggataaat ccgctctcct 600
ttcaaatcgt tccattcttt gaggcgctgt acagttacgc ccattttttc ggcgatatga 660
tgaagcgtat cccctttccg cactacatat gtaccggtct tcgattcatc gtcatgaagg 720
cggagtgttt ggccggcctt gagatttgaa tgtttcaacc cgtttattct catgatctcc 780
tcgatggata taccgctatc cttgctgatt ctccagagcg tgtccccttt ttgaacggtc 840
accgcaccgc tcattgtccc ggcgttttga taaacgtgga tagaattttg ccggaacgcc 900
tcctcacgaa gcaccgtcag cggattgatt gcatatcttt tatcttcagt ccatgaaccg 960
tgatgcattt caaaatgcag gtgggttccg gtcgatattc 1000
<210> 45
<211> 402
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 45
atgacgaact ttggacacca tttacgacaa ttaagggaac ggaaaaaact gaccgtcaat 60
caactggcga tgtattccgg cgtcagttcg gcaggcattt cgcgaatcga aaacggaaag 120
cgcggcgtgc cgaagccggc gacgatcaga aaactggcgg acgctttgaa agtcccgtat 180
gaggaactga tggcatctgc aggctatatc agcgcgtcta cagtccagga agcaagaagc 240
agctatgatt ccatttacga catcgtgtca cagtacgatt tagaggacct ttctctgttt 300
gacagcgaaa agtggaaggt gctttcaaaa aaagacatcg aaaacctgga caaatatttc 360
gactttctcg tgcaggaagc aagcagccga aacaaaaact ga 402
<210> 46
<211> 3
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 46
cgg 3
<210> 47
<211> 96
<212> RNA
<213> 人工序列
<220>
<223> 合成的
<400> 47
gaugccauca guuccucaua guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc 60
cguuaucaac uugaaaaagu ggcaccgagu cggugc 96
<210> 48
<211> 500
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 48
ttgatattca gcaccctgcg catttcgacc gggagaacga ctctgccgag ctcatcgatt 60
ctccggacaa tcccggtatt tttcacgttt gaaaagcctc cttttctcct ttctttattg 120
acttttgtca acatctttat aataaaagag atcttcaaat tttttgttga aatactgaat 180
catctttccg atcacaagtt gtccgggcct cctttcgcca tttaaaactc tgctgagtgt 240
cgccggggat acgccgattt caatggcaag ctgatttaag gagagattgt gttcaatcat 300
gtactggaga acaaaatctc ttttgatatg aatctttttt accatgatta ctcccctttc 360
taatctctta tgtttctttt tatctacatt gaacatatac gatttgttaa cttttgtcaa 420
tacttttacc atccatatgt ttcctatagg caatattcgt actaaaatat tttataataa 480
gagattgcga ggttttggcc 500
<210> 49
<211> 40
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 49
tgagtaaact tggtctgaca ttgatattca gcaccctgcg 40
<210> 50
<211> 38
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 50
tgtgccgcgg agaagtatgg ccaaaacctc gcaatctc 38
<210> 51
<211> 500
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 51
atacttctcc gcggcacact ctcctctcta tcattttcgt ctgtttacga tcctgctgtt 60
attttatccc ttatgttaac ttttgtcaat atttttcctg tctaagtatt tcctatagtc 120
aacatttgta ttaaaatgtt catatcatga atttgcgggg gggatggcga tgacaaggtt 180
cggcgagcgg ctcaaagagc tgagggaaca aagaagcctg tcggttaatc agcttgccat 240
gtatgccggt gtgagcgccg cagccatttc cagagccgca gccatttcca gaatcgaaaa 300
cggccaccgc ggcgttccca agcccgcgac gatcagaaaa ttggccgagg ctctgaaaat 360
gccgtacgag cagctcatgg atattgccgg ttatatgaga gctgacgaga ttcgcgaaca 420
gccgcgcggc tatgtcacga tgcaggagat cgcggccaag cacggcgtcg aagacctgtg 480
gctgtttaaa cccgagaaat 500
<210> 52
<211> 38
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 52
gagattgcga ggttttggcc atacttctcc gcggcaca 38
<210> 53
<211> 44
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 53
cagaagaaaa tggaggaatt catttctcgg gtttaaacag ccac 44
<210> 54
<211> 415
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 54
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt 60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg 120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca 180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggagatgcc atcagttcct 240
catagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa 300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca 360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaat 415
<210> 55
<211> 1000
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 55
ttgatattca gcaccctgcg catttcgacc gggagaacga ctctgccgag ctcatcgatt 60
ctccggacaa tcccggtatt tttcacgttt gaaaagcctc cttttctcct ttctttattg 120
acttttgtca acatctttat aataaaagag atcttcaaat tttttgttga aatactgaat 180
catctttccg atcacaagtt gtccgggcct cctttcgcca tttaaaactc tgctgagtgt 240
cgccggggat acgccgattt caatggcaag ctgatttaag gagagattgt gttcaatcat 300
gtactggaga acaaaatctc ttttgatatg aatctttttt accatgatta ctcccctttc 360
taatctctta tgtttctttt tatctacatt gaacatatac gatttgttaa cttttgtcaa 420
tacttttacc atccatatgt ttcctatagg caatattcgt actaaaatat tttataataa 480
gagattgcga ggttttggcc atacttctcc gcggcacact ctcctctcta tcattttcgt 540
ctgtttacga tcctgctgtt attttatccc ttatgttaac ttttgtcaat atttttcctg 600
tctaagtatt tcctatagtc aacatttgta ttaaaatgtt catatcatga atttgcgggg 660
gggatggcga tgacaaggtt cggcgagcgg ctcaaagagc tgagggaaca aagaagcctg 720
tcggttaatc agcttgccat gtatgccggt gtgagcgccg cagccatttc cagagccgca 780
gccatttcca gaatcgaaaa cggccaccgc ggcgttccca agcccgcgac gatcagaaaa 840
ttggccgagg ctctgaaaat gccgtacgag cagctcatgg atattgccgg ttatatgaga 900
gctgacgaga ttcgcgaaca gccgcgcggc tatgtcacga tgcaggagat cgcggccaag 960
cacggcgtcg aagacctgtg gctgtttaaa cccgagaaat 1000
<210> 56
<211> 1368
<212> PRT
<213> 人工序列
<220>
<223> 合成的
<400> 56
Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val
1 5 10 15
Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe
20 25 30
Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile
35 40 45
Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu
50 55 60
Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys
65 70 75 80
Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser
85 90 95
Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys
100 105 110
His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr
115 120 125
His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp
130 135 140
Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile His Leu Ala Leu Ala His
145 150 155 160
Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro
165 170 175
Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr
180 185 190
Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala
195 200 205
Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn
210 215 220
Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn
225 230 235 240
Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe
245 250 255
Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp
260 265 270
Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp
275 280 285
Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp
290 295 300
Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser
305 310 315 320
Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys
325 330 335
Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe
340 345 350
Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser
355 360 365
Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp
370 375 380
Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg
385 390 395 400
Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu
405 410 415
Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe
420 425 430
Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile
435 440 445
Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp
450 455 460
Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu
465 470 475 480
Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr
485 490 495
Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser
500 505 510
Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys
515 520 525
Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln
530 535 540
Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr
545 550 555 560
Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp
565 570 575
Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly
580 585 590
Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp
595 600 605
Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr
610 615 620
Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala
625 630 635 640
His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr
645 650 655
Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp
660 665 670
Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe
675 680 685
Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe
690 695 700
Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu
705 710 715 720
His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly
725 730 735
Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly
740 745 750
Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln
755 760 765
Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile
770 775 780
Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro
785 790 795 800
Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu
805 810 815
Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg
820 825 830
Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys
835 840 845
Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg
850 855 860
Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys
865 870 875 880
Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys
885 890 895
Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp
900 905 910
Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr
915 920 925
Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp
930 935 940
Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser
945 950 955 960
Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg
965 970 975
Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val
980 985 990
Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe
995 1000 1005
Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala
1010 1015 1020
Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe
1025 1030 1035
Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala
1040 1045 1050
Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu
1055 1060 1065
Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val
1070 1075 1080
Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr
1085 1090 1095
Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys
1100 1105 1110
Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro
1115 1120 1125
Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val
1130 1135 1140
Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys
1145 1150 1155
Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser
1160 1165 1170
Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys
1175 1180 1185
Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu
1190 1195 1200
Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly
1205 1210 1215
Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val
1220 1225 1230
Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser
1235 1240 1245
Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys
1250 1255 1260
His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys
1265 1270 1275
Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala
1280 1285 1290
Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn
1295 1300 1305
Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala
1310 1315 1320
Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser
1325 1330 1335
Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr
1340 1345 1350
Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp
1355 1360 1365
<210> 57
<211> 33
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 57
gatctgcgtt taatccatct tgcgttagcg cac 33
<210> 58
<211> 33
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 58
gtgcgctaac gcaagatgga ttaaacgcag atc 33
<210> 59
<211> 9724
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 59
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt 60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg 120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca 180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggactcgac ttcgaataca 240
tccagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa 300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca 360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatgtcga 420
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag 480
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag 540
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa 600
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta 660
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg 720
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt 780
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac 840
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca 900
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct 960
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt 1020
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt 1080
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa 1140
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat 1200
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg 1260
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt 1320
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca 1380
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa 1440
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg 1500
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc 1560
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt 1620
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg 1680
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg 1740
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata 1800
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg 1860
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga 1920
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg 1980
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt 2040
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc 2100
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat 2160
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg 2220
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg 2280
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa 2340
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa 2400
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta 2460
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga 2520
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt 2580
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc 2640
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt 2700
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct 2760
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt 2820
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc 2880
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 2940
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 3000
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 3060
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 3120
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 3180
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 3240
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 3300
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 3360
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 3420
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 3480
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 3540
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 3600
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 3660
tctgacaaat ggttctttcc cctgtcctaa acaaaaaacc cgctttattg aaaaagcggg 3720
gctgttttac agacaggtca aataaacgtt tgaaaatgtt catttcaaaa cgcgcggaac 3780
ctccatcttc tcccatccag actatactgt cggcttcgga atcgcaccga atcctgccca 3840
taaaaaggct cgcgggctta gagcgcttgc tcatcaccgc cggtagggaa tttcaccctg 3900
ccccgaagat tgatcttatt tatttttaat actgatatta ttataaatta attgtgaaaa 3960
aatgtacagg tgcaaagctt attgcgctgt tttgggacat cctgcacgat atttcggtaa 4020
actcactttt tccgcatact aaaaaccgca cattcacagt tatttcattt ttaattttcg 4080
tctttccgcg tgaaactcat tgacactctt tatggaatat ggtaaattat cagatattta 4140
tgacgcttat ttaggaggaa atcttacaca gaagctgcgg aacctgaaaa gaattccttt 4200
caggttccgt tttttttagg aattctccct gatctcaagc atctggcggg gataaatccg 4260
ctctcctttc aaatcgttcc attctttgag gcgctgtaca gttacgccca ttttttcggc 4320
gatatgatga agcgtatccc ctttccgcac tacatatgta ccggtcttcg attcatcgtc 4380
atgaaggcgg agtgtttggc cggccttgag atttgaatgt ttcaacccgt ttattctcat 4440
gatctcctcg atggatatac cgctatcctt gctgattctc cagagcgtgt cccctttttg 4500
aacggtcacc gcaccgctca ttgtcccggc gttttgataa acgtggatag aattttgccg 4560
gaacgcctcc tcacgaagca ccgtcagcgg attgattgca tatcttttat cttcagtcca 4620
tgaaccgtga tgcatttcaa aatgcaggtg ggttccggtc gatattcgaa ttcctccatt 4680
ttcttctgct atcaaaataa cagactcgtg attttccaaa cgagctttca aaaaagcctc 4740
tgccccttgc aaatcggatg cctgtctata aaattcccga tattggttaa acagcggcgc 4800
aatggcggcc gcatctgatg tctttgcttg gcgaatgttc atcttatttc ttcctccctc 4860
tcaataattt tttcattcta tcccttttct gtaaagttta tttttcagaa tacttttatc 4920
atcatgcttt gaaaaaatat cacgataata tccattgttc tcacggaagc acacgcaggt 4980
catttgaacg aattttttcg acaggaattt gccgggactc aggagcattt aacctaaaaa 5040
agcatgacat ttcagcataa tgaacattta ctcatgtcta ttttcgttct tttctgtatg 5100
aaaatagtta tttcgagtct ctacggaaat agcgagagat gatataccta aatagagata 5160
aaatcatctc aaaaaaatgg gtctactaaa atattattcc atctattaca ataaattcac 5220
agaatagtct tttaagtaag tctactctga atttttttaa aaggagaggg taactagtgg 5280
ccccaaaaaa gaaacgcaag gttatggata aaaaatacag cattggtctg gatatcggaa 5340
ccaacagcgt tgggtgggca gtaataacag atgaatacaa agtgccgtca aaaaaattta 5400
aggttctggg gaatacagat cgccacagca taaaaaagaa tctgattggg gcattgctgt 5460
ttgattcggg tgagacagct gaggccacgc gtctgaaacg tacagcaaga agacgttaca 5520
cacgtcgtaa aaatcgtatt tgctacttac aggaaatttt ttctaacgaa atggccaagg 5580
tagatgatag tttcttccat cgtctcgaag aatcttttct ggttgaggaa gataaaaaac 5640
acgaacgtca ccctatcttt ggcaatatcg tggatgaagt ggcctatcat gaaaaatacc 5700
ctacgattta tcatcttcgc aagaagttgg ttgatagtac ggacaaagcg gatctgcgtt 5760
taatccatct tgcgttagcg cacatgatca aatttcgtgg tcatttctta attgaaggtg 5820
atctgaatcc tgataactct gatgtggaca aattgtttat acaattagtg caaacctata 5880
atcagctgtt cgaggaaaac cccattaatg cctctggagt tgatgccaaa gcgattttaa 5940
gcgcgagact ttctaagtcc cggcgtctgg agaatctgat cgcccagtta ccaggggaaa 6000
agaaaaatgg tctgtttggt aatctgattg ccctcagtct ggggcttacc ccgaacttca 6060
aatccaattt tgacctggct gaggacgcaa agctgcagct gagcaaagat acttatgatg 6120
atgacctcga caatctgctc gcccagattg gtgaccaata tgcggatctg tttctggcag 6180
cgaagaatct ttcggatgct atcttgctgt cggatattct gcgtgttaat accgaaatca 6240
ccaaagcgcc tctgtctgca agtatgatca agagatacga cgagcaccac caggacctga 6300
ctcttcttaa ggcactggta cgccaacagc ttccggagaa atacaaagaa atattcttcg 6360
accagtccaa gaatggttac gcgggctaca tcgatggtgg tgcatcacag gaagagttct 6420
ataaatttat taaaccaatc cttgagaaaa tggatggcac ggaagagtta cttgttaaac 6480
ttaaccgcga agacttgctt agaaagcaac gtacattcga caacggctcc atcccacacc 6540
agattcattt aggtgaactt cacgccatct tgcgcagaca agaagatttc tatcccttct 6600
taaaagacaa tcgggagaaa atcgagaaga tcctgacgtt ccgcattccc tattatgtcg 6660
gtcccctggc acgtggtaat tctcggtttg cctggatgac gcgcaaaagt gaggaaacca 6720
tcaccccttg gaactttgaa gaagtcgtgg ataaaggtgc tagcgcgcag tcttttatag 6780
aaagaatgac gaacttcgat aaaaacttgc ccaacgaaaa agtcctgccc aagcactctc 6840
ttttatatga gtactttact gtgtacaacg aactgactaa agtgaaatac gttacggaag 6900
gtatgcgcaa acctgccttt cttagtggcg agcagaaaaa agcaattgtc gatcttctct 6960
ttaaaacgaa tcgcaaggta actgtaaaac agctgaagga agattatttc aaaaagatcg 7020
aatgctttga ttctgtcgag atctcgggtg tcgaagatcg tttcaacgct tccttaggga 7080
cctatcatga tttgctgaag ataataaaag acaaagactt tctcgacaat gaagaaaatg 7140
aagatattct ggaggatatt gttttgacct tgaccttatt cgaagataga gagatgatcg 7200
aggagcgctt aaaaacctat gcccacctgt ttgatgacaa agtcatgaag caattaaagc 7260
gccgcagata tacggggtgg ggccgcttga gccgcaagtt gattaacggt attagagaca 7320
agcagagcgg aaaaactatc ctggatttcc tcaaatctga cggatttgcg aaccgcaatt 7380
ttatgcagct tatacatgat gattcgctta cattcaaaga ggatattcag aaggctcagg 7440
tgtctgggca aggtgattca ctccacgaac atatagcaaa tttggccggc tctcctgcga 7500
ttaagaaggg gatcctgcaa acagttaaag ttgtggatga acttgtaaaa gtaatgggcc 7560
gccacaagcc ggagaatatc gtgatagaaa tggcgcgcga gaatcaaacg acacaaaaag 7620
gtcaaaagaa ctcaagagag agaatgaagc gcattgagga ggggataaag gaacttggat 7680
ctcaaattct gaaagaacat ccagttgaaa acactcagct gcaaaatgaa aaattgtacc 7740
tgtactacct gcagaatgga agagacatgt acgtggatca ggaattggat atcaatagac 7800
tctcggacta tgacgtagat cacattgtcc ctcagagctt cctcaaggat gattctatag 7860
ataataaagt acttacgaga tcggacaaaa atcgcggtaa atcggataac gtcccatcgg 7920
aggaagtcgt taaaaagatg aaaaactatt ggcgtcaact gctgaacgcc aagctgatca 7980
cacagcgtaa gtttgataat ctgactaaag ccgaacgcgg tggtcttagt gaactcgata 8040
aagcaggatt tataaaacgg cagttagtag aaacgcgcca aattacgaaa cacgtggctc 8100
agatcctcga ttctagaatg aatacaaagt acgatgaaaa cgataaactg atccgtgaag 8160
taaaagtcat taccttaaaa tctaaacttg tgtccgattt ccgcaaagat tttcagtttt 8220
acaaggtccg ggaaatcaat aactatcacc atgcacatga tgcatattta aatgcggttg 8280
taggcacggc ccttattaag aaatacccta aactcgaaag tgagtttgtt tatggggatt 8340
ataaagtgta tgacgttcgc aaaatgatcg cgaaatcaga acaggaaatc ggtaaggcta 8400
ccgctaaata ctttttttat tccaacatta tgaatttttt taagaccgaa ataactctcg 8460
cgaatggtga aatccgtaaa cggcctctta tagaaaccaa tggtgaaacg ggagaaatcg 8520
tttgggataa aggtcgtgac tttgccaccg ttcgtaaagt cctctcaatg ccgcaagtta 8580
acattgtcaa gaagacggaa gttcaaacag ggggattctc caaagaatct atcctgccga 8640
agcgtaacag tgataaactt attgccagaa aaaaagattg ggatccaaaa aaatacggag 8700
gctttgattc ccctaccgtc gcgtatagtg tgctggtggt tgctaaagtc gagaaaggga 8760
aaagcaagaa attgaaatca gttaaagaac tgctgggtat tacaattatg gaaagatcgt 8820
cctttgagaa aaatccgatc gactttttag aggccaaggg gtataaggaa gtgaaaaaag 8880
atctcatcat caaattaccg aagtatagtc tttttgagct ggaaaacggc agaaaaagaa 8940
tgctggcctc cgcgggcgag ttacagaagg gaaatgagct ggcgctgcct tccaaatatg 9000
ttaattttct gtaccttgcc agtcattatg agaaactgaa gggcagcccc gaagataacg 9060
aacagaaaca attattcgtg gaacagcata agcactattt agatgaaatt atagagcaaa 9120
ttagtgaatt ttctaagcgc gttatcctcg cggatgctaa tttagacaaa gtactgtcag 9180
cttataataa acatcgggat aagccgatta gagaacaggc cgaaaatatc attcatttgt 9240
ttaccttaac caaccttgga gcaccagctg ccttcaaata tttcgatacc acaattgatc 9300
gtaaacggta tacaagtaca aaagaagtct tggacgcaac cctcattcat caatctatta 9360
ctggattata tgagacacgc attgatcttt cacagctggg cggagacaag aagaaaaaac 9420
tgaaactgca ccatcatcac catcatcatc accatcattg ataactcgag aaagcttaca 9480
taaaaaaccg gccttggccc cgccggtttt ttattatttt tcttcctccg catgttcaat 9540
ccgctccata atcgacggat ggctccctct gaaaatttta acgagaaacg gcgggttgac 9600
ccggctcagt cccgtaacgg ccaagtcctg aaacgtctca atcgccgctt cccggtttcc 9660
ggtcagctca atgccgtaac ggtcggcggc gttttcctga taccgggaga cggcattcgt 9720
aatc 9724
<210> 60
<211> 5055
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 60
attcctccat tttcttctgc tatcaaaata acagactcgt gattttccaa acgagctttc 60
aaaaaagcct ctgccccttg caaatcggat gcctgtctat aaaattcccg atattggtta 120
aacagcggcg caatggcggc cgcatctgat gtctttgctt ggcgaatgtt catcttattt 180
cttcctccct ctcaataatt ttttcattct atcccttttc tgtaaagttt atttttcaga 240
atacttttat catcatgctt tgaaaaaata tcacgataat atccattgtt ctcacggaag 300
cacacgcagg tcatttgaac gaattttttc gacaggaatt tgccgggact caggagcatt 360
taacctaaaa aagcatgaca tttcagcata atgaacattt actcatgtct attttcgttc 420
ttttctgtat gaaaatagtt atttcgagtc tctacggaaa tagcgagaga tgatatacct 480
aaatagagat aaaatcatct caaaaaaatg ggtctactaa aatattattc catctattac 540
aataaattca cagaatagtc ttttaagtaa gtctactctg aattttttta aaaggagagg 600
gtaactagtg gccccaaaaa agaaacgcaa ggttatggat aaaaaataca gcattggtct 660
ggatatcgga accaacagcg ttgggtgggc agtaataaca gatgaataca aagtgccgtc 720
aaaaaaattt aaggttctgg ggaatacaga tcgccacagc ataaaaaaga atctgattgg 780
ggcattgctg tttgattcgg gtgagacagc tgaggccacg cgtctgaaac gtacagcaag 840
aagacgttac acacgtcgta aaaatcgtat ttgctactta caggaaattt tttctaacga 900
aatggccaag gtagatgata gtttcttcca tcgtctcgaa gaatcttttc tggttgagga 960
agataaaaaa cacgaacgtc accctatctt tggcaatatc gtggatgaag tggcctatca 1020
tgaaaaatac cctacgattt atcatcttcg caagaagttg gttgatagta cggacaaagc 1080
ggatctgcgt ttaatccatc ttgcgttagc gcacatgatc aaatttcgtg gtcatttctt 1140
aattgaaggt gatctgaatc ctgataactc tgatgtggac aaattgttta tacaattagt 1200
gcaaacctat aatcagctgt tcgaggaaaa ccccattaat gcctctggag ttgatgccaa 1260
agcgatttta agcgcgagac tttctaagtc ccggcgtctg gagaatctga tcgcccagtt 1320
accaggggaa aagaaaaatg gtctgtttgg taatctgatt gccctcagtc tggggcttac 1380
cccgaacttc aaatccaatt ttgacctggc tgaggacgca aagctgcagc tgagcaaaga 1440
tacttatgat gatgacctcg acaatctgct cgcccagatt ggtgaccaat atgcggatct 1500
gtttctggca gcgaagaatc tttcggatgc tatcttgctg tcggatattc tgcgtgttaa 1560
taccgaaatc accaaagcgc ctctgtctgc aagtatgatc aagagatacg acgagcacca 1620
ccaggacctg actcttctta aggcactggt acgccaacag cttccggaga aatacaaaga 1680
aatattcttc gaccagtcca agaatggtta cgcgggctac atcgatggtg gtgcatcaca 1740
ggaagagttc tataaattta ttaaaccaat ccttgagaaa atggatggca cggaagagtt 1800
acttgttaaa cttaaccgcg aagacttgct tagaaagcaa cgtacattcg acaacggctc 1860
catcccacac cagattcatt taggtgaact tcacgccatc ttgcgcagac aagaagattt 1920
ctatcccttc ttaaaagaca atcgggagaa aatcgagaag atcctgacgt tccgcattcc 1980
ctattatgtc ggtcccctgg cacgtggtaa ttctcggttt gcctggatga cgcgcaaaag 2040
tgaggaaacc atcacccctt ggaactttga agaagtcgtg gataaaggtg ctagcgcgca 2100
gtcttttata gaaagaatga cgaacttcga taaaaacttg cccaacgaaa aagtcctgcc 2160
caagcactct cttttatatg agtactttac tgtgtacaac gaactgacta aagtgaaata 2220
cgttacggaa ggtatgcgca aacctgcctt tcttagtggc gagcagaaaa aagcaattgt 2280
cgatcttctc tttaaaacga atcgcaaggt aactgtaaaa cagctgaagg aagattattt 2340
caaaaagatc gaatgctttg attctgtcga gatctcgggt gtcgaagatc gtttcaacgc 2400
ttccttaggg acctatcatg atttgctgaa gataataaaa gacaaagact ttctcgacaa 2460
tgaagaaaat gaagatattc tggaggatat tgttttgacc ttgaccttat tcgaagatag 2520
agagatgatc gaggagcgct taaaaaccta tgcccacctg tttgatgaca aagtcatgaa 2580
gcaattaaag cgccgcagat atacggggtg gggccgcttg agccgcaagt tgattaacgg 2640
tattagagac aagcagagcg gaaaaactat cctggatttc ctcaaatctg acggatttgc 2700
gaaccgcaat tttatgcagc ttatacatga tgattcgctt acattcaaag aggatattca 2760
gaaggctcag gtgtctgggc aaggtgattc actccacgaa catatagcaa atttggccgg 2820
ctctcctgcg attaagaagg ggatcctgca aacagttaaa gttgtggatg aacttgtaaa 2880
agtaatgggc cgccacaagc cggagaatat cgtgatagaa atggcgcgcg agaatcaaac 2940
gacacaaaaa ggtcaaaaga actcaagaga gagaatgaag cgcattgagg aggggataaa 3000
ggaacttgga tctcaaattc tgaaagaaca tccagttgaa aacactcagc tgcaaaatga 3060
aaaattgtac ctgtactacc tgcagaatgg aagagacatg tacgtggatc aggaattgga 3120
tatcaataga ctctcggact atgacgtaga tcacattgtc cctcagagct tcctcaagga 3180
tgattctata gataataaag tacttacgag atcggacaaa aatcgcggta aatcggataa 3240
cgtcccatcg gaggaagtcg ttaaaaagat gaaaaactat tggcgtcaac tgctgaacgc 3300
caagctgatc acacagcgta agtttgataa tctgactaaa gccgaacgcg gtggtcttag 3360
tgaactcgat aaagcaggat ttataaaacg gcagttagta gaaacgcgcc aaattacgaa 3420
acacgtggct cagatcctcg attctagaat gaatacaaag tacgatgaaa acgataaact 3480
gatccgtgaa gtaaaagtca ttaccttaaa atctaaactt gtgtccgatt tccgcaaaga 3540
ttttcagttt tacaaggtcc gggaaatcaa taactatcac catgcacatg atgcatattt 3600
aaatgcggtt gtaggcacgg cccttattaa gaaataccct aaactcgaaa gtgagtttgt 3660
ttatggggat tataaagtgt atgacgttcg caaaatgatc gcgaaatcag aacaggaaat 3720
cggtaaggct accgctaaat acttttttta ttccaacatt atgaattttt ttaagaccga 3780
aataactctc gcgaatggtg aaatccgtaa acggcctctt atagaaacca atggtgaaac 3840
gggagaaatc gtttgggata aaggtcgtga ctttgccacc gttcgtaaag tcctctcaat 3900
gccgcaagtt aacattgtca agaagacgga agttcaaaca gggggattct ccaaagaatc 3960
tatcctgccg aagcgtaaca gtgataaact tattgccaga aaaaaagatt gggatccaaa 4020
aaaatacgga ggctttgatt cccctaccgt cgcgtatagt gtgctggtgg ttgctaaagt 4080
cgagaaaggg aaaagcaaga aattgaaatc agttaaagaa ctgctgggta ttacaattat 4140
ggaaagatcg tcctttgaga aaaatccgat cgacttttta gaggccaagg ggtataagga 4200
agtgaaaaaa gatctcatca tcaaattacc gaagtatagt ctttttgagc tggaaaacgg 4260
cagaaaaaga atgctggcct ccgcgggcga gttacagaag ggaaatgagc tggcgctgcc 4320
ttccaaatat gttaattttc tgtaccttgc cagtcattat gagaaactga agggcagccc 4380
cgaagataac gaacagaaac aattattcgt ggaacagcat aagcactatt tagatgaaat 4440
tatagagcaa attagtgaat tttctaagcg cgttatcctc gcggatgcta atttagacaa 4500
agtactgtca gcttataata aacatcggga taagccgatt agagaacagg ccgaaaatat 4560
cattcatttg tttaccttaa ccaaccttgg agcaccagct gccttcaaat atttcgatac 4620
cacaattgat cgtaaacggt atacaagtac aaaagaagtc ttggacgcaa ccctcattca 4680
tcaatctatt actggattat atgagacacg cattgatctt tcacagctgg gcggagacaa 4740
gaagaaaaaa ctgaaactgc accatcatca ccatcatcat caccatcatt gataactcga 4800
gaaagcttac ataaaaaacc ggccttggcc ccgccggttt tttattattt ttcttcctcc 4860
gcatgttcaa tccgctccat aatcgacgga tggctccctc tgaaaatttt aacgagaaac 4920
ggcgggttga cccggctcag tcccgtaacg gccaagtcct gaaacgtctc aatcgccgct 4980
tcccggtttc cggtcagctc aatgccgtaa cggtcggcgg cgttttcctg ataccgggag 5040
acggcattcg taatc 5055
<210> 61
<211> 9724
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 61
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt 60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg 120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca 180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggagatgcc atcagttcct 240
catagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa 300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca 360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatgtcga 420
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag 480
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag 540
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa 600
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta 660
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg 720
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt 780
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac 840
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca 900
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct 960
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt 1020
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt 1080
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa 1140
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat 1200
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg 1260
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt 1320
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca 1380
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa 1440
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg 1500
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc 1560
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt 1620
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg 1680
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg 1740
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata 1800
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg 1860
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga 1920
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg 1980
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt 2040
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc 2100
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat 2160
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg 2220
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg 2280
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa 2340
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa 2400
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta 2460
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga 2520
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt 2580
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc 2640
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt 2700
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct 2760
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt 2820
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc 2880
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 2940
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 3000
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 3060
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 3120
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 3180
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 3240
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 3300
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 3360
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 3420
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 3480
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 3540
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 3600
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 3660
tctgacattg atattcagca ccctgcgcat ttcgaccggg agaacgactc tgccgagctc 3720
atcgattctc cggacaatcc cggtattttt cacgtttgaa aagcctcctt ttctcctttc 3780
tttattgact tttgtcaaca tctttataat aaaagagatc ttcaaatttt ttgttgaaat 3840
actgaatcat ctttccgatc acaagttgtc cgggcctcct ttcgccattt aaaactctgc 3900
tgagtgtcgc cggggatacg ccgatttcaa tggcaagctg atttaaggag agattgtgtt 3960
caatcatgta ctggagaaca aaatctcttt tgatatgaat cttttttacc atgattactc 4020
ccctttctaa tctcttatgt ttctttttat ctacattgaa catatacgat ttgttaactt 4080
ttgtcaatac ttttaccatc catatgtttc ctataggcaa tattcgtact aaaatatttt 4140
ataataagag attgcgaggt tttggccata cttctccgcg gcacactctc ctctctatca 4200
ttttcgtctg tttacgatcc tgctgttatt ttatccctta tgttaacttt tgtcaatatt 4260
tttcctgtct aagtatttcc tatagtcaac atttgtatta aaatgttcat atcatgaatt 4320
tgcggggggg atggcgatga caaggttcgg cgagcggctc aaagagctga gggaacaaag 4380
aagcctgtcg gttaatcagc ttgccatgta tgccggtgtg agcgccgcag ccatttccag 4440
agccgcagcc atttccagaa tcgaaaacgg ccaccgcggc gttcccaagc ccgcgacgat 4500
cagaaaattg gccgaggctc tgaaaatgcc gtacgagcag ctcatggata ttgccggtta 4560
tatgagagct gacgagattc gcgaacagcc gcgcggctat gtcacgatgc aggagatcgc 4620
ggccaagcac ggcgtcgaag acctgtggct gtttaaaccc gagaaatgaa ttcctccatt 4680
ttcttctgct atcaaaataa cagactcgtg attttccaaa cgagctttca aaaaagcctc 4740
tgccccttgc aaatcggatg cctgtctata aaattcccga tattggttaa acagcggcgc 4800
aatggcggcc gcatctgatg tctttgcttg gcgaatgttc atcttatttc ttcctccctc 4860
tcaataattt tttcattcta tcccttttct gtaaagttta tttttcagaa tacttttatc 4920
atcatgcttt gaaaaaatat cacgataata tccattgttc tcacggaagc acacgcaggt 4980
catttgaacg aattttttcg acaggaattt gccgggactc aggagcattt aacctaaaaa 5040
agcatgacat ttcagcataa tgaacattta ctcatgtcta ttttcgttct tttctgtatg 5100
aaaatagtta tttcgagtct ctacggaaat agcgagagat gatataccta aatagagata 5160
aaatcatctc aaaaaaatgg gtctactaaa atattattcc atctattaca ataaattcac 5220
agaatagtct tttaagtaag tctactctga atttttttaa aaggagaggg taactagtgg 5280
ccccaaaaaa gaaacgcaag gttatggata aaaaatacag cattggtctg gatatcggaa 5340
ccaacagcgt tgggtgggca gtaataacag atgaatacaa agtgccgtca aaaaaattta 5400
aggttctggg gaatacagat cgccacagca taaaaaagaa tctgattggg gcattgctgt 5460
ttgattcggg tgagacagct gaggccacgc gtctgaaacg tacagcaaga agacgttaca 5520
cacgtcgtaa aaatcgtatt tgctacttac aggaaatttt ttctaacgaa atggccaagg 5580
tagatgatag tttcttccat cgtctcgaag aatcttttct ggttgaggaa gataaaaaac 5640
acgaacgtca ccctatcttt ggcaatatcg tggatgaagt ggcctatcat gaaaaatacc 5700
ctacgattta tcatcttcgc aagaagttgg ttgatagtac ggacaaagcg gatctgcgtt 5760
taatccatct tgcgttagcg cacatgatca aatttcgtgg tcatttctta attgaaggtg 5820
atctgaatcc tgataactct gatgtggaca aattgtttat acaattagtg caaacctata 5880
atcagctgtt cgaggaaaac cccattaatg cctctggagt tgatgccaaa gcgattttaa 5940
gcgcgagact ttctaagtcc cggcgtctgg agaatctgat cgcccagtta ccaggggaaa 6000
agaaaaatgg tctgtttggt aatctgattg ccctcagtct ggggcttacc ccgaacttca 6060
aatccaattt tgacctggct gaggacgcaa agctgcagct gagcaaagat acttatgatg 6120
atgacctcga caatctgctc gcccagattg gtgaccaata tgcggatctg tttctggcag 6180
cgaagaatct ttcggatgct atcttgctgt cggatattct gcgtgttaat accgaaatca 6240
ccaaagcgcc tctgtctgca agtatgatca agagatacga cgagcaccac caggacctga 6300
ctcttcttaa ggcactggta cgccaacagc ttccggagaa atacaaagaa atattcttcg 6360
accagtccaa gaatggttac gcgggctaca tcgatggtgg tgcatcacag gaagagttct 6420
ataaatttat taaaccaatc cttgagaaaa tggatggcac ggaagagtta cttgttaaac 6480
ttaaccgcga agacttgctt agaaagcaac gtacattcga caacggctcc atcccacacc 6540
agattcattt aggtgaactt cacgccatct tgcgcagaca agaagatttc tatcccttct 6600
taaaagacaa tcgggagaaa atcgagaaga tcctgacgtt ccgcattccc tattatgtcg 6660
gtcccctggc acgtggtaat tctcggtttg cctggatgac gcgcaaaagt gaggaaacca 6720
tcaccccttg gaactttgaa gaagtcgtgg ataaaggtgc tagcgcgcag tcttttatag 6780
aaagaatgac gaacttcgat aaaaacttgc ccaacgaaaa agtcctgccc aagcactctc 6840
ttttatatga gtactttact gtgtacaacg aactgactaa agtgaaatac gttacggaag 6900
gtatgcgcaa acctgccttt cttagtggcg agcagaaaaa agcaattgtc gatcttctct 6960
ttaaaacgaa tcgcaaggta actgtaaaac agctgaagga agattatttc aaaaagatcg 7020
aatgctttga ttctgtcgag atctcgggtg tcgaagatcg tttcaacgct tccttaggga 7080
cctatcatga tttgctgaag ataataaaag acaaagactt tctcgacaat gaagaaaatg 7140
aagatattct ggaggatatt gttttgacct tgaccttatt cgaagataga gagatgatcg 7200
aggagcgctt aaaaacctat gcccacctgt ttgatgacaa agtcatgaag caattaaagc 7260
gccgcagata tacggggtgg ggccgcttga gccgcaagtt gattaacggt attagagaca 7320
agcagagcgg aaaaactatc ctggatttcc tcaaatctga cggatttgcg aaccgcaatt 7380
ttatgcagct tatacatgat gattcgctta cattcaaaga ggatattcag aaggctcagg 7440
tgtctgggca aggtgattca ctccacgaac atatagcaaa tttggccggc tctcctgcga 7500
ttaagaaggg gatcctgcaa acagttaaag ttgtggatga acttgtaaaa gtaatgggcc 7560
gccacaagcc ggagaatatc gtgatagaaa tggcgcgcga gaatcaaacg acacaaaaag 7620
gtcaaaagaa ctcaagagag agaatgaagc gcattgagga ggggataaag gaacttggat 7680
ctcaaattct gaaagaacat ccagttgaaa acactcagct gcaaaatgaa aaattgtacc 7740
tgtactacct gcagaatgga agagacatgt acgtggatca ggaattggat atcaatagac 7800
tctcggacta tgacgtagat cacattgtcc ctcagagctt cctcaaggat gattctatag 7860
ataataaagt acttacgaga tcggacaaaa atcgcggtaa atcggataac gtcccatcgg 7920
aggaagtcgt taaaaagatg aaaaactatt ggcgtcaact gctgaacgcc aagctgatca 7980
cacagcgtaa gtttgataat ctgactaaag ccgaacgcgg tggtcttagt gaactcgata 8040
aagcaggatt tataaaacgg cagttagtag aaacgcgcca aattacgaaa cacgtggctc 8100
agatcctcga ttctagaatg aatacaaagt acgatgaaaa cgataaactg atccgtgaag 8160
taaaagtcat taccttaaaa tctaaacttg tgtccgattt ccgcaaagat tttcagtttt 8220
acaaggtccg ggaaatcaat aactatcacc atgcacatga tgcatattta aatgcggttg 8280
taggcacggc ccttattaag aaatacccta aactcgaaag tgagtttgtt tatggggatt 8340
ataaagtgta tgacgttcgc aaaatgatcg cgaaatcaga acaggaaatc ggtaaggcta 8400
ccgctaaata ctttttttat tccaacatta tgaatttttt taagaccgaa ataactctcg 8460
cgaatggtga aatccgtaaa cggcctctta tagaaaccaa tggtgaaacg ggagaaatcg 8520
tttgggataa aggtcgtgac tttgccaccg ttcgtaaagt cctctcaatg ccgcaagtta 8580
acattgtcaa gaagacggaa gttcaaacag ggggattctc caaagaatct atcctgccga 8640
agcgtaacag tgataaactt attgccagaa aaaaagattg ggatccaaaa aaatacggag 8700
gctttgattc ccctaccgtc gcgtatagtg tgctggtggt tgctaaagtc gagaaaggga 8760
aaagcaagaa attgaaatca gttaaagaac tgctgggtat tacaattatg gaaagatcgt 8820
cctttgagaa aaatccgatc gactttttag aggccaaggg gtataaggaa gtgaaaaaag 8880
atctcatcat caaattaccg aagtatagtc tttttgagct ggaaaacggc agaaaaagaa 8940
tgctggcctc cgcgggcgag ttacagaagg gaaatgagct ggcgctgcct tccaaatatg 9000
ttaattttct gtaccttgcc agtcattatg agaaactgaa gggcagcccc gaagataacg 9060
aacagaaaca attattcgtg gaacagcata agcactattt agatgaaatt atagagcaaa 9120
ttagtgaatt ttctaagcgc gttatcctcg cggatgctaa tttagacaaa gtactgtcag 9180
cttataataa acatcgggat aagccgatta gagaacaggc cgaaaatatc attcatttgt 9240
ttaccttaac caaccttgga gcaccagctg ccttcaaata tttcgatacc acaattgatc 9300
gtaaacggta tacaagtaca aaagaagtct tggacgcaac cctcattcat caatctatta 9360
ctggattata tgagacacgc attgatcttt cacagctggg cggagacaag aagaaaaaac 9420
tgaaactgca ccatcatcac catcatcatc accatcattg ataactcgag aaagcttaca 9480
taaaaaaccg gccttggccc cgccggtttt ttattatttt tcttcctccg catgttcaat 9540
ccgctccata atcgacggat ggctccctct gaaaatttta acgagaaacg gcgggttgac 9600
ccggctcagt cccgtaacgg ccaagtcctg aaacgtctca atcgccgctt cccggtttcc 9660
ggtcagctca atgccgtaac ggtcggcggc gttttcctga taccgggaga cggcattcgt 9720
aatc 9724
<210> 62
<211> 8352
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 62
gaattcctcc attttcttct gctatcaaaa taacagactc gtgattttcc aaacgagctt 60
tcaaaaaagc ctctgcccct tgcaaatcgg atgcctgtct ataaaattcc cgatattggt 120
taaacagcgg cgcaatggcg gccgcatctg atgtctttgc ttggcgaatg ttcatcttat 180
ttcttcctcc ctctcaataa ttttttcatt ctatcccttt tctgtaaagt ttatttttca 240
gaatactttt atcatcatgc tttgaaaaaa tatcacgata atatccattg ttctcacgga 300
agcacacgca ggtcatttga acgaattttt tcgacaggaa tttgccggga ctcaggagca 360
tttaacctaa aaaagcatga catttcagca taatgaacat ttactcatgt ctattttcgt 420
tcttttctgt atgaaaatag ttatttcgag tctctacgga aatagcgaga gatgatatac 480
ctaaatagag ataaaatcat ctcaaaaaaa tgggtctact aaaatattat tccatctatt 540
acaataaatt cacagaatag tcttttaagt aagtctactc tgaatttttt taaaaggaga 600
gggtaactag tggccccaaa aaagaaacgc aaggttatgg ataaaaaata cagcattggt 660
ctggatatcg gaaccaacag cgttgggtgg gcagtaataa cagatgaata caaagtgccg 720
tcaaaaaaat ttaaggttct ggggaataca gatcgccaca gcataaaaaa gaatctgatt 780
ggggcattgc tgtttgattc gggtgagaca gctgaggcca cgcgtctgaa acgtacagca 840
agaagacgtt acacacgtcg taaaaatcgt atttgctact tacaggaaat tttttctaac 900
gaaatggcca aggtagatga tagtttcttc catcgtctcg aagaatcttt tctggttgag 960
gaagataaaa aacacgaacg tcaccctatc tttggcaata tcgtggatga agtggcctat 1020
catgaaaaat accctacgat ttatcatctt cgcaagaagt tggttgatag tacggacaaa 1080
gcggatctgc gtttaatcca tcttgcgtta gcgcacatga tcaaatttcg tggtcatttc 1140
ttaattgaag gtgatctgaa tcctgataac tctgatgtgg acaaattgtt tatacaatta 1200
gtgcaaacct ataatcagct gttcgaggaa aaccccatta atgcctctgg agttgatgcc 1260
aaagcgattt taagcgcgag actttctaag tcccggcgtc tggagaatct gatcgcccag 1320
ttaccagggg aaaagaaaaa tggtctgttt ggtaatctga ttgccctcag tctggggctt 1380
accccgaact tcaaatccaa ttttgacctg gctgaggacg caaagctgca gctgagcaaa 1440
gatacttatg atgatgacct cgacaatctg ctcgcccaga ttggtgacca atatgcggat 1500
ctgtttctgg cagcgaagaa tctttcggat gctatcttgc tgtcggatat tctgcgtgtt 1560
aataccgaaa tcaccaaagc gcctctgtct gcaagtatga tcaagagata cgacgagcac 1620
caccaggacc tgactcttct taaggcactg gtacgccaac agcttccgga gaaatacaaa 1680
gaaatattct tcgaccagtc caagaatggt tacgcgggct acatcgatgg tggtgcatca 1740
caggaagagt tctataaatt tattaaacca atccttgaga aaatggatgg cacggaagag 1800
ttacttgtta aacttaaccg cgaagacttg cttagaaagc aacgtacatt cgacaacggc 1860
tccatcccac accagattca tttaggtgaa cttcacgcca tcttgcgcag acaagaagat 1920
ttctatccct tcttaaaaga caatcgggag aaaatcgaga agatcctgac gttccgcatt 1980
ccctattatg tcggtcccct ggcacgtggt aattctcggt ttgcctggat gacgcgcaaa 2040
agtgaggaaa ccatcacccc ttggaacttt gaagaagtcg tggataaagg tgctagcgcg 2100
cagtctttta tagaaagaat gacgaacttc gataaaaact tgcccaacga aaaagtcctg 2160
cccaagcact ctcttttata tgagtacttt actgtgtaca acgaactgac taaagtgaaa 2220
tacgttacgg aaggtatgcg caaacctgcc tttcttagtg gcgagcagaa aaaagcaatt 2280
gtcgatcttc tctttaaaac gaatcgcaag gtaactgtaa aacagctgaa ggaagattat 2340
ttcaaaaaga tcgaatgctt tgattctgtc gagatctcgg gtgtcgaaga tcgtttcaac 2400
gcttccttag ggacctatca tgatttgctg aagataataa aagacaaaga ctttctcgac 2460
aatgaagaaa atgaagatat tctggaggat attgttttga ccttgacctt attcgaagat 2520
agagagatga tcgaggagcg cttaaaaacc tatgcccacc tgtttgatga caaagtcatg 2580
aagcaattaa agcgccgcag atatacgggg tggggccgct tgagccgcaa gttgattaac 2640
ggtattagag acaagcagag cggaaaaact atcctggatt tcctcaaatc tgacggattt 2700
gcgaaccgca attttatgca gcttatacat gatgattcgc ttacattcaa agaggatatt 2760
cagaaggctc aggtgtctgg gcaaggtgat tcactccacg aacatatagc aaatttggcc 2820
ggctctcctg cgattaagaa ggggatcctg caaacagtta aagttgtgga tgaacttgta 2880
aaagtaatgg gccgccacaa gccggagaat atcgtgatag aaatggcgcg cgagaatcaa 2940
acgacacaaa aaggtcaaaa gaactcaaga gagagaatga agcgcattga ggaggggata 3000
aaggaacttg gatctcaaat tctgaaagaa catccagttg aaaacactca gctgcaaaat 3060
gaaaaattgt acctgtacta cctgcagaat ggaagagaca tgtacgtgga tcaggaattg 3120
gatatcaata gactctcgga ctatgacgta gatcacattg tccctcagag cttcctcaag 3180
gatgattcta tagataataa agtacttacg agatcggaca aaaatcgcgg taaatcggat 3240
aacgtcccat cggaggaagt cgttaaaaag atgaaaaact attggcgtca actgctgaac 3300
gccaagctga tcacacagcg taagtttgat aatctgacta aagccgaacg cggtggtctt 3360
agtgaactcg ataaagcagg atttataaaa cggcagttag tagaaacgcg ccaaattacg 3420
aaacacgtgg ctcagatcct cgattctaga atgaatacaa agtacgatga aaacgataaa 3480
ctgatccgtg aagtaaaagt cattacctta aaatctaaac ttgtgtccga tttccgcaaa 3540
gattttcagt tttacaaggt ccgggaaatc aataactatc accatgcaca tgatgcatat 3600
ttaaatgcgg ttgtaggcac ggcccttatt aagaaatacc ctaaactcga aagtgagttt 3660
gtttatgggg attataaagt gtatgacgtt cgcaaaatga tcgcgaaatc agaacaggaa 3720
atcggtaagg ctaccgctaa atactttttt tattccaaca ttatgaattt ttttaagacc 3780
gaaataactc tcgcgaatgg tgaaatccgt aaacggcctc ttatagaaac caatggtgaa 3840
acgggagaaa tcgtttggga taaaggtcgt gactttgcca ccgttcgtaa agtcctctca 3900
atgccgcaag ttaacattgt caagaagacg gaagttcaaa cagggggatt ctccaaagaa 3960
tctatcctgc cgaagcgtaa cagtgataaa cttattgcca gaaaaaaaga ttgggatcca 4020
aaaaaatacg gaggctttga ttcccctacc gtcgcgtata gtgtgctggt ggttgctaaa 4080
gtcgagaaag ggaaaagcaa gaaattgaaa tcagttaaag aactgctggg tattacaatt 4140
atggaaagat cgtcctttga gaaaaatccg atcgactttt tagaggccaa ggggtataag 4200
gaagtgaaaa aagatctcat catcaaatta ccgaagtata gtctttttga gctggaaaac 4260
ggcagaaaaa gaatgctggc ctccgcgggc gagttacaga agggaaatga gctggcgctg 4320
ccttccaaat atgttaattt tctgtacctt gccagtcatt atgagaaact gaagggcagc 4380
cccgaagata acgaacagaa acaattattc gtggaacagc ataagcacta tttagatgaa 4440
attatagagc aaattagtga attttctaag cgcgttatcc tcgcggatgc taatttagac 4500
aaagtactgt cagcttataa taaacatcgg gataagccga ttagagaaca ggccgaaaat 4560
atcattcatt tgtttacctt aaccaacctt ggagcaccag ctgccttcaa atatttcgat 4620
accacaattg atcgtaaacg gtatacaagt acaaaagaag tcttggacgc aaccctcatt 4680
catcaatcta ttactggatt atatgagaca cgcattgatc tttcacagct gggcggagac 4740
aagaagaaaa aactgaaact gcaccatcat caccatcatc atcaccatca ttgataactc 4800
gagaaagctt acataaaaaa ccggccttgg ccccgccggt tttttattat ttttcttcct 4860
ccgcatgttc aatccgctcc ataatcgacg gatggctccc tctgaaaatt ttaacgagaa 4920
acggcgggtt gacccggctc agtcccgtaa cggccaagtc ctgaaacgtc tcaatcgccg 4980
cttcccggtt tccggtcagc tcaatgccgt aacggtcggc ggcgttttcc tgataccggg 5040
agacggcatt cgtaatcgaa ttcgcggccg cacgcgtcca tggggatccc cgcgggtcga 5100
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag 5160
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacactctt aaattcaaaa tctatcggtc 5220
agatttatac cgatttgatt ttatatattc ttgaataaca tacgccgagt tatcacataa 5280
aagcgggaac caatcataaa atttaaactt cattgcataa tccattaaac tcttaaattc 5340
tacgattcct tgttcatcaa taaactcaat catttcttta attaatttat atctatctgt 5400
tgttgttttc tttaataatt cattaacatc tacaccgcca taaactatca tatcttcttt 5460
ttgatattta aatttattag gatcgtccat gtgaagcata tatctcacaa gacctttcac 5520
acttcctgca atctgcggaa tagtcgcatt caattcttct gtaattattt ttatctgttc 5580
ataagattta ttaccctcat acatcactag aatatgataa tgctcttttt tcatcctacc 5640
ttctgtatca gtatccctat catgtaatgg agacactaca aattgaatgt gtaactcttt 5700
taaatactct aaccactcgg cttttgctga ttctggatat aaaacaaatg tccaattacg 5760
tcctcttgaa tttttcttgt tttcagtttc ttttattaca ttttcgctca tgatataata 5820
acggtgctaa tacacttaac aaaatttagt catagatagg cagcatgcca gtgctgtcta 5880
tctttttttg tttaaaatgc accgtattcc tcctttgcat atttttttat tagaataccg 5940
gttgcatctg atttgctaat attatatttt tctttgattc tatttaatat ctcattttct 6000
tctgttgtaa gtcttaaagt aacagcaact tttttctctt cttttctatc tacaactatc 6060
actgtacctc ccaacatctg tttttttcac tttaacataa aaaacaacct tttaacatta 6120
aaaacccaat atttatttat ttgtttggac aatggacact ggacacctag gggggaggtc 6180
gtagtacccc cctatgtttt ctcccctaaa taaccccaaa aatctaagaa aaaaagacct 6240
caaaaaggtc tttaattaac atctcaaatt tcgcatttat tccaatttcc tttttgcgtg 6300
tgatgcgagc tcatcggctc cgtcgatact atgttatacg ccaactttga aaacaacttt 6360
gaaaaagctg ttttctggta tttaaggttt tagaatgcaa ggaacagtga attggagttc 6420
gtcttgttat aattagcttc ttggggtatc tttaaatact gtagaaaaga ggaaggaaat 6480
aataaatggc taaaatgaga atatcaccgg aattgaaaaa actgatcgaa aaataccgct 6540
gcgtaaaaga tacggaagga atgtctcctg ctaaggtata taagctggtg ggagaaaatg 6600
aaaacctata tttaaaaatg acggacagcc ggtataaagg gaccacctat gatgtggaac 6660
gggaaaagga catgatgcta tggctggaag gaaagctgcc tgttccaaag gtcctgcact 6720
ttgaacggca tgatggctgg agcaatctgc tcatgagtga ggccgatggc gtcctttgct 6780
cggaagagta tgaagatgaa caaagccctg aaaagattat cgagctgtat gcggagtgca 6840
tcaggctctt tcactccatc gacatatcgg attgtcccta tacgaatagc ttagacagcc 6900
gcttagccga attggattac ttactgaata acgatctggc cgatgtggat tgcgaaaact 6960
gggaagaaga cactccattt aaagatccgc gcgagctgta tgatttttta aagacggaaa 7020
agcccgaaga ggaacttgtc ttttcccacg gcgacctggg agacagcaac atctttgtga 7080
aagatggcaa agtaagtggc tttattgatc ttgggagaag cggcagggcg gacaagtggt 7140
atgacattgc cttctgcgtc cggtcgatca gggaggatat cggggaagaa cagtatgtcg 7200
agctattttt tgacttactg gggatcaagc ctgattggga gaaaataaaa tattatattt 7260
tactggatga attgttttag tgactgcagt cgggaagatc tggtaatgac tctctagctt 7320
gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg 7380
tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ctctagctaa gcagaaggcc 7440
atcctgacgg atggcctttt tgcgtttcta caaactcttg ttaactctag agctgcctgc 7500
cgcgtttcgg tgatgaagat cttcccgatg attaattaat tcagaacgct cggttgccgc 7560
cgggcgtttt ttatgaagct tcgttgctgg cgtttttcca taggctccgc ccccctgacg 7620
agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 7680
accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 7740
ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 7800
gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaccccc 7860
ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 7920
gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 7980
taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 8040
tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 8100
gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 8160
cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 8220
agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 8280
cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 8340
cttggtctga ca 8352
<210> 63
<211> 498
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 63
cacgtcgtaa aaatcgtatt tgctacttac aggaaatttt ttctaacgaa atggccaagg 60
tagatgatag tttcttccat cgtctcgaag aatcttttct ggttgaggaa gataaaaaac 120
acgaacgtca ccctatcttt ggcaatatcg tggatgaagt ggcctatcat gaaaaatacc 180
ctacgattta tcatcttcgc aagaagttgg ttgatagtac ggacaaagcg gatctgcgtt 240
taatccatct tgcgttagcg cacatgatca aatttcgtgg tcatttctta attgaaggtg 300
atctgaatcc tgataactct gatgtggaca aattgtttat acaattagtg caaacctata 360
atcagctgtt cgaggaaaac cccattaatg cctctggagt tgatgccaaa gcgattttaa 420
gcgcgagact ttctaagtcc cggcgtctgg agaatctgat cgcccagtta ccaggggaaa 480
agaaaaatgg tctgtttg 498
<210> 64
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 64
cacgtcgtaa aaatcgtatt 20
<210> 65
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 65
caaacagacc atttttcttt 20
<210> 66
<211> 7888
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 66
aaagaaaaat ggtctgtttg gtaatctgat tgccctcagt ctggggctta ccccgaactt 60
caaatccaat tttgacctgg ctgaggacgc aaagctgcag ctgagcaaag atacttatga 120
tgatgacctc gacaatctgc tcgcccagat tggtgaccaa tatgcggatc tgtttctggc 180
agcgaagaat ctttcggatg ctatcttgct gtcggatatt ctgcgtgtta ataccgaaat 240
caccaaagcg cctctgtctg caagtatgat caagagatac gacgagcacc accaggacct 300
gactcttctt aaggcactgg tacgccaaca gcttccggag aaatacaaag aaatattctt 360
cgaccagtcc aagaatggtt acgcgggcta catcgatggt ggtgcatcac aggaagagtt 420
ctataaattt attaaaccaa tccttgagaa aatggatggc acggaagagt tacttgttaa 480
acttaaccgc gaagacttgc ttagaaagca acgtacattc gacaacggct ccatcccaca 540
ccagattcat ttaggtgaac ttcacgccat cttgcgcaga caagaagatt tctatccctt 600
cttaaaagac aatcgggaga aaatcgagaa gatcctgacg ttccgcattc cctattatgt 660
cggtcccctg gcacgtggta attctcggtt tgcctggatg acgcgcaaaa gtgaggaaac 720
catcacccct tggaactttg aagaagtcgt ggataaaggt gctagcgcgc agtcttttat 780
agaaagaatg acgaacttcg ataaaaactt gcccaacgaa aaagtcctgc ccaagcactc 840
tcttttatat gagtacttta ctgtgtacaa cgaactgact aaagtgaaat acgttacgga 900
aggtatgcgc aaacctgcct ttcttagtgg cgagcagaaa aaagcaattg tcgatcttct 960
ctttaaaacg aatcgcaagg taactgtaaa acagctgaag gaagattatt tcaaaaagat 1020
cgaatgcttt gattctgtcg agatctcggg tgtcgaagat cgtttcaacg cttccttagg 1080
gacctatcat gatttgctga agataataaa agacaaagac tttctcgaca atgaagaaaa 1140
tgaagatatt ctggaggata ttgttttgac cttgacctta ttcgaagata gagagatgat 1200
cgaggagcgc ttaaaaacct atgcccacct gtttgatgac aaagtcatga agcaattaaa 1260
gcgccgcaga tatacggggt ggggccgctt gagccgcaag ttgattaacg gtattagaga 1320
caagcagagc ggaaaaacta tcctggattt cctcaaatct gacggatttg cgaaccgcaa 1380
ttttatgcag cttatacatg atgattcgct tacattcaaa gaggatattc agaaggctca 1440
ggtgtctggg caaggtgatt cactccacga acatatagca aatttggccg gctctcctgc 1500
gattaagaag gggatcctgc aaacagttaa agttgtggat gaacttgtaa aagtaatggg 1560
ccgccacaag ccggagaata tcgtgataga aatggcgcgc gagaatcaaa cgacacaaaa 1620
aggtcaaaag aactcaagag agagaatgaa gcgcattgag gaggggataa aggaacttgg 1680
atctcaaatt ctgaaagaac atccagttga aaacactcag ctgcaaaatg aaaaattgta 1740
cctgtactac ctgcagaatg gaagagacat gtacgtggat caggaattgg atatcaatag 1800
actctcggac tatgacgtag atcacattgt ccctcagagc ttcctcaagg atgattctat 1860
agataataaa gtacttacga gatcggacaa aaatcgcggt aaatcggata acgtcccatc 1920
ggaggaagtc gttaaaaaga tgaaaaacta ttggcgtcaa ctgctgaacg ccaagctgat 1980
cacacagcgt aagtttgata atctgactaa agccgaacgc ggtggtctta gtgaactcga 2040
taaagcagga tttataaaac ggcagttagt agaaacgcgc caaattacga aacacgtggc 2100
tcagatcctc gattctagaa tgaatacaaa gtacgatgaa aacgataaac tgatccgtga 2160
agtaaaagtc attaccttaa aatctaaact tgtgtccgat ttccgcaaag attttcagtt 2220
ttacaaggtc cgggaaatca ataactatca ccatgcacat gatgcatatt taaatgcggt 2280
tgtaggcacg gcccttatta agaaataccc taaactcgaa agtgagtttg tttatgggga 2340
ttataaagtg tatgacgttc gcaaaatgat cgcgaaatca gaacaggaaa tcggtaaggc 2400
taccgctaaa tacttttttt attccaacat tatgaatttt tttaagaccg aaataactct 2460
cgcgaatggt gaaatccgta aacggcctct tatagaaacc aatggtgaaa cgggagaaat 2520
cgtttgggat aaaggtcgtg actttgccac cgttcgtaaa gtcctctcaa tgccgcaagt 2580
taacattgtc aagaagacgg aagttcaaac agggggattc tccaaagaat ctatcctgcc 2640
gaagcgtaac agtgataaac ttattgccag aaaaaaagat tgggatccaa aaaaatacgg 2700
aggctttgat tcccctaccg tcgcgtatag tgtgctggtg gttgctaaag tcgagaaagg 2760
gaaaagcaag aaattgaaat cagttaaaga actgctgggt attacaatta tggaaagatc 2820
gtcctttgag aaaaatccga tcgacttttt agaggccaag gggtataagg aagtgaaaaa 2880
agatctcatc atcaaattac cgaagtatag tctttttgag ctggaaaacg gcagaaaaag 2940
aatgctggcc tccgcgggcg agttacagaa gggaaatgag ctggcgctgc cttccaaata 3000
tgttaatttt ctgtaccttg ccagtcatta tgagaaactg aagggcagcc ccgaagataa 3060
cgaacagaaa caattattcg tggaacagca taagcactat ttagatgaaa ttatagagca 3120
aattagtgaa ttttctaagc gcgttatcct cgcggatgct aatttagaca aagtactgtc 3180
agcttataat aaacatcggg ataagccgat tagagaacag gccgaaaata tcattcattt 3240
gtttacctta accaaccttg gagcaccagc tgccttcaaa tatttcgata ccacaattga 3300
tcgtaaacgg tatacaagta caaaagaagt cttggacgca accctcattc atcaatctat 3360
tactggatta tatgagacac gcattgatct ttcacagctg ggcggagaca agaagaaaaa 3420
actgaaactg caccatcatc accatcatca tcaccatcat tgataactcg agaaagctta 3480
cataaaaaac cggccttggc cccgccggtt ttttattatt tttcttcctc cgcatgttca 3540
atccgctcca taatcgacgg atggctccct ctgaaaattt taacgagaaa cggcgggttg 3600
acccggctca gtcccgtaac ggccaagtcc tgaaacgtct caatcgccgc ttcccggttt 3660
ccggtcagct caatgccgta acggtcggcg gcgttttcct gataccggga gacggcattc 3720
gtaatcgaat tcgcggccgc acgcgtccat ggggatcccc gcgggtcgac ctcgagagtt 3780
acgctaggga taacagggta atataggagc tccagtcggc ttaaaccagt tttcgctggt 3840
gcgaaaaaag agtgtcttgt gacacctaaa ttcaaaatct atcggtcaga tttataccga 3900
tttgatttta tatattcttg aataacatac gccgagttat cacataaaag cgggaaccaa 3960
tcataaaatt taaacttcat tgcataatcc attaaactct taaattctac gattccttgt 4020
tcatcaataa actcaatcat ttctttaatt aatttatatc tatctgttgt tgttttcttt 4080
aataattcat taacatctac accgccataa actatcatat cttctttttg atatttaaat 4140
ttattaggat cgtccatgtg aagcatatat ctcacaagac ctttcacact tcctgcaatc 4200
tgcggaatag tcgcattcaa ttcttctgtt aattattttt atctgttcat aagatttatt 4260
accctcatac atcactagaa tatgataatg ctcttttttc atcctacctt ctgtatcagt 4320
atccctatca tgtaatggag acactacaaa ttgaatgtgt aactctttta aatactctaa 4380
ccactcggct tttgctgatt ctggatataa aacaaatgtc caattacgtc ctcttgaatt 4440
tttcttgttt tcagtttctt ttattacatt ttcgctcatg atataataac ggtgctaata 4500
cacttaacaa aatttagtca tagataggca gcatgccagt gctgtctatc tttttttgtt 4560
taaaatgcac cgtattcctc ctttgcatat ttttttatta gaataccggt tgcatctgat 4620
ttgctaatat tatatttttc tttgattcta tttaatatct cattttcttc tgttgtaagt 4680
cttaaagtaa cagcaacttt tttctcttct tttctatcta caactatcac tgtacctccc 4740
aacatctgtt tttttcactt taacataaaa aacaaccttt taacattaaa aacccaatat 4800
ttatttattt gtttggacaa tggacactgg acacctaggg gggaggtcgt agtacccccc 4860
tatgttttct cccctaaata accccaaaaa tctaagaaaa aaagacctca aaaaggtctt 4920
taattaacat ctcaaatttc gcatttattc caatttcctt tttgcgtgtg atgcgagctc 4980
atcggctccg tcgatactat gttatacgcc aactttcaaa acaactttga aaaagctgtt 5040
ttctggtatt taaggtttta gaatgcaagg aacagtgaat tggagttcgt cttgttataa 5100
ttagcttctt ggggtatctt taaatactgt agaaaagagg aaggaaataa taaatggcta 5160
aaatgagaat atcaccggaa ttgaaaaaac tgatcgaaaa ataccgctgc gtaaaagata 5220
cggaaggaat gtctcctgct aaggtatata agctggtggg agaaaatgaa aacctatatt 5280
taaaaatgac ggacagccgg tataaaggga ccacctatga tgtggaacgg gaaaaggaca 5340
tgatgctatg gctggaagga aagctgcctg ttccaaaggt cctgcacttt gaacggcatg 5400
atggctggag caatctgctc atgagtgagg ccgatggcgt cctttgctcg gaagagtatg 5460
aagatgaaca aagccctgaa aagattatcg agctgtatgc ggagtgcatc aggctctttc 5520
actccatcga catatcggat tgtccctata cgaatagctt agacagccgc ttagccgaat 5580
tggattactt actgaataac gatctggccg atgtggattg cgaaaactgg gaagaagaca 5640
ctccatttaa agatccgcgc gagctgtatg attttttaaa gacggaaaag cccgaagagg 5700
aacttgtctt ttcccacggc gacctgggag acagcaacat ctttgtgaaa gatggcaaag 5760
taagtggctt tattgatctt gggagaagcg gcagggcgga caagtggtat gacattgcct 5820
tctgcgtccg gtcgatcagg gaggatatcg gggaagaaca gtatgtcgag ctattttttg 5880
acttactggg gatcaagcct gattgggaga aaataaaata ttatatttta ctggatgaat 5940
tgttttagtg actgcagtga gatctggtaa tgactctcta gcttgaggca tcaaataaaa 6000
cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct 6060
ctcctgagta ggacaaatcc gccgctctag ctaagcagaa ggccatcctg acggatggcc 6120
tttttgcgtt tctacaaact cttgttaact ctagagctgc ctgccgcgtt tcggtgatga 6180
agatcttccc gatgattaat taattcagaa cgctcggttg ccgccgggcg ttttttatga 6240
agcttcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 6300
acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 6360
tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 6420
ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 6480
ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 6540
ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 6600
actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 6660
gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 6720
tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 6780
caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 6840
atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 6900
acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 6960
ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagaat 7020
tcctccattt tcttctgcta tcaaaataac agactcgtga ttttccaaac gagctttcaa 7080
aaaagcctct gccccttgca aatcggatgc ctgtctataa aattcccgat attggttaaa 7140
cagcggcgca atggcggccg catctgatgt ctttgcttgg cgaatgttca tcttatttct 7200
tcctccctct caataatttt ttcattctat cccttttctg taaagtttat ttttcagaat 7260
acttttatca tcatgctttg aaaaaatatc acgataatat ccattgttct cacggaagca 7320
cacgcaggtc atttgaacga attttttcga caggaatttg ccgggactca ggagcattta 7380
acctaaaaaa gcatgacatt tcagcataat gaacatttac tcatgtctat tttcgttctt 7440
ttctgtatga aaatagttat ttcgagtctc tacggaaata gcgagagatg atatacctaa 7500
atagagataa aatcatctca aaaaaatggg tctactaaaa tattattcca tctattacaa 7560
taaattcaca gaatagtctt ttaagtaagt ctactctgaa tttttttaaa aggagagggt 7620
aactagtggc cccaaaaaag aaacgcaagg ttatggataa aaaatacagc attggtctgg 7680
atatcggaac caacagcgtt gggtgggcag taataacaga tgaatacaaa gtgccgtcaa 7740
aaaaatttaa ggttctgggg aatacagatc gccacagcat aaaaaagaat ctgattgggg 7800
cattgctgtt tgattcgggt gagacagctg aggccacgcg tctgaaacgt acagcaagaa 7860
gacgttacac acgtcgtaaa aatcgtat 7888
<210> 67
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 67
aaagaaaaat ggtctgtttg 20
<210> 68
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 68
aatacgattt ttacgacgtg 20
<210> 69
<211> 9790
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 69
gaattcctcc attttcttct gctatcaaaa taacagactc gtgattttcc aaacgagctt 60
tcaaaaaagc ctctgcccct tgcaaatcgg atgcctgtct ataaaattcc cgatattggt 120
taaacagcgg cgcaatggcg gccgcatctg atgtctttgc ttggcgaatg ttcatcttat 180
ttcttcctcc ctctcaataa ttttttcatt ctatcccttt tctgtaaagt ttatttttca 240
gaatactttt atcatcatgc tttgaaaaaa tatcacgata atatccattg ttctcacgga 300
agcacacgca ggtcatttga acgaattttt tcgacaggaa tttgccggga ctcaggagca 360
tttaacctaa aaaagcatga catttcagca taatgaacat ttactcatgt ctattttcgt 420
tcttttctgt atgaaaatag ttatttcgag tctctacgga aatagcgaga gatgatatac 480
ctaaatagag ataaaatcat ctcaaaaaaa tgggtctact aaaatattat tccatctatt 540
acaataaatt cacagaatag tcttttaagt aagtctactc tgaatttttt taaaaggaga 600
gggtaactag tggccccaaa aaagaaacgc aaggttatgg ataaaaaata cagcattggt 660
ctggatatcg gaaccaacag cgttgggtgg gcagtaataa cagatgaata caaagtgccg 720
tcaaaaaaat ttaaggttct ggggaataca gatcgccaca gcataaaaaa gaatctgatt 780
ggggcattgc tgtttgattc gggtgagaca gctgaggcca cgcgtctgaa acgtacagca 840
agaagacgtt acacacgtcg taaaaatcgt atttgctact tacaggaaat tttttctaac 900
gaaatggcca aggtagatga tagtttcttc catcgtctcg aagaatcttt tctggttgag 960
gaagataaaa aacacgaacg tcaccctatc tttggcaata tcgtggatga agtggcctat 1020
catgaaaaat accctacgat ttatcatctt cgcaagaagt tggttgatag tacggacaaa 1080
gcggatctgc gtttaatcca tcttgcgtta gcgcacatga tcaaatttcg tggtcatttc 1140
ttaattgaag gtgatctgaa tcctgataac tctgatgtgg acaaattgtt tatacaatta 1200
gtgcaaacct ataatcagct gttcgaggaa aaccccatta atgcctctgg agttgatgcc 1260
aaagcgattt taagcgcgag actttctaag tcccggcgtc tggagaatct gatcgcccag 1320
ttaccagggg aaaagaaaaa tggtctgttt ggtaatctga ttgccctcag tctggggctt 1380
accccgaact tcaaatccaa ttttgacctg gctgaggacg caaagctgca gctgagcaaa 1440
gatacttatg atgatgacct cgacaatctg ctcgcccaga ttggtgacca atatgcggat 1500
ctgtttctgg cagcgaagaa tctttcggat gctatcttgc tgtcggatat tctgcgtgtt 1560
aataccgaaa tcaccaaagc gcctctgtct gcaagtatga tcaagagata cgacgagcac 1620
caccaggacc tgactcttct taaggcactg gtacgccaac agcttccgga gaaatacaaa 1680
gaaatattct tcgaccagtc caagaatggt tacgcgggct acatcgatgg tggtgcatca 1740
caggaagagt tctataaatt tattaaacca atccttgaga aaatggatgg cacggaagag 1800
ttacttgtta aacttaaccg cgaagacttg cttagaaagc aacgtacatt cgacaacggc 1860
tccatcccac accagattca tttaggtgaa cttcacgcca tcttgcgcag acaagaagat 1920
ttctatccct tcttaaaaga caatcgggag aaaatcgaga agatcctgac gttccgcatt 1980
ccctattatg tcggtcccct ggcacgtggt aattctcggt ttgcctggat gacgcgcaaa 2040
agtgaggaaa ccatcacccc ttggaacttt gaagaagtcg tggataaagg tgctagcgcg 2100
cagtctttta tagaaagaat gacgaacttc gataaaaact tgcccaacga aaaagtcctg 2160
cccaagcact ctcttttata tgagtacttt actgtgtaca acgaactgac taaagtgaaa 2220
tacgttacgg aaggtatgcg caaacctgcc tttcttagtg gcgagcagaa aaaagcaatt 2280
gtcgatcttc tctttaaaac gaatcgcaag gtaactgtaa aacagctgaa ggaagattat 2340
ttcaaaaaga tcgaatgctt tgattctgtc gagatctcgg gtgtcgaaga tcgtttcaac 2400
gcttccttag ggacctatca tgatttgctg aagataataa aagacaaaga ctttctcgac 2460
aatgaagaaa atgaagatat tctggaggat attgttttga ccttgacctt attcgaagat 2520
agagagatga tcgaggagcg cttaaaaacc tatgcccacc tgtttgatga caaagtcatg 2580
aagcaattaa agcgccgcag atatacgggg tggggccgct tgagccgcaa gttgattaac 2640
ggtattagag acaagcagag cggaaaaact atcctggatt tcctcaaatc tgacggattt 2700
gcgaaccgca attttatgca gcttatacat gatgattcgc ttacattcaa agaggatatt 2760
cagaaggctc aggtgtctgg gcaaggtgat tcactccacg aacatatagc aaatttggcc 2820
ggctctcctg cgattaagaa ggggatcctg caaacagtta aagttgtgga tgaacttgta 2880
aaagtaatgg gccgccacaa gccggagaat atcgtgatag aaatggcgcg cgagaatcaa 2940
acgacacaaa aaggtcaaaa gaactcaaga gagagaatga agcgcattga ggaggggata 3000
aaggaacttg gatctcaaat tctgaaagaa catccagttg aaaacactca gctgcaaaat 3060
gaaaaattgt acctgtacta cctgcagaat ggaagagaca tgtacgtgga tcaggaattg 3120
gatatcaata gactctcgga ctatgacgta gatcacattg tccctcagag cttcctcaag 3180
gatgattcta tagataataa agtacttacg agatcggaca aaaatcgcgg taaatcggat 3240
aacgtcccat cggaggaagt cgttaaaaag atgaaaaact attggcgtca actgctgaac 3300
gccaagctga tcacacagcg taagtttgat aatctgacta aagccgaacg cggtggtctt 3360
agtgaactcg ataaagcagg atttataaaa cggcagttag tagaaacgcg ccaaattacg 3420
aaacacgtgg ctcagatcct cgattctaga atgaatacaa agtacgatga aaacgataaa 3480
ctgatccgtg aagtaaaagt cattacctta aaatctaaac ttgtgtccga tttccgcaaa 3540
gattttcagt tttacaaggt ccgggaaatc aataactatc accatgcaca tgatgcatat 3600
ttaaatgcgg ttgtaggcac ggcccttatt aagaaatacc ctaaactcga aagtgagttt 3660
gtttatgggg attataaagt gtatgacgtt cgcaaaatga tcgcgaaatc agaacaggaa 3720
atcggtaagg ctaccgctaa atactttttt tattccaaca ttatgaattt ttttaagacc 3780
gaaataactc tcgcgaatgg tgaaatccgt aaacggcctc ttatagaaac caatggtgaa 3840
acgggagaaa tcgtttggga taaaggtcgt gactttgcca ccgttcgtaa agtcctctca 3900
atgccgcaag ttaacattgt caagaagacg gaagttcaaa cagggggatt ctccaaagaa 3960
tctatcctgc cgaagcgtaa cagtgataaa cttattgcca gaaaaaaaga ttgggatcca 4020
aaaaaatacg gaggctttga ttcccctacc gtcgcgtata gtgtgctggt ggttgctaaa 4080
gtcgagaaag ggaaaagcaa gaaattgaaa tcagttaaag aactgctggg tattacaatt 4140
atggaaagat cgtcctttga gaaaaatccg atcgactttt tagaggccaa ggggtataag 4200
gaagtgaaaa aagatctcat catcaaatta ccgaagtata gtctttttga gctggaaaac 4260
ggcagaaaaa gaatgctggc ctccgcgggc gagttacaga agggaaatga gctggcgctg 4320
ccttccaaat atgttaattt tctgtacctt gccagtcatt atgagaaact gaagggcagc 4380
cccgaagata acgaacagaa acaattattc gtggaacagc ataagcacta tttagatgaa 4440
attatagagc aaattagtga attttctaag cgcgttatcc tcgcggatgc taatttagac 4500
aaagtactgt cagcttataa taaacatcgg gataagccga ttagagaaca ggccgaaaat 4560
atcattcatt tgtttacctt aaccaacctt ggagcaccag ctgccttcaa atatttcgat 4620
accacaattg atcgtaaacg gtatacaagt acaaaagaag tcttggacgc aaccctcatt 4680
catcaatcta ttactggatt atatgagaca cgcattgatc tttcacagct gggcggagac 4740
aagaagaaaa aactgaaact gcaccatcat caccatcatc atcaccatca ttgataactc 4800
gagaaagctt acataaaaaa ccggccttgg ccccgccggt tttttattat ttttcttcct 4860
ccgcatgttc aatccgctcc ataatcgacg gatggctccc tctgaaaatt ttaacgagaa 4920
acggcgggtt gacccggctc agtcccgtaa cggccaagtc ctgaaacgtc tcaatcgccg 4980
cttcccggtt tccggtcagc tcaatgccgt aacggtcggc ggcgttttcc tgataccggg 5040
agacggcatt cgtaatcggg tgaagtggtc aagacctcac taggcacctt aaaaatagcg 5100
caccctgaag aagatttatt tgaggtagcc cttgcctacc tagcttccaa gaaagatatc 5160
ctaacagcac aagagcggaa agatgttttg ttctacatcc agaacaacct ctgctaaaat 5220
tcctgaaaaa ttttgcaaaa agttgttgac tttatctaca aggtgtggca taatgtgtgg 5280
aagaatcgaa aacggccacc ggttttagag ctagaaatag caagttaaaa taaggctagt 5340
ccgttatcaa cttgaaaaag tggcaccgag tcggtgcgac tcctgttgat agatccagta 5400
atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt 5460
attggtgaga atcgcgtcta cagtccagga agcaagaagc agctatgatt ccatttacga 5520
catcgtgtca cagtacgatt tagaggacct ttctctgttt gacagcgaaa agtggaaggt 5580
gctttcaaaa aaagacatcg aaaacctgga caaatatttc gactttctcg tgcaggaagc 5640
aagcagccga aacaaaaact gaatacttct ccgcggcaca ctctcctctc tatcattttc 5700
gtctgtttac gatcctgctg ttattttatc ccttatgtta acttttgtca atatttttcc 5760
tgtctaagta tttcctatag tcaacatttg tattaaaatg ttcatatcat gaatttgcgg 5820
gggggatggc gatgacaagg ttcggcgagc ggctcaaaga gctgagggaa caaagaagcc 5880
tgtcggttaa tcagcttgcc atgtatgccg gtgtgagcgc cgcagccatt tccagaatcg 5940
aaaacggcca ccgctaagtt cccaagcccg cgacgatcag aaaattggcc tgataactga 6000
aaatgccgta cgagcagctc atggatattg ccggttatat gagagctgac gagattcgcg 6060
aacagccgcg cggctatgtc acgatgcagg agatcgcggc caagcacggc gtcgaagacc 6120
tgtggctgtt taaacccgag aaatgggact gtttgtcccg cgaagacctg ctcaacctcg 6180
aacagtattt tcattttttg gttaatgaag cgaagaagcg ccaatcataa aaagccgaat 6240
ttccctttta ggagaagttc ggcttttttc ggctgcctta agcggcatcc ggattcggcg 6300
tcttgccttt atgatgctta acggggctca gcgcacgctc gagccatccc atgaacagat 6360
cggcgatgat cgccatcagc gccgtcggga tcgcgcctgc tagaatgatc gctgttccgt 6420
tggtcgcgtt tgatcccctg acaatgatat ccccgaggcc gcctgcgccg acaaacgtgc 6480
cgatggccgt aatgcgaatt cgcggccgca cgcgtccatg gggatccccg cgggtcgacc 6540
tcgagagtta cgctagggat aacagggtaa tataggagct ccagtcggct taaaccagtt 6600
ttcgctggtg cgaaaaaaga gtgtcttgtg acactcttaa attcaaaatc tatcggtcag 6660
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa 6720
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta 6780
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg 6840
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt 6900
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac 6960
ttcctgcaat ctgcggaata gtcgcattca attcttctgt aattattttt atctgttcat 7020
aagatttatt accctcatac atcactagaa tatgataatg ctcttttttc atcctacctt 7080
ctgtatcagt atccctatca tgtaatggag acactacaaa ttgaatgtgt aactctttta 7140
aatactctaa ccactcggct tttgctgatt ctggatataa aacaaatgtc caattacgtc 7200
ctcttgaatt tttcttgttt tcagtttctt ttattacatt ttcgctcatg atataataac 7260
ggtgctaata cacttaacaa aatttagtca tagataggca gcatgccagt gctgtctatc 7320
tttttttgtt taaaatgcac cgtattcctc ctttgcatat ttttttatta gaataccggt 7380
tgcatctgat ttgctaatat tatatttttc tttgattcta tttaatatct cattttcttc 7440
tgttgtaagt cttaaagtaa cagcaacttt tttctcttct tttctatcta caactatcac 7500
tgtacctccc aacatctgtt tttttcactt taacataaaa aacaaccttt taacattaaa 7560
aacccaatat ttatttattt gtttggacaa tggacactgg acacctaggg gggaggtcgt 7620
agtacccccc tatgttttct cccctaaata accccaaaaa tctaagaaaa aaagacctca 7680
aaaaggtctt taattaacat ctcaaatttc gcatttattc caatttcctt tttgcgtgtg 7740
atgcgagctc atcggctccg tcgatactat gttatacgcc aactttgaaa acaactttga 7800
aaaagctgtt ttctggtatt taaggtttta gaatgcaagg aacagtgaat tggagttcgt 7860
cttgttataa ttagcttctt ggggtatctt taaatactgt agaaaagagg aaggaaataa 7920
taaatggcta aaatgagaat atcaccggaa ttgaaaaaac tgatcgaaaa ataccgctgc 7980
gtaaaagata cggaaggaat gtctcctgct aaggtatata agctggtggg agaaaatgaa 8040
aacctatatt taaaaatgac ggacagccgg tataaaggga ccacctatga tgtggaacgg 8100
gaaaaggaca tgatgctatg gctggaagga aagctgcctg ttccaaaggt cctgcacttt 8160
gaacggcatg atggctggag caatctgctc atgagtgagg ccgatggcgt cctttgctcg 8220
gaagagtatg aagatgaaca aagccctgaa aagattatcg agctgtatgc ggagtgcatc 8280
aggctctttc actccatcga catatcggat tgtccctata cgaatagctt agacagccgc 8340
ttagccgaat tggattactt actgaataac gatctggccg atgtggattg cgaaaactgg 8400
gaagaagaca ctccatttaa agatccgcgc gagctgtatg attttttaaa gacggaaaag 8460
cccgaagagg aacttgtctt ttcccacggc gacctgggag acagcaacat ctttgtgaaa 8520
gatggcaaag taagtggctt tattgatctt gggagaagcg gcagggcgga caagtggtat 8580
gacattgcct tctgcgtccg gtcgatcagg gaggatatcg gggaagaaca gtatgtcgag 8640
ctattttttg acttactggg gatcaagcct gattgggaga aaataaaata ttatatttta 8700
ctggatgaat tgttttagtg actgcagtcg ggaagatctg gtaatgactc tctagcttga 8760
ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt 8820
tgtcggtgaa cgctctcctg agtaggacaa atccgccgct ctagctaagc agaaggccat 8880
cctgacggat ggcctttttg cgtttctaca aactcttgtt aactctagag ctgcctgccg 8940
cgtttcggtg atgaagatct tcccgatgat taattaattc agaacgctcg gttgccgccg 9000
ggcgtttttt atgaagcttc gttgctggcg tttttccata ggctccgccc ccctgacgag 9060
catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 9120
caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 9180
ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 9240
aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaccccccc 9300
gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 9360
cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 9420
ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 9480
tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 9540
tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 9600
cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 9660
tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 9720
tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 9780
tggtctgaca 9790
<210> 70
<211> 399
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 70
atgacaaggt tcggcgagcg gctcaaagag ctgagggaac aaagaagcct gtcggttaat 60
cagcttgcca tgtatgccgg tgtgagcgcc gcagccattt ccagaatcga aaacggccac 120
cgcggcgttc ccaagcccgc gacgatcaga aaattggccg aggctctgaa aatgccgtac 180
gagcagctca tggatattgc cggttatatg agagctgacg agattcgcga acagccgcgc 240
ggctatgtca cgatgcagga gatcgcggcc aagcacggcg tcgaagacct gtggctgttt 300
aaacccgaga aatgggactg tttgtcccgc gaagacctgc tcaacctcga acagtatttt 360
cattttttgg ttaatgaagc gaagaagcgc caatcataa 399
<210> 71
<211> 1438
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 71
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt 60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg 120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca 180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggaagaatc gaaaacggcc 240
accggtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa 300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca 360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatcgcgt 420
ctacagtcca ggaagcaaga agcagctatg attccattta cgacatcgtg tcacagtacg 480
atttagagga cctttctctg tttgacagcg aaaagtggaa ggtgctttca aaaaaagaca 540
tcgaaaacct ggacaaatat ttcgactttc tcgtgcagga agcaagcagc cgaaacaaaa 600
actgaatact tctccgcggc acactctcct ctctatcatt ttcgtctgtt tacgatcctg 660
ctgttatttt atcccttatg ttaacttttg tcaatatttt tcctgtctaa gtatttccta 720
tagtcaacat ttgtattaaa atgttcatat catgaatttg cgggggggat ggcgatgaca 780
aggttcggcg agcggctcaa agagctgagg gaacaaagaa gcctgtcggt taatcagctt 840
gccatgtatg ccggtgtgag cgccgcagcc atttccagaa tcgaaaacgg ccaccgctaa 900
gttcccaagc ccgcgacgat cagaaaattg gcctgataac tgaaaatgcc gtacgagcag 960
ctcatggata ttgccggtta tatgagagct gacgagattc gcgaacagcc gcgcggctat 1020
gtcacgatgc aggagatcgc ggccaagcac ggcgtcgaag acctgtggct gtttaaaccc 1080
gagaaatggg actgtttgtc ccgcgaagac ctgctcaacc tcgaacagta ttttcatttt 1140
ttggttaatg aagcgaagaa gcgccaatca taaaaagccg aatttccctt ttaggagaag 1200
ttcggctttt ttcggctgcc ttaagcggca tccggattcg gcgtcttgcc tttatgatgc 1260
ttaacggggc tcagcgcacg ctcgagccat cccatgaaca gatcggcgat gatcgccatc 1320
agcgccgtcg ggatcgcgcc tgctagaatg atcgctgttc cgttggtcgc gtttgatccc 1380
ctgacaatga tatccccgag gccgcctgcg ccgacaaacg tgccgatggc cgtaatgc 1438
<210> 72
<211> 1023
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 72
cgcgtctaca gtccaggaag caagaagcag ctatgattcc atttacgaca tcgtgtcaca 60
gtacgattta gaggaccttt ctctgtttga cagcgaaaag tggaaggtgc tttcaaaaaa 120
agacatcgaa aacctggaca aatatttcga ctttctcgtg caggaagcaa gcagccgaaa 180
caaaaactga atacttctcc gcggcacact ctcctctcta tcattttcgt ctgtttacga 240
tcctgctgtt attttatccc ttatgttaac ttttgtcaat atttttcctg tctaagtatt 300
tcctatagtc aacatttgta ttaaaatgtt catatcatga atttgcgggg gggatggcga 360
tgacaaggtt cggcgagcgg ctcaaagagc tgagggaaca aagaagcctg tcggttaatc 420
agcttgccat gtatgccggt gtgagcgccg cagccatttc cagaatcgaa aacggccacc 480
gctaagttcc caagcccgcg acgatcagaa aattggcctg ataactgaaa atgccgtacg 540
agcagctcat ggatattgcc ggttatatga gagctgacga gattcgcgaa cagccgcgcg 600
gctatgtcac gatgcaggag atcgcggcca agcacggcgt cgaagacctg tggctgttta 660
aacccgagaa atgggactgt ttgtcccgcg aagacctgct caacctcgaa cagtattttc 720
attttttggt taatgaagcg aagaagcgcc aatcataaaa agccgaattt cccttttagg 780
agaagttcgg cttttttcgg ctgccttaag cggcatccgg attcggcgtc ttgcctttat 840
gatgcttaac ggggctcagc gcacgctcga gccatcccat gaacagatcg gcgatgatcg 900
ccatcagcgc cgtcgggatc gcgcctgcta gaatgatcgc tgttccgttg gtcgcgtttg 960
atcccctgac aatgatatcc ccgaggccgc ctgcgccgac aaacgtgccg atggccgtaa 1020
tgc 1023
<210> 73
<211> 415
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 73
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt 60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg 120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca 180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggaagaatc gaaaacggcc 240
accggtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa 300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca 360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaat 415
<210> 74
<211> 17
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 74
cgtgcggccg cgaattc 17
<210> 75
<211> 29
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 75
cctgataccg ggagacggca ttcgtaatc 29
<210> 76
<211> 17
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 76
gaattcgcgg ccgcacg 17
<210> 77
<211> 29
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 77
gattacgaat gccgtctccc ggtatcagg 29
<210> 78
<211> 9738
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 78
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt 60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg 120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca 180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggagcgagc ggctcaaaga 240
gctggtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa 300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca 360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatgtcga 420
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag 480
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag 540
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa 600
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta 660
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg 720
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt 780
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac 840
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca 900
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct 960
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt 1020
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt 1080
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa 1140
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat 1200
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg 1260
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt 1320
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca 1380
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa 1440
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg 1500
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc 1560
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt 1620
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg 1680
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg 1740
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata 1800
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg 1860
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga 1920
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg 1980
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt 2040
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc 2100
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat 2160
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg 2220
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg 2280
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa 2340
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa 2400
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta 2460
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga 2520
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt 2580
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc 2640
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt 2700
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct 2760
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt 2820
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc 2880
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 2940
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 3000
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 3060
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 3120
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 3180
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 3240
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 3300
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 3360
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 3420
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 3480
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 3540
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 3600
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 3660
tctgacacgt cagttcggca ggcatttcgc gaatcgaaaa cggaaagcgc ggcgtgccga 3720
agccggcgac gatcagaaaa ctggcggacg ctttgaaagt cccgtatgag gaactgatgg 3780
catctgcagg ctatatcagc gcgtctacag tccaggaagc aagaagcagc tatgattcca 3840
tttacgacat cgtgtcacag tacgatttag aggacctttc tctgtttgac agcgaaaagt 3900
ggaaggtgct ttcaaaaaaa gacatcgaaa acctggacaa atatttcgac tttctcgtgc 3960
aggaagcaag cagccgaaac aaaaactgaa tacttctccg cggcacactc tcctctctat 4020
cattttcgtc tgtttacgat cctgctgtta ttttatccct tatgttaact tttgtcaata 4080
tttttcctgt ctaagtattt cctatagtca acatttgtat taaaatgttc atatcatgaa 4140
tttgcggggg ggatggcgat gacaaggcaa tcataaaaag ccgaatttcc cttttaggag 4200
aagttcggct tttttcggct gccttaagcg gcatccggat tcggcgtctt gcctttatga 4260
tgcttaacgg ggctcagcgc acgctcgagc catcccatga acagatcggc gatgatcgcc 4320
atcagcgccg tcgggatcgc gcctgctaga atgatcgctg ttccgttggt cgcgtttgat 4380
cccctgacaa tgatatcccc gaggccgcct gcgccgacaa acgtgccgat ggccgtaatg 4440
ccgatcgcga tgacgagcgc ggttctgagc cccgccataa tgaccgacaa ggcgagggga 4500
agctccacca tccggagcac ttgaaatttc gtcatgccca tcgccttccc tgattcaaga 4560
taggcatgct cgatgctggc gattcccgta tatgtgtttc gaatgatcgg caacagcgaa 4620
tacaaaaaca atgaaagaat caccgtgttt gcgccgagcc ccatgacaag catcaagacg 4680
ggaattcctc cattttcttc tgctatcaaa ataacagact cgtgattttc caaacgagct 4740
ttcaaaaaag cctctgcccc ttgcaaatcg gatgcctgtc tataaaattc ccgatattgg 4800
ttaaacagcg gcgcaatggc ggccgcatct gatgtctttg cttggcgaat gttcatctta 4860
tttcttcctc cctctcaata attttttcat tctatccctt ttctgtaaag tttatttttc 4920
agaatacttt tatcatcatg ctttgaaaaa atatcacgat aatatccatt gttctcacgg 4980
aagcacacgc aggtcatttg aacgaatttt ttcgacagga atttgccggg actcaggagc 5040
atttaaccta aaaaagcatg acatttcagc ataatgaaca tttactcatg tctattttcg 5100
ttcttttctg tatgaaaata gttatttcga gtctctacgg aaatagcgag agatgatata 5160
cctaaataga gataaaatca tctcaaaaaa atgggtctac taaaatatta ttccatctat 5220
tacaataaat tcacagaata gtcttttaag taagtctact ctgaattttt ttaaaaggag 5280
agggtaacta gtggccccaa aaaagaaacg caaggttatg gataaaaaat acagcattgg 5340
tctggatatc ggaaccaaca gcgttgggtg ggcagtaata acagatgaat acaaagtgcc 5400
gtcaaaaaaa tttaaggttc tggggaatac agatcgccac agcataaaaa agaatctgat 5460
tggggcattg ctgtttgatt cgggtgagac agctgaggcc acgcgtctga aacgtacagc 5520
aagaagacgt tacacacgtc gtaaaaatcg tatttgctac ttacaggaaa ttttttctaa 5580
cgaaatggcc aaggtagatg atagtttctt ccatcgtctc gaagaatctt ttctggttga 5640
ggaagataaa aaacacgaac gtcaccctat ctttggcaat atcgtggatg aagtggccta 5700
tcatgaaaaa taccctacga tttatcatct tcgcaagaag ttggttgata gtacggacaa 5760
agcggatctg cgtttaatcc atcttgcgtt agcgcacatg atcaaatttc gtggtcattt 5820
cttaattgaa ggtgatctga atcctgataa ctctgatgtg gacaaattgt ttatacaatt 5880
agtgcaaacc tataatcagc tgttcgagga aaaccccatt aatgcctctg gagttgatgc 5940
caaagcgatt ttaagcgcga gactttctaa gtcccggcgt ctggagaatc tgatcgccca 6000
gttaccaggg gaaaagaaaa atggtctgtt tggtaatctg attgccctca gtctggggct 6060
taccccgaac ttcaaatcca attttgacct ggctgaggac gcaaagctgc agctgagcaa 6120
agatacttat gatgatgacc tcgacaatct gctcgcccag attggtgacc aatatgcgga 6180
tctgtttctg gcagcgaaga atctttcgga tgctatcttg ctgtcggata ttctgcgtgt 6240
taataccgaa atcaccaaag cgcctctgtc tgcaagtatg atcaagagat acgacgagca 6300
ccaccaggac ctgactcttc ttaaggcact ggtacgccaa cagcttccgg agaaatacaa 6360
agaaatattc ttcgaccagt ccaagaatgg ttacgcgggc tacatcgatg gtggtgcatc 6420
acaggaagag ttctataaat ttattaaacc aatccttgag aaaatggatg gcacggaaga 6480
gttacttgtt aaacttaacc gcgaagactt gcttagaaag caacgtacat tcgacaacgg 6540
ctccatccca caccagattc atttaggtga acttcacgcc atcttgcgca gacaagaaga 6600
tttctatccc ttcttaaaag acaatcggga gaaaatcgag aagatcctga cgttccgcat 6660
tccctattat gtcggtcccc tggcacgtgg taattctcgg tttgcctgga tgacgcgcaa 6720
aagtgaggaa accatcaccc cttggaactt tgaagaagtc gtggataaag gtgctagcgc 6780
gcagtctttt atagaaagaa tgacgaactt cgataaaaac ttgcccaacg aaaaagtcct 6840
gcccaagcac tctcttttat atgagtactt tactgtgtac aacgaactga ctaaagtgaa 6900
atacgttacg gaaggtatgc gcaaacctgc ctttcttagt ggcgagcaga aaaaagcaat 6960
tgtcgatctt ctctttaaaa cgaatcgcaa ggtaactgta aaacagctga aggaagatta 7020
tttcaaaaag atcgaatgct ttgattctgt cgagatctcg ggtgtcgaag atcgtttcaa 7080
cgcttcctta gggacctatc atgatttgct gaagataata aaagacaaag actttctcga 7140
caatgaagaa aatgaagata ttctggagga tattgttttg accttgacct tattcgaaga 7200
tagagagatg atcgaggagc gcttaaaaac ctatgcccac ctgtttgatg acaaagtcat 7260
gaagcaatta aagcgccgca gatatacggg gtggggccgc ttgagccgca agttgattaa 7320
cggtattaga gacaagcaga gcggaaaaac tatcctggat ttcctcaaat ctgacggatt 7380
tgcgaaccgc aattttatgc agcttataca tgatgattcg cttacattca aagaggatat 7440
tcagaaggct caggtgtctg ggcaaggtga ttcactccac gaacatatag caaatttggc 7500
cggctctcct gcgattaaga aggggatcct gcaaacagtt aaagttgtgg atgaacttgt 7560
aaaagtaatg ggccgccaca agccggagaa tatcgtgata gaaatggcgc gcgagaatca 7620
aacgacacaa aaaggtcaaa agaactcaag agagagaatg aagcgcattg aggaggggat 7680
aaaggaactt ggatctcaaa ttctgaaaga acatccagtt gaaaacactc agctgcaaaa 7740
tgaaaaattg tacctgtact acctgcagaa tggaagagac atgtacgtgg atcaggaatt 7800
ggatatcaat agactctcgg actatgacgt agatcacatt gtccctcaga gcttcctcaa 7860
ggatgattct atagataata aagtacttac gagatcggac aaaaatcgcg gtaaatcgga 7920
taacgtccca tcggaggaag tcgttaaaaa gatgaaaaac tattggcgtc aactgctgaa 7980
cgccaagctg atcacacagc gtaagtttga taatctgact aaagccgaac gcggtggtct 8040
tagtgaactc gataaagcag gatttataaa acggcagtta gtagaaacgc gccaaattac 8100
gaaacacgtg gctcagatcc tcgattctag aatgaataca aagtacgatg aaaacgataa 8160
actgatccgt gaagtaaaag tcattacctt aaaatctaaa cttgtgtccg atttccgcaa 8220
agattttcag ttttacaagg tccgggaaat caataactat caccatgcac atgatgcata 8280
tttaaatgcg gttgtaggca cggcccttat taagaaatac cctaaactcg aaagtgagtt 8340
tgtttatggg gattataaag tgtatgacgt tcgcaaaatg atcgcgaaat cagaacagga 8400
aatcggtaag gctaccgcta aatacttttt ttattccaac attatgaatt tttttaagac 8460
cgaaataact ctcgcgaatg gtgaaatccg taaacggcct cttatagaaa ccaatggtga 8520
aacgggagaa atcgtttggg ataaaggtcg tgactttgcc accgttcgta aagtcctctc 8580
aatgccgcaa gttaacattg tcaagaagac ggaagttcaa acagggggat tctccaaaga 8640
atctatcctg ccgaagcgta acagtgataa acttattgcc agaaaaaaag attgggatcc 8700
aaaaaaatac ggaggctttg attcccctac cgtcgcgtat agtgtgctgg tggttgctaa 8760
agtcgagaaa gggaaaagca agaaattgaa atcagttaaa gaactgctgg gtattacaat 8820
tatggaaaga tcgtcctttg agaaaaatcc gatcgacttt ttagaggcca aggggtataa 8880
ggaagtgaaa aaagatctca tcatcaaatt accgaagtat agtctttttg agctggaaaa 8940
cggcagaaaa agaatgctgg cctccgcggg cgagttacag aagggaaatg agctggcgct 9000
gccttccaaa tatgttaatt ttctgtacct tgccagtcat tatgagaaac tgaagggcag 9060
ccccgaagat aacgaacaga aacaattatt cgtggaacag cataagcact atttagatga 9120
aattatagag caaattagtg aattttctaa gcgcgttatc ctcgcggatg ctaatttaga 9180
caaagtactg tcagcttata ataaacatcg ggataagccg attagagaac aggccgaaaa 9240
tatcattcat ttgtttacct taaccaacct tggagcacca gctgccttca aatatttcga 9300
taccacaatt gatcgtaaac ggtatacaag tacaaaagaa gtcttggacg caaccctcat 9360
tcatcaatct attactggat tatatgagac acgcattgat ctttcacagc tgggcggaga 9420
caagaagaaa aaactgaaac tgcaccatca tcaccatcat catcaccatc attgataact 9480
cgagaaagct tacataaaaa accggccttg gccccgccgg ttttttatta tttttcttcc 9540
tccgcatgtt caatccgctc cataatcgac ggatggctcc ctctgaaaat tttaacgaga 9600
aacggcgggt tgacccggct cagtcccgta acggccaagt cctgaaacgt ctcaatcgcc 9660
gcttcccggt ttccggtcag ctcaatgccg taacggtcgg cggcgttttc ctgataccgg 9720
gagacggcat tcgtaatc 9738
<210> 79
<211> 23
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 79
gcgagcggct caaagagctg agg 23
<210> 80
<211> 1014
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 80
cgtcagttcg gcaggcattt cgcgaatcga aaacggaaag cgcggcgtgc cgaagccggc 60
gacgatcaga aaactggcgg acgctttgaa agtcccgtat gaggaactga tggcatctgc 120
aggctatatc agcgcgtcta cagtccagga agcaagaagc agctatgatt ccatttacga 180
catcgtgtca cagtacgatt tagaggacct ttctctgttt gacagcgaaa agtggaaggt 240
gctttcaaaa aaagacatcg aaaacctgga caaatatttc gactttctcg tgcaggaagc 300
aagcagccga aacaaaaact gaatacttct ccgcggcaca ctctcctctc tatcattttc 360
gtctgtttac gatcctgctg ttattttatc ccttatgtta acttttgtca atatttttcc 420
tgtctaagta tttcctatag tcaacatttg tattaaaatg ttcatatcat gaatttgcgg 480
gggggatggc gatgacaagg caatcataaa aagccgaatt tcccttttag gagaagttcg 540
gcttttttcg gctgccttaa gcggcatccg gattcggcgt cttgccttta tgatgcttaa 600
cggggctcag cgcacgctcg agccatccca tgaacagatc ggcgatgatc gccatcagcg 660
ccgtcgggat cgcgcctgct agaatgatcg ctgttccgtt ggtcgcgttt gatcccctga 720
caatgatatc cccgaggccg cctgcgccga caaacgtgcc gatggccgta atgccgatcg 780
cgatgacgag cgcggttctg agccccgcca taatgaccga caaggcgagg ggaagctcca 840
ccatccggag cacttgaaat ttcgtcatgc ccatcgcctt ccctgattca agataggcat 900
gctcgatgct ggcgattccc gtatatgtgt ttcgaatgat cggcaacagc gaatacaaaa 960
acaatgaaag aatcaccgtg tttgcgccga gccccatgac aagcatcaag acgg 1014
<210> 81
<211> 9724
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 81
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt 60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg 120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca 180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggagagctt ctttttcttg 240
agcagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa 300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca 360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatgtcga 420
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag 480
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag 540
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa 600
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta 660
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg 720
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt 780
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac 840
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca 900
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct 960
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt 1020
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt 1080
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa 1140
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat 1200
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg 1260
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt 1320
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca 1380
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa 1440
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg 1500
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc 1560
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt 1620
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg 1680
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg 1740
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata 1800
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg 1860
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga 1920
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg 1980
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt 2040
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc 2100
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat 2160
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg 2220
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg 2280
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa 2340
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa 2400
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta 2460
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga 2520
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt 2580
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc 2640
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt 2700
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct 2760
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt 2820
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc 2880
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 2940
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 3000
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 3060
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 3120
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 3180
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 3240
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 3300
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 3360
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 3420
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 3480
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 3540
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 3600
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 3660
tctgacaacc tgtcagccaa agcttgcaga gacagcggat cgcgataaga ttcctcgatg 3720
atttccacca cacgctgtgc cagctcttca tccggcagca gcgccccggc cggattgcag 3780
cgtttgcagg ggcggtaccc ttctgataaa gcatcttttg cattgaaaaa gatctgcaca 3840
ttgtcgattt gcggaactct cgatttgcag gaagggcggc aaaatatgcc ggtcgttttg 3900
accgcgtaat aaaaaactcc gtcataggcg gaatcgtttt ccgtaatcgc ccgccacatt 3960
tcaggcgtca atcgtgattt gctgttcata tcttcacccc gatctatgtc agtataacct 4020
atatgacagc cggaggtgga gaggcggaga acggcacagc aagaagacaa agaagaagag 4080
agactgttgc ctggacctcc gaaacgcgct acaattcatt tacaacacag gatggggtga 4140
gaatattgcc ggaatcagtg aagcaggtga cagaggaaag gccgatatga ttcggccttt 4200
tttatatgta cttcttagcg ggtctctaaa ataaccaaaa agcaaggact gctgaaaggg 4260
ctgacataag ccttttgccg gcggtccttt tttaattctg atttttcaaa cttagccccc 4320
tcgagaaaac cgcttgagta tcctccgctc atgttattga cctccttttc ttgatcaata 4380
tatttgtatg agcggaaaga aaatttgact agacggatgc ccctctgttg aaaatctttt 4440
tacagcatgt cagaatatga tatgatacaa ttcaaaggaa agtttaaact gttatgatta 4500
ggagtgtttg catttatgaa gaagattgca attgcggcga ttacagcgac aagcgtgctg 4560
gctctcagcg catgcagcgg gggagattct gaggttgttg cggaaacaaa agctggaaat 4620
attacaaaag aagaccttta tcaaacatta aaagacaatg ccggagcgaa ttcctccatt 4680
ttcttctgct atcaaaataa cagactcgtg attttccaaa cgagctttca aaaaagcctc 4740
tgccccttgc aaatcggatg cctgtctata aaattcccga tattggttaa acagcggcgc 4800
aatggcggcc gcatctgatg tctttgcttg gcgaatgttc atcttatttc ttcctccctc 4860
tcaataattt tttcattcta tcccttttct gtaaagttta tttttcagaa tacttttatc 4920
atcatgcttt gaaaaaatat cacgataata tccattgttc tcacggaagc acacgcaggt 4980
catttgaacg aattttttcg acaggaattt gccgggactc aggagcattt aacctaaaaa 5040
agcatgacat ttcagcataa tgaacattta ctcatgtcta ttttcgttct tttctgtatg 5100
aaaatagtta tttcgagtct ctacggaaat agcgagagat gatataccta aatagagata 5160
aaatcatctc aaaaaaatgg gtctactaaa atattattcc atctattaca ataaattcac 5220
agaatagtct tttaagtaag tctactctga atttttttaa aaggagaggg taactagtgg 5280
ccccaaaaaa gaaacgcaag gttatggata aaaaatacag cattggtctg gatatcggaa 5340
ccaacagcgt tgggtgggca gtaataacag atgaatacaa agtgccgtca aaaaaattta 5400
aggttctggg gaatacagat cgccacagca taaaaaagaa tctgattggg gcattgctgt 5460
ttgattcggg tgagacagct gaggccacgc gtctgaaacg tacagcaaga agacgttaca 5520
cacgtcgtaa aaatcgtatt tgctacttac aggaaatttt ttctaacgaa atggccaagg 5580
tagatgatag tttcttccat cgtctcgaag aatcttttct ggttgaggaa gataaaaaac 5640
acgaacgtca ccctatcttt ggcaatatcg tggatgaagt ggcctatcat gaaaaatacc 5700
ctacgattta tcatcttcgc aagaagttgg ttgatagtac ggacaaagcg gatctgcgtt 5760
taatccatct tgcgttagcg cacatgatca aatttcgtgg tcatttctta attgaaggtg 5820
atctgaatcc tgataactct gatgtggaca aattgtttat acaattagtg caaacctata 5880
atcagctgtt cgaggaaaac cccattaatg cctctggagt tgatgccaaa gcgattttaa 5940
gcgcgagact ttctaagtcc cggcgtctgg agaatctgat cgcccagtta ccaggggaaa 6000
agaaaaatgg tctgtttggt aatctgattg ccctcagtct ggggcttacc ccgaacttca 6060
aatccaattt tgacctggct gaggacgcaa agctgcagct gagcaaagat acttatgatg 6120
atgacctcga caatctgctc gcccagattg gtgaccaata tgcggatctg tttctggcag 6180
cgaagaatct ttcggatgct atcttgctgt cggatattct gcgtgttaat accgaaatca 6240
ccaaagcgcc tctgtctgca agtatgatca agagatacga cgagcaccac caggacctga 6300
ctcttcttaa ggcactggta cgccaacagc ttccggagaa atacaaagaa atattcttcg 6360
accagtccaa gaatggttac gcgggctaca tcgatggtgg tgcatcacag gaagagttct 6420
ataaatttat taaaccaatc cttgagaaaa tggatggcac ggaagagtta cttgttaaac 6480
ttaaccgcga agacttgctt agaaagcaac gtacattcga caacggctcc atcccacacc 6540
agattcattt aggtgaactt cacgccatct tgcgcagaca agaagatttc tatcccttct 6600
taaaagacaa tcgggagaaa atcgagaaga tcctgacgtt ccgcattccc tattatgtcg 6660
gtcccctggc acgtggtaat tctcggtttg cctggatgac gcgcaaaagt gaggaaacca 6720
tcaccccttg gaactttgaa gaagtcgtgg ataaaggtgc tagcgcgcag tcttttatag 6780
aaagaatgac gaacttcgat aaaaacttgc ccaacgaaaa agtcctgccc aagcactctc 6840
ttttatatga gtactttact gtgtacaacg aactgactaa agtgaaatac gttacggaag 6900
gtatgcgcaa acctgccttt cttagtggcg agcagaaaaa agcaattgtc gatcttctct 6960
ttaaaacgaa tcgcaaggta actgtaaaac agctgaagga agattatttc aaaaagatcg 7020
aatgctttga ttctgtcgag atctcgggtg tcgaagatcg tttcaacgct tccttaggga 7080
cctatcatga tttgctgaag ataataaaag acaaagactt tctcgacaat gaagaaaatg 7140
aagatattct ggaggatatt gttttgacct tgaccttatt cgaagataga gagatgatcg 7200
aggagcgctt aaaaacctat gcccacctgt ttgatgacaa agtcatgaag caattaaagc 7260
gccgcagata tacggggtgg ggccgcttga gccgcaagtt gattaacggt attagagaca 7320
agcagagcgg aaaaactatc ctggatttcc tcaaatctga cggatttgcg aaccgcaatt 7380
ttatgcagct tatacatgat gattcgctta cattcaaaga ggatattcag aaggctcagg 7440
tgtctgggca aggtgattca ctccacgaac atatagcaaa tttggccggc tctcctgcga 7500
ttaagaaggg gatcctgcaa acagttaaag ttgtggatga acttgtaaaa gtaatgggcc 7560
gccacaagcc ggagaatatc gtgatagaaa tggcgcgcga gaatcaaacg acacaaaaag 7620
gtcaaaagaa ctcaagagag agaatgaagc gcattgagga ggggataaag gaacttggat 7680
ctcaaattct gaaagaacat ccagttgaaa acactcagct gcaaaatgaa aaattgtacc 7740
tgtactacct gcagaatgga agagacatgt acgtggatca ggaattggat atcaatagac 7800
tctcggacta tgacgtagat cacattgtcc ctcagagctt cctcaaggat gattctatag 7860
ataataaagt acttacgaga tcggacaaaa atcgcggtaa atcggataac gtcccatcgg 7920
aggaagtcgt taaaaagatg aaaaactatt ggcgtcaact gctgaacgcc aagctgatca 7980
cacagcgtaa gtttgataat ctgactaaag ccgaacgcgg tggtcttagt gaactcgata 8040
aagcaggatt tataaaacgg cagttagtag aaacgcgcca aattacgaaa cacgtggctc 8100
agatcctcga ttctagaatg aatacaaagt acgatgaaaa cgataaactg atccgtgaag 8160
taaaagtcat taccttaaaa tctaaacttg tgtccgattt ccgcaaagat tttcagtttt 8220
acaaggtccg ggaaatcaat aactatcacc atgcacatga tgcatattta aatgcggttg 8280
taggcacggc ccttattaag aaatacccta aactcgaaag tgagtttgtt tatggggatt 8340
ataaagtgta tgacgttcgc aaaatgatcg cgaaatcaga acaggaaatc ggtaaggcta 8400
ccgctaaata ctttttttat tccaacatta tgaatttttt taagaccgaa ataactctcg 8460
cgaatggtga aatccgtaaa cggcctctta tagaaaccaa tggtgaaacg ggagaaatcg 8520
tttgggataa aggtcgtgac tttgccaccg ttcgtaaagt cctctcaatg ccgcaagtta 8580
acattgtcaa gaagacggaa gttcaaacag ggggattctc caaagaatct atcctgccga 8640
agcgtaacag tgataaactt attgccagaa aaaaagattg ggatccaaaa aaatacggag 8700
gctttgattc ccctaccgtc gcgtatagtg tgctggtggt tgctaaagtc gagaaaggga 8760
aaagcaagaa attgaaatca gttaaagaac tgctgggtat tacaattatg gaaagatcgt 8820
cctttgagaa aaatccgatc gactttttag aggccaaggg gtataaggaa gtgaaaaaag 8880
atctcatcat caaattaccg aagtatagtc tttttgagct ggaaaacggc agaaaaagaa 8940
tgctggcctc cgcgggcgag ttacagaagg gaaatgagct ggcgctgcct tccaaatatg 9000
ttaattttct gtaccttgcc agtcattatg agaaactgaa gggcagcccc gaagataacg 9060
aacagaaaca attattcgtg gaacagcata agcactattt agatgaaatt atagagcaaa 9120
ttagtgaatt ttctaagcgc gttatcctcg cggatgctaa tttagacaaa gtactgtcag 9180
cttataataa acatcgggat aagccgatta gagaacaggc cgaaaatatc attcatttgt 9240
ttaccttaac caaccttgga gcaccagctg ccttcaaata tttcgatacc acaattgatc 9300
gtaaacggta tacaagtaca aaagaagtct tggacgcaac cctcattcat caatctatta 9360
ctggattata tgagacacgc attgatcttt cacagctggg cggagacaag aagaaaaaac 9420
tgaaactgca ccatcatcac catcatcatc accatcattg ataactcgag aaagcttaca 9480
taaaaaaccg gccttggccc cgccggtttt ttattatttt tcttcctccg catgttcaat 9540
ccgctccata atcgacggat ggctccctct gaaaatttta acgagaaacg gcgggttgac 9600
ccggctcagt cccgtaacgg ccaagtcctg aaacgtctca atcgccgctt cccggtttcc 9660
ggtcagctca atgccgtaac ggtcggcggc gttttcctga taccgggaga cggcattcgt 9720
aatc 9724
<210> 82
<211> 23
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 82
gagcttcttt ttcttgagca cgg 23
<210> 83
<211> 1000
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 83
acctgtcagc caaagcttgc agagacagcg gatcgcgata agattcctcg atgatttcca 60
ccacacgctg tgccagctct tcatccggca gcagcgcccc ggccggattg cagcgtttgc 120
aggggcggta cccttctgat aaagcatctt ttgcattgaa aaagatctgc acattgtcga 180
tttgcggaac tctcgatttg caggaagggc ggcaaaatat gccggtcgtt ttgaccgcgt 240
aataaaaaac tccgtcatag gcggaatcgt tttccgtaat cgcccgccac atttcaggcg 300
tcaatcgtga tttgctgttc atatcttcac cccgatctat gtcagtataa cctatatgac 360
agccggaggt ggagaggcgg agaacggcac agcaagaaga caaagaagaa gagagactgt 420
tgcctggacc tccgaaacgc gctacaattc atttacaaca caggatgggg tgagaatatt 480
gccggaatca gtgaagcagg tgacagagga aaggccgata tgattcggcc ttttttatat 540
gtacttctta gcgggtctct aaaataacca aaaagcaagg actgctgaaa gggctgacat 600
aagccttttg ccggcggtcc ttttttaatt ctgatttttc aaacttagcc ccctcgagaa 660
aaccgcttga gtatcctccg ctcatgttat tgacctcctt ttcttgatca atatatttgt 720
atgagcggaa agaaaatttg actagacgga tgcccctctg ttgaaaatct ttttacagca 780
tgtcagaata tgatatgata caattcaaag gaaagtttaa actgttatga ttaggagtgt 840
ttgcatttat gaagaagatt gcaattgcgg cgattacagc gacaagcgtg ctggctctca 900
gcgcatgcag cgggggagat tctgaggttg ttgcggaaac aaaagctgga aatattacaa 960
aagaagacct ttatcaaaca ttaaaagaca atgccggagc 1000
<210> 84
<211> 9754
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 84
gggtgaagtg gtcaagacct cactaggcac cttaaaaata gcgcaccctg aagaagattt 60
atttgaggta gcccttgcct acctagcttc caagaaagat atcctaacag cacaagagcg 120
gaaagatgtt ttgttctaca tccagaacaa cctctgctaa aattcctgaa aaattttgca 180
aaaagttgtt gactttatct acaaggtgtg gcataatgtg tggatccagt tgacgtatcg 240
attcgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa 300
aagtggcacc gagtcggtgc gactcctgtt gatagatcca gtaatgacct cagaactcca 360
tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatgtcga 420
cctcgagagt tacgctaggg ataacagggt aatataggag ctccagtcgg cttaaaccag 480
ttttcgctgg tgcgaaaaaa gagtgtcttg tgacacctaa attcaaaatc tatcggtcag 540
atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa 600
gcgggaacca atcataaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta 660
cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg 720
ttgttttctt taataattca ttaacatcta caccgccata aactatcata tcttcttttt 780
gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac 840
ttcctgcaat ctgcggaata gtcgcattca attcttctgt taattatttt tatctgttca 900
taagatttat taccctcata catcactaga atatgataat gctctttttt catcctacct 960
tctgtatcag tatccctatc atgtaatgga gacactacaa attgaatgtg taactctttt 1020
aaatactcta accactcggc ttttgctgat tctggatata aaacaaatgt ccaattacgt 1080
cctcttgaat ttttcttgtt ttcagtttct tttattacat tttcgctcat gatataataa 1140
cggtgctaat acacttaaca aaatttagtc atagataggc agcatgccag tgctgtctat 1200
ctttttttgt ttaaaatgca ccgtattcct cctttgcata tttttttatt agaataccgg 1260
ttgcatctga tttgctaata ttatattttt ctttgattct atttaatatc tcattttctt 1320
ctgttgtaag tcttaaagta acagcaactt ttttctcttc ttttctatct acaactatca 1380
ctgtacctcc caacatctgt ttttttcact ttaacataaa aaacaacctt ttaacattaa 1440
aaacccaata tttatttatt tgtttggaca atggacactg gacacctagg ggggaggtcg 1500
tagtaccccc ctatgttttc tcccctaaat aaccccaaaa atctaagaaa aaaagacctc 1560
aaaaaggtct ttaattaaca tctcaaattt cgcatttatt ccaatttcct ttttgcgtgt 1620
gatgcgagct catcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg 1680
aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg 1740
tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata 1800
ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg 1860
cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga 1920
aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg 1980
ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt 2040
tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc 2100
ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat 2160
caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg 2220
cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg 2280
ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa 2340
gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa 2400
agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta 2460
tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga 2520
gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt 2580
actggatgaa ttgttttagt gactgcagtg agatctggta atgactctct agcttgaggc 2640
atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt 2700
cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gctaagcaga aggccatcct 2760
gacggatggc ctttttgcgt ttctacaaac tcttgttaac tctagagctg cctgccgcgt 2820
ttcggtgatg aagatcttcc cgatgattaa ttaattcaga acgctcggtt gccgccgggc 2880
gttttttatg aagcttcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 2940
cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 3000
gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 3060
tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 3120
tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 3180
cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 3240
gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 3300
ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 3360
ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 3420
ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 3480
agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 3540
aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 3600
atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 3660
tctgacactc taatagcttc gctttgattg ttgtattatt tattttgctg atcattgtgg 3720
gctccgcttt tgtaaagtat taattcattc gcgcacgaag ccttcttccg aaaaactgca 3780
ggttcatatc gaaaggcgct ccgcttttca tatgagcctg tttttttgta atttaaaaag 3840
aatttatttc cacaataaca aaaaatgttt ataataggat aggctctgag ctgacggctg 3900
acggcgctcg gaaataaaaa ctctcccaca aatgaaactt ttatacaaca ttttcgtctt 3960
atactgttgt taaggcaatt caataacgga aaggaaaaga tcaatgttaa ataagctaag 4020
aatgttgtat gaaaaaacaa caatgaagtg gctgctcaac acttcctatt actttctcat 4080
tctggtgctg ctgtttttaa tttacggatt ccatacagca aatacaggtt catatattta 4140
taatgacttt taatggagaa gaaaacaatg aaactgattg aaacaattaa aaaataccac 4200
aaatgtgctt gatggatccg tcttattcgg aagagctgct tccggagctg agcttgttta 4260
tgttctgcgg agaaacgctt cctgcgtcag tcgccagaca gcttaaagaa agatttccaa 4320
aggctcgtgt ttttaacaca tacggcccga ccgaagcgac cgttgccgtc acttccatcg 4380
aagtaacaga cgacgtatta aacaagtatt cttctcttcc cgtcggttct gaaaagcccg 4440
aaaccgaaat cgtaatcatc aatgaagacg gaaaagccgt ccaagacggg gaaaaaggcg 4500
agatcataat taccggagca agcgtcagca aaggctattt aggcgaaaaa gcgctgacgg 4560
aaaaagcgtt tttctcttac aacggctccc ctgcttaccg caccggtgac gcaggttaca 4620
aagaaaacgg ccagctcttc ttcctcggaa gactcgattt ccaaatcaaa ctgcacggct 4680
accggattga gcttgaagaa ttcctccatt ttcttctgct atcaaaataa cagactcgtg 4740
attttccaaa cgagctttca aaaaagcctc tgccccttgc aaatcggatg cctgtctata 4800
aaattcccga tattggttaa acagcggcgc aatggcggcc gcatctgatg tctttgcttg 4860
gcgaatgttc atcttatttc ttcctccctc tcaataattt tttcattcta tcccttttct 4920
gtaaagttta tttttcagaa tacttttatc atcatgcttt gaaaaaatat cacgataata 4980
tccattgttc tcacggaagc acacgcaggt catttgaacg aattttttcg acaggaattt 5040
gccgggactc aggagcattt aacctaaaaa agcatgacat ttcagcataa tgaacattta 5100
ctcatgtcta ttttcgttct tttctgtatg aaaatagtta tttcgagtct ctacggaaat 5160
agcgagagat gatataccta aatagagata aaatcatctc aaaaaaatgg gtctactaaa 5220
atattattcc atctattaca ataaattcac agaatagtct tttaagtaag tctactctga 5280
atttttttaa aaggagaggg taactagtgg ccccaaaaaa gaaacgcaag gttatggata 5340
aaaaatacag cattggtctg gatatcggaa ccaacagcgt tgggtgggca gtaataacag 5400
atgaatacaa agtgccgtca aaaaaattta aggttctggg gaatacagat cgccacagca 5460
taaaaaagaa tctgattggg gcattgctgt ttgattcggg tgagacagct gaggccacgc 5520
gtctgaaacg tacagcaaga agacgttaca cacgtcgtaa aaatcgtatt tgctacttac 5580
aggaaatttt ttctaacgaa atggccaagg tagatgatag tttcttccat cgtctcgaag 5640
aatcttttct ggttgaggaa gataaaaaac acgaacgtca ccctatcttt ggcaatatcg 5700
tggatgaagt ggcctatcat gaaaaatacc ctacgattta tcatcttcgc aagaagttgg 5760
ttgatagtac ggacaaagcg gatctgcgtt taatccatct tgcgttagcg cacatgatca 5820
aatttcgtgg tcatttctta attgaaggtg atctgaatcc tgataactct gatgtggaca 5880
aattgtttat acaattagtg caaacctata atcagctgtt cgaggaaaac cccattaatg 5940
cctctggagt tgatgccaaa gcgattttaa gcgcgagact ttctaagtcc cggcgtctgg 6000
agaatctgat cgcccagtta ccaggggaaa agaaaaatgg tctgtttggt aatctgattg 6060
ccctcagtct ggggcttacc ccgaacttca aatccaattt tgacctggct gaggacgcaa 6120
agctgcagct gagcaaagat acttatgatg atgacctcga caatctgctc gcccagattg 6180
gtgaccaata tgcggatctg tttctggcag cgaagaatct ttcggatgct atcttgctgt 6240
cggatattct gcgtgttaat accgaaatca ccaaagcgcc tctgtctgca agtatgatca 6300
agagatacga cgagcaccac caggacctga ctcttcttaa ggcactggta cgccaacagc 6360
ttccggagaa atacaaagaa atattcttcg accagtccaa gaatggttac gcgggctaca 6420
tcgatggtgg tgcatcacag gaagagttct ataaatttat taaaccaatc cttgagaaaa 6480
tggatggcac ggaagagtta cttgttaaac ttaaccgcga agacttgctt agaaagcaac 6540
gtacattcga caacggctcc atcccacacc agattcattt aggtgaactt cacgccatct 6600
tgcgcagaca agaagatttc tatcccttct taaaagacaa tcgggagaaa atcgagaaga 6660
tcctgacgtt ccgcattccc tattatgtcg gtcccctggc acgtggtaat tctcggtttg 6720
cctggatgac gcgcaaaagt gaggaaacca tcaccccttg gaactttgaa gaagtcgtgg 6780
ataaaggtgc tagcgcgcag tcttttatag aaagaatgac gaacttcgat aaaaacttgc 6840
ccaacgaaaa agtcctgccc aagcactctc ttttatatga gtactttact gtgtacaacg 6900
aactgactaa agtgaaatac gttacggaag gtatgcgcaa acctgccttt cttagtggcg 6960
agcagaaaaa agcaattgtc gatcttctct ttaaaacgaa tcgcaaggta actgtaaaac 7020
agctgaagga agattatttc aaaaagatcg aatgctttga ttctgtcgag atctcgggtg 7080
tcgaagatcg tttcaacgct tccttaggga cctatcatga tttgctgaag ataataaaag 7140
acaaagactt tctcgacaat gaagaaaatg aagatattct ggaggatatt gttttgacct 7200
tgaccttatt cgaagataga gagatgatcg aggagcgctt aaaaacctat gcccacctgt 7260
ttgatgacaa agtcatgaag caattaaagc gccgcagata tacggggtgg ggccgcttga 7320
gccgcaagtt gattaacggt attagagaca agcagagcgg aaaaactatc ctggatttcc 7380
tcaaatctga cggatttgcg aaccgcaatt ttatgcagct tatacatgat gattcgctta 7440
cattcaaaga ggatattcag aaggctcagg tgtctgggca aggtgattca ctccacgaac 7500
atatagcaaa tttggccggc tctcctgcga ttaagaaggg gatcctgcaa acagttaaag 7560
ttgtggatga acttgtaaaa gtaatgggcc gccacaagcc ggagaatatc gtgatagaaa 7620
tggcgcgcga gaatcaaacg acacaaaaag gtcaaaagaa ctcaagagag agaatgaagc 7680
gcattgagga ggggataaag gaacttggat ctcaaattct gaaagaacat ccagttgaaa 7740
acactcagct gcaaaatgaa aaattgtacc tgtactacct gcagaatgga agagacatgt 7800
acgtggatca ggaattggat atcaatagac tctcggacta tgacgtagat cacattgtcc 7860
ctcagagctt cctcaaggat gattctatag ataataaagt acttacgaga tcggacaaaa 7920
atcgcggtaa atcggataac gtcccatcgg aggaagtcgt taaaaagatg aaaaactatt 7980
ggcgtcaact gctgaacgcc aagctgatca cacagcgtaa gtttgataat ctgactaaag 8040
ccgaacgcgg tggtcttagt gaactcgata aagcaggatt tataaaacgg cagttagtag 8100
aaacgcgcca aattacgaaa cacgtggctc agatcctcga ttctagaatg aatacaaagt 8160
acgatgaaaa cgataaactg atccgtgaag taaaagtcat taccttaaaa tctaaacttg 8220
tgtccgattt ccgcaaagat tttcagtttt acaaggtccg ggaaatcaat aactatcacc 8280
atgcacatga tgcatattta aatgcggttg taggcacggc ccttattaag aaatacccta 8340
aactcgaaag tgagtttgtt tatggggatt ataaagtgta tgacgttcgc aaaatgatcg 8400
cgaaatcaga acaggaaatc ggtaaggcta ccgctaaata ctttttttat tccaacatta 8460
tgaatttttt taagaccgaa ataactctcg cgaatggtga aatccgtaaa cggcctctta 8520
tagaaaccaa tggtgaaacg ggagaaatcg tttgggataa aggtcgtgac tttgccaccg 8580
ttcgtaaagt cctctcaatg ccgcaagtta acattgtcaa gaagacggaa gttcaaacag 8640
ggggattctc caaagaatct atcctgccga agcgtaacag tgataaactt attgccagaa 8700
aaaaagattg ggatccaaaa aaatacggag gctttgattc ccctaccgtc gcgtatagtg 8760
tgctggtggt tgctaaagtc gagaaaggga aaagcaagaa attgaaatca gttaaagaac 8820
tgctgggtat tacaattatg gaaagatcgt cctttgagaa aaatccgatc gactttttag 8880
aggccaaggg gtataaggaa gtgaaaaaag atctcatcat caaattaccg aagtatagtc 8940
tttttgagct ggaaaacggc agaaaaagaa tgctggcctc cgcgggcgag ttacagaagg 9000
gaaatgagct ggcgctgcct tccaaatatg ttaattttct gtaccttgcc agtcattatg 9060
agaaactgaa gggcagcccc gaagataacg aacagaaaca attattcgtg gaacagcata 9120
agcactattt agatgaaatt atagagcaaa ttagtgaatt ttctaagcgc gttatcctcg 9180
cggatgctaa tttagacaaa gtactgtcag cttataataa acatcgggat aagccgatta 9240
gagaacaggc cgaaaatatc attcatttgt ttaccttaac caaccttgga gcaccagctg 9300
ccttcaaata tttcgatacc acaattgatc gtaaacggta tacaagtaca aaagaagtct 9360
tggacgcaac cctcattcat caatctatta ctggattata tgagacacgc attgatcttt 9420
cacagctggg cggagacaag aagaaaaaac tgaaactgca ccatcatcac catcatcatc 9480
accatcattg ataactcgag aaagcttaca taaaaaaccg gccttggccc cgccggtttt 9540
ttattatttt tcttcctccg catgttcaat ccgctccata atcgacggat ggctccctct 9600
gaaaatttta acgagaaacg gcgggttgac ccggctcagt cccgtaacgg ccaagtcctg 9660
aaacgtctca atcgccgctt cccggtttcc ggtcagctca atgccgtaac ggtcggcggc 9720
gttttcctga taccgggaga cggcattcgt aatc 9754
<210> 85
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 85
tccagttgac gtatcgattc cgg 23
<210> 86
<211> 1030
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 86
ctctaatagc ttcgctttga ttgttgtatt atttattttg ctgatcattg tgggctccgc 60
ttttgtaaag tattaattca ttcgcgcacg aagccttctt ccgaaaaact gcaggttcat 120
atcgaaaggc gctccgcttt tcatatgagc ctgttttttt gtaatttaaa aagaatttat 180
ttccacaata acaaaaaatg tttataatag gataggctct gagctgacgg ctgacggcgc 240
tcggaaataa aaactctccc acaaatgaaa cttttataca acattttcgt cttatactgt 300
tgttaaggca attcaataac ggaaaggaaa agatcaatgt taaataagct aagaatgttg 360
tatgaaaaaa caacaatgaa gtggctgctc aacacttcct attactttct cattctggtg 420
ctgctgtttt taatttacgg attccataca gcaaatacag gttcatatat ttataatgac 480
ttttaatgga gaagaaaaca atgaaactga ttgaaacaat taaaaaatac cacaaatgtg 540
cttgatggat ccgtcttatt cggaagagct gcttccggag ctgagcttgt ttatgttctg 600
cggagaaacg cttcctgcgt cagtcgccag acagcttaaa gaaagatttc caaaggctcg 660
tgtttttaac acatacggcc cgaccgaagc gaccgttgcc gtcacttcca tcgaagtaac 720
agacgacgta ttaaacaagt attcttctct tcccgtcggt tctgaaaagc ccgaaaccga 780
aatcgtaatc atcaatgaag acggaaaagc cgtccaagac ggggaaaaag gcgagatcat 840
aattaccgga gcaagcgtca gcaaaggcta tttaggcgaa aaagcgctga cggaaaaagc 900
gtttttctct tacaacggct cccctgctta ccgcaccggt gacgcaggtt acaaagaaaa 960
cggccagctc ttcttcctcg gaagactcga tttccaaatc aaactgcacg gctaccggat 1020
tgagcttgaa 1030
<210> 87
<211> 1320
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 87
ttgtttttac acggtactag cagacaaaat gaaagagggc acctcgaaat cggcggtgtc 60
gatgttctat cattggcaga aagatacgga acacctcttt atgtatacga tgtcgcgctg 120
attagagagc gcgcccgaaa attccagaag gcattcaagg aagccggttt aaaagcgcag 180
gtagcgtatg caagcaaggc gttttcatcg gttgccatga ttcagcttgc cgaacaagag 240
gggctgtctc tggatgtggt atcgggagga gagcttttca ctgcgatcaa agcagggttc 300
ccagctgagc ggattcattt tcacggaaac aataagagcc ctgaagaact agccatggcg 360
ctggagcatc aaatcggctg catcgtgctc gataactttc acgagatcgc cattacagaa 420
gatctttgca agcgatcagg acaaactgta gacgttttgc tcagaatcac tccgggagtt 480
gaagcgcaca cgcacgatta tattacgacg gggcaggaag attccaaatt cggttttgat 540
ctgcataatg gacaggtcga acaagccatc gaacaagtcc tccgctcgtc tgcgtttaag 600
ctcctcggcg tgcactgcca catcggttcg caaatttttg atacggcagg atttgtcctt 660
gcagcagaca agattttcga gaagcttgcg gaatggcggg agacttactc tttcattccg 720
gaagtgctca atcttggcgg gggcttcggc atccgctata caaaagacga cgagccgctt 780
gcagctgatg tttatgttga aaaaatcatc gaggcggtca aagcaaatgc cgagcatttc 840
ggctttgaca tccctgagat ttggatcgaa ccaggccggt ctctcgtcgg tgatgcgggg 900
actacgctgt acacgatcgg ttctcaaaaa gaggtgccgg gcattcgcaa atatgtagcc 960
atcgacggcg gcatgagcga taatatcagg ccggcgcttt atgaggcaaa atatgaagca 1020
gccgtcgcca acaggatgaa cgatgcttgt catgataccg catcaatcgc aggaaaatgc 1080
tgcgaaagcg gagatatgct gatttgggat ttggaaatcc ccgaagttcg cgacggagat 1140
gtgctcgccg ttttctgcac cggtgcgtac ggctacagca tggccaacaa ctacaaccgc 1200
attccgcgcc cggccgtcgt ctttgtcgag gacggggaag cgcagctcgt cattcagaga 1260
gagacgtatg aggatatcgt caagctggat ctgccgctga aatcgaaagt caaacaataa 1320
<210> 88
<211> 6393
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 88
aagcttcata tgcaagggtt tattgttttc taaaatctga ttaccaatta gaatgaatat 60
ttcccaaata ttaaataata aaacaaaaaa attgaaaaaa gtgtttccac cattttttca 120
atttttttat aattttttta atctgttatt taaatagttt atagttaaat ttacattttc 180
attagtccat tcaatattct ctccaagata actacgaact gctaacaaaa ttctctccct 240
atgttctaat ggagaagatt cagccactgc atttcccgca atatcttttg gtatgatttt 300
acccgtgtcc atagttaaaa tcatacggca taaagttaat atagagttgg tttcatcatc 360
ctgataatta tctattaatt cctctgacga atccataatg gctcttctca catcagaaaa 420
tggaatatca ggtagtaatt cctctaagtc ataatttccg tatattcttt tattttttcg 480
ttttgcttgg taaagcatta tggttaaatc tgaatttaat tccttctgag gaatgtatcc 540
ttgttcataa agctcttgta accattctcc ataaataaat tcttgtttgg gaggatgatt 600
ccacggtacc atttcttgct gaataataat tgttaattca atatatcgta agttgctttt 660
atctcctatt ttttttgaaa taggtctaat tttttgtata agtatttctt tactttgatc 720
tgtcaatggt tcagatacga cgactaaaaa gtcaagatca ctatttggtt ttagtccact 780
ctcaactcct gatccaaaca tgtaagtacc aataaggtta ttttttaaat gtttccgaag 840
tatttttttc actttattaa tttgttcgta tgtattcaaa tatatcctcc tcactatttt 900
gattagtacc tattttatat ccatagttgt taattaaata aacttaattt agtttattta 960
tggatttcat tggcttctaa attttttatc tagataataa ttattttagt taattttatt 1020
ctagattata tatgatatga tctttcattt ccataaaact aaagtaagtg taaacctatt 1080
cattgtttta aaaatatctc ttgccagtca cgttacgtta ttagttatag ttattataac 1140
atgtattcac gaacgggcgc gccggtatcc gcgcttcttg agcactattt attcaaagcc 1200
gctccagatc aatagcgctt tttcagctcc ctgaggatga attcgtatat cagctgattc 1260
cggtcttctt tcggatagag cataaattcc tgtttcttct gcatggggtt tccttcaatc 1320
ctgtcgataa attttgttct cagccatgcc gttcggtaaa cctggttttc gaaagatgag 1380
atggatacgg gcagctccag cgtttccccg ttgacaaacg tgacaaacgt gttgtcatac 1440
tttgccgcgc aaaactcgtg aacatgcgca tgggaaagcc acccgcactg aggacgagtt 1500
gaggaaaatg tggggaaaag aaaaatgttg tttgagtgat ccaccatgat cggcggttta 1560
tgggaaactt taatgacttc atatgtgccc gcttttcttc ccgcatagct cgatccgaaa 1620
tagcggcagc ttctttcgat aatttgaaac ggcttcatat tgacgcggaa agtcctgtcg 1680
gtctcaagta tttttgaggc ggatttctcc ccctcaccca gaggcaggac agccattgtc 1740
gaactgttta cttcatacgt atcctttgtc atatcctctg tgctcatgtg atttccccct 1800
taaaaataaa ttcattcaaa tacagatgca ttttatttca tatagtaagt acatcaccta 1860
ttagtttgtt gtttaaacaa actaacttat tttcatctta tataacctcg tcagtatttt 1920
caatattttt tttagttttt tatgaacaca ttagatttaa taaagggaag attcgctatg 1980
tactatgttg atacttaatt taaagattaa acaaatggag tggatgaagt ggatatcgct 2040
gatcaaacct ttgtcaaaaa agtaaatcaa aagttattat taaaagaaat ccttaaaaat 2100
tcacctattt caagagcaaa attatctgaa atgactggat taaataaatc aactgtctca 2160
tcacaggtaa acacgttaat gaaagaaagt atggtatttg aaataggtca aggacaatca 2220
agtggcggaa gaagacctgt catgcttgtt tttaataaaa aggcaggata ctccgttgga 2280
atagatgttg gtgtggatta tattaatggc attttaacag accttgaagg aacaatcgtt 2340
cttgatcaat accgccattt ggaatccaat tctccagaaa taacgaaaga cattttgatt 2400
gatatgattc atcactttat tacgcaaatg ccccaatctc cgtacgggtt tattggtata 2460
ggtacttgcg tgcctggact cattgataaa gatcaaaaaa ttgttttcac tccgaactcc 2520
aactggagag atattgactt aaaatcttcg atacaagaga agtacaatgt gtctgttttt 2580
attgaaaatg aggcaaatgc tggcgcatat ggagaaaaac tatttggagc tgcaaaaaat 2640
cacgataaca ttatttacgt aagtatcagc acaggaatag ggatcggtgt tattatcaac 2700
aatcatttat atagaggagt aagcggcttc tctggagaaa tgggacatat gacaatagac 2760
tttaatggtc ctaaatgcag ttgcggaaac cgaggatgct gggaattgta tgcttcagag 2820
aaggctttat ttaaatctct tcagaccaaa gagaaaaaac tgtcctatca agatatcata 2880
aacctcgccc atctgaatga tatcggaacc ttaaatgcat tacaaaattt tggattctat 2940
ttaggaatag gccttaccaa tattctaaat actctcaacc cacaagccgt aattttaaga 3000
aatagcataa ttgaatcgca tcctatggtt ttaaattcaa tgagaagtga agtatcatca 3060
agggtttatt cccaattagg caatagctat gaattattgc catcttcctt aggacagaat 3120
gcaccggcat taggaatgtc ctccattgtg attgatcatt ttctggacat gattacaatg 3180
taatttttta tggaatggac agctcatctt taaagatgag tttttttatt ctaggagtat 3240
ttctgaagca atagtgacat ggcaccttct catatgaaaa aggagttcta aaataaaaat 3300
ctcctttttc atgtgcaaat tatttttctt tataacgaaa atatctaaat gacaatgcat 3360
atgcaagagg ggatcacata aatatatatt ttaaaaatat cccactttat ccaattttcg 3420
tttgttgaac taatgggtgc tttagttgaa gaataaaaga ccacattaaa aaatgtggtc 3480
ttttgtgttt ttttaaagga tttgagcgta gcgaaaaatc cttttctttc ttatcttgat 3540
actatataga aacaacatca tttttcaaaa ttaggtcaaa gccttgtgta tcaagggttt 3600
gatggttctt tgacaggtaa aaactccttc tgctattatt aaatactata tagaaacaac 3660
atcatttttc aaaattaggt caaagccttg tgtatcaagg gtttgatggt tctttgacag 3720
gtaaaaactc cttctgctat tattaaggtg tcgaatcaaa ataatagaat gctagagaac 3780
tagctcagaa ggagtttttt tgttgattta ttcatctgaa aatgattata gcatcctcga 3840
agataaaacc gcaacaggta aaaagcggga ttggaagggg aaaaagagac ggacgaacct 3900
catggcggag cattacgaag cgttagagag taagattggg gcaccttact atggcaaaaa 3960
ggctgaaaaa ctaattagtt gtgcagagta tctttcgttt aagagagacc cggagacggg 4020
caagttaaaa ctgtatcaag cccatttttg taaagtgagg ttatgtccga tgtgtgcgtg 4080
gcgcaggtcg ttaaaaattg cttatcacaa taagttgatc gtagaggaag ccaatagaca 4140
gtacggctgc ggatggattt ttctcacgct gacgattcga aatgtaaagg gagaacggct 4200
gaagccacaa atttctgcga tgatggaagg ctttaggaaa ctgttccagt acaaaaaagt 4260
aaaaacttcg gttcttggat ttttcagagc tttagagatt accaaaaatc atgaagaaga 4320
tacatatcat cctcattttc atgtgttgat accagtaagg aaaaattatt ttgggaaaaa 4380
ctatattaag caggcggagt ggacgagcct ttggaaaaag gcgatgaaat tggattacac 4440
tccaattgtc gatattcgtc gagtgaaagg taaagctaag attgacgctg aacagattga 4500
aaacgatgtg cggaacgcaa tgatggagca aaaagctgtt ctcgaaatct ctaaatatcc 4560
ggttaaggat acggatgttg tgcgcggtaa taaggtgact gaagacaatc tgaacacggt 4620
gctttacttg gatgatgcgt tggcagctcg aaggttaatt ggatacggtg gcattttgaa 4680
ggagatacat aaagagctga atcttggtga tgcggaggac ggcgatctgg tcaagattga 4740
ggaagaagat gacgaggttg caaatggtgc atttgaggtt atggcttatt ggcatcctgg 4800
cattaaaaat tacataatca aataaaaaaa gcagaccttt agaaggcctg cttttttaac 4860
taacccattt gtattgtgtt gaaatatgtt ttgtatggtg cactctcagt acaatctgct 4920
ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac 4980
gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca 5040
tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac 5100
gcctattttt ataggttaat gtcatgataa taatggtttc ttagcgattc acaaaaaata 5160
ggcacacgaa aaacaagtta agggatgcag tttatgcatc ccttaactta aaatactaaa 5220
aatgcccata ttttttcctc cttataaaat tagtataatt atagcacgag atctaaaagg 5280
atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 5340
ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 5400
ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 5460
ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 5520
ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 5580
ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 5640
tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 5700
tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 5760
tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 5820
tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 5880
gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 5940
tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 6000
ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct 6060
gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc 6120
gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc 6180
cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg 6240
ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta 6300
cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca 6360
ggaaacagct atgaccatga ttacgccgga tcc 6393
<210> 89
<211> 765
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 89
gtgaggagga tatatttgaa tacatacgaa caaattaata aagtgaaaaa aatacttcgg 60
aaacatttaa aaaataacct tattggtact tacatgtttg gatcaggagt tgagagtgga 120
ctaaaaccaa atagtgatct tgacttttta gtcgtcgtat ctgaaccatt gacagatcaa 180
agtaaagaaa tacttataca aaaaattaga cctatttcaa aaaaaatagg agataaaagc 240
aacttacgat atattgaatt aacaattatt attcagcaag aaatggtacc gtggaatcat 300
cctcccaaac aagaatttat ttatggagaa tggttacaag agctttatga acaaggatac 360
attcctcaga aggaattaaa ttcagattta accataatgc tttaccaagc aaaacgaaaa 420
aataaaagaa tatacggaaa ttatgactta gaggaattac tacctgatat tccattttct 480
gatgtgagaa gagccattat ggattcgtca gaggaattaa tagataatta tcaggatgat 540
gaaaccaact ctatattaac tttatgccgt atgattttaa ctatggacac gggtaaaatc 600
ataccaaaag atattgcggg aaatgcagtg gctgaatctt ctccattaga acatagggag 660
agaattttgt tagcagttcg tagttatctt ggagagaata ttgaatggac taatgaaaat 720
gtaaatttaa ctataaacta tttaaataac agattaaaaa aatta 765
<210> 90
<211> 1161
<212> DNA
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 90
gtggatgaag tggatatcgc tgatcaaacc tttgtcaaaa aagtaaatca aaagttatta 60
ttaaaagaaa tccttaaaaa ttcacctatt tcaagagcaa aattatctga aatgactgga 120
ttaaataaat caactgtctc atcacaggta aacacgttaa tgaaagaaag tatggtattt 180
gaaataggtc aaggacaatc aagtggcgga agaagacctg tcatgcttgt ttttaataaa 240
aaggcaggat actccgttgg aatagatgtt ggtgtggatt atattaatgg cattttaaca 300
gaccttgaag gaacaatcgt tcttgatcaa taccgccatt tggaatccaa ttctccagaa 360
ataacgaaag acattttgat tgatatgatt catcacttta ttacgcaaat gccccaatct 420
ccgtacgggt ttattggtat aggtacttgc gtgcctggac tcattgataa agatcaaaaa 480
attgttttca ctccgaactc caactggaga gatattgact taaaatcttc gatacaagag 540
aagtacaatg tgtctgtttt tattgaaaat gaggcaaatg ctggcgcata tggagaaaaa 600
ctatttggag ctgcaaaaaa tcacgataac attatttacg taagtatcag cacaggaata 660
gggatcggtg ttattatcaa caatcattta tatagaggag taagcggctt ctctggagaa 720
atgggacata tgacaataga ctttaatggt cctaaatgca gttgcggaaa ccgaggatgc 780
tgggaattgt atgcttcaga gaaggcttta tttaaatctc ttcagaccaa agagaaaaaa 840
ctgtcctatc aagatatcat aaacctcgcc catctgaatg atatcggaac cttaaatgca 900
ttacaaaatt ttggattcta tttaggaata ggccttacca atattctaaa tactctcaac 960
ccacaagccg taattttaag aaatagcata attgaatcgc atcctatggt tttaaattca 1020
atgagaagtg aagtatcatc aagggtttat tcccaattag gcaatagcta tgaattattg 1080
ccatcttcct taggacagaa tgcaccggca ttaggaatgt cctccattgt gattgatcat 1140
tttctggaca tgattacaat g 1161
<210> 91
<211> 66
<212> DNA
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 91
tgtacttact atatgaaata aaatgcatct gtatttgaat gaatttattt ttaaggggga 60
aatcac 66
<210> 92
<211> 576
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 92
atgagcacag aggatatgac aaaggatacg tatgaagtaa acagttcgac aatggctgtc 60
ctgcctctgg gtgaggggga gaaatccgcc tcaaaaatac ttgagaccga caggactttc 120
cgcgtcaata tgaagccgtt tcaaattatc gaaagaagct gccgctattt cggatcgagc 180
tatgcgggaa gaaaagcggg cacatatgaa gtcattaaag tttcccataa accgccgatc 240
atggtggatc actcaaacaa catttttctt ttccccacat tttcctcaac tcgtcctcag 300
tgcgggtggc tttcccatgc gcatgttcac gagttttgcg cggcaaagta tgacaacacg 360
tttgtcacgt ttgtcaacgg ggaaacgctg gagctgcccg tatccatctc atctttcgaa 420
aaccaggttt accgaacggc atggctgaga acaaaattta tcgacaggat tgaaggaaac 480
cccatgcaga agaaacagga atttatgctc tatccgaaag aagaccggaa tcagctgata 540
tacgaattca tcctcaggga gctgaaaaag cgctat 576
<210> 93
<211> 4946
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 93
tcgcggacaa gtacggcaag acaccggccc aagtcatttt gcgctgggat ttgcaaaacg 60
gggtcgttac gattccgaag tcgactaaag cggagcggat tgcccaaaac gcggacatat 120
ttgattttga actgaccacc gaggaaatga agcaaattga cgcgctgaat gaaaacaccc 180
gtgtcggccc tgatcccgat aactttgact tttaacaaaa cggccccgtt cgacattcga 240
acggggcttt aattgaattg tgcggttaca ccgccggact ccatcatcat cagttctttt 300
ttcatatcca atccgccccg gtatcccgtg agctgcccgc ttttaccgat aacccgatgg 360
caaggcacca ccattaacag cggatttgcg ccgatcgccg cgcctactgc ccgcacagcg 420
gcctgctttt caatatgctc ggcgatatcg gaataggagc aagtgctgcc gtaagggatt 480
tcggagagcg ccttccacac tgccagctga aaaggcgtgc cggcaaggtc gacaggaaag 540
ctgaaatgag ttcgcttgcc gttcaaatac gcctgcagct gctcggcgta ttctgccaat 600
cctttgtcat cccgaatgaa aactggctgt gtaaatcttt tttcagccca agcggccaaa 660
tcctcgaagc cttgattcca tccccctgta aaacagagcc cgcgggcagt cgccccaatg 720
tgaatctgcc aacctcggca aataagcgta cgccagtata cgatttgatc gtccatatgt 780
ttacctccgt ttcatttgcc ggtacgacgt cggcgattgc ccagtcttct ttttaaacaa 840
agaggcaaaa tattccgcat tcgcaatgcc taccattgaa gcgatttctg cgatcgatcg 900
ttctgaatga gcaagcaaat cgaccgcttt ctcaatcctt ttctgcagga tgtattctgc 960
cggcgagacg cctttgattc gtttaaatgt ccgctgcagg tgaaaagggc tgatatggca 1020
cctgtcagcc aaagcttgca gagacagcgg atcgcgataa gattcctcga tgatttccac 1080
cacacgctgt gccagctctt catccggcag cagcgccccg gccggattgc agcgtttgca 1140
ggggcggtac ccttctgata aagcatcttt tgcattgaaa aagatctgca cattgtcgat 1200
ttgcggaact ctcgatttgc aggaagggcg gcaaaatatg ccggtcgttt tgaccgcgta 1260
ataaaaaact ccgtcatagg cggaatcgtt ttccgtaatc gcccgccaca tttcaggcgt 1320
caatcgtgat ttgctgttca tatcttcacc ccgatctatg tcagtataac ctatatgaca 1380
gccggaggtg gagaggcgga gaacggcaca gcaagaagac aaagaagaag agagactgtt 1440
gcctggacct ccgaaacgcg ctacaattca tttacaacac aggatggggt gagaatattg 1500
ccggaatcag tgaagcaggc ctcctaaaat aaaaatctat attttaggag gtaaaacatg 1560
aattttcaaa caatcgagct tgacacatgg tatagaaaat cttattttga ccattacatg 1620
aaggaagcga aatgttcttt cagcatcacg gcaaacgtca atgtgacaaa tttgctcgcc 1680
gtgctcaaga aaaagaagct caagctgtat ccggctttta tttatatcgt atcaagggtc 1740
attcattcgc gccctgagtt tagaacaacg tttgatgaca aaggacagct gggttattgg 1800
gaacaaatgc atccgtgcta tgcgattttt catcaggacg accaaacgtt ttccgccctc 1860
tggacggaat actcagacga tttttcgcag ttttatcatc aatatcttct ggacgccgag 1920
cgctttggag acaaaagggg cctttgggct aagccggaca tcccgcccaa tacgttttca 1980
gtttcttcta ttccatgggt gcgcttttca aacttcaatt taaaccttga taacagcgaa 2040
cacttgctgc cgattattac aaacgggaaa tacttttcag aaggcaggga aacatttttg 2100
cccgtttcct tgcaagttca ccatgcagtg tgtgacggct atcatgccgg cgcttttata 2160
aacgagttgg aacggcttgc cgccgattgt gaggagtggc ttgtgtgaca gaggaaaggc 2220
cgatatgatt cggccttttt tatatgtact tcttagcggg tctctaaaat aaccaaaaag 2280
caaggactgc tgaaagggct gacataagcc ttttgccggc ggtccttttt taattctgat 2340
ttttcaaact tagccccctc gagaaaaccg cttgagtatc ctccgctcat gttattgacc 2400
tccttttctt gatcaatata tttgtatgag cggaaagaaa atttgactag acggatgccc 2460
ctctgttgaa aatcttttta cagcatgtca gaatatgata tgatacaatt caaaggaaag 2520
tttaaactgt tatgattagg agtgtttgca tttatgaaga agattgcaat tgcggcgatt 2580
acagcgacaa gcgtgctggc tctcagcgca tgcagcgggg gagattctga ggttgttgcg 2640
gaaacaaaag ctggaaatat tacaaaagaa gacctttatc aaacattaaa agacaatgcc 2700
ggagcggacg cactgaacat gcttgttcag caaaaagtac tcgatgataa atacgatgtc 2760
tccgacaaag aaatcgacaa aaagctgaac gagtacaaaa aatcaatggg tgaccagctc 2820
aaccagctca ttgaccaaaa aggcgaagac ttcgtcaaag aacagatcaa atacgaactt 2880
ctgatgcaaa aagccgcaaa ggataacata aaagtaaccg atgatgacgt aaaagaatat 2940
tatgacggcc tgaaaggcaa aatccactta agccacattc ttgtgaaaga aaagaaaacg 3000
gctgaagaag ttgagaaaaa gctgaaaaaa ggcgaaaaat tcgaagacct tgcaaaagag 3060
tattcaactg acggtacagc cgaaaaaggc ggcgacctcg gctgggtcgg caaagacgat 3120
aacatggaca aggatttcgt caaagcggca tttgctttga aaaccggcga aatcagcgga 3180
cctgtgaaat cccaattcgg ctatcacatc attaaaaaag acgaagaacg cggcaaatat 3240
gaagacatga aaaaagagct taaaaaagaa gtccaagaac aaaagcaaaa tgatcaaact 3300
gaactgcaat ccgtcattga caaacttgtc aaagatgctg atttaaaagt aaaagacaaa 3360
gagttgaaaa aacaagtcga ccagcgtcaa gctcagacaa gcagcagcag ctgaaagagc 3420
agagaggacg gatttcctga aggaaatccg tttttttatt tttaacatct ctcactgctg 3480
tgtgatttta ctcacggcat ttggaacgcc ggctctcaac aaactttctg tagtgaaaat 3540
catgaaccaa acggatcgtc ggcctgatta acagctgaaa gctgccgatc acaaacatcc 3600
atagtcccgc cggcttcagt tcctcggaga aaaagcagaa gctcccgaca aggaataaaa 3660
ggccgatgag aaaatcgttt aatgtatgta gaactttgta tctttttttg aaaaagagtt 3720
catatcgatt gttattgttt tgcggcattg cttgatcact ccaatccttt tatttaccct 3780
gccggaagcc ggagtgaaac gccggtatac ataggattta tgaattagga aaacatatgg 3840
ggaaataaac catccaggag tgaaaaatat gcggttattc atatgtgcat cgtgcctgtt 3900
cggcttgatt gttccgtcat ttgaaacgaa agcgctgacg tttgaagaat tgccggttaa 3960
acaagcttca aaacaatggg aagttcaaat cggtaaagcc gaagccggaa acggaatggc 4020
gaaaccggaa aaaggagcgt ttcatactta tgctgtcgaa atcaaaaaca ttggacacga 4080
tgtggcttcg gcggaaattt ttgtctatcg gaacgagcct aattcttcaa cgaaattttc 4140
gctttggaac attcctcacg aaaatccggt ttctttagcc aaaagcttaa atcacggaag 4200
ctctgtcaag caccgcaatc tgcttatggc agagaatgcg accgaattgg aagtggacat 4260
gatttggacg gaaaaaggaa gcgaaggcag acttttaaag gaaacgttca ttttcaaggg 4320
agatgaatca tgaagaaaaa atggccgttc atcgtcaacg gtcttttttt aatgacttag 4380
gcagccgatc gttcggccat acgatatcga agcgacctcg aaccagcaga gctcgtcaca 4440
aaacatttgc atttaaagaa aaatacagga tgttttcacc aatatttttc tcaatgatga 4500
tacactattg acaagctgct actttgggag ggtgtttcca tagatgccga tgaagcaaaa 4560
acaccaaatg tgtcatgaga gctctctcta atcgatataa aagtagggtg aaccggggtt 4620
gtcaatctgt aaaagatctt tttttatccc gtgatacgct tttggaattc tgaatcttca 4680
agaaagtccc cagccttttg ctgatcaatc gagaacaaag gatgatacat atgaaaagaa 4740
tagataaaat ctaccatcag ctgctggata attttcgcga aaagaatatc aatcagcttt 4800
taaagataca agggaattcg gctaaagaaa tcgccgggca gctgcaaatg gagcgttcca 4860
atgtcagctt tgaattaaac aatctcgttc gggccaaaaa ggtgatcaag attaaaacgt 4920
tccccgtccg ctacatcccg gtggaa 4946
<210> 94
<211> 1341
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 94
tcgcggacaa gtacggcaag acaccggccc aagtcatttt gcgctgggat ttgcaaaacg 60
gggtcgttac gattccgaag tcgactaaag cggagcggat tgcccaaaac gcggacatat 120
ttgattttga actgaccacc gaggaaatga agcaaattga cgcgctgaat gaaaacaccc 180
gtgtcggccc tgatcccgat aactttgact tttaacaaaa cggccccgtt cgacattcga 240
acggggcttt aattgaattg tgcggttaca ccgccggact ccatcatcat cagttctttt 300
ttcatatcca atccgccccg gtatcccgtg agctgcccgc ttttaccgat aacccgatgg 360
caaggcacca ccattaacag cggatttgcg ccgatcgccg cgcctactgc ccgcacagcg 420
gcctgctttt caatatgctc ggcgatatcg gaataggagc aagtgctgcc gtaagggatt 480
tcggagagcg ccttccacac tgccagctga aaaggcgtgc cggcaaggtc gacaggaaag 540
ctgaaatgag ttcgcttgcc gttcaaatac gcctgcagct gctcggcgta ttctgccaat 600
cctttgtcat cccgaatgaa aactggctgt gtaaatcttt tttcagccca agcggccaaa 660
tcctcgaagc cttgattcca tccccctgta aaacagagcc cgcgggcagt cgccccaatg 720
tgaatctgcc aacctcggca aataagcgta cgccagtata cgatttgatc gtccatatgt 780
ttacctccgt ttcatttgcc ggtacgacgt cggcgattgc ccagtcttct ttttaaacaa 840
agaggcaaaa tattccgcat tcgcaatgcc taccattgaa gcgatttctg cgatcgatcg 900
ttctgaatga gcaagcaaat cgaccgcttt ctcaatcctt ttctgcagga tgtattctgc 960
cggcgagacg cctttgattc gtttaaatgt ccgctgcagg tgaaaagggc tgatatggca 1020
cctgtcagcc aaagcttgca gagacagcgg atcgcgataa gattcctcga tgatttccac 1080
cacacgctgt gccagctctt catccggcag cagcgccccg gccggattgc agcgtttgca 1140
ggggcggtac ccttctgata aagcatcttt tgcattgaaa aagatctgca cattgtcgat 1200
ttgcggaact ctcgatttgc aggaagggcg gcaaaatatg ccggtcgttt tgaccgcgta 1260
ataaaaaact ccgtcatagg cggaatcgtt ttccgtaatc gcccgccaca tttcaggcgt 1320
caatcgtgat ttgctgttca t 1341
<210> 95
<211> 216
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 95
atcttcaccc cgatctatgt cagtataacc tatatgacag ccggaggtgg agaggcggag 60
aacggcacag caagaagaca aagaagaaga gagactgttg cctggacctc cgaaacgcgc 120
tacaattcat ttacaacaca ggatggggtg agaatattgc cggaatcagt gaagcaggcc 180
tcctaaaata aaaatctata ttttaggagg taaaac 216
<210> 96
<211> 648
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 96
atgaattttc aaacaatcga gcttgacaca tggtatagaa aatcttattt tgaccattac 60
atgaaggaag cgaaatgttc tttcagcatc acggcaaacg tcaatgtgac aaatttgctc 120
gccgtgctca agaaaaagaa gctcaagctg tatccggctt ttatttatat cgtatcaagg 180
gtcattcatt cgcgccctga gtttagaaca acgtttgatg acaaaggaca gctgggttat 240
tgggaacaaa tgcatccgtg ctatgcgatt tttcatcagg acgaccaaac gttttccgcc 300
ctctggacgg aatactcaga cgatttttcg cagttttatc atcaatatct tctggacgcc 360
gagcgctttg gagacaaaag gggcctttgg gctaagccgg acatcccgcc caatacgttt 420
tcagtttctt ctattccatg ggtgcgcttt tcaaacttca atttaaacct tgataacagc 480
gaacacttgc tgccgattat tacaaacggg aaatactttt cagaaggcag ggaaacattt 540
ttgcccgttt ccttgcaagt tcaccatgca gtgtgtgacg gctatcatgc cggcgctttt 600
ataaacgagt tggaacggct tgccgccgat tgtgaggagt ggcttgtg 648
<210> 97
<211> 149
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 97
aaaggccgat atgattcggc cttttttata tgtacttctt agcgggtctc taaaataacc 60
aaaaagcaag gactgctgaa agggctgaca taagcctttt gccggcggtc cttttttaat 120
tctgattttt caaacttagc cccctcgag 149
<210> 98
<211> 60
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 98
tgacagagga aaggccgata tgattcggcc ttttttatat gtacttctta gcgggtctct 60
<210> 99
<211> 98
<212> DNA
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 99
aaaataacca aaaagcaagg actgctgaaa gggctgacat aagccttttg ccggcggtcc 60
ttttttaatt ctgatttttc aaacttagcc ccctcgag 98
<210> 100
<211> 190
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 100
aaaaccgctt gagtatcctc cgctcatgtt attgacctcc ttttcttgat caatatattt 60
gtatgagcgg aaagaaaatt tgactagacg gatgcccctc tgttgaaaat ctttttacag 120
catgtcagaa tatgatatga tacaattcaa aggaaagttt aaactgttat gattaggagt 180
gtttgcattt 190
<210> 101
<211> 861
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 101
atgaagaaga ttgcaattgc ggcgattaca gcgacaagcg tgctggctct cagcgcatgc 60
agcgggggag attctgaggt tgttgcggaa acaaaagctg gaaatattac aaaagaagac 120
ctttatcaaa cattaaaaga caatgccgga gcggacgcac tgaacatgct tgttcagcaa 180
aaagtactcg atgataaata cgatgtctcc gacaaagaaa tcgacaaaaa gctgaacgag 240
tacaaaaaat caatgggtga ccagctcaac cagctcattg accaaaaagg cgaagacttc 300
gtcaaagaac agatcaaata cgaacttctg atgcaaaaag ccgcaaagga taacataaaa 360
gtaaccgatg atgacgtaaa agaatattat gacggcctga aaggcaaaat ccacttaagc 420
cacattcttg tgaaagaaaa gaaaacggct gaagaagttg agaaaaagct gaaaaaaggc 480
gaaaaattcg aagaccttgc aaaagagtat tcaactgacg gtacagccga aaaaggcggc 540
gacctcggct gggtcggcaa agacgataac atggacaagg atttcgtcaa agcggcattt 600
gctttgaaaa ccggcgaaat cagcggacct gtgaaatccc aattcggcta tcacatcatt 660
aaaaaagacg aagaacgcgg caaatatgaa gacatgaaaa aagagcttaa aaaagaagtc 720
caagaacaaa agcaaaatga tcaaactgaa ctgcaatccg tcattgacaa acttgtcaaa 780
gatgctgatt taaaagtaaa agacaaagag ttgaaaaaac aagtcgacca gcgtcaagct 840
cagacaagca gcagcagctg a 861
<210> 102
<211> 34
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 102
cggatttcct gaaggaaatc cgttttttta tttt 34
<210> 103
<211> 1484
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 103
taacatctct cactgctgtg tgattttact cacggcattt ggaacgccgg ctctcaacaa 60
actttctgta gtgaaaatca tgaaccaaac ggatcgtcgg cctgattaac agctgaaagc 120
tgccgatcac aaacatccat agtcccgccg gcttcagttc ctcggagaaa aagcagaagc 180
tcccgacaag gaataaaagg ccgatgagaa aatcgtttaa tgtatgtaga actttgtatc 240
tttttttgaa aaagagttca tatcgattgt tattgttttg cggcattgct tgatcactcc 300
aatcctttta tttaccctgc cggaagccgg agtgaaacgc cggtatacat aggatttatg 360
aattaggaaa acatatgggg aaataaacca tccaggagtg aaaaatatgc ggttattcat 420
atgtgcatcg tgcctgttcg gcttgattgt tccgtcattt gaaacgaaag cgctgacgtt 480
tgaagaattg ccggttaaac aagcttcaaa acaatgggaa gttcaaatcg gtaaagccga 540
agccggaaac ggaatggcga aaccggaaaa aggagcgttt catacttatg ctgtcgaaat 600
caaaaacatt ggacacgatg tggcttcggc ggaaattttt gtctatcgga acgagcctaa 660
ttcttcaacg aaattttcgc tttggaacat tcctcacgaa aatccggttt ctttagccaa 720
aagcttaaat cacggaagct ctgtcaagca ccgcaatctg cttatggcag agaatgcgac 780
cgaattggaa gtggacatga tttggacgga aaaaggaagc gaaggcagac ttttaaagga 840
aacgttcatt ttcaagggag atgaatcatg aagaaaaaat ggccgttcat cgtcaacggt 900
ctttttttaa tgacttaggc agccgatcgt tcggccatac gatatcgaag cgacctcgaa 960
ccagcagagc tcgtcacaaa acatttgcat ttaaagaaaa atacaggatg ttttcaccaa 1020
tatttttctc aatgatgata cactattgac aagctgctac tttgggaggg tgtttccata 1080
gatgccgatg aagcaaaaac accaaatgtg tcatgagagc tctctctaat cgatataaaa 1140
gtagggtgaa ccggggttgt caatctgtaa aagatctttt tttatcccgt gatacgcttt 1200
tggaattctg aatcttcaag aaagtcccca gccttttgct gatcaatcga gaacaaagga 1260
tgatacatat gaaaagaata gataaaatct accatcagct gctggataat tttcgcgaaa 1320
agaatatcaa tcagctttta aagatacaag ggaattcggc taaagaaatc gccgggcagc 1380
tgcaaatgga gcgttccaat gtcagctttg aattaaacaa tctcgttcgg gccaaaaagg 1440
tgatcaagat taaaacgttc cccgtccgct acatcccggt ggaa 1484
<210> 104
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 104
tcgttctgaa tgagcaagca 20
<210> 105
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 105
tgttaatcag gccgacgatc 20
<210> 106
<211> 2676
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 106
tcgttctgaa tgagcaagca aatcgaccgc tttctcaatc cttttctgca ggatgtattc 60
tgccggcgag acgcctttga ttcgtttaaa tgtccgctgc aggtgaaaag ggctgatatg 120
gcacctgtca gccaaagctt gcagagacag cggatcgcga taagattcct cgatgatttc 180
caccacacgc tgtgccagct cttcatccgg cagcagcgcc ccggccggat tgcagcgttt 240
gcaggggcgg tacccttctg ataaagcatc ttttgcattg aaaaagatct gcacattgtc 300
gatttgcgga actctcgatt tgcaggaagg gcggcaaaat atgccggtcg ttttgaccgc 360
gtaataaaaa actccgtcat aggcggaatc gttttccgta atcgcccgcc acatttcagg 420
cgtcaatcgt gatttgctgt tcatatcttc accccgatct atgtcagtat aacctatatg 480
acagccggag gtggagaggc ggagaacggc acagcaagaa gacaaagaag aagagagact 540
gttgcctgga cctccgaaac gcgctacaat tcatttacaa cacaggatgg ggtgagaata 600
ttgccggaat cagtgaagca ggcctcctaa aataaaaatc tatattttag gaggtaaaac 660
atgaattttc aaacaatcga gcttgacaca tggtatagaa aatcttattt tgaccattac 720
atgaaggaag cgaaatgttc tttcagcatc acggcaaacg tcaatgtgac aaatttgctc 780
gccgtgctca agaaaaagaa gctcaagctg tatccggctt ttatttatat cgtatcaagg 840
gtcattcatt cgcgccctga gtttagaaca acgtttgatg acaaaggaca gctgggttat 900
tgggaacaaa tgcatccgtg ctatgcgatt tttcatcagg acgaccaaac gttttccgcc 960
ctctggacgg aatactcaga cgatttttcg cagttttatc atcaatatct tctggacgcc 1020
gagcgctttg gagacaaaag gggcctttgg gctaagccgg acatcccgcc caatacgttt 1080
tcagtttctt ctattccatg ggtgcgcttt tcaaacttca atttaaacct tgataacagc 1140
gaacacttgc tgccgattat tacaaacggg aaatactttt cagaaggcag ggaaacattt 1200
ttgcccgttt ccttgcaagt tcaccatgca gtgtgtgacg gctatcatgc cggcgctttt 1260
ataaacgagt tggaacggct tgccgccgat tgtgaggagt ggcttgtgtg acagaggaaa 1320
ggccgatatg attcggcctt ttttatatgt acttcttagc gggtctctaa aataaccaaa 1380
aagcaaggac tgctgaaagg gctgacataa gccttttgcc ggcggtcctt ttttaattct 1440
gatttttcaa acttagcccc ctcgagaaaa ccgcttgagt atcctccgct catgttattg 1500
acctcctttt cttgatcaat atatttgtat gagcggaaag aaaatttgac tagacggatg 1560
cccctctgtt gaaaatcttt ttacagcatg tcagaatatg atatgataca attcaaagga 1620
aagtttaaac tgttatgatt aggagtgttt gcatttatga agaagattgc aattgcggcg 1680
attacagcga caagcgtgct ggctctcagc gcatgcagcg ggggagattc tgaggttgtt 1740
gcggaaacaa aagctggaaa tattacaaaa gaagaccttt atcaaacatt aaaagacaat 1800
gccggagcgg acgcactgaa catgcttgtt cagcaaaaag tactcgatga taaatacgat 1860
gtctccgaca aagaaatcga caaaaagctg aacgagtaca aaaaatcaat gggtgaccag 1920
ctcaaccagc tcattgacca aaaaggcgaa gacttcgtca aagaacagat caaatacgaa 1980
cttctgatgc aaaaagccgc aaaggataac ataaaagtaa ccgatgatga cgtaaaagaa 2040
tattatgacg gcctgaaagg caaaatccac ttaagccaca ttcttgtgaa agaaaagaaa 2100
acggctgaag aagttgagaa aaagctgaaa aaaggcgaaa aattcgaaga ccttgcaaaa 2160
gagtattcaa ctgacggtac agccgaaaaa ggcggcgacc tcggctgggt cggcaaagac 2220
gataacatgg acaaggattt cgtcaaagcg gcatttgctt tgaaaaccgg cgaaatcagc 2280
ggacctgtga aatcccaatt cggctatcac atcattaaaa aagacgaaga acgcggcaaa 2340
tatgaagaca tgaaaaaaga gcttaaaaaa gaagtccaag aacaaaagca aaatgatcaa 2400
actgaactgc aatccgtcat tgacaaactt gtcaaagatg ctgatttaaa agtaaaagac 2460
aaagagttga aaaaacaagt cgaccagcgt caagctcaga caagcagcag cagctgaaag 2520
agcagagagg acggatttcc tgaaggaaat ccgttttttt atttttaaca tctctcactg 2580
ctgtgtgatt ttactcacgg catttggaac gccggctctc aacaaacttt ctgtagtgaa 2640
aatcatgaac caaacggatc gtcggcctga ttaaca 2676
<210> 107
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 107
aacctatatg acagccggag 20
<210> 108
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 108
ggcaaaatcc acttaagcca c 21
<210> 109
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 109
aacgagttgg aacggcttgc 20
<210> 110
<211> 686
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 110
cctcctaaaa taaaaatcta tattttagga ggtaaaacat gaattttcaa acaatcgagc 60
ttgacacatg gtatagaaaa tcttattttg accattacat gaaggaagcg aaatgttctt 120
tcagcatcac ggcaaacgtc aatgtgacaa atttgctcgc cgtgctcaag aaaaagaagc 180
tcaagctgta tccggctttt atttatatcg tatcaagggt cattcattcg cgccctgagt 240
ttagaacaac gtttgatgac aaaggacagc tgggttattg ggaacaaatg catccgtgct 300
atgcgatttt tcatcaggac gaccaaacgt tttccgccct ctggacggaa tactcagacg 360
atttttcgca gttttatcat caatatcttc tggacgccga gcgctttgga gacaaaaggg 420
gcctttgggc taagccggac atcccgccca atacgttttc agtttcttct attccatggg 480
tgcgcttttc aaacttcaat ttaaaccttg ataacagcga acacttgctg ccgattatta 540
caaacgggaa atacttttca gaaggcaggg aaacattttt gcccgtttcc ttgcaagttc 600
accatgcagt gtgtgacggc tatcatgccg gcgcttttat aaacgagttg gaacggcttg 660
ccgccgattg tgaggagtgg cttgtg 686
<210> 111
<211> 4260
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 111
tcgcggacaa gtacggcaag acaccggccc aagtcatttt gcgctgggat ttgcaaaacg 60
gggtcgttac gattccgaag tcgactaaag cggagcggat tgcccaaaac gcggacatat 120
ttgattttga actgaccacc gaggaaatga agcaaattga cgcgctgaat gaaaacaccc 180
gtgtcggccc tgatcccgat aactttgact tttaacaaaa cggccccgtt cgacattcga 240
acggggcttt aattgaattg tgcggttaca ccgccggact ccatcatcat cagttctttt 300
ttcatatcca atccgccccg gtatcccgtg agctgcccgc ttttaccgat aacccgatgg 360
caaggcacca ccattaacag cggatttgcg ccgatcgccg cgcctactgc ccgcacagcg 420
gcctgctttt caatatgctc ggcgatatcg gaataggagc aagtgctgcc gtaagggatt 480
tcggagagcg ccttccacac tgccagctga aaaggcgtgc cggcaaggtc gacaggaaag 540
ctgaaatgag ttcgcttgcc gttcaaatac gcctgcagct gctcggcgta ttctgccaat 600
cctttgtcat cccgaatgaa aactggctgt gtaaatcttt tttcagccca agcggccaaa 660
tcctcgaagc cttgattcca tccccctgta aaacagagcc cgcgggcagt cgccccaatg 720
tgaatctgcc aacctcggca aataagcgta cgccagtata cgatttgatc gtccatatgt 780
ttacctccgt ttcatttgcc ggtacgacgt cggcgattgc ccagtcttct ttttaaacaa 840
agaggcaaaa tattccgcat tcgcaatgcc taccattgaa gcgatttctg cgatcgatcg 900
ttctgaatga gcaagcaaat cgaccgcttt ctcaatcctt ttctgcagga tgtattctgc 960
cggcgagacg cctttgattc gtttaaatgt ccgctgcagg tgaaaagggc tgatatggca 1020
cctgtcagcc aaagcttgca gagacagcgg atcgcgataa gattcctcga tgatttccac 1080
cacacgctgt gccagctctt catccggcag cagcgccccg gccggattgc agcgtttgca 1140
ggggcggtac ccttctgata aagcatcttt tgcattgaaa aagatctgca cattgtcgat 1200
ttgcggaact ctcgatttgc aggaagggcg gcaaaatatg ccggtcgttt tgaccgcgta 1260
ataaaaaact ccgtcatagg cggaatcgtt ttccgtaatc gcccgccaca tttcaggcgt 1320
caatcgtgat ttgctgttca tatcttcacc ccgatctatg tcagtataac ctatatgaca 1380
gccggaggtg gagaggcgga gaacggcaca gcaagaagac aaagaagaag agagactgtt 1440
gcctggacct ccgaaacgcg ctacaattca tttacaacac aggatggggt gagaatattg 1500
ccggaatcag tgaagcaggt gacagaggaa aggccgatat gattcggcct tttttatatg 1560
tacttcttag cgggtctcta aaataaccaa aaagcaagga ctgctgaaag ggctgacata 1620
agccttttgc cggcggtcct tttttaattc tgatttttca aacttagccc cctcgagaaa 1680
accgcttgag tatcctccgc tcatgttatt gacctccttt tcttgatcaa tatatttgta 1740
tgagcggaaa gaaaatttga ctagacggat gcccctctgt tgaaaatctt tttacagcat 1800
gtcagaatat gatatgatac aattcaaagg aaagtttaaa ctgttatgat taggagtgtt 1860
tgcatttatg aagaagattg caattgcggc gattacagcg acaagcgtgc tggctctcag 1920
cgcatgcagc gggggagatt ctgaggttgt tgcggaaaca aaagctggaa atattacaaa 1980
agaagacctt tatcaaacat taaaagacaa tgccggagcg gacgcactga acatgcttgt 2040
tcagcaaaaa gtactcgatg ataaatacga tgtctccgac aaagaaatcg acaaaaagct 2100
gaacgagtac aaaaaatcaa tgggtgacca gctcaaccag ctcattgacc aaaaaggcga 2160
agacttcgtc aaagaacaga tcaaatacga acttctgatg caaaaagccg caaaggataa 2220
cataaaagta accgatgatg acgtaaaaga atattatgac ggcctgaaag gcaaaatcca 2280
cttaagccac attcttgtga aagaaaagaa aacggctgaa gaagttgaga aaaagctgaa 2340
aaaaggcgaa aaattcgaag accttgcaaa agagtattca actgacggta cagccgaaaa 2400
aggcggcgac ctcggctggg tcggcaaaga cgataacatg gacaaggatt tcgtcaaagc 2460
ggcatttgct ttgaaaaccg gcgaaatcag cggacctgtg aaatcccaat tcggctatca 2520
catcattaaa aaagacgaag aacgcggcaa atatgaagac atgaaaaaag agcttaaaaa 2580
agaagtccaa gaacaaaagc aaaatgatca aactgaactg caatccgtca ttgacaaact 2640
tgtcaaagat gctgatttaa aagtaaaaga caaagagttg aaaaaacaag tcgaccagcg 2700
tcaagctcag acaagcagca gcagctgaaa gagcagagag gacggatttc ctgaaggaaa 2760
tccgtttttt tatttttaac atctctcact gctgtgtgat tttactcacg gcatttggaa 2820
cgccggctct caacaaactt tctgtagtga aaatcatgaa ccaaacggat cgtcggcctg 2880
attaacagct gaaagctgcc gatcacaaac atccatagtc ccgccggctt cagttcctcg 2940
gagaaaaagc agaagctccc gacaaggaat aaaaggccga tgagaaaatc gtttaatgta 3000
tgtagaactt tgtatctttt tttgaaaaag agttcatatc gattgttatt gttttgcggc 3060
attgcttgat cactccaatc cttttattta ccctgccgga agccggagtg aaacgccggt 3120
atacatagga tttatgaatt aggaaaacat atggggaaat aaaccatcca ggagtgaaaa 3180
atatgcggtt attcatatgt gcatcgtgcc tgttcggctt gattgttccg tcatttgaaa 3240
cgaaagcgct gacgtttgaa gaattgccgg ttaaacaagc ttcaaaacaa tgggaagttc 3300
aaatcggtaa agccgaagcc ggaaacggaa tggcgaaacc ggaaaaagga gcgtttcata 3360
cttatgctgt cgaaatcaaa aacattggac acgatgtggc ttcggcggaa atttttgtct 3420
atcggaacga gcctaattct tcaacgaaat tttcgctttg gaacattcct cacgaaaatc 3480
cggtttcttt agccaaaagc ttaaatcacg gaagctctgt caagcaccgc aatctgctta 3540
tggcagagaa tgcgaccgaa ttggaagtgg acatgatttg gacggaaaaa ggaagcgaag 3600
gcagactttt aaaggaaacg ttcattttca agggagatga atcatgaaga aaaaatggcc 3660
gttcatcgtc aacggtcttt ttttaatgac ttaggcagcc gatcgttcgg ccatacgata 3720
tcgaagcgac ctcgaaccag cagagctcgt cacaaaacat ttgcatttaa agaaaaatac 3780
aggatgtttt caccaatatt tttctcaatg atgatacact attgacaagc tgctactttg 3840
ggagggtgtt tccatagatg ccgatgaagc aaaaacacca aatgtgtcat gagagctctc 3900
tctaatcgat ataaaagtag ggtgaaccgg ggttgtcaat ctgtaaaaga tcttttttta 3960
tcccgtgata cgcttttgga attctgaatc ttcaagaaag tccccagcct tttgctgatc 4020
aatcgagaac aaaggatgat acatatgaaa agaatagata aaatctacca tcagctgctg 4080
gataattttc gcgaaaagaa tatcaatcag cttttaaaga tacaagggaa ttcggctaaa 4140
gaaatcgccg ggcagctgca aatggagcgt tccaatgtca gctttgaatt aaacaatctc 4200
gttcgggcca aaaaggtgat caagattaaa acgttccccg tccgctacat cccggtggaa 4260
<210> 112
<211> 1990
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 112
tcgttctgaa tgagcaagca aatcgaccgc tttctcaatc cttttctgca ggatgtattc 60
tgccggcgag acgcctttga ttcgtttaaa tgtccgctgc aggtgaaaag ggctgatatg 120
gcacctgtca gccaaagctt gcagagacag cggatcgcga taagattcct cgatgatttc 180
caccacacgc tgtgccagct cttcatccgg cagcagcgcc ccggccggat tgcagcgttt 240
gcaggggcgg tacccttctg ataaagcatc ttttgcattg aaaaagatct gcacattgtc 300
gatttgcgga actctcgatt tgcaggaagg gcggcaaaat atgccggtcg ttttgaccgc 360
gtaataaaaa actccgtcat aggcggaatc gttttccgta atcgcccgcc acatttcagg 420
cgtcaatcgt gatttgctgt tcatatcttc accccgatct atgtcagtat aacctatatg 480
acagccggag gtggagaggc ggagaacggc acagcaagaa gacaaagaag aagagagact 540
gttgcctgga cctccgaaac gcgctacaat tcatttacaa cacaggatgg ggtgagaata 600
ttgccggaat cagtgaagca ggtgacagag gaaaggccga tatgattcgg ccttttttat 660
atgtacttct tagcgggtct ctaaaataac caaaaagcaa ggactgctga aagggctgac 720
ataagccttt tgccggcggt ccttttttaa ttctgatttt tcaaacttag ccccctcgag 780
aaaaccgctt gagtatcctc cgctcatgtt attgacctcc ttttcttgat caatatattt 840
gtatgagcgg aaagaaaatt tgactagacg gatgcccctc tgttgaaaat ctttttacag 900
catgtcagaa tatgatatga tacaattcaa aggaaagttt aaactgttat gattaggagt 960
gtttgcattt atgaagaaga ttgcaattgc ggcgattaca gcgacaagcg tgctggctct 1020
cagcgcatgc agcgggggag attctgaggt tgttgcggaa acaaaagctg gaaatattac 1080
aaaagaagac ctttatcaaa cattaaaaga caatgccgga gcggacgcac tgaacatgct 1140
tgttcagcaa aaagtactcg atgataaata cgatgtctcc gacaaagaaa tcgacaaaaa 1200
gctgaacgag tacaaaaaat caatgggtga ccagctcaac cagctcattg accaaaaagg 1260
cgaagacttc gtcaaagaac agatcaaata cgaacttctg atgcaaaaag ccgcaaagga 1320
taacataaaa gtaaccgatg atgacgtaaa agaatattat gacggcctga aaggcaaaat 1380
ccacttaagc cacattcttg tgaaagaaaa gaaaacggct gaagaagttg agaaaaagct 1440
gaaaaaaggc gaaaaattcg aagaccttgc aaaagagtat tcaactgacg gtacagccga 1500
aaaaggcggc gacctcggct gggtcggcaa agacgataac atggacaagg atttcgtcaa 1560
agcggcattt gctttgaaaa ccggcgaaat cagcggacct gtgaaatccc aattcggcta 1620
tcacatcatt aaaaaagacg aagaacgcgg caaatatgaa gacatgaaaa aagagcttaa 1680
aaaagaagtc caagaacaaa agcaaaatga tcaaactgaa ctgcaatccg tcattgacaa 1740
acttgtcaaa gatgctgatt taaaagtaaa agacaaagag ttgaaaaaac aagtcgacca 1800
gcgtcaagct cagacaagca gcagcagctg aaagagcaga gaggacggat ttcctgaagg 1860
aaatccgttt ttttattttt aacatctctc actgctgtgt gattttactc acggcatttg 1920
gaacgccggc tctcaacaaa ctttctgtag tgaaaatcat gaaccaaacg gatcgtcggc 1980
ctgattaaca 1990
<210> 113
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 113
gggtacctcc atggtaaagt 20
<210> 114
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 114
acgtattaat gcagtagccg 20
<210> 115
<211> 2067
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 115
gggtacctcc atggtaaagt ttcgacttaa agcggagatc aaaaaatagc ctggtttacc 60
attttccaca tttttaaatt tatacccctt tcaaaagtca taaaactatc atattctgta 120
ataccacttt tcggaggtga gaatatgtca tactctaata gcttcgcttt gattgttgta 180
ttatttattt tgctgatcat tgtgggctcc gcttttgtaa agtattaatt cattcgcgca 240
cgaagccttc ttccgaaaaa ctgcaggttc atatcgaaag gcgctccgct tttcatatga 300
gcctgttttt ttgtaattta aaaagaattt atttccacaa taacaaaaaa tgtttataat 360
aggataggct ctgagctgac ggctgacggc gctcggaaat aaaaactctc ccacaaatga 420
aacttttata caacattttc gtcttatact gttgttaagg caattcaata acggaaagga 480
aaagatcaat gttaaataag ctaagaatgt tgtatgaaaa aacaacaatg aagtggctgc 540
tcaacacttc ctattacttt ctcattctgg tgctgctgtt tttaatttac ggattccata 600
cagcaaatac aggttcatat atttataatg acttttaatg gagaagaaaa caatgaaact 660
gattgaaaca attaaaaaat accacaaatg tgcttgatgg atccgtctta ttcggaagag 720
ctgcttccgg agctgagctt gtttatgttc tgcggagaaa cgcttcctgc gtcagtcgcc 780
agacagctta aagaaagatt tccaaaggct cgtgttttta acacatacgg cccgaccgaa 840
gcgaccgttg ccgtcacttc catcgaagta acagacgacg tattaaacaa gtattcttct 900
cttcccgtcg gttctgaaaa gcccgaaacc gaaatcgtaa tcatcaatga agacggaaaa 960
gccgtccaag acggggaaaa aggcgagatc ataattaccg gagcaagcgt cagcaaaggc 1020
tatttaggcg aaaaagcgct gacggaaaaa gcgtttttct cttacaacgg ctcccctgct 1080
taccgcaccg gtgacgcagg ttacaaagaa aacggccagc tcttcttcct cggaagactc 1140
gatttccaaa tcaaactgca cggctaccgg attgagcttg aagaaatcga atatcaaatc 1200
aatcagtccc gatatgtgca gtcggcagtc gttattccat tctaccgcga ggagaaaatc 1260
gagtatctga tcgccatgat tgtgcctgcc gaacacgatt tcgaaaagga ataccagtta 1320
accagcgcga ttaagaaaga tttgggcagc aagctgccgg cctatatgat tccgagaaaa 1380
ttcatgtatc aaaaagagat tccgatgaca gcaaacggta aaatcgaccg caagagatta 1440
aaagaagagg taaccgtatg acgccttatg gttcatttct tttcttcatt atattaggaa 1500
ttttactggc gccgaccatc attctcggat tgaacggaaa aagcttccgc ttatataata 1560
tggcggtttc cgttctggtt ctggcgctta ttttttcgaa cagcctgcac gggctgatca 1620
tgctgtgcct gtttacgctt tggcagacgg ttctgatcaa aggctatatc gcttaccgtc 1680
taaaagcgaa cagcggcatc gtattttgtc tggctgcagc agcttctatc ctgcctctgg 1740
cactgtcaaa gctgctgccg tttttcgccg ttgacaactg ggcaacattt ctcggaatct 1800
cgtatttaac ttttaaaggg gttcagctca tcattgaaac ccgcgacggt ctcattaaaa 1860
agcagctccc aatcagcaga ctgctttact ttattctctt ttttccaacc atctcgtcag 1920
gtccgatcga caggtaccgc cgttttgaaa aagacgatca gacggtttgg acaaaggagc 1980
aatacgaaga actgctttac aaaggaatca ataaaatttt tctcggcttt ttgtacaaat 2040
tcattatcgg ctactgcatt aatacgt 2067
<210> 116
<211> 2767
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 116
gggtacctcc atggtaaagt ttcgacttaa agcggagatc aaaaaatagc ctggtttacc 60
attttccaca tttttaaatt tatacccctt tcaaaagtca taaaactatc atattctgta 120
ataccacttt tcggaggtga gaatatgtca tactctaata gcttcgcttt gattgttgta 180
ttatttattt tgctgatcat tgtgggctcc gcttttgtaa agtattaatt cattcgcgca 240
cgaagccttc ttccgaaaaa ctgcaggttc atatcgaaag gcgctccgct tttcatatga 300
gcctgttttt ttgtaattta aaaagaattt atttccacaa taacaaaaaa tgtttataat 360
aggataggct ctgagctgac ggctgacggc gctcggaaat aaaaactctc ccacaaatga 420
aacttttata caacattttc gtcttatact gttgttaagg caattcaata acggaaagga 480
aaagatcaat gttaaataag ctaagaatgt tgtatgaaaa aacaacaatg aagtggctgc 540
tcaacacttc ctattacttt ctcattctgg tgctgctgtt tttaatttac ggattccata 600
cagcaaatac aggttcatat atttataatg acttttaatg gagaagaaaa caatgaaact 660
gattgaaaca attaaaaaat acgcgcaaac acaacccgat acccttgctt ttgtgaatga 720
agaagaaaag ctgacatacg gagagctttg gtcacaatct gagcgccttg ccgcaaggat 780
tcaaagcgaa gctttaacag atgcttctcc gatcattgtc tacggacata tgaaaccggt 840
tatggcggtt tccttcctgg cttgtgtaaa agccgggcat ccgtacattc cagttgacgt 900
atcgattccg gcagaccgga ttctgaaaat catcaacagt tcaaaagccg agcttctgct 960
caataattca ggcacttcgg ttgacacggg agatgccctc atttccgtag tggaaccggg 1020
cgttcttgaa ggagacggga tgccggagac agatcccgga cgctgggtac atggggaaga 1080
tactttttac attatctaca catccggaag cacgggaaat ccaaaaggcg tgcaaatctc 1140
agccgataac cttcagagct tcacagattg gatcacgaat gactttccgg tggaaagcgg 1200
ccaagtgttc ttaaatcaag ccccgttttc ctttgattta tcggtaatgg acctttatcc 1260
gtgcctgcag tccggaggca ccttatggac ggttacgaaa gacatgatta atcggccgaa 1320
actattgttt gaagcactga aacagtccaa cgtcaacgtc tggacatcga cgccttcatt 1380
cgcacaaatg tgcttgatgg atccgtctta ttcggaagag ctgcttccgg agctgagctt 1440
gtttatgttc tgcggagaaa cgcttcctgc gtcagtcgcc agacagctta aagaaagatt 1500
tccaaaggct cgtgttttta acacatacgg cccgaccgaa gcgaccgttg ccgtcacttc 1560
catcgaagta acagacgacg tattaaacaa gtattcttct cttcccgtcg gttctgaaaa 1620
gcccgaaacc gaaatcgtaa tcatcaatga agacggaaaa gccgtccaag acggggaaaa 1680
aggcgagatc ataattaccg gagcaagcgt cagcaaaggc tatttaggcg aaaaagcgct 1740
gacggaaaaa gcgtttttct cttacaacgg ctcccctgct taccgcaccg gtgacgcagg 1800
ttacaaagaa aacggccagc tcttcttcct cggaagactc gatttccaaa tcaaactgca 1860
cggctaccgg attgagcttg aagaaatcga atatcaaatc aatcagtccc gatatgtgca 1920
gtcggcagtc gttattccat tctaccgcga ggagaaaatc gagtatctga tcgccatgat 1980
tgtgcctgcc gaacacgatt tcgaaaagga ataccagtta accagcgcga ttaagaaaga 2040
tttgggcagc aagctgccgg cctatatgat tccgagaaaa ttcatgtatc aaaaagagat 2100
tccgatgaca gcaaacggta aaatcgaccg caagagatta aaagaagagg taaccgtatg 2160
acgccttatg gttcatttct tttcttcatt atattaggaa ttttactggc gccgaccatc 2220
attctcggat tgaacggaaa aagcttccgc ttatataata tggcggtttc cgttctggtt 2280
ctggcgctta ttttttcgaa cagcctgcac gggctgatca tgctgtgcct gtttacgctt 2340
tggcagacgg ttctgatcaa aggctatatc gcttaccgtc taaaagcgaa cagcggcatc 2400
gtattttgtc tggctgcagc agcttctatc ctgcctctgg cactgtcaaa gctgctgccg 2460
tttttcgccg ttgacaactg ggcaacattt ctcggaatct cgtatttaac ttttaaaggg 2520
gttcagctca tcattgaaac ccgcgacggt ctcattaaaa agcagctccc aatcagcaga 2580
ctgctttact ttattctctt ttttccaacc atctcgtcag gtccgatcga caggtaccgc 2640
cgttttgaaa aagacgatca gacggtttgg acaaaggagc aatacgaaga actgctttac 2700
aaaggaatca ataaaatttt tctcggcttt ttgtacaaat tcattatcgg ctactgcatt 2760
aatacgt 2767
<210> 117
<211> 21
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 117
ggatacgccg atttcaatgg c 21
<210> 118
<211> 20
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 118
ggctatgtgc tgggggaatt 20
<210> 119
<211> 1523
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 119
ggatacgccg atttcaatgg caagctgatt taaggagaga ttgtgttcaa tcatgtactg 60
gagaacaaaa tctcttttga tatgaatctt ttttaccatg attactcccc tttctaatct 120
cttatgtttc tttttatcta cattgaacat atacgatttg ttaacttttg tcaatacttt 180
taccatccat atgtttccta taggcaatat tcgtactaaa atattttata ataagagatt 240
gcgaggtttt ggccatgacg aactttggac accatttacg acaattaagg gaacggaaaa 300
aactgaccgt caatcaactg gcgatgtatt ccggcgtcag ttcggcaggc atttcgcgaa 360
tcgaaaacgg aaagcgcggc gtgccgaagc cggcgacgat cagaaaactg gcggacgctt 420
tgaaagtccc gtatgaggaa ctgatggcat ctgcaggcta tatcagcgcg tctacagtcc 480
aggaagcaag aagcagctat gattccattt acgacatcgt gtcacagtac gatttagagg 540
acctttctct gtttgacagc gaaaagtgga aggtgctttc aaaaaaagac atcgaaaacc 600
tggacaaata tttcgacttt ctcgtgcagg aagcaagcag ccgaaacaaa aactgaatac 660
ttctccgcgg cacactctcc tctctatcat tttcgtctgt ttacgatcct gctgttattt 720
tatcccttat gttaactttt gtcaatattt ttcctgtcta agtatttcct atagtcaaca 780
tttgtattaa aatgttcata tcatgaattt gcggggggga tggcgatgac aaggcaatca 840
taaaaagccg aatttccctt ttaggagaag ttcggctttt ttcggctgcc ttaagcggca 900
tccggattcg gcgtcttgcc tttatgatgc ttaacggggc tcagcgcacg ctcgagccat 960
cccatgaaca gatcggcgat gatcgccatc agcgccgtcg ggatcgcgcc tgctagaatg 1020
atcgctgttc cgttggtcgc gtttgatccc ctgacaatga tatccccgag gccgcctgcg 1080
ccgacaaacg tgccgatggc cgtaatgccg atcgcgatga cgagcgcggt tctgagcccc 1140
gccataatga ccgacaaggc gaggggaagc tccaccatcc ggagcacttg aaatttcgtc 1200
atgcccatcg ccttccctga ttcaagatag gcatgctcga tgctggcgat tcccgtatat 1260
gtgtttcgaa tgatcggcaa cagcgaatac aaaaacaatg aaagaatcac cgtgtttgcg 1320
ccgagcccca tgacaagcat caagacggcg agcatcgcca gcgccggaac cgtttgaatg 1380
acattagtga tggaaaagac ccatttgctg attttacggt atctggcgat gaaaatgccg 1440
gccgggatgc cgacgacggc ggcgaacaat acgccgtatg ccgacattaa aaagtggcgg 1500
taaaattccc ccagcacata gcc 1523
<210> 120
<211> 1922
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 120
ggatacgccg atttcaatgg caagctgatt taaggagaga ttgtgttcaa tcatgtactg 60
gagaacaaaa tctcttttga tatgaatctt ttttaccatg attactcccc tttctaatct 120
cttatgtttc tttttatcta cattgaacat atacgatttg ttaacttttg tcaatacttt 180
taccatccat atgtttccta taggcaatat tcgtactaaa atattttata ataagagatt 240
gcgaggtttt ggccatgacg aactttggac accatttacg acaattaagg gaacggaaaa 300
aactgaccgt caatcaactg gcgatgtatt ccggcgtcag ttcggcaggc atttcgcgaa 360
tcgaaaacgg aaagcgcggc gtgccgaagc cggcgacgat cagaaaactg gcggacgctt 420
tgaaagtccc gtatgaggaa ctgatggcat ctgcaggcta tatcagcgcg tctacagtcc 480
aggaagcaag aagcagctat gattccattt acgacatcgt gtcacagtac gatttagagg 540
acctttctct gtttgacagc gaaaagtgga aggtgctttc aaaaaaagac atcgaaaacc 600
tggacaaata tttcgacttt ctcgtgcagg aagcaagcag ccgaaacaaa aactgaatac 660
ttctccgcgg cacactctcc tctctatcat tttcgtctgt ttacgatcct gctgttattt 720
tatcccttat gttaactttt gtcaatattt ttcctgtcta agtatttcct atagtcaaca 780
tttgtattaa aatgttcata tcatgaattt gcggggggga tggcgatgac aaggttcggc 840
gagcggctca aagagctgag ggaacaaaga agcctgtcgg ttaatcagct tgccatgtat 900
gccggtgtga gcgccgcagc catttccaga gccgcagcca tttccagaat cgaaaacggc 960
caccgcggcg ttcccaagcc cgcgacgatc agaaaattgg ccgaggctct gaaaatgccg 1020
tacgagcagc tcatggatat tgccggttat atgagagctg acgagattcg cgaacagccg 1080
cgcggctatg tcacgatgca ggagatcgcg gccaagcacg gcgtcgaaga cctgtggctg 1140
tttaaacccg agaaatggga ctgtttgtcc cgcgaagacc tgctcaacct cgaacagtat 1200
tttcattttt tggttaatga agcgaagaag cgccaatcat aaaaagccga atttcccttt 1260
taggagaagt tcggcttttt tcggctgcct taagcggcat ccggattcgg cgtcttgcct 1320
ttatgatgct taacggggct cagcgcacgc tcgagccatc ccatgaacag atcggcgatg 1380
atcgccatca gcgccgtcgg gatcgcgcct gctagaatga tcgctgttcc gttggtcgcg 1440
tttgatcccc tgacaatgat atccccgagg ccgcctgcgc cgacaaacgt gccgatggcc 1500
gtaatgccga tcgcgatgac gagcgcggtt ctgagccccg ccataatgac cgacaaggcg 1560
aggggaagct ccaccatccg gagcacttga aatttcgtca tgcccatcgc cttccctgat 1620
tcaagatagg catgctcgat gctggcgatt cccgtatatg tgtttcgaat gatcggcaac 1680
agcgaataca aaaacaatga aagaatcacc gtgtttgcgc cgagccccat gacaagcatc 1740
aagacggcga gcatcgccag cgccggaacc gtttgaatga cattagtgat ggaaaagacc 1800
catttgctga ttttacggta tctggcgatg aaaatgccgg ccgggatgcc gacgacggcg 1860
gcgaacaata cgccgtatgc cgacattaaa aagtggcggt aaaattcccc cagcacatag 1920
cc 1922
<210> 121
<211> 1413
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 121
cgtcagttcg gcaggcattt cgcgaatcga aaacggaaag cgcggcgtgc cgaagccggc 60
gacgatcaga aaactggcgg acgctttgaa agtcccgtat gaggaactga tggcatctgc 120
aggctatatc agcgcgtcta cagtccagga agcaagaagc agctatgatt ccatttacga 180
catcgtgtca cagtacgatt tagaggacct ttctctgttt gacagcgaaa agtggaaggt 240
gctttcaaaa aaagacatcg aaaacctgga caaatatttc gactttctcg tgcaggaagc 300
aagcagccga aacaaaaact gaatacttct ccgcggcaca ctctcctctc tatcattttc 360
gtctgtttac gatcctgctg ttattttatc ccttatgtta acttttgtca atatttttcc 420
tgtctaagta tttcctatag tcaacatttg tattaaaatg ttcatatcat gaatttgcgg 480
gggggatggc gatgacaagg ttcggcgagc ggctcaaaga gctgagggaa caaagaagcc 540
tgtcggttaa tcagcttgcc atgtatgccg gtgtgagcgc cgcagccatt tccagagccg 600
cagccatttc cagaatcgaa aacggccacc gcggcgttcc caagcccgcg acgatcagaa 660
aattggccga ggctctgaaa atgccgtacg agcagctcat ggatattgcc ggttatatga 720
gagctgacga gattcgcgaa cagccgcgcg gctatgtcac gatgcaggag atcgcggcca 780
agcacggcgt cgaagacctg tggctgttta aacccgagaa atgggactgt ttgtcccgcg 840
aagacctgct caacctcgaa cagtattttc attttttggt taatgaagcg aagaagcgcc 900
aatcataaaa agccgaattt cccttttagg agaagttcgg cttttttcgg ctgccttaag 960
cggcatccgg attcggcgtc ttgcctttat gatgcttaac ggggctcagc gcacgctcga 1020
gccatcccat gaacagatcg gcgatgatcg ccatcagcgc cgtcgggatc gcgcctgcta 1080
gaatgatcgc tgttccgttg gtcgcgtttg atcccctgac aatgatatcc ccgaggccgc 1140
ctgcgccgac aaacgtgccg atggccgtaa tgccgatcgc gatgacgagc gcggttctga 1200
gccccgccat aatgaccgac aaggcgaggg gaagctccac catccggagc acttgaaatt 1260
tcgtcatgcc catcgccttc cctgattcaa gataggcatg ctcgatgctg gcgattcccg 1320
tatatgtgtt tcgaatgatc ggcaacagcg aatacaaaaa caatgaaaga atcaccgtgt 1380
ttgcgccgag ccccatgaca agcatcaaga cgg 1413
<210> 122
<211> 2009
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 122
ctctaatagc ttcgctttga ttgttgtatt atttattttg ctgatcattg tgggctccgc 60
ttttgtaaag tattaattca ttcgcgcacg aagccttctt ccgaaaaact gcaggttcat 120
atcgaaaggc gctccgcttt tcatatgagc ctgttttttt gtaatttaaa aagaatttat 180
ttccacaata acaaaaaatg tttataatag gataggctct gagctgacgg ctgacggcgc 240
tcggaaataa aaactctccc acaaatgaaa cttttataca acattttcgt cttatactgt 300
tgttaaggca attcaataac ggaaaggaaa agatcaatgt taaataagct aagaatgttg 360
tatgaaaaaa caacaatgaa gtggctgctc aacacttcct attactttct cattctggtg 420
ctgctgtttt taatttacgg attccataca gcaaatacag gttcatatat ttataatgac 480
ttttaatgga gaagaaaaca atgaaactga ttgaaacaat taaaaaatac gcgcaaacac 540
aacccgatac ccttgctttt gtgaatgaag aagaaaagct gacatacgga gagctttggt 600
cacaatctga gcgccttgcc gcaaggattc aaagcgaagc tttaacagat gcttctccga 660
tcattgtcta cggacatatg aaaccggtta tggcggtttc cttcctggct tgtgtaaaag 720
ccgggcatcc gtacattcca gttgacgtat cgattccggc agaccggatt ctgaaaatca 780
tcaacagttc aaaagccgag cttctgctca ataattcagg cacttcggtt gacacgggag 840
atgccctcat ttccgtagtg gaaccgggcg ttcttgaagg agacgggatg ccggagacag 900
atcccggacg ctgggtacat ggggaagata ctttttacat tatctacaca tccggaagca 960
cgggaaatcc aaaaggcgtg caaatctcag ccgataacct tcagagcttc acagattgga 1020
tcacgaatga ctttccggtg gaaagcggcc aagtgttctt aaatcaagcc ccgttttcct 1080
ttgatttatc ggtaatggac ctttatccgt gcctgcagtc cggaggcacc ttatggacgg 1140
ttacgaaaga catgattaat cggccgaaac tattgtttga agcactgaaa cagtccaacg 1200
tcaacgtctg gacatcgacg ccttcattcg cacaaatgtg cttgatggat ccgtcttatt 1260
cggaagagct gcttccggag ctgagcttgt ttatgttctg cggagaaacg cttcctgcgt 1320
cagtcgccag acagcttaaa gaaagatttc caaaggctcg tgtttttaac acatacggcc 1380
cgaccgaagc gaccgttgcc gtcacttcca tcgaagtaac agacgacgta ttaaacaagt 1440
attcttctct tcccgtcggt tctgaaaagc ccgaaaccga aatcgtaatc atcaatgaag 1500
acggaaaagc cgtccaagac ggggaaaaag gcgagatcat aattaccgga gcaagcgtca 1560
gcaaaggcta tttaggcgaa aaagcgctga cggaaaaagc gtttttctct tacaacggct 1620
cccctgctta ccgcaccggt gacgcaggtt acaaagaaaa cggccagctc ttcttcctcg 1680
gaagactcga tttccaaatc aaactgcacg gctaccggat tgagcttgaa gaaatcgaat 1740
atcaaatcaa tcagtcccga tatgtgcagt cggcagtcgt tattccattc taccgcgagg 1800
agaaaatcga gtatctgatc gccatgattg tgcctgccga acacgatttc gaaaaggaat 1860
accagttaac cagcgcgatt aagaaagatt tgggcagcaa gctgccggcc tatatgattc 1920
cgagaaaatt catgtatcaa aaagagattc cgatgacagc aaacggtaaa atcgaccgca 1980
agagattaaa agaagaggta accgtatga 2009
<210> 123
<211> 1557
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 123
ttaattgaat tgtgcggtta caccgccgga ctccatcatc atcagttctt ttttcatatc 60
caatccgccc cggtatcccg tgagctgccc gcttttaccg ataacccgat ggcaaggcac 120
caccattaac agcggatttg cgccgatcgc cgcgcctact gcccgcacag cggcctgctt 180
ttcaatatgc tcggcgatat cggaatagga gcaagtgctg ccgtaaggga tttcggagag 240
cgccttccac actgccagct gaaaaggcgt gccggcaagg tcgacaggaa agctgaaatg 300
agttcgcttg ccgttcaaat acgcctgcag ctgctcggcg tattctgcca atcctttgtc 360
atcccgaatg aaaactggct gtgtaaatct tttttcagcc caagcggcca aatcctcgaa 420
gccttgattc catccccctg taaaacagag cccgcgggca gtcgccccaa tgtgaatctg 480
ccaacctcgg caaataagcg tacgccagta tacgatttga tcgtccatat gtttacctcc 540
gtttcatttg ccggtacgac gtcggcgatt gcccagtctt ctttttaaac aaagaggcaa 600
aatattccgc attcgcaatg cctaccattg aagcgatttc tgcgatcgat cgttctgaat 660
gagcaagcaa atcgaccgct ttctcaatcc ttttctgcag gatgtattct gccggcgaga 720
cgcctttgat tcgtttaaat gtccgctgca ggtgaaaagg gctgatatgg cacctgtcag 780
ccaaagcttg cagagacagc ggatcgcgat aagattcctc gatgatttcc accacacgct 840
gtgccagctc ttcatccggc agcagcgccc cggccggatt gcagcgtttg caggggcggt 900
acccttctga taaagcatct tttgcattga aaaagatctg cacattgtcg atttgcggaa 960
ctctcgattt gcaggaaggg cggcaaaata tgccggtcgt tttgaccgcg taataaaaaa 1020
ctccgtcata ggcggaatcg ttttccgtaa tcgcccgcca catttcaggc gtcaatcgtg 1080
atttgctgtt catatcttca ccccgatcta tgtcagtata acctatatga cagccggagg 1140
tggagaggcg gagaacggca cagcaagaag acaaagaaga agagagactg ttgcctggac 1200
ctccgaaacg cgctacaatt catttacaac acaggatggg gtgagaatat tgccggaatc 1260
agtgaagcag gtaacatctc tcactgctgt gtgattttac tcacggcatt tggaacgccg 1320
gctctcaaca aactttctgt agtgaaaatc atgaaccaaa cggatcgtcg gcctgattaa 1380
cagctgaaag ctgccgatca caaacatcca tagtcccgcc ggcttcagtt cctcggagaa 1440
aaagcagaag ctcccgacaa ggaataaaag gccgatgaga aaatcgttta atgtatgtag 1500
aactttgtat ctttttttga aaaagagttc atatcgattg ttattgtttt gcggcat 1557
<210> 124
<211> 2814
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 124
ttaattgaat tgtgcggtta caccgccgga ctccatcatc atcagttctt ttttcatatc 60
caatccgccc cggtatcccg tgagctgccc gcttttaccg ataacccgat ggcaaggcac 120
caccattaac agcggatttg cgccgatcgc cgcgcctact gcccgcacag cggcctgctt 180
ttcaatatgc tcggcgatat cggaatagga gcaagtgctg ccgtaaggga tttcggagag 240
cgccttccac actgccagct gaaaaggcgt gccggcaagg tcgacaggaa agctgaaatg 300
agttcgcttg ccgttcaaat acgcctgcag ctgctcggcg tattctgcca atcctttgtc 360
atcccgaatg aaaactggct gtgtaaatct tttttcagcc caagcggcca aatcctcgaa 420
gccttgattc catccccctg taaaacagag cccgcgggca gtcgccccaa tgtgaatctg 480
ccaacctcgg caaataagcg tacgccagta tacgatttga tcgtccatat gtttacctcc 540
gtttcatttg ccggtacgac gtcggcgatt gcccagtctt ctttttaaac aaagaggcaa 600
aatattccgc attcgcaatg cctaccattg aagcgatttc tgcgatcgat cgttctgaat 660
gagcaagcaa atcgaccgct ttctcaatcc ttttctgcag gatgtattct gccggcgaga 720
cgcctttgat tcgtttaaat gtccgctgca ggtgaaaagg gctgatatgg cacctgtcag 780
ccaaagcttg cagagacagc ggatcgcgat aagattcctc gatgatttcc accacacgct 840
gtgccagctc ttcatccggc agcagcgccc cggccggatt gcagcgtttg caggggcggt 900
acccttctga taaagcatct tttgcattga aaaagatctg cacattgtcg atttgcggaa 960
ctctcgattt gcaggaaggg cggcaaaata tgccggtcgt tttgaccgcg taataaaaaa 1020
ctccgtcata ggcggaatcg ttttccgtaa tcgcccgcca catttcaggc gtcaatcgtg 1080
atttgctgtt catatcttca ccccgatcta tgtcagtata acctatatga cagccggagg 1140
tggagaggcg gagaacggca cagcaagaag acaaagaaga agagagactg ttgcctggac 1200
ctccgaaacg cgctacaatt catttacaac acaggatggg gtgagaatat tgccggaatc 1260
agtgaagcag gtgacagagg aaaggccgat atgattcggc cttttttata tgtacttctt 1320
agcgggtctc taaaataacc aaaaagcaag gactgctgaa agggctgaca taagcctttt 1380
gccggcggtc cttttttaat tctgattttt caaacttagc cccctcgaga aaaccgcttg 1440
agtatcctcc gctcatgtta ttgacctcct tttcttgatc aatatatttg tatgagcgga 1500
aagaaaattt gactagacgg atgcccctct gttgaaaatc tttttacagc atgtcagaat 1560
atgatatgat acaattcaaa ggaaagttta aactgttatg attaggagtg tttgcattta 1620
tgaagaagat tgcaattgcg gcgattacag cgacaagcgt gctggctctc agcgcatgca 1680
gcgggggaga ttctgaggtt gttgcggaaa caaaagctgg aaatattaca aaagaagacc 1740
tttatcaaac attaaaagac aatgccggag cggacgcact gaacatgctt gttcagcaaa 1800
aagtactcga tgataaatac gatgtctccg acaaagaaat cgacaaaaag ctgaacgagt 1860
acaaaaaatc aatgggtgac cagctcaacc agctcattga ccaaaaaggc gaagacttcg 1920
tcaaagaaca gatcaaatac gaacttctga tgcaaaaagc cgcaaaggat aacataaaag 1980
taaccgatga tgacgtaaaa gaatattatg acggcctgaa aggcaaaatc cacttaagcc 2040
acattcttgt gaaagaaaag aaaacggctg aagaagttga gaaaaagctg aaaaaaggcg 2100
aaaaattcga agaccttgca aaagagtatt caactgacgg tacagccgaa aaaggcggcg 2160
acctcggctg ggtcggcaaa gacgataaca tggacaagga tttcgtcaaa gcggcatttg 2220
ctttgaaaac cggcgaaatc agcggacctg tgaaatccca attcggctat cacatcatta 2280
aaaaagacga agaacgcggc aaatatgaag acatgaaaaa agagcttaaa aaagaagtcc 2340
aagaacaaaa gcaaaatgat caaactgaac tgcaatccgt cattgacaaa cttgtcaaag 2400
atgctgattt aaaagtaaaa gacaaagagt tgaaaaaaca agtcgaccag cgtcaagctc 2460
agacaagcag cagcagctga aagagcagag aggacggatt tcctgaagga aatccgtttt 2520
tttattttta acatctctca ctgctgtgtg attttactca cggcatttgg aacgccggct 2580
ctcaacaaac tttctgtagt gaaaatcatg aaccaaacgg atcgtcggcc tgattaacag 2640
ctgaaagctg ccgatcacaa acatccatag tcccgccggc ttcagttcct cggagaaaaa 2700
gcagaagctc ccgacaagga ataaaaggcc gatgagaaaa tcgtttaatg tatgtagaac 2760
tttgtatctt tttttgaaaa agagttcata tcgattgtta ttgttttgcg gcat 2814
<210> 125
<211> 1309
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 125
ctctaatagc ttcgctttga ttgttgtatt atttattttg ctgatcattg tgggctccgc 60
ttttgtaaag tattaattca ttcgcgcacg aagccttctt ccgaaaaact gcaggttcat 120
atcgaaaggc gctccgcttt tcatatgagc ctgttttttt gtaatttaaa aagaatttat 180
ttccacaata acaaaaaatg tttataatag gataggctct gagctgacgg ctgacggcgc 240
tcggaaataa aaactctccc acaaatgaaa cttttataca acattttcgt cttatactgt 300
tgttaaggca attcaataac ggaaaggaaa agatcaatgt taaataagct aagaatgttg 360
tatgaaaaaa caacaatgaa gtggctgctc aacacttcct attactttct cattctggtg 420
ctgctgtttt taatttacgg attccataca gcaaatacag gttcatatat ttataatgac 480
ttttaatgga gaagaaaaca atgaaactga ttgaaacaat taaaaaatac cacaaatgtg 540
cttgatggat ccgtcttatt cggaagagct gcttccggag ctgagcttgt ttatgttctg 600
cggagaaacg cttcctgcgt cagtcgccag acagcttaaa gaaagatttc caaaggctcg 660
tgtttttaac acatacggcc cgaccgaagc gaccgttgcc gtcacttcca tcgaagtaac 720
agacgacgta ttaaacaagt attcttctct tcccgtcggt tctgaaaagc ccgaaaccga 780
aatcgtaatc atcaatgaag acggaaaagc cgtccaagac ggggaaaaag gcgagatcat 840
aattaccgga gcaagcgtca gcaaaggcta tttaggcgaa aaagcgctga cggaaaaagc 900
gtttttctct tacaacggct cccctgctta ccgcaccggt gacgcaggtt acaaagaaaa 960
cggccagctc ttcttcctcg gaagactcga tttccaaatc aaactgcacg gctaccggat 1020
tgagcttgaa gaaatcgaat atcaaatcaa tcagtcccga tatgtgcagt cggcagtcgt 1080
tattccattc taccgcgagg agaaaatcga gtatctgatc gccatgattg tgcctgccga 1140
acacgatttc gaaaaggaat accagttaac cagcgcgatt aagaaagatt tgggcagcaa 1200
gctgccggcc tatatgattc cgagaaaatt catgtatcaa aaagagattc cgatgacagc 1260
aaacggtaaa atcgaccgca agagattaaa agaagaggta accgtatga 1309
<210> 126
<211> 483
<212> PRT
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 126
Ala Asn Leu Asn Gly Thr Leu Met Gln Tyr Phe Glu Trp Tyr Met Pro
1 5 10 15
Asn Asp Gly Gln His Trp Lys Arg Leu Gln Asn Asp Ser Ala Tyr Leu
20 25 30
Ala Glu His Gly Ile Thr Ala Val Trp Ile Pro Pro Ala Tyr Lys Gly
35 40 45
Thr Ser Gln Ala Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr Asp Leu
50 55 60
Gly Glu Phe His Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly Thr Lys
65 70 75 80
Gly Glu Leu Gln Ser Ala Ile Lys Ser Leu His Ser Arg Asp Ile Asn
85 90 95
Val Tyr Gly Asp Val Val Ile Asn His Lys Gly Gly Ala Asp Ala Thr
100 105 110
Glu Asp Val Thr Ala Val Glu Val Asp Pro Ala Asp Arg Asn Arg Val
115 120 125
Ile Ser Gly Glu His Leu Ile Lys Ala Trp Thr His Phe His Phe Pro
130 135 140
Gly Arg Gly Ser Thr Tyr Ser Asp Phe Lys Trp His Trp Tyr His Phe
145 150 155 160
Asp Gly Thr Asp Trp Asp Glu Ser Arg Lys Leu Asn Arg Ile Tyr Lys
165 170 175
Phe Gln Gly Lys Ala Trp Asp Trp Glu Val Ser Asn Glu Asn Gly Asn
180 185 190
Tyr Asp Tyr Leu Met Tyr Ala Asp Ile Asp Tyr Asp His Pro Asp Val
195 200 205
Ala Ala Glu Ile Lys Arg Trp Gly Thr Trp Tyr Ala Asn Glu Leu Gln
210 215 220
Leu Asp Gly Phe Arg Leu Asp Ala Val Lys His Ile Lys Phe Ser Phe
225 230 235 240
Leu Arg Asp Trp Val Asn His Val Arg Glu Lys Thr Gly Lys Glu Met
245 250 255
Phe Thr Val Ala Glu Tyr Trp Gln Asn Asp Leu Gly Ala Leu Glu Asn
260 265 270
Tyr Leu Asn Lys Thr Asn Phe Asn His Ser Val Phe Asp Val Pro Leu
275 280 285
His Tyr Gln Phe His Ala Ala Ser Thr Gln Gly Gly Gly Tyr Asp Met
290 295 300
Arg Lys Leu Leu Asn Gly Thr Val Val Ser Lys His Pro Leu Lys Ser
305 310 315 320
Val Thr Phe Val Asp Asn His Asp Thr Gln Pro Gly Gln Ser Leu Glu
325 330 335
Ser Thr Val Gln Thr Trp Phe Lys Pro Leu Ala Tyr Ala Phe Ile Leu
340 345 350
Thr Arg Glu Ser Gly Tyr Pro Gln Val Phe Tyr Gly Asp Met Tyr Gly
355 360 365
Thr Lys Gly Asp Ser Gln Arg Glu Ile Pro Ala Leu Lys His Lys Ile
370 375 380
Glu Pro Ile Leu Lys Ala Arg Lys Gln Tyr Ala Tyr Gly Ala Gln His
385 390 395 400
Asp Tyr Phe Asp His His Asp Ile Val Gly Trp Thr Arg Glu Gly Asp
405 410 415
Ser Ser Val Ala Asn Ser Gly Leu Ala Ala Leu Ile Thr Asp Gly Pro
420 425 430
Gly Gly Ala Lys Arg Met Tyr Val Gly Arg Gln Asn Ala Gly Glu Thr
435 440 445
Trp His Asp Ile Thr Gly Asn Arg Ser Glu Pro Val Val Ile Asn Ser
450 455 460
Glu Gly Trp Gly Glu Phe His Val Asn Gly Gly Ser Val Ser Ile Tyr
465 470 475 480
Val Gln Arg
<210> 127
<211> 9514
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 127
ctgggagctg atagcacagc gatttcccgc ctcctgtcgg aagcatcgcc accgtgtccc 60
gtccctccag cacgcttttg atgatcgttt cctggccttt tttaaaagca tgataaccaa 120
aatacctgcg gagcgcctga tgaagtttat ccattcctga tcaccgtttt cgacagggcc 180
aggcggattt taaaataact gtatcggccg ccgagtccct ctttaatttg tctgatttta 240
ttggtttggt tcgctatcgc ataatcggca ataattttct gttcctcttt cgacacgtac 300
tgatcgatgg aaaaagacgg atcatgaatg gcgatttcca caatgtgatc ctcgatcgtc 360
gccaatttca gctttctaat ttttgcgatc cggtctattg tatacccttc ttttataaga 420
aagagcgttt ttctcgtcga ttgtgtcaga ccatcattca aaggaatgtc atgaatgagc 480
gtttgaaaaa gcggactttc accattttgg gcggattgaa taaaatggtg cagaacatcc 540
caaaacagtg catatatgta ccactcatca aggttcattt tttcggaaag ctgtctgaat 600
gtatagcccg cttttgttct ggatgtcagg gagtgaacaa aaatagcggc ctgttcatca 660
tgatttaaaa cagacagctt ctctttcagc tcttggtgaa actgggccgc tgtttcagca 720
gcattccggt ttcttaaata ctgtttaacc cagttctgaa tctgataatc ttttacgatc 780
ggcaaataca cgcgttcgcg gtaaagcttg ttggagagca cttggatcaa aagcgacatc 840
cttgcccaca tcactttggc tgccgcctga taatagccgc cgtgaaaatg gcggggccac 900
ggataaaggg caaaaaagcc ggcaagttcc gcttctcctt tttcggtgac ggtgtaagcg 960
ccgctttccg ctttttctct gaccaaagac tcctgcttca atcgttgaac gctggctgcc 1020
acctgctctc tcgacagagc cgaacaaaag ccgaagtatt ttgaaacggc aaataaaccg 1080
gcgtcctgta tcgtctgtga cgaccttttt ccttttaata aatgatagac cgcgcttgga 1140
gaacgctcac ccttcatgga tgacagaatg tcaagcacaa tcgcgtcaaa aaaatgaacc 1200
ggcatatcat cacctgcaat cttccggcaa cattcgatca tttcttcctt ttattttaac 1260
agattttgcg gagaaatcga cgtttaaact catataaaag gggtatgtta gcagtagaac 1320
ccttgtgtga taagcattct caatattttt gagttgaaat gtaagattaa caccattaca 1380
ataaggaatg ggaataggtt tcatatcgga tagatagagg gttaaaccat ttgttccaac 1440
gaagaacaat ctgggaggtt ttttattcat gccaaaatat acaattgtag acaaagatac 1500
gtgcatcgca tgcggagctt gtggtgctgc ggctcctgat atttatgatt acgacgatga 1560
gggaatcgca tttgtcaccc ttgacgacaa tcagggtgtc gtcgaagtcc ctgacgtctt 1620
agaagaagac atgatggacg cgtttgaagg ctgtcctaca gattcgatca aagttgcgga 1680
tgagccgttc gaaggcgacc cgcttaaaca cgaataaagc caaaaaacat ccggtgcaca 1740
aagtgccgga tgttttttta tgagataagc acggctttac caacaagcaa aaagaagccg 1800
gctaaagaca tccggcttct tctgcagctg acaatatccg ggaacatgca cccgatattg 1860
tcatgtttat ttatttggcc atgcggacgt tttccttcag ccgcggtttc agcgaaagga 1920
aaatcggcgt ggacacgagg gccacagcga tgcctttaat gaaattaaaa ggcaggattc 1980
cggccagaac tgttgtcttg agcgcctctc cagtcagcgc tggagcattt aaaaaccaag 2040
tgtaggcagg cagaaacagc agataattta aaatgctcat cgaaacggcc atcacaagcg 2100
tccctgcgaa aagagctgtg acaaaccctt tggcagaact tgattttttc agcagtacag 2160
ctgccggcag gataaacaat gttccggcaa tgaagttagc cgcctgatca atcggaacgc 2220
ccgaggcgct tcctgcaata aagtaattca gcacgttttt gatcgcttca acggcaatcc 2280
cggctcccgg accgtacaaa ataacagcga gcaatgccgg gatatcactg aaatcgattt 2340
ttaaatacgg gaatgccccc aggatcggaa agctcagcat cattaaaata aatgcgatgc 2400
tgctcagcat gctgatagag acgagacgtc tcaccttgtt gtgtttcatt ttgtcactct 2460
ctccttttcg atcacatctc acgaaaagag gaatggttct ttcccctgtc ctaaacaaaa 2520
aacccgcttt attgaaaaag cggggctgtt ttacagacag gtcaaataaa cgtttgaaaa 2580
tgttcatttc aaaacgcgcg gaacctccat cttctcccat ccagactata ctgtcggctt 2640
cggaatcgca ccgaatcctg cccataaaaa ggctcgcggg cttagagcgc ttgctcatca 2700
ccgccggtag ggaatttcac cctgccccga agattgatct tatttatttt taatactgat 2760
attattataa attaattgtg aaaaaatgta caggtgcaaa gcttattgcg ctgttttggg 2820
acatcctgca cgatatttcg gtaaactcac tttttccgca tactaaaaac cgcacattca 2880
cagttatttc atttttaatt ttcgtctttc cgcgtgaaac tcattgacac tctttatgga 2940
atatggtaaa ttatcagata tttatgacgc ttatttagga ggaaatctta catgtttcga 3000
gtattggtct cagataaaat gtccagcgac ggcctcaaac cattaatgga agcagatttt 3060
attgaaattg tagaaaagaa tgttgcggaa gcggaagacg agcttcatac gtttgacgcg 3120
ctcttggtgc ggagcgccac gaaggtaacc gaagagctgt ttaaaaagat gacttcgctg 3180
aaaatcgtcg ccagagcagg tgtcggcgtc gacaatatcg atattgacga ggcgacaaaa 3240
cacggtgtta tcgtcgtaaa cgcgccaaac gggaatacaa tttcaaccgc tgaacatacc 3300
tttgcaatgt tttcagcgtt aatgagacat attccgcagg caaacatctc cgtgaaatca 3360
agggagtgga atcgttcggc ttacgtcggt tcagagcttt acggaaaaac gctcggcatc 3420
atcggaatgg gccgcatcgg aagcgaaatc gcgagccgcg caaaagcatt cggtatgacc 3480
gttcatgtat ttgacccgtt cctgacccaa gaaagggcaa gcaagctcgg cgttaacgcg 3540
aacagctttg aagaagttct ggcatgcgcc gacatcatta cggttcatac cccgctcacg 3600
aaagaaacga agggactttt gaacaaagaa accatcgcaa aaacgaaaaa aggcgttcgt 3660
ctcgttaact gtgcaagagg cggcatcatc gatgaagcag cgcttttgga agctctggaa 3720
agcggacatg tcgctggcgc tgccttggat gtattcgaag tcgagcctcc ggtcgattca 3780
aaactgatcg atcatccgct tgtagtcgcg actcctcact tgggcgcctc aacaaaagaa 3840
gcccagctga atgtcgctgc acaagtgtcc gaagaagtcc ttcagtatgc gcaaggaaac 3900
cctgtgatgt ccgcgatcaa ccttccggcc atgacaaagg attcattcga aaaaatccag 3960
ccttatcatc agtttgccaa tacgatcgga aaccttgtgt ctcagtgcat gaatgagcct 4020
gttcaagatg tagccatcca atatgaaggc tccatcgcca aacttgaaac gtcatttatt 4080
acgaaaagcc ttttggccgg atttctgaag ccgagggtcg cggctaccgt taacgaagtg 4140
aatgccggca ccgttgcgaa agagcgcggc atcagcttca gcgaaaaaat ttcttccaat 4200
gagtcaggct atgaaaactg catctctgtg actgtcacgg gagatgtaac aacattctct 4260
ttaagagcga cgtacattcc gcacttcggc ggacgcatcg ttgccttaaa cggctttgat 4320
attgattttt atccggctgg acaccttgtc tacattcacc accaggataa accaggggct 4380
atcggccatg tcggacgaat tttaggagac catgacatca atatcgccac tatgcaggta 4440
ggccgaaaag aaaaaggcgg agaagcgatc atgatgcttt cctttgaccg ccaccttgag 4500
gacgatattt tagctgagct gaaaaacatc ccggatatcg tgtctgttaa agccatcgac 4560
cttccttaag tcgctgataa acagctgaca tcaatatcct attttttcaa aaaatatttt 4620
aaaaagttgt tgacttaaaa gaagctaaat gttatagtaa taaaacagaa tagtctttta 4680
agtaagtcta ctctgaattt ttttaaaagg agagggtaaa gatgaaacaa caaaaacggc 4740
tttacgcccg attgctgacg ctgttatttg cgctcatctt cttgctgcct cattctgcag 4800
ctagcgcagc aaatcttaat gggacgctga tgcagtattt tgaatggtac atgcccaatg 4860
acggccaaca ttggaagcgt ttgcaaaacg actcggcata tttggctgaa cacggtatta 4920
ctgccgtctg gattcccccg gcatataagg gaacgagcca agcggatgtg ggctacggtg 4980
cttacgacct ttatgattta ggggagtttc atcaaaaagg gacggttcgg acaaagtacg 5040
gcacaaaagg agagctgcaa tctgcgatca aaagtcttca ttcccgcgac attaacgttt 5100
acggggatgt ggtcatcaac cacaaaggcg gcgctgatgc gaccgaagat gtaaccgcgg 5160
ttgaagtcga tcccgctgac cgcaaccgcg taatttcagg agaacaccta attaaagcct 5220
ggacacattt tcattttccg gggcgcggca gcacatacag cgattttaaa tggcattggt 5280
accattttga cggaaccgat tgggacgagt cccgaaagct gaaccgcatc tataagtttc 5340
aaggaaaggc ttgggattgg gaagtttcca atgaaaacgg caactatgat tatttgatgt 5400
atgccgacat cgattatgac catcctgatg tcgcagcaga aattaagaga tggggcactt 5460
ggtatgccaa tgaactgcaa ttggacggtt tccgtcttga tgctgtcaaa cacattaaat 5520
tttctttttt gcgggattgg gttaatcatg tcagggaaaa aacggggaag gaaatgttta 5580
cggtagctga atattggcag aatgacttgg gcgcgctgga aaactatttg aacaaaacaa 5640
attttaatca ttcagtgttt gacgtgccgc ttcattatca gttccatgct gcatcgacac 5700
agggaggcgg ctatgatatg aggaaattgc tgaacggtac ggtcgtttcc aagcatccgt 5760
tgaaatcggt tacatttgtc gataaccatg atacacagcc ggggcaatcg cttgagtcga 5820
ctgtccaaac atggtttaag ccgcttgctt acgcttttat tctcacaagg gaatctggat 5880
accctcaggt tttctacggg gatatgtacg ggacgaaagg agactcccag cgcgaaattc 5940
ctgccttgaa acacaaaatt gaaccgatct taaaagcgag aaaacagtat gcgtacggag 6000
cacagcatga ttatttcgac caccatgaca ttgtcggctg gacaagggaa ggcgacagct 6060
cggttgcaaa ttcaggtttg gcggcattaa taacagacgg acccggtggg gcaaagcgaa 6120
tgtatgtcgg ccggcaaaac gccggtgaga catggcatga cattaccgga aaccgttcgg 6180
agccggttgt catcaattcg gaaggctggg gagagtttca cgtaaacggc gggtcggttt 6240
caatttatgt tcaaagatag aagcttctcg aggttaacag aggacggatt tcctgaagga 6300
aatccgtttt tttattttac agaagctgcg gaacctgaaa agaattcctt tcaggttccg 6360
ttttttttag gaattctccc tgatctcaag catctggcgg ggataaatcc gctctccttt 6420
caaatcgttc cattctttga ggcgctgtac agttacgccc attttttcgg cgatatgatg 6480
aagcgtatcc cctttccgca ctacatatgt accggtcttc gattcatcgt catgaaggcg 6540
gagtgtttgg ccggccttga gatttgaatg tttcaacccg tttattctca tgatctcctc 6600
gatggatata ccgctatcct tgctgattct ccagagcgtg tccccttttt gaacggtcac 6660
cgcaccgctc attgtcccgg cgttttgata aacgtggata gaattttgcc ggaacgcctc 6720
ctcacgaagc accgtcagcg gattgattgc atatctttta tcttcagtcc atgaaccgtg 6780
atgcatttca aaatgcaggt gggttccggt cgatattccc gtattgccga tgattccgat 6840
ttgctcgcct tttttcaccc gctccttttc ctttttcagg cgtttgctta agtgggcata 6900
aacggtttca tatccgttgt catgtttaat aaatatcact tggccgtagg agtcggattg 6960
atacgatttg cttatcgttc cgtctgcggc tgccgctact gcttcccctt cgggagcagc 7020
gatgtcaagc cccttatgct ttccgcctct cgtaccgaat tgatctgtga tctctccttt 7080
aatcggttca atccactctg aggcttccgc ccccggggca ttgacgaaaa gcgccaatcc 7140
cgaaagccat gcgatcgcga acaggaagtt ttgatgtctg agtttcttca aggttttcca 7200
tatcctccta ttacatgcat cttcggtaaa attgccccct attcggagac agcttagtat 7260
acttccaaat caatacaatt tatacattaa aaaaagactc cgcacaggga gtcttttagt 7320
tttctatcgt catcggattc ggtgcgtacg gaacctgtac agatttcgac aggtcatagg 7380
cgccgacctt ggttatggat gcgtttttaa atttcacttt tgtgaagccg aaatctttcg 7440
cggtcaatag aaggccttcc accatcaaga catcttcggg tttattttca atattcgcgg 7500
aggaagaaaa ttgaatgatc agttcttttc cattcttttg aatatcttca atcggcgtat 7560
catcggataa aatgggtttt aaatgagtgc cgctttcttc gtttttcatc atcttaatcg 7620
cttcctgcac cgattcgtaa gattcgcttg aaggtgcaag gaaccggcgc ccgtctgagc 7680
tttcatataa atagtagcat ttttgcgtct ggtgcataat cgccatatcg gcgagcattc 7740
cgaatgtttc aaattcaaca cccgatttat cattggaaat aaacagaaca gaatcatacg 7800
atccccattt aaaggtttcg ttgatcacat ttttcagccg ttcgaaatct tcgactgata 7860
gctccggtat tttctcatca acttgaatct tcagtttttt attgtttttc tgctctttga 7920
acttcacctt atcaaggtaa gctgtgtcaa atgatgtaaa ctggtccact ccaagccggc 7980
tgtaagcgtg aagcgcatct tcaagatttg tcatgccagt gcttttctcg aggcttaccg 8040
ggacaacgac agacttggac tcgtcaagga aagcgaaggt gatatagtcg tctttttgat 8100
tctgtgagac gacaaacgta tttgcaggtt cagacttggc agcatcagcc tccgtctgca 8160
ccaattttcc gtcagaagaa atgttggcgt cggcgctgtt ttgagatctg atctgttcga 8220
ttaactgggg agtgatcagc atcagaagaa agagaaccaa aactgtagca gcaaatgcgc 8280
cgacccgttt tttcggtgat ttacgctttg gtgcgagaat cagcttttga tagatctgat 8340
ttgccgaacg attatcctta accgttggaa gttggcttag taacgccttc agccgttctt 8400
cgttccattc tgacttcttc attctttgga tcctccttca aaagctccat ctgtttacga 8460
agcactttca gaccgcggtg ctgagtggtt tttaccttgc tttcggaaaa attcaaggct 8520
tttgctgttt cactgatcga atatccttga ataaaacgca agacgataac tgatctttgg 8580
tcaagcgtac acttgtctag ggcctcgaaa atttccttta ggttttcatt ttgcatcacg 8640
atgtcctcag gcagaggctt gcggtctttt acatcttgtt tctcccagtc aaacgtcccc 8700
aaaatccgct ggcggatcgt ctgctgcttt ctgaaccagt cgatcgcaac gtgccgcgca 8760
atcgaaagaa gccaggtttt ttcgctgctc ctgccttcaa atgtttcgta agaatgcagg 8820
acgcggatgt atacttcctg aactaagtct tccgcctgat ttttgtcttt taccatataa 8880
aataaaaact gaaataaatc ctgatgatac tgatcatata ttttctgaaa ggtttcttcc 8940
acctgaaacc cctccgttca atttattgtc gtttgtcaat cttaaaaggt tacattacaa 9000
ctattacaac tatattacga acatatgaaa atggaaaggg ggttttgcga aagttaagct 9060
taattttaac ttaacaagca caaaagcacc cgttctaaat gaacaggtgc caaggttata 9120
ggagcccaca ttttcactaa gctgtgccct tacaaggctt tcgttctcct gaccggagcg 9180
ttgcggatcc gctgaaatga actaatttca atccgtttat gactttaagt ccaattgttg 9240
gcgaagcttt ttggaaatct ccattctctt ttcgtcagtc actaggtgat accataagcc 9300
gtcaactttt ttgtcttcac cttcaatttc tacctgctta atatctttag cagcttcttt 9360
ataagaactt tgaatatcga tcatcttgtc cagtgttaag tttgttttga tgtttttttc 9420
taatgctgcc aaaatctctt gatagtttgt taatgatttt aattgtgcac cttcatgaat 9480
aatttcgttg ataatctcac gctgacgctc ttga 9514
<210> 128
<211> 95
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 128
gtcgctgata aacagctgac atcaatatcc tattttttca aaaaatattt taaaaagttg 60
ttgacttaaa agaagctaaa tgttatagta ataaa 95
<210> 129
<211> 57
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 129
acagaatagt cttttaagta agtctactct gaattttttt aaaaggagag ggtaaag 57
<210> 130
<211> 87
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 130
atgaaacaac aaaaacggct ttacgcccga ttgctgacgc tgttatttgc gctcatcttc 60
ttgctgcctc attctgcagc tagcgca 87
<210> 131
<211> 1452
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 131
gcaaatctta atgggacgct gatgcagtat tttgaatggt acatgcccaa tgacggccaa 60
cattggaagc gtttgcaaaa cgactcggca tatttggctg aacacggtat tactgccgtc 120
tggattcccc cggcatataa gggaacgagc caagcggatg tgggctacgg tgcttacgac 180
ctttatgatt taggggagtt tcatcaaaaa gggacggttc ggacaaagta cggcacaaaa 240
ggagagctgc aatctgcgat caaaagtctt cattcccgcg acattaacgt ttacggggat 300
gtggtcatca accacaaagg cggcgctgat gcgaccgaag atgtaaccgc ggttgaagtc 360
gatcccgctg accgcaaccg cgtaatttca ggagaacacc taattaaagc ctggacacat 420
tttcattttc cggggcgcgg cagcacatac agcgatttta aatggcattg gtaccatttt 480
gacggaaccg attgggacga gtcccgaaag ctgaaccgca tctataagtt tcaaggaaag 540
gcttgggatt gggaagtttc caatgaaaac ggcaactatg attatttgat gtatgccgac 600
atcgattatg accatcctga tgtcgcagca gaaattaaga gatggggcac ttggtatgcc 660
aatgaactgc aattggacgg tttccgtctt gatgctgtca aacacattaa attttctttt 720
ttgcgggatt gggttaatca tgtcagggaa aaaacgggga aggaaatgtt tacggtagct 780
gaatattggc agaatgactt gggcgcgctg gaaaactatt tgaacaaaac aaattttaat 840
cattcagtgt ttgacgtgcc gcttcattat cagttccatg ctgcatcgac acagggaggc 900
ggctatgata tgaggaaatt gctgaacggt acggtcgttt ccaagcatcc gttgaaatcg 960
gttacatttg tcgataacca tgatacacag ccggggcaat cgcttgagtc gactgtccaa 1020
acatggttta agccgcttgc ttacgctttt attctcacaa gggaatctgg ataccctcag 1080
gttttctacg gggatatgta cgggacgaaa ggagactccc agcgcgaaat tcctgccttg 1140
aaacacaaaa ttgaaccgat cttaaaagcg agaaaacagt atgcgtacgg agcacagcat 1200
gattatttcg accaccatga cattgtcggc tggacaaggg aaggcgacag ctcggttgca 1260
aattcaggtt tggcggcatt aataacagac ggacccggtg gggcaaagcg aatgtatgtc 1320
ggccggcaaa acgccggtga gacatggcat gacattaccg gaaaccgttc ggagccggtt 1380
gtcatcaatt cggaaggctg gggagagttt cacgtaaacg gcgggtcggt ttcaatttat 1440
gttcaaagat ag 1452
<210> 132
<211> 6218
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 132
ggtagaggaa ttcctggtcg gccaaggcta caatccgttt ccgcttgtca ggtttacgga 60
aagggcagac gtagccgcaa gccatatttt agaggggcat gtcatcgtga tcgtcgatac 120
gtcgccaagc gtcatcatca caccgaccac tttgtttcac catgttcagc atgctgagga 180
atacagacag acgccggctg ttgggacgtt tttaaggtgg gtgcggtttt tcggtatttt 240
ggcctccacc tttttgctgc cgctttggct gctgtttgtc attcatccgt cgctcttgcc 300
tgataattta tcgtttatcg ggttgaataa agacacccat attccgatta tcatgcagat 360
tttcctggcg gatctcggcg tcgaattttt aagaatggcc gccattcata cgccgacggc 420
gctttcgact gcaatgggcc tgatcgccgc tgtattgatc ggcgatatcg cgatcaatgt 480
cggcttgttt tctcccgaag tcattttata cgtttccctc tcggcaatcg gagcctacac 540
gacaccaagc tacgagctga gcctggcgaa taaaatggtg aagctgttta tgctgatatt 600
ggtggcgctt tttaaagtgg agggatttgt catcggatta acgatcttaa ctatagtgat 660
gacttcgatc aggtcattgc gaacgcctta cttatggcct ctcctcccgt tcaatggaaa 720
agcgttttgg catgttctcg tgcgcacgtc cgttccaggg ggaaaagtca ggccgagcat 780
cgttcatccg agaaaccgct ccagacagcc gtgaagccgg cattcgaaga ggcttttccc 840
cggggaaaag cctctttttc aataatcgaa ttccggtctt tgagtaccga tgcctctgta 900
ttcattggca gagatcgcga ctgcccggag gctgcagatg ttgttctgtc ttctgatcgg 960
atagacgaca tacagcattt cgcggccgta cgggtcaatc gttgacgaat gaaggaaaac 1020
ctcagttcct ctccgccaaa atctcgtatt cgccggagct gtaataatct gcccttcata 1080
aggctcataa attctctgtt cataatgcgc agccggctga taaggggcgt atacatcttc 1140
aggtgcatag ccgggagcgg gggtgtaggg atagcgattt ggatacatat gataacctct 1200
ttcccacttc gttttttggt tttcatcttt aagattatat tcaggtaaat gcctatttgt 1260
atgggcgaaa atctcagctt ttcggctctt tttttattga atggacgttg tgtatgccta 1320
tttctatcaa gcgctgtttt ctgttattct ataatcaata gaatggatta gttgtttagg 1380
gaatcatttc ctttataaat caagaaaatt tggacaaatg gtggtttagt ttttaaaacg 1440
aaatgttata atacaacata agaatcgcac tatcatgaag ccggaagatg catcgggcag 1500
caaccggagc gccccttgca cctttgtcga tagagaaaga gggaatgaca attgttttta 1560
cacggtacta gcagacaaaa tgaaagaggg cacctcgaaa tcggcggtgt cgatgttcta 1620
tcattggcag aaagatacgg aacacctctt tatgtatacg atgtcgcgct gattagagag 1680
cgcgcccgaa aattccagaa ggcattcaag gaagccggtt taaaagcgca ggtagcgtat 1740
gcaagcaagg cgttttcatc ggttgccatg attcagcttg ccgaacaaga ggggctgtct 1800
ctggatgtgg tatcgggagg agagcttttc actgcgatca aagcagggtt cccagctgag 1860
cggattcatt ttcacggaaa caataagagc cctgaagaac tagccatggc gctggagcat 1920
caaatcggct gcatcgtgct cgataacttt cacgagatcg ccattacaga agatctttgc 1980
aagcgatcag gacaaactgt agacgttttg ctcagaatca ctccgggagt tgaagcgcac 2040
acgcacgatt atattacgac ggggcaggaa gattccaaat tcggttttga tctgcataat 2100
ggacaggtcg aacaagccat cgaacaagtc ctccgctcgt ctgcgtttaa gctcctcggc 2160
gtgcactgcc acatcggttc gcaaattttt gatacggcag gatttgtcct tgcagcagac 2220
aagattttcg agaagcttgc ggaatggcgg gagacttact ctttcattcc ggaagtgctc 2280
aatcttggcg ggggcttcgg catccgctat acaaaagacg acgagccgct tgcagctgat 2340
gtttatgttg aaaaaatcat cgaggcggtc aaagcaaatg ccgagcattt cggctttgac 2400
atccctgaga tttggatcga accaggccgg tctctcgtcg gtgatgcggg gactacgctg 2460
tacacgatcg gttctcaaaa agaggtgccg ggcattcgca aatatgtagc catcgacggc 2520
ggcatgagcg ataatatcag gccggcgctt tatgaggcaa aatatgaagc agccgtcgcc 2580
aacaggatga acgatgcttg tcatgatacc gcatcaatcg caggaaaatg ctgcgaaagc 2640
ggagatatgc tgatttggga tttggaaatc cccgaagttc gcgacggaga tgtgctcgcc 2700
gttttctgca ccggtgcgta cggctacagc atggccaaca actacaaccg cattccgcgc 2760
ccggccgtcg tctttgtcga ggacggggaa gcgcagctcg tcattcagag agagacgtat 2820
gaggatatcg tcaagctgga tctgccgctg aaatcgaaag tcaaacaata aaaaaatgga 2880
gattccctaa gaggggggtc tccattttta attcaagctg ataaacagct gacatcaact 2940
aaaagtttca ttaaatactt tgaaaaaagt tgttgactta aaagaagcta aatgttatag 3000
taattgtaca gaatagtctt ttaagtaagt ctactctgaa tttttttaaa aggagagggt 3060
aaagaatgaa acaacaaaaa cggctttacg cccgattgct gacgctgtta tttgcgctca 3120
tcttcttgct gcctcattct gcagctagcg cagcaaatct taatgggacg ctgatgcagt 3180
attttgaatg gtacatgccc aatgacggcc aacattggaa gcgtttgcaa aacgactcgg 3240
catatttggc tgaacacggt attactgccg tctggattcc cccggcatat aagggaacga 3300
gccaagcgga tgtgggctac ggtgcttacg acctttatga tttaggggag tttcatcaaa 3360
aagggacggt tcggacaaag tacggcacaa aaggagagct gcaatctgcg atcaaaagtc 3420
ttcattcccg cgacattaac gtttacgggg atgtggtcat caaccacaaa ggcggcgctg 3480
atgcgaccga agatgtaacc gcggttgaag tcgatcccgc tgaccgcaac cgcgtaattt 3540
caggagaaca cctaattaaa gcctggacac attttcattt tccggggcgc ggcagcacat 3600
acagcgattt taaatggcat tggtaccatt ttgacggaac cgattgggac gagtcccgaa 3660
agctgaaccg catctataag tttcaaggaa aggcttggga ttgggaagtt tccaatgaaa 3720
acggcaacta tgattatttg atgtatgccg acatcgatta tgaccatcct gatgtcgcag 3780
cagaaattaa gagatggggc acttggtatg ccaatgaact gcaattggac ggtttccgtc 3840
ttgatgctgt caaacacatt aaattttctt ttttgcggga ttgggttaat catgtcaggg 3900
aaaaaacggg gaaggaaatg tttacggtag ctgaatattg gcagaatgac ttgggcgcgc 3960
tggaaaacta tttgaacaaa acaaatttta atcattcagt gtttgacgtg ccgcttcatt 4020
atcagttcca tgctgcatcg acacagggag gcggctatga tatgaggaaa ttgctgaacg 4080
gtacggtcgt ttccaagcat ccgttgaaat cggttacatt tgtcgataac catgatacac 4140
agccggggca atcgcttgag tcgactgtcc aaacatggtt taagccgctt gcttacgctt 4200
ttattctcac aagggaatct ggataccctc aggttttcta cggggatatg tacgggacga 4260
aaggagactc ccagcgcgaa attcctgcct tgaaacacaa aattgaaccg atcttaaaag 4320
cgagaaaaca gtatgcgtac ggagcacagc atgattattt cgaccaccat gacattgtcg 4380
gctggacaag ggaaggcgac agctcggttg caaattcagg tttggcggca ttaataacag 4440
acggacccgg tggggcaaag cgaatgtatg tcggccggca aaacgccggt gagacatggc 4500
atgacattac cggaaaccgt tcggagccgg ttgtcatcaa ttcggaaggc tggggagagt 4560
ttcacgtaaa cggcgggtcg gtttcaattt atgttcaaag atagaagctt ctcgaggtta 4620
acagaggacg gatttcctga aggaaatccg tttttttatt ttcaagcacg aaaaacactt 4680
cccggtgatc gggaggtgtt ttttgttaaa aagatcatga catgcataga acagcgaccg 4740
ggctaattgt atataatatt gtgaatttaa caaaaaattt acaaaggaga tgataaaggc 4800
aatgaccagg gtgaaaagga tgagatttgc tgatttgttg gatttagagg cggagtagat 4860
gaaaccggcc aaagtatccc tactccaccg attgctccag tgcctgaagc aatgtgttga 4920
ttgtaacaca gtaaatcgtt ttacagcaat aaacattttt gtgaatattt tattgatttc 4980
ggctgtgatc tcattcccat attctgctgc ggcccatggc gcaacacagt ccggcgatca 5040
atattcaagc tttgaagaat tggagcggaa tgaagatcca gcttcttacc gaattacgga 5100
gaagaacgca agagtgccga tgctcatcat ggccatccat ggaggcggca tcgaacccgg 5160
aacgagcgaa atcgccaatg aagtgtccaa aaactattcc ctgtacttgt ttgaagggct 5220
gaaatcatca ggcaatacgg accttcacat tacaagcacg cgttttgacg agccagcggc 5280
gctcgcaatt actgcaagcc accagtatgt catgtcgctc cacggctatt acagtgaaga 5340
ccgcgatatt aaagtaggcg gcacagaccg cgctaaaatc agaatattgg ttgatgagct 5400
gaaccgctcg gggtttgccg ctgaaatgct ggggacagat gacaagtatg ccggaaccca 5460
tccgaataac atcgccaaca agtcgctttc cgggctgagc attcagcttg aaatgagcac 5520
gggtttccgc aaatctttat tcgaccggtt tacactaaaa gacagggcgg cgacgcaaaa 5580
cgaaacgttt taccgattta caaagctgct gacagatttt attcatgaaa actatgaaga 5640
agacggaggg gatttcccct ctgcaaaaat aaaacacccc cttcaagtga aaaaggaggt 5700
gtttcggcgg ttgtgttaac cgttggactc tgaggtgccg ccgccggtga atacggaaac 5760
gatggcgttc cacagagaca caaagaagtc gatcagtttt tgaagaaagt tttgtccttc 5820
ttcagaatcc aagaatttcg tgattttatc ctttgctttg tcaagctggt ctccaacctg 5880
gttccagtcg atattaatat ttttcatgtt attaaataaa gatataagag agtttttctg 5940
atcttctgtg agtgtcacgc caagttcgga agcagccgaa tcaatcgttt tctccaattc 6000
ctcttttgac tcgggaactc cgtttttcga gatttcttcc ttgactttgg ccatcagcgc 6060
tgacgcgttt tcactgccga ttttctcgcc aagctctgaa gtggtgacaa gctcttcatt 6120
cgcgaccttt ttcacatctt cggaaatttt ttcgcccgaa gtcgtttcat acgctttcat 6180
caatccggtt aaagcggctg tgcctgacac ttcaaacg 6218
<210> 133
<211> 1000
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 133
tcgttccggg ttcgatgccg cctccatgga tggccatgat gagcatcggc actcttgcgt 60
tcttctccgt aattcggtaa gaagctggat cttcattccg ctccaattct tcaaagcttg 120
aatattgatc gccggactgt gttgcgccat gggccgcagc agaatatggg aatgagatca 180
cagccgaaat caataaaata ttcacaaaaa tgtttattgc tgtaaaacga tttactgtgt 240
tacaatcaac acattgcttc aggcactgga gcaatcggtg gagtagggat actttggccg 300
gtttcatcta ctccgcctct aaatccaaca aatcagcaaa tctcatcctt ttcaccctgg 360
tcattgcctt tatcatctcc tttgtaaatt ttttgttaaa ttcacaatat tatatacaat 420
tagcccggtc gctgttctat gcatgtcatg atctttttaa caaaaaacac ctcccgatca 480
ccgggaagtg tttttcgtgc ttgtcattcc ctctttctct atcgacaaag gtgcaagggg 540
cgctccggtt gctgcccgat gcatcttccg gcttcatgat agtgcgattc ttatgttgta 600
ttataacatt tcgttttaaa aactaaacca ccatttgtcc aaattttctt gatttataaa 660
ggaaatgatt ccctaaacaa ctaatccatt ctattgatta tagaataaca gaaaacagcg 720
cttgatagaa ataggcatac acaacgtcca ttcaataaaa aaagagccga aaagctgaga 780
ttttcgccca tacaaatagg catttacctg aatataatct taaagatgaa aaccaaaaaa 840
cgaagtggga aagaggttat catatgtatc caaatcgcta tccctacacc cccgctcccg 900
gctatgcacc tgaagatgta tacgcccctt atcagccggc tgcgcattat gaacagagaa 960
tttatgagcc ttatgaaggg cagattatta cagctccggc 1000
<210> 134
<211> 1320
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 134
ttgtttttac acggtactag cagacaaaat gaaagagggc acctcgaaat cggcggtgtc 60
gatgttctat cattggcaga aagatacgga acacctcttt atgtatacga tgtcgcgctg 120
attagagagc gcgcccgaaa attccagaag gcattcaagg aagccggttt aaaagcgcag 180
gtagcgtatg caagcaaggc gttttcatcg gttgccatga ttcagcttgc cgaacaagag 240
gggctgtctc tggatgtggt atcgggagga gagcttttca ctgcgatcaa agcagggttc 300
ccagctgagc ggattcattt tcacggaaac aataagagcc ctgaagaact agccatggcg 360
ctggagcatc aaatcggctg catcgtgctc gataactttc acgagatcgc cattacagaa 420
gatctttgca agcgatcagg acaaactgta gacgttttgc tcagaatcac tccgggagtt 480
gaagcgcaca cgcacgatta tattacgacg gggcaggaag attccaaatt cggttttgat 540
ctgcataatg gacaggtcga acaagccatc gaacaagtcc tccgctcgtc tgcgtttaag 600
ctcctcggcg tgcactgcca catcggttcg caaatttttg atacggcagg atttgtcctt 660
gcagcagaca agattttcga gaagcttgcg gaatggcggg agacttactc tttcattccg 720
gaagtgctca atcttggcgg gggcttcggc atccgctata caaaagacga cgagccgctt 780
gcagctgatg tttatgttga aaaaatcatc gaggcggtca aagcaaatgc cgagcatttc 840
ggctttgaca tccctgagat ttggatcgaa ccaggccggt ctctcgtcgg tgatgcgggg 900
actacgctgt acacgatcgg ttctcaaaaa gaggtgccgg gcattcgcaa atatgtagcc 960
atcgacggcg gcatgagcga taatatcagg ccggcgcttt atgaggcaaa atatgaagca 1020
gccgtcgcca acaggatgaa cgatgcttgt catgataccg catcaatcgc aggaaaatgc 1080
tgcgaaagcg gagatatgct gatttgggat ttggaaatcc ccgaagttcg cgacggagat 1140
gtgctcgccg ttttctgcac cggtgcgtac ggctacagca tggccaacaa ctacaaccgc 1200
attccgcgcc cggccgtcgt ctttgtcgag gacggggaag cgcagctcgt cattcagaga 1260
gagacgtatg aggatatcgt caagctggat ctgccgctga aatcgaaagt caaacaataa 1320
<210> 135
<211> 91
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 135
gctgataaac agctgacatc aactaaaagt ttcattaaat actttgaaaa aagttgttga 60
cttaaaagaa gctaaatgtt atagtaattg t 91
<210> 136
<211> 484
<212> PRT
<213> 人工序列
<220>
<223> 合成的
<400> 136
His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp His
1 5 10 15
Leu Pro Asn Asp Gly Gln His Trp Asn Arg Leu Arg Asn Asp Ala Ala
20 25 30
Asn Leu Lys Asn Leu Gly Ile Thr Ala Val Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Thr Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Ile Arg Thr Lys Tyr Gly
65 70 75 80
Thr Arg Ser Gln Leu Gln Ser Ala Ile Ala Ser Leu Gln Asn Asn Gly
85 90 95
Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Gly Thr Glu Trp Val Gln Ala Val Glu Val Asn Pro Ser Asn Arg Asn
115 120 125
Gln Glu Val Thr Gly Glu Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp
130 135 140
Phe Pro Gly Arg Gly Asn Thr His Ser Ser Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Thr Asp Trp Asp Gln Ser Arg Arg Leu Asn Asn Arg
165 170 175
Ile Tyr Lys Phe Thr Gly Lys Gly Trp Asp Trp Glu Val Asp Thr Glu
180 185 190
Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Val Asp Met Asp His
195 200 205
Pro Glu Val Ile Asn Glu Leu Arg Arg Trp Gly Val Trp Tyr Thr Asn
210 215 220
Thr Leu Asn Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His Ile Lys
225 230 235 240
Tyr Ser Phe Thr Arg Asp Trp Leu Asn His Val Arg Ser Thr Thr Gly
245 250 255
Lys Asn Asn Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu Gly
260 265 270
Ala Ile Glu Asn Tyr Leu His Lys Thr Asn Trp Asn His Ser Val Phe
275 280 285
Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Lys Ser Gly Gly
290 295 300
Asn Tyr Asp Met Arg Gln Ile Leu Asn Gly Thr Val Val Ser Lys His
305 310 315 320
Pro Met His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro Ala
325 330 335
Glu Ala Leu Glu Ser Phe Val Glu Ala Trp Phe Lys Pro Leu Ala Tyr
340 345 350
Ala Leu Ile Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr Gly
355 360 365
Asp Tyr Tyr Gly Ile Pro Thr His Gly Val Ala Ala Met Lys Gly Lys
370 375 380
Ile Asp Pro Ile Leu Glu Ala Arg Gln Lys Tyr Ala Tyr Gly Thr Gln
385 390 395 400
His Asp Tyr Leu Asp His His Asn Ile Ile Gly Trp Thr Arg Glu Gly
405 410 415
Asn Ser Ala His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp Gly
420 425 430
Pro Gly Gly Ser Lys Trp Met Tyr Val Gly Arg His Lys Ala Gly Gln
435 440 445
Val Trp Arg Asp Ile Thr Gly Asn Arg Thr Gly Thr Val Thr Ile Asn
450 455 460
Ala Asp Gly Trp Gly Asn Phe Ser Val Asn Gly Gly Ser Val Ser Ile
465 470 475 480
Trp Val Asn Lys
<210> 137
<211> 9545
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 137
ctgggagctg atagcacagc gatttcccgc ctcctgtcgg aagcatcgcc accgtgtccc 60
gtccctccag cacgcttttg atgatcgttt cctggccttt tttaaaagca tgataaccaa 120
aatacctgcg gagcgcctga tgaagtttat ccattcctga tcaccgtttt cgacagggcc 180
aggcggattt taaaataact gtatcggccg ccgagtccct ctttaatttg tctgatttta 240
ttggtttggt tcgctatcgc ataatcggca ataattttct gttcctcttt cgacacgtac 300
tgatcgatgg aaaaagacgg atcatgaatg gcgatttcca caatgtgatc ctcgatcgtc 360
gccaatttca gctttctaat ttttgcgatc cggtctattg tatacccttc ttttataaga 420
aagagcgttt ttctcgtcga ttgtgtcaga ccatcattca aaggaatgtc atgaatgagc 480
gtttgaaaaa gcggactttc accattttgg gcggattgaa taaaatggtg cagaacatcc 540
caaaacagtg catatatgta ccactcatca aggttcattt tttcggaaag ctgtctgaat 600
gtatagcccg cttttgttct ggatgtcagg gagtgaacaa aaatagcggc ctgttcatca 660
tgatttaaaa cagacagctt ctctttcagc tcttggtgaa actgggccgc tgtttcagca 720
gcattccggt ttcttaaata ctgtttaacc cagttctgaa tctgataatc ttttacgatc 780
ggcaaataca cgcgttcgcg gtaaagcttg ttggagagca cttggatcaa aagcgacatc 840
cttgcccaca tcactttggc tgccgcctga taatagccgc cgtgaaaatg gcggggccac 900
ggataaaggg caaaaaagcc ggcaagttcc gcttctcctt tttcggtgac ggtgtaagcg 960
ccgctttccg ctttttctct gaccaaagac tcctgcttca atcgttgaac gctggctgcc 1020
acctgctctc tcgacagagc cgaacaaaag ccgaagtatt ttgaaacggc aaataaaccg 1080
gcgtcctgta tcgtctgtga cgaccttttt ccttttaata aatgatagac cgcgcttgga 1140
gaacgctcac ccttcatgga tgacagaatg tcaagcacaa tcgcgtcaaa aaaatgaacc 1200
ggcatatcat cacctgcaat cttccggcaa cattcgatca tttcttcctt ttattttaac 1260
agattttgcg gagaaatcga cgtttaaact catataaaag gggtatgtta gcagtagaac 1320
ccttgtgtga taagcattct caatattttt gagttgaaat gtaagattaa caccattaca 1380
ataaggaatg ggaataggtt tcatatcgga tagatagagg gttaaaccat ttgttccaac 1440
gaagaacaat ctgggaggtt ttttattcat gccaaaatat acaattgtag acaaagatac 1500
gtgcatcgca tgcggagctt gtggtgctgc ggctcctgat atttatgatt acgacgatga 1560
gggaatcgca tttgtcaccc ttgacgacaa tcagggtgtc gtcgaagtcc ctgacgtctt 1620
agaagaagac atgatggacg cgtttgaagg ctgtcctaca gattcgatca aagttgcgga 1680
tgagccgttc gaaggcgacc cgcttaaaca cgaataaagc caaaaaacat ccggtgcaca 1740
aagtgccgga tgttttttta tgagataagc acggctttac caacaagcaa aaagaagccg 1800
gctaaagaca tccggcttct tctgcagctg acaatatccg ggaacatgca cccgatattg 1860
tcatgtttat ttatttggcc atgcggacgt tttccttcag ccgcggtttc agcgaaagga 1920
aaatcggcgt ggacacgagg gccacagcga tgcctttaat gaaattaaaa ggcaggattc 1980
cggccagaac tgttgtcttg agcgcctctc cagtcagcgc tggagcattt aaaaaccaag 2040
tgtaggcagg cagaaacagc agataattta aaatgctcat cgaaacggcc atcacaagcg 2100
tccctgcgaa aagagctgtg acaaaccctt tggcagaact tgattttttc agcagtacag 2160
ctgccggcag gataaacaat gttccggcaa tgaagttagc cgcctgatca atcggaacgc 2220
ccgaggcgct tcctgcaata aagtaattca gcacgttttt gatcgcttca acggcaatcc 2280
cggctcccgg accgtacaaa ataacagcga gcaatgccgg gatatcactg aaatcgattt 2340
ttaaatacgg gaatgccccc aggatcggaa agctcagcat cattaaaata aatgcgatgc 2400
tgctcagcat gctgatagag acgagacgtc tcaccttgtt gtgtttcatt ttgtcactct 2460
ctccttttcg atcacatctc acgaaaagag gaatggttct ttcccctgtc ctaaacaaaa 2520
aacccgcttt attgaaaaag cggggctgtt ttacagacag gtcaaataaa cgtttgaaaa 2580
tgttcatttc aaaacgcgcg gaacctccat cttctcccat ccagactata ctgtcggctt 2640
cggaatcgca ccgaatcctg cccataaaaa ggctcgcggg cttagagcgc ttgctcatca 2700
ccgccggtag ggaatttcac cctgccccga agattgatct tatttatttt taatactgat 2760
attattataa attaattgtg aaaaaatgta caggtgcaaa gcttattgcg ctgttttggg 2820
acatcctgca cgatatttcg gtaaactcac tttttccgag ctctcgctga taaacagctg 2880
acatcaacta aaagcttcat taaatacttt gaaaaaagtt gttgacttaa aagaagctaa 2940
atgttatagt aataaaacag aatagtcttt taagtaagtc tactctgaat ttttttaaaa 3000
ggagagggta aagaatgaaa caacaaaaac ggctttacgc ccgattgctg acgctgttat 3060
ttgcgctcat cttcttgctg cctcattctg cagctagcgc acaccacaac ggaacaaacg 3120
gcacaatgat gcagtatttt gaatggcacc tgccgaatga tggacagcat tggaatagac 3180
tgagaaacga cgcagcgaac ctgaagaacc ttggcatcac ggcagtctgg attccgcctg 3240
cgtggaaagg cacatcacaa aatgatgtcg gctatggcgc atatgacctg tacgacctgg 3300
gagagttcaa ccagaaggga acaatcagaa cgaaatatgg aacaagatca caactgcaaa 3360
gcgctatcgc gagcctgcaa aataatggca tccaagttta tggcgacgtg gtcatgaacc 3420
acaaaggcgg agcagacgga acggaatggg ttcaagcggt cgaggtgaat ccgagcaaca 3480
gaaaccaaga ggttacgggc gaatacacga tcgaagcctg gacaaagttc gactttccgg 3540
gcagaggcaa tacacactca agcttcaagt ggagatggta tcactttgac ggcacggact 3600
gggatcaaag cagacgtctg aataacagaa tctataagtt tacaggcaaa ggttgggatt 3660
gggaggtgga cacagaaaac ggaaactatg actatctgat gtatgctgac gtcgacatgg 3720
atcatccgga ggtcatcaat gagctgagaa gatggggcgt ttggtacacg aacacactga 3780
acctggatgg attcagaatt gacgcagtca aacacatcaa gtactcattt acaagagact 3840
ggcttaacca cgtgagatca acaacgggaa agaacaatat gttcgccgtt gcggagtttt 3900
ggaagaatga tctgggcgca atcgagaact atcttcacaa gacgaactgg aatcatagcg 3960
tcttcgatgt cccgctgcac tacaatctgt ataatgcatc aaagtcaggc ggcaactacg 4020
atatgagaca aatcctgaat ggaacggtcg tctcaaaaca cccgatgcac gccgtcacgt 4080
ttgtcgataa tcacgattca caaccggcag aagcccttga gagctttgtt gaggcatggt 4140
tcaaaccgct ggcctatgct ctgatcctga caagagagca aggctatccg tcagtgtttt 4200
acggcgatta ctacggcatt ccgacacatg gcgtcgcagc catgaaagga aagattgatc 4260
cgattctgga ggctagacag aaatacgcct acggaacgca acacgattac cttgatcacc 4320
ataacatcat cggatggacg agagagggaa actcagcaca tcctaactca ggccttgcaa 4380
caattatgtc agatggaccg ggaggcagca aatggatgta cgttggcaga cacaaggcag 4440
gacaggtttg gagagatatc acgggcaata gaacaggaac agttacaatc aacgctgacg 4500
gctggggcaa ttttagcgtt aatggcggct cagttagcat ctgggtgaat aaatgaaaga 4560
gcagagagga cggatttcct gaaggaaatc cgttttttta ttttgcggcc gcatattccg 4620
cattcgcaat gcctaccgca tactaaaaac cgcacattca cagttatttc atttttaatt 4680
ttcgtctttc cgcgtgaaac tcattgacac tctttatgga atatggtaaa ttatcagata 4740
tttatgacgc ttatttagga ggaaatctta catgtttcga gtattggtct cagataaaat 4800
gtccagcgac ggcctcaaac cattaatgga agcagatttt attgaaattg tagaaaagaa 4860
tgttgcggaa gcggaagacg agcttcatac gtttgacgcg ctcttggtgc ggagcgccac 4920
gaaggtaacc gaagagctgt ttaaaaagat gacttcgctg aaaatcgtcg ccagagcagg 4980
tgtcggcgtc gacaatatcg atattgacga ggcgacaaaa cacggtgtta tcgtcgtaaa 5040
cgcgccaaac gggaatacaa tttcaaccgc tgaacatacc tttgcaatgt tttcagcgtt 5100
aatgagacat attccgcagg caaacatctc cgtgaaatca agggagtgga atcgttcggc 5160
ttacgtcggt tcagagcttt acggaaaaac gctcggcatc atcggaatgg gccgcatcgg 5220
aagcgaaatc gcgagccgcg caaaagcatt cggtatgacc gttcatgtat ttgacccgtt 5280
cctgacccaa gaaagggcaa gcaagctcgg cgttaacgcg aacagctttg aagaagttct 5340
ggcatgcgcc gacatcatta cggttcatac cccgctcacg aaagaaacga agggactttt 5400
gaacaaagaa accatcgcaa aaacgaaaaa aggcgttcgt ctcgttaact gtgcaagagg 5460
cggcatcatc gatgaagcag cgcttttgga agctctggaa agcggacatg tcgctggcgc 5520
tgccttggat gtattcgaag tcgagcctcc ggtcgattca aaactgatcg atcatccgct 5580
tgtagtcgcg actcctcact tgggcgcctc aacaaaagaa gcccagctga atgtcgctgc 5640
acaagtgtcc gaagaagtcc ttcagtatgc gcaaggaaac cctgtgatgt ccgcgatcaa 5700
ccttccggcc atgacaaagg attcattcga aaaaatccag ccttatcatc agtttgccaa 5760
tacgatcgga aaccttgtgt ctcagtgcat gaatgagcct gttcaagatg tagccatcca 5820
atatgaaggc tccatcgcca aacttgaaac gtcatttatt acgaaaagcc ttttggccgg 5880
atttctgaag ccgagggtcg cggctaccgt taacgaagtg aatgccggca ccgttgcgaa 5940
agagcgcggc atcagcttca gcgaaaaaat ttcttccaat gagtcaggct atgaaaactg 6000
catctctgtg actgtcacgg gagatgtaac aacattctct ttaagagcga cgtacattcc 6060
gcacttcggc ggacgcatcg ttgccttaaa cggctttgat attgattttt atccggctgg 6120
acaccttgtc tacattcacc accaggataa accaggggct atcggccatg tcggacgaat 6180
tttaggagac catgacatca atatcgccac tatgcaggta ggccgaaaag aaaaaggcgg 6240
agaagcgatc atgatgcttt cctttgaccg ccaccttgag gacgatattt tagctgagct 6300
gaaaaacatc ccggatatcg tgtctgttaa agccatcgac cttccttaaa cagaagctgc 6360
ggaacctgaa aagaattcct ttcaggttcc gtttttttta ggaattctcc ctgatctcaa 6420
gcatctggcg gggataaatc cgctctcctt tcaaatcgtt ccattctttg aggcgctgta 6480
cagttacgcc cattttttcg gcgatatgat gaagcgtatc ccctttccgc actacatatg 6540
taccggtctt cgattcatcg tcatgaaggc ggagtgtttg gccggccttg agatttgaat 6600
gtttcaaccc gtttattctc atgatctcct cgatggatat accgctatcc ttgctgattc 6660
tccagagcgt gtcccctttt tgaacggtca ccgcaccgct cattgtcccg gcgttttgat 6720
aaacgtggat agaattttgc cggaacgcct cctcacgaag caccgtcagc ggattgattg 6780
catatctttt atcttcagtc catgaaccgt gatgcatttc aaaatgcagg tgggttccgg 6840
tcgatattcc cgtattgccg atgattccga tttgctcgcc ttttttcacc cgctcctttt 6900
cctttttcag gcgtttgctt aagtgggcat aaacggtttc atatccgttg tcatgtttaa 6960
taaatatcac ttggccgtag gagtcggatt gatacgattt gcttatcgtt ccgtctgcgg 7020
ctgccgctac tgcttcccct tcgggagcag cgatgtcaag ccccttatgc tttccgcctc 7080
tcgtaccgaa ttgatctgtg atctctcctt taatcggttc aatccactct gaggcttccg 7140
cccccggggc attgacgaaa agcgccaatc ccgaaagcca tgcgatcgcg aacaggaagt 7200
tttgatgtct gagtttcttc aaggttttcc atatcctcct attacatgca tcttcggtaa 7260
aattgccccc tattcggaga cagcttagta tacttccaaa tcaatacaat ttatacatta 7320
aaaaaagact ccgcacaggg agtcttttag ttttctatcg tcatcggatt cggtgcgtac 7380
ggaacctgta cagatttcga caggtcatag gcgccgacct tggttatgga tgcgttttta 7440
aatttcactt ttgtgaagcc gaaatctttc gcggtcaata gaaggccttc caccatcaag 7500
acatcttcgg gtttattttc aatattcgcg gaggaagaaa attgaatgat cagttctttt 7560
ccattctttt gaatatcttc aatcggcgta tcatcggata aaatgggttt taaatgagtg 7620
ccgctttctt cgtttttcat catcttaatc gcttcctgca ccgattcgta agattcgctt 7680
gaaggtgcaa ggaaccggcg cccgtctgag ctttcatata aatagtagca tttttgcgtc 7740
tggtgcataa tcgccatatc ggcgagcatt ccgaatgttt caaattcaac acccgattta 7800
tcattggaaa taaacagaac agaatcatac gatccccatt taaaggtttc gttgatcaca 7860
tttttcagcc gttcgaaatc ttcgactgat agctccggta ttttctcatc aacttgaatc 7920
ttcagttttt tattgttttt ctgctctttg aacttcacct tatcaaggta agctgtgtca 7980
aatgatgtaa actggtccac tccaagccgg ctgtaagcgt gaagcgcatc ttcaagattt 8040
gtcatgccag tgcttttctc gaggcttacc gggacaacga cagacttgga ctcgtcaagg 8100
aaagcgaagg tgatatagtc gtctttttga ttctgtgaga cgacaaacgt atttgcaggt 8160
tcagacttgg cagcatcagc ctccgtctgc accaattttc cgtcagaaga aatgttggcg 8220
tcggcgctgt tttgagatct gatctgttcg attaactggg gagtgatcag catcagaaga 8280
aagagaacca aaactgtagc agcaaatgcg ccgacccgtt ttttcggtga tttacgcttt 8340
ggtgcgagaa tcagcttttg atagatctga tttgccgaac gattatcctt aaccgttgga 8400
agttggctta gtaacgcctt cagccgttct tcgttccatt ctgacttctt cattctttgg 8460
atcctccttc aaaagctcca tctgtttacg aagcactttc agaccgcggt gctgagtggt 8520
ttttaccttg ctttcggaaa aattcaaggc ttttgctgtt tcactgatcg aatatccttg 8580
aataaaacgc aagacgataa ctgatctttg gtcaagcgta cacttgtcta gggcctcgaa 8640
aatttccttt aggttttcat tttgcatcac gatgtcctca ggcagaggct tgcggtcttt 8700
tacatcttgt ttctcccagt caaacgtccc caaaatccgc tggcggatcg tctgctgctt 8760
tctgaaccag tcgatcgcaa cgtgccgcgc aatcgaaaga agccaggttt tttcgctgct 8820
cctgccttca aatgtttcgt aagaatgcag gacgcggatg tatacttcct gaactaagtc 8880
ttccgcctga tttttgtctt ttaccatata aaataaaaac tgaaataaat cctgatgata 8940
ctgatcatat attttctgaa aggtttcttc cacctgaaac ccctccgttc aatttattgt 9000
cgtttgtcaa tcttaaaagg ttacattaca actattacaa ctatattacg aacatatgaa 9060
aatggaaagg gggttttgcg aaagttaagc ttaattttaa cttaacaagc acaaaagcac 9120
ccgttctaaa tgaacaggtg ccaaggttat aggagcccac attttcacta agctgtgccc 9180
ttacaaggct ttcgttctcc tgaccggagc gttgcggatc cgctgaaatg aactaatttc 9240
aatccgttta tgactttaag tccaattgtt ggcgaagctt tttggaaatc tccattctct 9300
tttcgtcagt cactaggtga taccataagc cgtcaacttt tttgtcttca ccttcaattt 9360
ctacctgctt aatatcttta gcagcttctt tataagaact ttgaatatcg atcatcttgt 9420
ccagtgttaa gtttgttttg atgttttttt ctaatgctgc caaaatctct tgatagtttg 9480
ttaatgattt taattgtgca ccttcatgaa taatttcgtt gataatctca cgctgacgct 9540
cttga 9545
<210> 138
<211> 93
<212> DNA
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 138
tcgctgataa acagctgaca tcaactaaaa gcttcattaa atactttgaa aaaagttgtt 60
gacttaaaag aagctaaatg ttatagtaat aaa 93
<210> 139
<211> 58
<212> DNA
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 139
acagaatagt cttttaagta agtctactct gaattttttt aaaaggagag ggtaaaga 58
<210> 140
<211> 1455
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 140
caccacaacg gaacaaacgg cacaatgatg cagtattttg aatggcacct gccgaatgat 60
ggacagcatt ggaatagact gagaaacgac gcagcgaacc tgaagaacct tggcatcacg 120
gcagtctgga ttccgcctgc gtggaaaggc acatcacaaa atgatgtcgg ctatggcgca 180
tatgacctgt acgacctggg agagttcaac cagaagggaa caatcagaac gaaatatgga 240
acaagatcac aactgcaaag cgctatcgcg agcctgcaaa ataatggcat ccaagtttat 300
ggcgacgtgg tcatgaacca caaaggcgga gcagacggaa cggaatgggt tcaagcggtc 360
gaggtgaatc cgagcaacag aaaccaagag gttacgggcg aatacacgat cgaagcctgg 420
acaaagttcg actttccggg cagaggcaat acacactcaa gcttcaagtg gagatggtat 480
cactttgacg gcacggactg ggatcaaagc agacgtctga ataacagaat ctataagttt 540
acaggcaaag gttgggattg ggaggtggac acagaaaacg gaaactatga ctatctgatg 600
tatgctgacg tcgacatgga tcatccggag gtcatcaatg agctgagaag atggggcgtt 660
tggtacacga acacactgaa cctggatgga ttcagaattg acgcagtcaa acacatcaag 720
tactcattta caagagactg gcttaaccac gtgagatcaa caacgggaaa gaacaatatg 780
ttcgccgttg cggagttttg gaagaatgat ctgggcgcaa tcgagaacta tcttcacaag 840
acgaactgga atcatagcgt cttcgatgtc ccgctgcact acaatctgta taatgcatca 900
aagtcaggcg gcaactacga tatgagacaa atcctgaatg gaacggtcgt ctcaaaacac 960
ccgatgcacg ccgtcacgtt tgtcgataat cacgattcac aaccggcaga agcccttgag 1020
agctttgttg aggcatggtt caaaccgctg gcctatgctc tgatcctgac aagagagcaa 1080
ggctatccgt cagtgtttta cggcgattac tacggcattc cgacacatgg cgtcgcagcc 1140
atgaaaggaa agattgatcc gattctggag gctagacaga aatacgccta cggaacgcaa 1200
cacgattacc ttgatcacca taacatcatc ggatggacga gagagggaaa ctcagcacat 1260
cctaactcag gccttgcaac aattatgtca gatggaccgg gaggcagcaa atggatgtac 1320
gttggcagac acaaggcagg acaggtttgg agagatatca cgggcaatag aacaggaaca 1380
gttacaatca acgctgacgg ctggggcaat tttagcgtta atggcggctc agttagcatc 1440
tgggtgaata aatga 1455
<210> 141
<211> 3113
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 141
ttgtttttac acggtactag cagacaaaat gaaagagggc acctcgaaat cggcggtgtc 60
gatgttctat cattggcaga aagatacgga acacctcttt atgtatacga tgtcgcgctg 120
attagagagc gcgcccgaaa attccagaag gcattcaagg aagccggttt aaaagcgcag 180
gtagcgtatg caagcaaggc gttttcatcg gttgccatga ttcagcttgc cgaacaagag 240
gggctgtctc tggatgtggt atcgggagga gagcttttca ctgcgatcaa agcagggttc 300
ccagctgagc ggattcattt tcacggaaac aataagagcc ctgaagaact agccatggcg 360
ctggagcatc aaatcggctg catcgtgctc gataactttc acgagatcgc cattacagaa 420
gatctttgca agcgatcagg acaaactgta gacgttttgc tcagaatcac tccgggagtt 480
gaagcgcaca cgcacgatta tattacgacg gggcaggaag attccaaatt cggttttgat 540
ctgcataatg gacaggtcga acaagccatc gaacaagtcc tccgctcgtc tgcgtttaag 600
ctcctcggcg tgcactgcca catcggttcg caaatttttg atacggcagg atttgtcctt 660
gcagcagaca agattttcga gaagcttgcg gaatggcggg agacttactc tttcattccg 720
gaagtgctca atcttggcgg gggcttcggc atccgctata caaaagacga cgagccgctt 780
gcagctgatg tttatgttga aaaaatcatc gaggcggtca aagcaaatgc cgagcatttc 840
ggctttgaca tccctgagat ttggatcgaa ccaggccggt ctctcgtcgg tgatgcgggg 900
actacgctgt acacgatcgg ttctcaaaaa gaggtgccgg gcattcgcaa atatgtagcc 960
atcgacggcg gcatgagcga taatatcagg ccggcgcttt atgaggcaaa atatgaagca 1020
gccgtcgcca acaggatgaa cgatgcttgt catgataccg catcaatcgc aggaaaatgc 1080
tgcgaaagcg gagatatgct gatttgggat ttggaaatcc ccgaagttcg cgacggagat 1140
gtgctcgccg ttttctgcac cggtgcgtac ggctacagca tggccaacaa ctacaaccgc 1200
attccgcgcc cggccgtcgt ctttgtcgag gacggggaag cgcagctcgt cattcagaga 1260
gagacgtatg aggatatcgt caagctggat ctgccgctga aatcgaaagt caaacaataa 1320
aaaaatggag attccctaag aggggggtct ccatttttaa ttcagagctc gtcgctgata 1380
aacagctgac atcaatatcc tattttttca aaaaatattt taaaaagttg ttgacttaaa 1440
agaagctaaa tgttatagta ataaaacaga atagtctttt aagtaagtct actctgaatt 1500
tttttaaaag gagagggtaa agaatgaaac aacaaaaacg gctttacgcc cgattgctga 1560
cgctgttatt tgcgctcatc ttcttgctgc ctcattctgc agctagcgca caccacaacg 1620
gaacaaacgg cacaatgatg cagtattttg aatggcacct gccgaatgat ggacagcatt 1680
ggaatagact gagaaacgac gcagcgaacc tgaagaacct tggcatcacg gcagtctgga 1740
ttccgcctgc gtggaaaggc acatcacaaa atgatgtcgg ctatggcgca tatgacctgt 1800
acgacctggg agagttcaac cagaagggaa caatcagaac gaaatatgga acaagatcac 1860
aactgcaaag cgctatcgcg agcctgcaaa ataatggcat ccaagtttat ggcgacgtgg 1920
tcatgaacca caaaggcgga gcagacggaa cggaatgggt tcaagcggtc gaggtgaatc 1980
cgagcaacag aaaccaagag gttacgggcg aatacacgat cgaagcctgg acaaagttcg 2040
actttccggg cagaggcaat acacactcaa gcttcaagtg gagatggtat cactttgacg 2100
gcacggactg ggatcaaagc agacgtctga ataacagaat ctataagttt acaggcaaag 2160
gttgggattg ggaggtggac acagaaaacg gaaactatga ctatctgatg tatgctgacg 2220
tcgacatgga tcatccggag gtcatcaatg agctgagaag atggggcgtt tggtacacga 2280
acacactgaa cctggatgga ttcagaattg acgcagtcaa acacatcaag tactcattta 2340
caagagactg gcttaaccac gtgagatcaa caacgggaaa gaacaatatg ttcgccgttg 2400
cggagttttg gaagaatgat ctgggcgcaa tcgagaacta tcttcacaag acgaactgga 2460
atcatagcgt cttcgatgtc ccgctgcact acaatctgta taatgcatca aagtcaggcg 2520
gcaactacga tatgagacaa atcctgaatg gaacggtcgt ctcaaaacac ccgatgcacg 2580
ccgtcacgtt tgtcgataat cacgattcac aaccggcaga agcccttgag agctttgttg 2640
aggcatggtt caaaccgctg gcctatgctc tgatcctgac aagagagcaa ggctatccgt 2700
cagtgtttta cggcgattac tacggcattc cgacacatgg cgtcgcagcc atgaaaggaa 2760
agattgatcc gattctggag gctagacaga aatacgccta cggaacgcaa cacgattacc 2820
ttgatcacca taacatcatc ggatggacga gagagggaaa ctcagcacat cctaactcag 2880
gccttgcaac aattatgtca gatggaccgg gaggcagcaa atggatgtac gttggcagac 2940
acaaggcagg acaggtttgg agagatatca cgggcaatag aacaggaaca gttacaatca 3000
acgctgacgg ctggggcaat tttagcgtta atggcggctc agttagcatc tgggtgaata 3060
aatgaaagag cagagaggac ggatttcctg aaggaaatcc gtttttttat ttt 3113
<210> 142
<211> 2231
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 142
gtgttgtgct gaatcaatac atagaggccg gcggcgtatc gggtacaaca tctgtcaaga 60
cgtttaacgc tgatttaaag cagaaaaagc tgctgtccct tcaagatctg tttgaagaga 120
attcagattt tctgaacagg atttcaagca ttgcctatca ggaattgaaa aatcggaatc 180
cgtctgctga catggctttt ttaaaagaag ggacgagccc tcaggaagaa catttcagcc 240
gctttgcgct tcttgaaaac gaggtggaat tttattttga gaaaaaacaa gccggtcttg 300
aacagtttgt aaaaataaaa aaagaatggg taaaagatat tttaaaagac cgatatcagg 360
atatgaaaaa gaatcgtctt caggccaaac ctgatcagga gcctgttccg cttccgaagc 420
aagcgaaaat taatcccgat gaaaaagtga ttgccctcac atttgatgac ggtccgaatc 480
ccgctacaac gaataaaata ttaaacgctt tacagaagca tgaagggcat gcgaccttct 540
ttgtgcttgg aagcagagcc caatattatc ccgaaacgat aaaacggatg ctgaaggaag 600
gaaacgaagt cggcaaccat tcctgggacc atccgttatt gacaaggctg tcaaacgaaa 660
aagcgtatca ggagattaac gacacgcaag aaatgatcga aaaaatcagc ggacacctgc 720
ctgtacactt gcgtcctcca tacggcggga tcaatgattc cgtccgctcg ctttccaatc 780
tgaaggtttc attgtgggat gttgatccgg aagattggaa gtacaaaaat aagcaaaaga 840
ttgtcaatca tgtcatgagc catgcgggag acggaaaaat cgtcttaatg cacgatattt 900
atgcaacgtc cgcagatgct gctgaagaga ttattaaaaa gctgaaagca aaaggctatc 960
aattggtaac tgtatctcag cttgaagaag tgaagaagca gagaggctat tgaataaatg 1020
agtagaaagc gccatatcgg cgcttttctt ttggaagaaa atatagggaa aatggtactt 1080
gttaaaaatt cggaatattt atacaatatc atatgtttca cattgaaagg ggaggagaat 1140
catgaaacaa caaaaacggc tttacgcccg attgctgacg ctgttaggat cccacgtaaa 1200
cggcgggtcg gtttcaattt atgttcaaag atagaagagc aggctgacag tttgaatcgc 1260
ataggtaagg cggggatgaa atggcaacgt tatctgatgt agcaaagaaa gcaaatgtgt 1320
cgaaaatgac ggtatcgcgg gtgatcaatc atcctgagac tgtgacggat gaattgaaaa 1380
agcttgttca ttccgcaatg aaggagctca attatatacc gaactatgca gcaagagcgc 1440
tcgttcaaaa cagaacacag gtcgtcaagc tgctcatact ggaagaaatg gatacaacag 1500
aaccttatta tatgaatctg ttaacgggaa tcagccgcga gctggaccgt catcattatg 1560
ctttgcagct tgtcacaagg aaatctctca atatcggcca gtgcgacggc attattgcga 1620
cggggttgag aaaagccgat tttgaagggc tcatcaaggt ttttgaaaag cctgtcgttg 1680
tattcgggca aaatgaaatg ggctacgatt ttattgatgt taacaatgaa aaaggaacct 1740
atatggcaac acgtcacgtc attggtctgg gcgtccgcaa tgtcgtcttt tttgggatcg 1800
atttggatga gccctttgaa cgctcaaggg aaaaaggcta tcttcaggcg atggaaggca 1860
gtctgaaaaa agcagcgatt ttccggatgg aaaacagttc aaaaaaaagt gaagcacgcg 1920
cgcgggaagt gcttgcatcc tttgacgcac ctgcagcggt tgtttgcgct tcggaccgaa 1980
tcgcgctcgg ggttatccgc gcggtgcaat cgcttggtaa aagaattccg gaagatgtcg 2040
cggtcaccgg ctatgacggg gtgtttctcg accggatcgc ttcgcctcgc ctgacaaccg 2100
tcagacagcc tgttgttgaa atgggagagg cttgcgcgag aatcctgctg aaaaaaatca 2160
atgaagacgg agcgccgcaa ggcaatcaat tttttgagcc ggagcttatt gtccgcgaat 2220
cgactttgta g 2231
<210> 143
<211> 483
<212> PRT
<213> 人工序列
<220>
<223> 合成的
<400> 143
Ala Ala Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr Val Pro
1 5 10 15
Asn Asp Gly Gln Gln Trp Asn Arg Leu Arg Thr Asp Ala Pro Tyr Leu
20 25 30
Ser Ser Val Gly Ile Thr Ala Val Trp Thr Pro Pro Ala Tyr Lys Gly
35 40 45
Thr Ser Gln Ala Asp Val Gly Tyr Gly Pro Tyr Asp Leu Tyr Asp Leu
50 55 60
Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly Thr Lys
65 70 75 80
Gly Glu Leu Lys Ser Ala Val Asn Thr Leu His Ser Asn Gly Ile Gln
85 90 95
Val Tyr Gly Asp Val Val Met Asn His Lys Ala Gly Ala Asp Tyr Thr
100 105 110
Glu Asn Val Thr Ala Val Glu Val Asn Pro Ser Asn Arg Tyr Gln Glu
115 120 125
Thr Ser Gly Glu Tyr Asn Ile Gln Ala Trp Thr Gly Phe Asn Phe Pro
130 135 140
Gly Arg Gly Thr Thr Tyr Ser Asn Trp Lys Trp Gln Trp Phe His Phe
145 150 155 160
Asp Gly Thr Asp Trp Asp Gln Ser Arg Ser Leu Ser Arg Ile Phe Lys
165 170 175
Phe His Gly Lys Ala Trp Asp Trp Pro Val Ser Ser Glu Asn Gly Asn
180 185 190
Tyr Asp Tyr Leu Met Tyr Ala Asp Tyr Asp Tyr Asp His Pro Asp Val
195 200 205
Val Asn Glu Met Lys Lys Trp Gly Val Trp Tyr Ala Asn Glu Val Gly
210 215 220
Leu Asp Gly Tyr Arg Leu Asp Ala Val Lys His Ile Lys Phe Ser Phe
225 230 235 240
Leu Lys Asp Trp Val Asp Asn Ala Arg Ala Ala Thr Gly Lys Glu Met
245 250 255
Phe Thr Val Gly Glu Tyr Trp Gln Asn Asp Leu Gly Ala Leu Asn Asn
260 265 270
Tyr Leu Ala Lys Val Asn Tyr Asn Gln Ser Leu Phe Asp Ala Pro Leu
275 280 285
His Tyr Asn Phe Tyr Ala Ala Ser Thr Gly Gly Gly Ala Tyr Asp Met
290 295 300
Arg Asn Ile Leu Asn Asn Thr Leu Val Ala Ser Asn Pro Thr Lys Ala
305 310 315 320
Val Thr Leu Val Glu Asn His Asp Thr Gln Pro Gly Gln Ser Leu Glu
325 330 335
Ser Thr Val Gln Pro Trp Phe Lys Pro Leu Ala Tyr Ala Phe Ile Leu
340 345 350
Thr Arg Ser Gly Gly Tyr Pro Ala Val Phe Tyr Gly Asp Met Tyr Gly
355 360 365
Thr Lys Gly Thr Thr Thr Tyr Glu Ile Pro Ala Leu Lys Ser Lys Ile
370 375 380
Glu Pro Leu Leu Lys Ala Arg Lys Asp Tyr Ala Tyr Gly Thr Gln Arg
385 390 395 400
Asp Tyr Ile Asp Asn Pro Asp Val Ile Gly Trp Thr Arg Glu Gly Asp
405 410 415
Ser Thr Lys Ala Lys Ser Gly Leu Ala Thr Val Ile Thr Asp Gly Pro
420 425 430
Gly Gly Ser Lys Arg Met Tyr Val Gly Thr Ser Asn Ala Gly Glu Ile
435 440 445
Trp Tyr Asp Leu Thr Gly Asn Arg Thr Asp Lys Ile Thr Ile Gly Ser
450 455 460
Asp Gly Tyr Ala Thr Phe Pro Val Asn Gly Gly Ser Val Ser Val Trp
465 470 475 480
Val Gln Gln
<210> 144
<211> 9515
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 144
ctgggagctg atagcacagc gatttcccgc ctcctgtcgg aagcatcgcc accgtgtccc 60
gtccctccag cacgcttttg atgatcgttt cctggccttt tttaaaagca tgataaccaa 120
aatacctgcg gagcgcctga tgaagtttat ccattcctga tcaccgtttt cgacagggcc 180
aggcggattt taaaataact gtatcggccg ccgagtccct ctttaatttg tctgatttta 240
ttggtttggt tcgctatcgc ataatcggca ataattttct gttcctcttt cgacacgtac 300
tgatcgatgg aaaaagacgg atcatgaatg gcgatttcca caatgtgatc ctcgatcgtc 360
gccaatttca gctttctaat ttttgcgatc cggtctattg tatacccttc ttttataaga 420
aagagcgttt ttctcgtcga ttgtgtcaga ccatcattca aaggaatgtc atgaatgagc 480
gtttgaaaaa gcggactttc accattttgg gcggattgaa taaaatggtg cagaacatcc 540
caaaacagtg catatatgta ccactcatca aggttcattt tttcggaaag ctgtctgaat 600
gtatagcccg cttttgttct ggatgtcagg gagtgaacaa aaatagcggc ctgttcatca 660
tgatttaaaa cagacagctt ctctttcagc tcttggtgaa actgggccgc tgtttcagca 720
gcattccggt ttcttaaata ctgtttaacc cagttctgaa tctgataatc ttttacgatc 780
ggcaaataca cgcgttcgcg gtaaagcttg ttggagagca cttggatcaa aagcgacatc 840
cttgcccaca tcactttggc tgccgcctga taatagccgc cgtgaaaatg gcggggccac 900
ggataaaggg caaaaaagcc ggcaagttcc gcttctcctt tttcggtgac ggtgtaagcg 960
ccgctttccg ctttttctct gaccaaagac tcctgcttca atcgttgaac gctggctgcc 1020
acctgctctc tcgacagagc cgaacaaaag ccgaagtatt ttgaaacggc aaataaaccg 1080
gcgtcctgta tcgtctgtga cgaccttttt ccttttaata aatgatagac cgcgcttgga 1140
gaacgctcac ccttcatgga tgacagaatg tcaagcacaa tcgcgtcaaa aaaatgaacc 1200
ggcatatcat cacctgcaat cttccggcaa cattcgatca tttcttcctt ttattttaac 1260
agattttgcg gagaaatcga cgtttaaact catataaaag gggtatgtta gcagtagaac 1320
ccttgtgtga taagcattct caatattttt gagttgaaat gtaagattaa caccattaca 1380
ataaggaatg ggaataggtt tcatatcgga tagatagagg gttaaaccat ttgttccaac 1440
gaagaacaat ctgggaggtt ttttattcat gccaaaatat acaattgtag acaaagatac 1500
gtgcatcgca tgcggagctt gtggtgctgc ggctcctgat atttatgatt acgacgatga 1560
gggaatcgca tttgtcaccc ttgacgacaa tcagggtgtc gtcgaagtcc ctgacgtctt 1620
agaagaagac atgatggacg cgtttgaagg ctgtcctaca gattcgatca aagttgcgga 1680
tgagccgttc gaaggcgacc cgcttaaaca cgaataaagc caaaaaacat ccggtgcaca 1740
aagtgccgga tgttttttta tgagataagc acggctttac caacaagcaa aaagaagccg 1800
gctaaagaca tccggcttct tctgcagctg acaatatccg ggaacatgca cccgatattg 1860
tcatgtttat ttatttggcc atgcggacgt tttccttcag ccgcggtttc agcgaaagga 1920
aaatcggcgt ggacacgagg gccacagcga tgcctttaat gaaattaaaa ggcaggattc 1980
cggccagaac tgttgtcttg agcgcctctc cagtcagcgc tggagcattt aaaaaccaag 2040
tgtaggcagg cagaaacagc agataattta aaatgctcat cgaaacggcc atcacaagcg 2100
tccctgcgaa aagagctgtg acaaaccctt tggcagaact tgattttttc agcagtacag 2160
ctgccggcag gataaacaat gttccggcaa tgaagttagc cgcctgatca atcggaacgc 2220
ccgaggcgct tcctgcaata aagtaattca gcacgttttt gatcgcttca acggcaatcc 2280
cggctcccgg accgtacaaa ataacagcga gcaatgccgg gatatcactg aaatcgattt 2340
ttaaatacgg gaatgccccc aggatcggaa agctcagcat cattaaaata aatgcgatgc 2400
tgctcagcat gctgatagag acgagacgtc tcaccttgtt gtgtttcatt ttgtcactct 2460
ctccttttcg atcacatctc acgaaaagag gaatggttct ttcccctgtc ctaaacaaaa 2520
aacccgcttt attgaaaaag cggggctgtt ttacagacag gtcaaataaa cgtttgaaaa 2580
tgttcatttc aaaacgcgcg gaacctccat cttctcccat ccagactata ctgtcggctt 2640
cggaatcgca ccgaatcctg cccataaaaa ggctcgcggg cttagagcgc ttgctcatca 2700
ccgccggtag ggaatttcac cctgccccga agattgatct tatttatttt taatactgat 2760
attattataa attaattgtg aaaaaatgta caggtgcaaa gcttattgcg ctgttttggg 2820
acatcctgca cgatatttcg gtaaactcac tttttccgca tactaaaaac cgcacattca 2880
cagttatttc atttttaatt ttcgtctttc cgcgtgaaac tcattgacac tctttatgga 2940
atatggtaaa ttatcagata tttatgacgc ttatttagga ggaaatctta catgtttcga 3000
gtattggtct cagataaaat gtccagcgac ggcctcaaac cattaatgga agcagatttt 3060
attgaaattg tagaaaagaa tgttgcggaa gcggaagacg agcttcatac gtttgacgcg 3120
ctcttggtgc ggagcgccac gaaggtaacc gaagagctgt ttaaaaagat gacttcgctg 3180
aaaatcgtcg ccagagcagg tgtcggcgtc gacaatatcg atattgacga ggcgacaaaa 3240
cacggtgtta tcgtcgtaaa cgcgccaaac gggaatacaa tttcaaccgc tgaacatacc 3300
tttgcaatgt tttcagcgtt aatgagacat attccgcagg caaacatctc cgtgaaatca 3360
agggagtgga atcgttcggc ttacgtcggt tcagagcttt acggaaaaac gctcggcatc 3420
atcggaatgg gccgcatcgg aagcgaaatc gcgagccgcg caaaagcatt cggtatgacc 3480
gttcatgtat ttgacccgtt cctgacccaa gaaagggcaa gcaagctcgg cgttaacgcg 3540
aacagctttg aagaagttct ggcatgcgcc gacatcatta cggttcatac cccgctcacg 3600
aaagaaacga agggactttt gaacaaagaa accatcgcaa aaacgaaaaa aggcgttcgt 3660
ctcgttaact gtgcaagagg cggcatcatc gatgaagcag cgcttttgga agctctggaa 3720
agcggacatg tcgctggcgc tgccttggat gtattcgaag tcgagcctcc ggtcgattca 3780
aaactgatcg atcatccgct tgtagtcgcg actcctcact tgggcgcctc aacaaaagaa 3840
gcccagctga atgtcgctgc acaagtgtcc gaagaagtcc ttcagtatgc gcaaggaaac 3900
cctgtgatgt ccgcgatcaa ccttccggcc atgacaaagg attcattcga aaaaatccag 3960
ccttatcatc agtttgccaa tacgatcgga aaccttgtgt ctcagtgcat gaatgagcct 4020
gttcaagatg tagccatcca atatgaaggc tccatcgcca aacttgaaac gtcatttatt 4080
acgaaaagcc ttttggccgg atttctgaag ccgagggtcg cggctaccgt taacgaagtg 4140
aatgccggca ccgttgcgaa agagcgcggc atcagcttca gcgaaaaaat ttcttccaat 4200
gagtcaggct atgaaaactg catctctgtg actgtcacgg gagatgtaac aacattctct 4260
ttaagagcga cgtacattcc gcacttcggc ggacgcatcg ttgccttaaa cggctttgat 4320
attgattttt atccggctgg acaccttgtc tacattcacc accaggataa accaggggct 4380
atcggccatg tcggacgaat tttaggagac catgacatca atatcgccac tatgcaggta 4440
ggccgaaaag aaaaaggcgg agaagcgatc atgatgcttt cctttgaccg ccaccttgag 4500
gacgatattt tagctgagct gaaaaacatc ccggatatcg tgtctgttaa agccatcgac 4560
cttccttaag tcgctgataa acagctgaca tcaatatcct attttttcaa aaaatatttt 4620
aaaaagttgt tgacttaaaa gaagctaaat gttatagtaa taaaacagaa tagtctttta 4680
agtaagtcta ctctgaattt ttttaaaagg agagggtaaa gatgaaacaa caaaaacggc 4740
tttacgcccg attgctgacg ctgttatttg cgctcatctt cttgctgcct cattctgcag 4800
ctagcgcagc agcgacaaac ggaacaatga tgcagtattt cgagtggtat gtacctaacg 4860
acggccagca atggaacaga ctgagaacag atgcccctta cttgtcatct gttggtatta 4920
cagcagtatg gacaccgccg gcttataagg gcacgtctca agcagatgtg gggtacggcc 4980
cgtacgatct gtatgattta ggcgagttta atcaaaaagg tacagtcaga acgaagtatg 5040
gcacaaaagg agaacttaaa tctgctgtca acacgctgca ttcaaatgga atccaagtgt 5100
atggtgatgt cgtgatgaat cataaagcag gtgctgatta tacagaaaac gtaacggcgg 5160
tggaggtgaa tccgtctaat agatatcagg aaacgagcgg cgaatataat attcaggcat 5220
ggacaggctt caactttccg ggcagaggaa caacgtattc taactggaaa tggcagtggt 5280
tccattttga tggaacggat tgggaccaga gcagaagcct ctctagaatc ttcaaattcc 5340
atggaaaggc gtgggactgg ccggtttctt cagaaaacgg aaattatgac tatctgatgt 5400
acgcggacta tgattatgac catccggatg tcgtgaatga aatgaaaaag tggggcgtct 5460
ggtatgccaa cgaagttggg ttagatggat acagacttga cgcggtcaaa catattaaat 5520
ttagctttct caaagactgg gtggataacg caagagcagc gacgggaaaa gaaatgttta 5580
cggttggcga atattggcaa aatgatttag gggccctgaa taactacctg gcaaaggtaa 5640
attacaacca atctcttttt gatgcgccgt tgcattacaa cttttacgct gcctcaacag 5700
ggggtggagc gtacgatatg agaaatattc ttaataacac gttagtcgca agcaatccga 5760
caaaggctgt tacgttagtt gagaatcatg acacacagcc tggacaatca ctggaatcaa 5820
cagtccaacc gtggtttaaa ccgttagcct acgcgtttat tctcacgaga agcggaggct 5880
atcctgcggt attttatgga gatatgtacg gtacaaaagg aacgacaaca tatgagatcc 5940
ctgctcttaa atctaaaatc gaacctttgc ttaaggctag aaaagactat gcttatggaa 6000
cacagagaga ctatattgat aacccggatg tcattggctg gacgagagaa ggggactcaa 6060
cgaaagccaa gagcggtctg gccacagtga ttacagatgg gccgggcggt tcaaaaagaa 6120
tgtatgttgg cacgagcaat gcgggtgaaa tctggtatga tttgacaggg aatagaacag 6180
ataaaatcac gattggaagc gatggctatg caacatttcc tgtcaatggg ggctcagttt 6240
cagtatgggt gcagcaatga aagcttctcg aggttaacag aggacggatt tcctgaagga 6300
aatccgtttt tttattttac agaagctgcg gaacctgaaa agaattcctt tcaggttccg 6360
ttttttttag gaattctccc tgatctcaag catctggcgg ggataaatcc gctctccttt 6420
caaatcgttc cattctttga ggcgctgtac agttacgccc attttttcgg cgatatgatg 6480
aagcgtatcc cctttccgca ctacatatgt accggtcttc gattcatcgt catgaaggcg 6540
gagtgtttgg ccggccttga gatttgaatg tttcaacccg tttattctca tgatctcctc 6600
gatggatata ccgctatcct tgctgattct ccagagcgtg tccccttttt gaacggtcac 6660
cgcaccgctc attgtcccgg cgttttgata aacgtggata gaattttgcc ggaacgcctc 6720
ctcacgaagc accgtcagcg gattgattgc atatctttta tcttcagtcc atgaaccgtg 6780
atgcatttca aaatgcaggt gggttccggt cgatattccc gtattgccga tgattccgat 6840
ttgctcgcct tttttcaccc gctccttttc ctttttcagg cgtttgctta agtgggcata 6900
aacggtttca tatccgttgt catgtttaat aaatatcact tggccgtagg agtcggattg 6960
atacgatttg cttatcgttc cgtctgcggc tgccgctact gcttcccctt cgggagcagc 7020
gatgtcaagc cccttatgct ttccgcctct cgtaccgaat tgatctgtga tctctccttt 7080
aatcggttca atccactctg aggcttccgc ccccggggca ttgacgaaaa gcgccaatcc 7140
cgaaagccat gcgatcgcga acaggaagtt ttgatgtctg agtttcttca aggttttcca 7200
tatcctccta ttacatgcat cttcggtaaa attgccccct attcggagac agcttagtat 7260
acttccaaat caatacaatt tatacattaa aaaaagactc cgcacaggga gtcttttagt 7320
tttctatcgt catcggattc ggtgcgtacg gaacctgtac agatttcgac aggtcatagg 7380
cgccgacctt ggttatggat gcgtttttaa atttcacttt tgtgaagccg aaatctttcg 7440
cggtcaatag aaggccttcc accatcaaga catcttcggg tttattttca atattcgcgg 7500
aggaagaaaa ttgaatgatc agttcttttc cattcttttg aatatcttca atcggcgtat 7560
catcggataa aatgggtttt aaatgagtgc cgctttcttc gtttttcatc atcttaatcg 7620
cttcctgcac cgattcgtaa gattcgcttg aaggtgcaag gaaccggcgc ccgtctgagc 7680
tttcatataa atagtagcat ttttgcgtct ggtgcataat cgccatatcg gcgagcattc 7740
cgaatgtttc aaattcaaca cccgatttat cattggaaat aaacagaaca gaatcatacg 7800
atccccattt aaaggtttcg ttgatcacat ttttcagccg ttcgaaatct tcgactgata 7860
gctccggtat tttctcatca acttgaatct tcagtttttt attgtttttc tgctctttga 7920
acttcacctt atcaaggtaa gctgtgtcaa atgatgtaaa ctggtccact ccaagccggc 7980
tgtaagcgtg aagcgcatct tcaagatttg tcatgccagt gcttttctcg aggcttaccg 8040
ggacaacgac agacttggac tcgtcaagga aagcgaaggt gatatagtcg tctttttgat 8100
tctgtgagac gacaaacgta tttgcaggtt cagacttggc agcatcagcc tccgtctgca 8160
ccaattttcc gtcagaagaa atgttggcgt cggcgctgtt ttgagatctg atctgttcga 8220
ttaactgggg agtgatcagc atcagaagaa agagaaccaa aactgtagca gcaaatgcgc 8280
cgacccgttt tttcggtgat ttacgctttg gtgcgagaat cagcttttga tagatctgat 8340
ttgccgaacg attatcctta accgttggaa gttggcttag taacgccttc agccgttctt 8400
cgttccattc tgacttcttc attctttgga tcctccttca aaagctccat ctgtttacga 8460
agcactttca gaccgcggtg ctgagtggtt tttaccttgc tttcggaaaa attcaaggct 8520
tttgctgttt cactgatcga atatccttga ataaaacgca agacgataac tgatctttgg 8580
tcaagcgtac acttgtctag ggcctcgaaa atttccttta ggttttcatt ttgcatcacg 8640
atgtcctcag gcagaggctt gcggtctttt acatcttgtt tctcccagtc aaacgtcccc 8700
aaaatccgct ggcggatcgt ctgctgcttt ctgaaccagt cgatcgcaac gtgccgcgca 8760
atcgaaagaa gccaggtttt ttcgctgctc ctgccttcaa atgtttcgta agaatgcagg 8820
acgcggatgt atacttcctg aactaagtct tccgcctgat ttttgtcttt taccatataa 8880
aataaaaact gaaataaatc ctgatgatac tgatcatata ttttctgaaa ggtttcttcc 8940
acctgaaacc cctccgttca atttattgtc gtttgtcaat cttaaaaggt tacattacaa 9000
ctattacaac tatattacga acatatgaaa atggaaaggg ggttttgcga aagttaagct 9060
taattttaac ttaacaagca caaaagcacc cgttctaaat gaacaggtgc caaggttata 9120
ggagcccaca ttttcactaa gctgtgccct tacaaggctt tcgttctcct gaccggagcg 9180
ttgcggatcc gctgaaatga actaatttca atccgtttat gactttaagt ccaattgttg 9240
gcgaagcttt ttggaaatct ccattctctt ttcgtcagtc actaggtgat accataagcc 9300
gtcaactttt ttgtcttcac cttcaatttc tacctgctta atatctttag cagcttcttt 9360
ataagaactt tgaatatcga tcatcttgtc cagtgttaag tttgttttga tgtttttttc 9420
taatgctgcc aaaatctctt gatagtttgt taatgatttt aattgtgcac cttcatgaat 9480
aatttcgttg ataatctcac gctgacgctc ttgac 9515
<210> 145
<211> 1452
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 145
gcagcgacaa acggaacaat gatgcagtat ttcgagtggt atgtacctaa cgacggccag 60
caatggaaca gactgagaac agatgcccct tacttgtcat ctgttggtat tacagcagta 120
tggacaccgc cggcttataa gggcacgtct caagcagatg tggggtacgg cccgtacgat 180
ctgtatgatt taggcgagtt taatcaaaaa ggtacagtca gaacgaagta tggcacaaaa 240
ggagaactta aatctgctgt caacacgctg cattcaaatg gaatccaagt gtatggtgat 300
gtcgtgatga atcataaagc aggtgctgat tatacagaaa acgtaacggc ggtggaggtg 360
aatccgtcta atagatatca ggaaacgagc ggcgaatata atattcaggc atggacaggc 420
ttcaactttc cgggcagagg aacaacgtat tctaactgga aatggcagtg gttccatttt 480
gatggaacgg attgggacca gagcagaagc ctctctagaa tcttcaaatt ccatggaaag 540
gcgtgggact ggccggtttc ttcagaaaac ggaaattatg actatctgat gtacgcggac 600
tatgattatg accatccgga tgtcgtgaat gaaatgaaaa agtggggcgt ctggtatgcc 660
aacgaagttg ggttagatgg atacagactt gacgcggtca aacatattaa atttagcttt 720
ctcaaagact gggtggataa cgcaagagca gcgacgggaa aagaaatgtt tacggttggc 780
gaatattggc aaaatgattt aggggccctg aataactacc tggcaaaggt aaattacaac 840
caatctcttt ttgatgcgcc gttgcattac aacttttacg ctgcctcaac agggggtgga 900
gcgtacgata tgagaaatat tcttaataac acgttagtcg caagcaatcc gacaaaggct 960
gttacgttag ttgagaatca tgacacacag cctggacaat cactggaatc aacagtccaa 1020
ccgtggttta aaccgttagc ctacgcgttt attctcacga gaagcggagg ctatcctgcg 1080
gtattttatg gagatatgta cggtacaaaa ggaacgacaa catatgagat ccctgctctt 1140
aaatctaaaa tcgaaccttt gcttaaggct agaaaagact atgcttatgg aacacagaga 1200
gactatattg ataacccgga tgtcattggc tggacgagag aaggggactc aacgaaagcc 1260
aagagcggtc tggccacagt gattacagat gggccgggcg gttcaaaaag aatgtatgtt 1320
ggcacgagca atgcgggtga aatctggtat gatttgacag ggaatagaac agataaaatc 1380
acgattggaa gcgatggcta tgcaacattt cctgtcaatg ggggctcagt ttcagtatgg 1440
gtgcagcaat ga 1452
<210> 146
<211> 9214
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 146
catcggacag ctcttgcttg atatcttcaa aatgacgccg gctcatgtca tgtcaacttt 60
tgtcgtatct ggagcgatcc ttgacggatt cggcatttac gaccgtttta tcgaatttgc 120
cggtgccggg gctacagtcc cgattgtcag cttcggccac tctcttttgc acggcgcgat 180
gcaccaggct gagaaacatg gctttatcgg aatcggcatg gggatatttg aactgacatc 240
tgccggtata tctgccgcta tcttgttcgc ttttcttgtt gccgtgattt ttaaaccgaa 300
aggataaagg aaaatgccag caaaacgcaa ggtcattttg gtcacagacg gcgatatata 360
cgctgcaaaa gcaatcgaat atgcagcaag aaaaacgggt ggccgctgca tttcccaatc 420
ggcggggaat ccgagcgtta aaacaggacc ggagcttgta accatgatcc tgcaaacccc 480
tcatgatcct gtattcgtca tgtttgatga ttccggactt caaggtgaag gcccgggaga 540
gacagctatg aaatatgtag cgatgcatcc cgatatcgag gtgctcggag tcatcgccgt 600
cgcttcaaaa actcattatg cagagtggac gagagtcgat gtatcaatcg atgcagaagg 660
cgaactgaca gagtacggcg tcgataaaca cggggtcaaa gagttcgatg tcaaacgaat 720
gaatggtgat acagtctatt gccttgacca gctggatgtt ccgatcattg tcggaatcgg 780
tgatatcggt aagatgaaca gaaaagacga tgtggaaaaa ggttcgccga ttacaatgaa 840
agcggtcgag ctcattttag aaaggagcgg gtatcatgag tgctcaaaag caagagaaga 900
cgaacgtatt ccttgatcct tctaagaatg aagcgtattt caagaagcgg gtcggcatgg 960
gagaaagctt tgaccttggc gtacggaagg tctttattct cggacatgaa gttcagcttt 1020
attatgtcaa cggattgtgc gacacacaat acatcattca cctgttaaga gaactggtgc 1080
atctgaatga taaagaaaaa gaatcgggcg aggtcgaaga catcgtcgaa aacaggcttt 1140
tgaaccagca ggtttcaaaa gcggaaacgc ttgatgaagc tgtcgaccaa gtgttgtcag 1200
gactggttgc catcatcgtc gaagatgcgg gctttgcttt tatcatcgat gtcagaagct 1260
acccgggcag aacgccggaa gaacctgata cagaaaaagt cgtacgcggt gcaagggacg 1320
gactcgtcga gaacatcatc gtcaacacag ccctgattag acgccggatc agagatgagc 1380
gcttgcgcta caaaatgctt catatcggtg aacgctctaa aacagacatc tgcctctgct 1440
atttggaaga cgttgcagat cccgatcttg ttgaagtatt aaaaaaagaa attgaagatg 1500
tgaagatcga cgggctgccg atgtcggata aatcggtaga ggaattcctg gtcggccaag 1560
gctacaatcc gtttccgctt gtcaggttta cggaaagggc agacgtagcc gcaagccata 1620
ttttagaggg gcatgtcatc gtgatcgtcg atacgtcgcc aagcgtcatc atcacaccga 1680
ccactttgtt tcaccatgtt cagcatgctg aggaatacag acagacgccg gctgttggga 1740
cgtttttaag gtgggtgcgg tttttcggta ttttggcctc cacctttttg ctgccgcttt 1800
ggctgctgtt tgtcattcat ccgtcgctct tgcctgataa tttatcgttt atcgggttga 1860
ataaagacac ccatattccg attatcatgc agattttcct ggcggatctc ggcgtcgaat 1920
ttttaagaat ggccgccatt catacgccga cggcgctttc gactgcaatg ggcctgatcg 1980
ccgctgtatt gatcggcgat atcgcgatca atgtcggctt gttttctccc gaagtcattt 2040
tatacgtttc cctctcggca atcggagcct acacgacacc aagctacgag ctgagcctgg 2100
cgaataaaat ggtgaagctg tttatgctga tattggtggc gctttttaaa gtggagggat 2160
ttgtcatcgg attaacgatc ttaactatag tgatgacttc gatcaggtca ttgcgaacgc 2220
cttacttatg gcctctcctc ccgttcaatg gaaaagcgtt ttggcatgtt ctcgtgcgca 2280
cgtccgttcc agggggaaaa gtcaggccga gcatcgttca tccgagaaac cgctccagac 2340
agccgtgaag ccggcattcg aagaggcttt tccccgggga aaagcctctt tttcaataat 2400
cgaattccgg tctttgagta ccgatgcctc tgtattcatt ggcagagatc gcgactgccc 2460
ggaggctgca gatgttgttc tgtcttctga tcggatagac gacatacagc atttcgcggc 2520
cgtacgggtc aatcgttgac gaatgaagga aaacctcagt tcctctccgc caaaatctcg 2580
tattcgccgg agctgtaata atctgccctt cataaggctc ataaattctc tgttcataat 2640
gcgcagccgg ctgataaggg gcgtatacat cttcaggtgc atagccggga gcgggggtgt 2700
agggatagcg atttggatac atatgataac ctctttccca cttcgttttt tggttttcat 2760
ctttaagatt atattcaggt aaatgcctat ttgtatgggc gaaaatctca gcttttcggc 2820
tcttttttta ttgaatggac gttgtgtatg cctatttcta tcaagcgctg ttttctgtta 2880
ttctataatc aatagaatgg attagttgtt tagggaatca tttcctttat aaatcaagaa 2940
aatttggaca aatggtggtt tagtttttaa aacgaaatgt tataatacaa cataagaatc 3000
gcactatcat gaagccggaa gatgcatcgg gcagcaaccg gagcgcccct tgcacctttg 3060
tcgatagaga aagagggaat gacaattgtt tttacacggt actagcagac aaaatgaaag 3120
agggcacctc gaaatcggcg gtgtcgatgt tctatcattg gcagaaagat acggaacacc 3180
tctttatgta tacgatgtcg cgctgattag agagcgcgcc cgaaaattcc agaaggcatt 3240
caaggaagcc ggtttaaaag cgcaggtagc gtatgcaagc aaggcgtttt catcggttgc 3300
catgattcag cttgccgaac aagaggggct gtctctggat gtggtatcgg gaggagagct 3360
tttcactgcg atcaaagcag ggttcccagc tgagcggatt cattttcacg gaaacaataa 3420
gagccctgaa gaactagcca tggcgctgga gcatcaaatc ggctgcatcg tgctcgataa 3480
ctttcacgag atcgccatta cagaagatct ttgcaagcga tcaggacaaa ctgtagacgt 3540
tttgctcaga atcactccgg gagttgaagc gcacacgcac gattatatta cgacggggca 3600
ggaagattcc aaattcggtt ttgatctgca taatggacag gtcgaacaag ccatcgaaca 3660
agtcctccgc tcgtctgcgt ttaagctcct cggcgtgcac tgccacatcg gttcgcaaat 3720
ttttgatacg gcaggatttg tccttgcagc agacaagatt ttcgagaagc ttgcggaatg 3780
gcgggagact tactctttca ttccggaagt gctcaatctt ggcgggggct tcggcatccg 3840
ctatacaaaa gacgacgagc cgcttgcagc tgatgtttat gttgaaaaaa tcatcgaggc 3900
ggtcaaagca aatgccgagc atttcggctt tgacatccct gagatttgga tcgaaccagg 3960
ccggtctctc gtcggtgatg cggggactac gctgtacacg atcggttctc aaaaagaggt 4020
gccgggcatt cgcaaatatg tagccatcga cggcggcatg agcgataata tcaggccggc 4080
gctttatgag gcaaaatatg aagcagccgt cgccaacagg atgaacgatg cttgtcatga 4140
taccgcatca atcgcaggaa aatgctgcga aagcggagat atgctgattt gggatttgga 4200
aatccccgaa gttcgcgacg gagatgtgct cgccgttttc tgcaccggtg cgtacggcta 4260
cagcatggcc aacaactaca accgcattcc gcgcccggcc gtcgtctttg tcgaggacgg 4320
ggaagcgcag ctcgtcattc agagagagac gtatgaggat atcgtcaagc tggatctgcc 4380
gctgaaatcg aaagtcaaac aataaaaaaa tggagattcc ctaagagggg ggtctccatt 4440
tttaattcaa gctgataaac agctgacatc aactaaaagt ttcattaaat actttgaaaa 4500
aagttgttga cttaaaagaa gctaaatgtt atagtaattg tacagaatag tcttttaagt 4560
aagtctactc tgaatttttt taaaaggaga gggtaaagat gaaacaacaa aaacggcttt 4620
acgcccgatt gctgacgctg ttatttgcgc tcatcttctt gctgcctcat tctgcagcta 4680
gcgcagcagc gacaaacgga acaatgatgc agtatttcga gtggtatgta cctaacgacg 4740
gccagcaatg gaacagactg agaacagatg ccccttactt gtcatctgtt ggtattacag 4800
cagtatggac accgccggct tataagggca cgtctcaagc agatgtgggg tacggcccgt 4860
acgatctgta tgatttaggc gagtttaatc aaaaaggtac agtcagaacg aagtatggca 4920
caaaaggaga acttaaatct gctgtcaaca cgctgcattc aaatggaatc caagtgtatg 4980
gtgatgtcgt gatgaatcat aaagcaggtg ctgattatac agaaaacgta acggcggtgg 5040
aggtgaatcc gtctaataga tatcaggaaa cgagcggcga atataatatt caggcatgga 5100
caggcttcaa ctttccgggc agaggaacaa cgtattctaa ctggaaatgg cagtggttcc 5160
attttgatgg aacggattgg gaccagagca gaagcctctc tagaatcttc aaattccatg 5220
gaaaggcgtg ggactggccg gtttcttcag aaaacggaaa ttatgactat ctgatgtacg 5280
cggactatga ttatgaccat ccggatgtcg tgaatgaaat gaaaaagtgg ggcgtctggt 5340
atgccaacga agttgggtta gatggataca gacttgacgc ggtcaaacat attaaattta 5400
gctttctcaa agactgggtg gataacgcaa gagcagcgac gggaaaagaa atgtttacgg 5460
ttggcgaata ttggcaaaat gatttagggg ccctgaataa ctacctggca aaggtaaatt 5520
acaaccaatc tctttttgat gcgccgttgc attacaactt ttacgctgcc tcaacagggg 5580
gtggagcgta cgatatgaga aatattctta ataacacgtt agtcgcaagc aatccgacaa 5640
aggctgttac gttagttgag aatcatgaca cacagcctgg acaatcactg gaatcaacag 5700
tccaaccgtg gtttaaaccg ttagcctacg cgtttattct cacgagaagc ggaggctatc 5760
ctgcggtatt ttatggagat atgtacggta caaaaggaac gacaacatat gagatccctg 5820
ctcttaaatc taaaatcgaa cctttgctta aggctagaaa agactatgct tatggaacac 5880
agagagacta tattgataac ccggatgtca ttggctggac gagagaaggg gactcaacga 5940
aagccaagag cggtctggcc acagtgatta cagatgggcc gggcggttca aaaagaatgt 6000
atgttggcac gagcaatgcg ggtgaaatct ggtatgattt gacagggaat agaacagata 6060
aaatcacgat tggaagcgat ggctatgcaa catttcctgt caatgggggc tcagtttcag 6120
tatgggtgca gcaatgaaag cttctcgagg ttaacagagg acggatttcc tgaaggaaat 6180
ccgttttttt attttcaagc acgaaaaaca cttcccggtg atcgggaggt gttttttgtt 6240
aaaaagatca tgacatgcat agaacagcga ccgggctaat tgtatataat attgtgaatt 6300
taacaaaaaa tttacaaagg agatgataaa ggcaatgacc agggtgaaaa ggatgagatt 6360
tgctgatttg ttggatttag aggcggagta gatgaaaccg gccaaagtat ccctactcca 6420
ccgattgctc cagtgcctga agcaatgtgt tgattgtaac acagtaaatc gttttacagc 6480
aataaacatt tttgtgaata ttttattgat ttcggctgtg atctcattcc catattctgc 6540
tgcggcccat ggcgcaacac agtccggcga tcaatattca agctttgaag aattggagcg 6600
gaatgaagat ccagcttctt accgaattac ggagaagaac gcaagagtgc cgatgctcat 6660
catggccatc catggaggcg gcatcgaacc cggaacgagc gaaatcgcca atgaagtgtc 6720
caaaaactat tccctgtact tgtttgaagg gctgaaatca tcaggcaata cggaccttca 6780
cattacaagc acgcgttttg acgagccagc ggcgctcgca attactgcaa gccaccagta 6840
tgtcatgtcg ctccacggct attacagtga agaccgcgat attaaagtag gcggcacaga 6900
ccgcgctaaa atcagaatat tggttgatga gctgaaccgc tcggggtttg ccgctgaaat 6960
gctggggaca gatgacaagt atgccggaac ccatccgaat aacatcgcca acaagtcgct 7020
ttccgggctg agcattcagc ttgaaatgag cacgggtttc cgcaaatctt tattcgaccg 7080
gtttacacta aaagacaggg cggcgacgca aaacgaaacg ttttaccgat ttacaaagct 7140
gctgacagat tttattcatg aaaactatga agaagacgga ggggatttcc cctctgcaaa 7200
aataaaacac ccccttcaag tgaaaaagga ggtgtttcgg cggttgtgtt aaccgttgga 7260
ctctgaggtg ccgccgccgg tgaatacgga aacgatggcg ttccacagag acacaaagaa 7320
gtcgatcagt ttttgaagaa agttttgtcc ttcttcagaa tccaagaatt tcgtgatttt 7380
atcctttgct ttgtcaagct ggtctccaac ctggttccag tcgatattaa tatttttcat 7440
gttattaaat aaagatataa gagagttttt ctgatcttct gtgagtgtca cgccaagttc 7500
ggaagcagcc gaatcaatcg ttttctccaa ttcctctttt gactcgggaa ctccgttttt 7560
cgagatttct tccttgactt tggccatcag cgctgacgcg ttttcactgc cgattttctc 7620
gccaagctct gaagtggtga caagctcttc attcgcgacc tttttcacat cttcggaaat 7680
tttttcgccc gaagtcgttt catacgcttt catcaatccg gttaaagcgg ctgtgcctga 7740
cacttcaaac ggagcggtga catagacttt ggcgtctttt acaccggccg tcatcagcgc 7800
gttcaaatac atctcatctg taattctgct gatattgtgt gtctgaactt ccaaaccggt 7860
gccttttttc gctacggtaa ttgaagaaga agaaatcgct cttgttccga tttgtgcttt 7920
cggtatataa tcccctaaat atttatgctc ctcatcattt gtcacctcga tgatggtcgc 7980
attttcaggc gcattcattt cttttaatac tttttgtctg tcctggcttg acaagtcttt 8040
ccccagcgtg acgatgacat cacccactgc ggcgtcagcg aagctgacct gcgggaaaat 8100
gagcagacac aatgctgtaa agattcctag tatcgatttt ttcaagctca atgccctcct 8160
taaaaatgca ggcttcaggc agaattgctg tacttttaaa gaagcctgcc ggaacggaaa 8220
taatgcgttc cgaaatatag acggatgaaa gatgagtgag gtttcaaaga aaaaaagaga 8280
gaattttctc ttcaagtcaa atgccctccc ggcatcgtat ctcgccgctc ttttatcatt 8340
catgattttc acaggcgatt caaccttttt ttaaaatttt ttacaaaaac gatacaagag 8400
cggcgtttat ttcggtcgat tggctctctg cttcttcaat atgatataat gacccttgtg 8460
aaatgaaagg agagaatcaa gatggctaaa aaaggataca tacaactgac aaacggcaaa 8520
aaaatcgagt ttgaactata tccggatgcg gcgccgggaa ctgtcgccaa ctttgaaaaa 8580
cttgcaaacg aagggttcta tgacgggctg aagttccacc gcgtcatccc gggcttcgtc 8640
agccagggag gctgcccgca cggcaccgga acaggcggac ctggatatac gattaaatgc 8700
gagacagaag ggaatccgca caaacacgaa gccggttctc tctcaatggc tcacgcagga 8760
aaagataccg gaggcagcca attttttatc gtccatgagc ctcagccgca cttgaacggc 8820
gttcacaccg ttttcggaaa ggtcacatca ggccttgatg ccgtcacttc aatggagcag 8880
ggacaaggca tggaaaaagt cgaagtattt gatgcataat cagagagcgc aaaaaacagc 8940
ccgcttagcc gggctgtttt tttgtctgta acggtgttta ttttccaggt gcaacaggac 9000
ttgaggccga ttcttcgtcc acatcctgat aggaaataac gatgctaata aataaaataa 9060
ttgtgaaaaa atgacccttt atgtaaaata tattcaagtg aagagctaga tagagaacgc 9120
aatctgtaaa aaaggaaggg gcgtaagggg tgagcgtaaa aatcccatcg acggcagtcg 9180
gcgtaaaaat taatgactgg tataacgcga tacg 9214
<210> 147
<211> 483
<212> PRT
<213> 人工序列
<220>
<223> 合成的
<400> 147
Ala Ala Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr Val Pro
1 5 10 15
Asn Asp Gly Gln Gln Trp Asn Arg Leu Arg Thr Asp Ala Pro Tyr Leu
20 25 30
Ser Ser Val Gly Ile Thr Ala Val Trp Thr Pro Pro Ala Tyr Lys Gly
35 40 45
Thr Ser Gln Ala Asp Val Gly Tyr Gly Pro Tyr Asp Leu Tyr Asp Leu
50 55 60
Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly Thr Lys
65 70 75 80
Gly Glu Leu Lys Ser Ala Val Asn Thr Leu His Ser Asn Gly Ile Gln
85 90 95
Val Tyr Gly Asp Val Val Met Asn His Lys Ala Gly Ala Asp Tyr Thr
100 105 110
Glu Asn Val Thr Ala Val Glu Val Asn Pro Ser Asn Arg Asn Gln Glu
115 120 125
Thr Ser Gly Glu Tyr Asn Ile Gln Ala Trp Thr Gly Phe Asn Phe Pro
130 135 140
Gly Arg Gly Thr Thr Tyr Ser Asn Phe Lys Trp Gln Trp Phe His Phe
145 150 155 160
Asp Gly Thr Asp Trp Asp Gln Ser Arg Ser Leu Ser Arg Ile Phe Lys
165 170 175
Phe Thr Gly Lys Ala Trp Asp Trp Pro Val Ser Ser Glu Asn Gly Asn
180 185 190
Tyr Asp Tyr Leu Met Tyr Ala Asp Tyr Asp Tyr Asp His Pro Asp Val
195 200 205
Val Asn Glu Met Lys Lys Trp Gly Val Trp Tyr Ala Asn Glu Val Gly
210 215 220
Leu Asp Gly Tyr Arg Leu Asp Ala Val Lys His Ile Lys Phe Ser Phe
225 230 235 240
Leu Lys Asp Trp Val Asp Asn Ala Arg Ala Ala Thr Gly Lys Glu Met
245 250 255
Phe Thr Val Gly Glu Tyr Trp Gln Asn Asp Leu Gly Ala Leu Asn Asn
260 265 270
Tyr Leu Ala Lys Val Asn Tyr Asn Gln Ser Leu Phe Asp Ala Pro Leu
275 280 285
His Tyr Asn Phe Tyr Ala Ala Ser Thr Gly Gly Gly Tyr Tyr Asp Met
290 295 300
Arg Asn Ile Leu Asn Asn Thr Leu Val Ala Ser Asn Pro Thr Lys Ala
305 310 315 320
Val Thr Leu Val Glu Asn His Asp Thr Gln Pro Gly Gln Ser Leu Glu
325 330 335
Ser Thr Val Gln Pro Trp Phe Lys Pro Leu Ala Tyr Ala Phe Ile Leu
340 345 350
Thr Arg Ser Gly Gly Tyr Pro Ser Val Phe Tyr Gly Asp Met Tyr Gly
355 360 365
Thr Lys Gly Thr Thr Thr Arg Glu Ile Pro Ala Leu Lys Ser Lys Ile
370 375 380
Glu Pro Leu Leu Lys Ala Arg Lys Asp Tyr Ala Tyr Gly Thr Gln Arg
385 390 395 400
Asp Tyr Ile Asp Asn Pro Asp Val Ile Gly Trp Thr Arg Glu Gly Asp
405 410 415
Ser Thr Lys Ala Lys Ser Gly Leu Ala Thr Val Ile Thr Asp Gly Pro
420 425 430
Gly Gly Ser Lys Arg Met Tyr Val Gly Thr Ser Asn Ala Gly Glu Ile
435 440 445
Trp Tyr Asp Leu Thr Gly Asn Asn Ser Thr Lys Ile Thr Ile Gly Ser
450 455 460
Asp Gly Tyr Ala Thr Phe Pro Val Asn Lys Gly Ser Val Ser Val Trp
465 470 475 480
Val Gln Gln
<210> 148
<211> 9516
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 148
ctgggagctg atagcacagc gatttcccgc ctcctgtcgg aagcatcgcc accgtgtccc 60
gtccctccag cacgcttttg atgatcgttt cctggccttt tttaaaagca tgataaccaa 120
aatacctgcg gagcgcctga tgaagtttat ccattcctga tcaccgtttt cgacagggcc 180
aggcggattt taaaataact gtatcggccg ccgagtccct ctttaatttg tctgatttta 240
ttggtttggt tcgctatcgc ataatcggca ataattttct gttcctcttt cgacacgtac 300
tgatcgatgg aaaaagacgg atcatgaatg gcgatttcca caatgtgatc ctcgatcgtc 360
gccaatttca gctttctaat ttttgcgatc cggtctattg tatacccttc ttttataaga 420
aagagcgttt ttctcgtcga ttgtgtcaga ccatcattca aaggaatgtc atgaatgagc 480
gtttgaaaaa gcggactttc accattttgg gcggattgaa taaaatggtg cagaacatcc 540
caaaacagtg catatatgta ccactcatca aggttcattt tttcggaaag ctgtctgaat 600
gtatagcccg cttttgttct ggatgtcagg gagtgaacaa aaatagcggc ctgttcatca 660
tgatttaaaa cagacagctt ctctttcagc tcttggtgaa actgggccgc tgtttcagca 720
gcattccggt ttcttaaata ctgtttaacc cagttctgaa tctgataatc ttttacgatc 780
ggcaaataca cgcgttcgcg gtaaagcttg ttggagagca cttggatcaa aagcgacatc 840
cttgcccaca tcactttggc tgccgcctga taatagccgc cgtgaaaatg gcggggccac 900
ggataaaggg caaaaaagcc ggcaagttcc gcttctcctt tttcggtgac ggtgtaagcg 960
ccgctttccg ctttttctct gaccaaagac tcctgcttca atcgttgaac gctggctgcc 1020
acctgctctc tcgacagagc cgaacaaaag ccgaagtatt ttgaaacggc aaataaaccg 1080
gcgtcctgta tcgtctgtga cgaccttttt ccttttaata aatgatagac cgcgcttgga 1140
gaacgctcac ccttcatgga tgacagaatg tcaagcacaa tcgcgtcaaa aaaatgaacc 1200
ggcatatcat cacctgcaat cttccggcaa cattcgatca tttcttcctt ttattttaac 1260
agattttgcg gagaaatcga cgtttaaact catataaaag gggtatgtta gcagtagaac 1320
ccttgtgtga taagcattct caatattttt gagttgaaat gtaagattaa caccattaca 1380
ataaggaatg ggaataggtt tcatatcgga tagatagagg gttaaaccat ttgttccaac 1440
gaagaacaat ctgggaggtt ttttattcat gccaaaatat acaattgtag acaaagatac 1500
gtgcatcgca tgcggagctt gtggtgctgc ggctcctgat atttatgatt acgacgatga 1560
gggaatcgca tttgtcaccc ttgacgacaa tcagggtgtc gtcgaagtcc ctgacgtctt 1620
agaagaagac atgatggacg cgtttgaagg ctgtcctaca gattcgatca aagttgcgga 1680
tgagccgttc gaaggcgacc cgcttaaaca cgaataaagc caaaaaacat ccggtgcaca 1740
aagtgccgga tgttttttta tgagataagc acggctttac caacaagcaa aaagaagccg 1800
gctaaagaca tccggcttct tctgcagctg acaatatccg ggaacatgca cccgatattg 1860
tcatgtttat ttatttggcc atgcggacgt tttccttcag ccgcggtttc agcgaaagga 1920
aaatcggcgt ggacacgagg gccacagcga tgcctttaat gaaattaaaa ggcaggattc 1980
cggccagaac tgttgtcttg agcgcctctc cagtcagcgc tggagcattt aaaaaccaag 2040
tgtaggcagg cagaaacagc agataattta aaatgctcat cgaaacggcc atcacaagcg 2100
tccctgcgaa aagagctgtg acaaaccctt tggcagaact tgattttttc agcagtacag 2160
ctgccggcag gataaacaat gttccggcaa tgaagttagc cgcctgatca atcggaacgc 2220
ccgaggcgct tcctgcaata aagtaattca gcacgttttt gatcgcttca acggcaatcc 2280
cggctcccgg accgtacaaa ataacagcga gcaatgccgg gatatcactg aaatcgattt 2340
ttaaatacgg gaatgccccc aggatcggaa agctcagcat cattaaaata aatgcgatgc 2400
tgctcagcat gctgatagag acgagacgtc tcaccttgtt gtgtttcatt ttgtcactct 2460
ctccttttcg atcacatctc acgaaaagag gaatggttct ttcccctgtc ctaaacaaaa 2520
aacccgcttt attgaaaaag cggggctgtt ttacagacag gtcaaataaa cgtttgaaaa 2580
tgttcatttc aaaacgcgcg gaacctccat cttctcccat ccagactata ctgtcggctt 2640
cggaatcgca ccgaatcctg cccataaaaa ggctcgcggg cttagagcgc ttgctcatca 2700
ccgccggtag ggaatttcac cctgccccga agattgatct tatttatttt taatactgat 2760
attattataa attaattgtg aaaaaatgta caggtgcaaa gcttattgcg ctgttttggg 2820
acatcctgca cgatatttcg gtaaactcac tttttccgca tactaaaaac cgcacattca 2880
cagttatttc atttttaatt ttcgtctttc cgcgtgaaac tcattgacac tctttatgga 2940
atatggtaaa ttatcagata tttatgacgc ttatttagga ggaaatctta catgtttcga 3000
gtattggtct cagataaaat gtccagcgac ggcctcaaac cattaatgga agcagatttt 3060
attgaaattg tagaaaagaa tgttgcggaa gcggaagacg agcttcatac gtttgacgcg 3120
ctcttggtgc ggagcgccac gaaggtaacc gaagagctgt ttaaaaagat gacttcgctg 3180
aaaatcgtcg ccagagcagg tgtcggcgtc gacaatatcg atattgacga ggcgacaaaa 3240
cacggtgtta tcgtcgtaaa cgcgccaaac gggaatacaa tttcaaccgc tgaacatacc 3300
tttgcaatgt tttcagcgtt aatgagacat attccgcagg caaacatctc cgtgaaatca 3360
agggagtgga atcgttcggc ttacgtcggt tcagagcttt acggaaaaac gctcggcatc 3420
atcggaatgg gccgcatcgg aagcgaaatc gcgagccgcg caaaagcatt cggtatgacc 3480
gttcatgtat ttgacccgtt cctgacccaa gaaagggcaa gcaagctcgg cgttaacgcg 3540
aacagctttg aagaagttct ggcatgcgcc gacatcatta cggttcatac cccgctcacg 3600
aaagaaacga agggactttt gaacaaagaa accatcgcaa aaacgaaaaa aggcgttcgt 3660
ctcgttaact gtgcaagagg cggcatcatc gatgaagcag cgcttttgga agctctggaa 3720
agcggacatg tcgctggcgc tgccttggat gtattcgaag tcgagcctcc ggtcgattca 3780
aaactgatcg atcatccgct tgtagtcgcg actcctcact tgggcgcctc aacaaaagaa 3840
gcccagctga atgtcgctgc acaagtgtcc gaagaagtcc ttcagtatgc gcaaggaaac 3900
cctgtgatgt ccgcgatcaa ccttccggcc atgacaaagg attcattcga aaaaatccag 3960
ccttatcatc agtttgccaa tacgatcgga aaccttgtgt ctcagtgcat gaatgagcct 4020
gttcaagatg tagccatcca atatgaaggc tccatcgcca aacttgaaac gtcatttatt 4080
acgaaaagcc ttttggccgg atttctgaag ccgagggtcg cggctaccgt taacgaagtg 4140
aatgccggca ccgttgcgaa agagcgcggc atcagcttca gcgaaaaaat ttcttccaat 4200
gagtcaggct atgaaaactg catctctgtg actgtcacgg gagatgtaac aacattctct 4260
ttaagagcga cgtacattcc gcacttcggc ggacgcatcg ttgccttaaa cggctttgat 4320
attgattttt atccggctgg acaccttgtc tacattcacc accaggataa accaggggct 4380
atcggccatg tcggacgaat tttaggagac catgacatca atatcgccac tatgcaggta 4440
ggccgaaaag aaaaaggcgg agaagcgatc atgatgcttt cctttgaccg ccaccttgag 4500
gacgatattt tagctgagct gaaaaacatc ccggatatcg tgtctgttaa agccatcgac 4560
cttccttaag tcgctgataa acagctgaca tcaatatcct attttttcaa aaaatatttt 4620
aaaaagttgt tgacttaaaa gaagctaaat gttatagtaa taaaacagaa tagtctttta 4680
agtaagtcta ctctgaattt ttttaaaagg agagggtaaa gaatgaaaca acaaaaacgg 4740
ctttacgccc gattgctgac gctgttattt gcgctcatct tcttgctgcc tcattctgca 4800
gctagcgcag cagcgacaaa cggaacaatg atgcagtatt tcgagtggta tgtacctaac 4860
gacggccagc aatggaacag actgagaaca gatgcccctt acttgtcatc tgttggtatt 4920
acagcagtat ggacaccgcc ggcttataag ggcacgtctc aagcagatgt ggggtacggc 4980
ccgtacgatc tgtatgattt aggcgagttt aatcaaaaag gtacagtcag aacgaagtat 5040
ggcacaaaag gagaacttaa atctgctgtc aacacgctgc attcaaatgg aatccaagtg 5100
tatggtgatg tcgtgatgaa tcataaagca ggtgctgatt atacagaaaa cgtaacggcg 5160
gtggaggtga atccgtctaa tagaaatcag gaaacgagcg gcgaatataa tattcaggca 5220
tggacaggct tcaactttcc gggcagagga acaacgtatt ctaacttcaa atggcagtgg 5280
ttccattttg atggaacgga ttgggaccag agcagaagcc tctctagaat cttcaaattc 5340
acgggaaagg cgtgggactg gccggtttct tcagaaaacg gaaattatga ctatctgatg 5400
tacgcggact acgattatga ccatccggat gtcgtgaatg aaatgaaaaa gtggggcgtc 5460
tggtatgcca acgaagttgg gttagatgga tacagacttg acgcggtcaa acatattaaa 5520
tttagctttc tcaaagactg ggtggataac gcaagagcag cgacgggaaa agaaatgttt 5580
acggttggcg aatattggca aaatgattta ggggccctga ataactacct ggcaaaggta 5640
aattacaacc aatctctttt tgatgcgccg ttgcattaca acttttacgc tgcctcaaca 5700
gggggtggat attacgatat gagaaatatt cttaataaca cgttagtcgc aagcaatccg 5760
acaaaggctg ttacgttagt tgagaatcat gacacacagc ctggacaatc actggaatca 5820
acagtccaac cgtggtttaa accgttagcc tacgcgttta ttctcacgag aagcggaggc 5880
tatccttctg tattttatgg agatatgtac ggtacaaaag gaacgacaac aagagagatc 5940
cctgctctta aatctaaaat cgaacctttg cttaaggcta gaaaagacta tgcttatgga 6000
acacagagag actatattga taacccggat gtcattggct ggacgagaga aggggactca 6060
acgaaagcca agagcggtct ggccacagtg attacagatg ggccgggcgg ttcaaaaaga 6120
atgtatgttg gcacgagcaa tgcgggtgaa atctggtatg atttgacagg gaataactca 6180
acaaaaatca cgattggaag cgatggctat gcaacatttc ctgtcaataa aggctcagtt 6240
tcagtatggg tgcagcaata gaagcttctc gaggttaaca gaggacggat ttcctgaagg 6300
aaatccgttt ttttatttta cagaagctgc ggaacctgaa aagaattcct ttcaggttcc 6360
gtttttttta ggaattctcc ctgatctcaa gcatctggcg gggataaatc cgctctcctt 6420
tcaaatcgtt ccattctttg aggcgctgta cagttacgcc cattttttcg gcgatatgat 6480
gaagcgtatc ccctttccgc actacatatg taccggtctt cgattcatcg tcatgaaggc 6540
ggagtgtttg gccggccttg agatttgaat gtttcaaccc gtttattctc atgatctcct 6600
cgatggatat accgctatcc ttgctgattc tccagagcgt gtcccctttt tgaacggtca 6660
ccgcaccgct cattgtcccg gcgttttgat aaacgtggat agaattttgc cggaacgcct 6720
cctcacgaag caccgtcagc ggattgattg catatctttt atcttcagtc catgaaccgt 6780
gatgcatttc aaaatgcagg tgggttccgg tcgatattcc cgtattgccg atgattccga 6840
tttgctcgcc ttttttcacc cgctcctttt cctttttcag gcgtttgctt aagtgggcat 6900
aaacggtttc atatccgttg tcatgtttaa taaatatcac ttggccgtag gagtcggatt 6960
gatacgattt gcttatcgtt ccgtctgcgg ctgccgctac tgcttcccct tcgggagcag 7020
cgatgtcaag ccccttatgc tttccgcctc tcgtaccgaa ttgatctgtg atctctcctt 7080
taatcggttc aatccactct gaggcttccg cccccggggc attgacgaaa agcgccaatc 7140
ccgaaagcca tgcgatcgcg aacaggaagt tttgatgtct gagtttcttc aaggttttcc 7200
atatcctcct attacatgca tcttcggtaa aattgccccc tattcggaga cagcttagta 7260
tacttccaaa tcaatacaat ttatacatta aaaaaagact ccgcacaggg agtcttttag 7320
ttttctatcg tcatcggatt cggtgcgtac ggaacctgta cagatttcga caggtcatag 7380
gcgccgacct tggttatgga tgcgttttta aatttcactt ttgtgaagcc gaaatctttc 7440
gcggtcaata gaaggccttc caccatcaag acatcttcgg gtttattttc aatattcgcg 7500
gaggaagaaa attgaatgat cagttctttt ccattctttt gaatatcttc aatcggcgta 7560
tcatcggata aaatgggttt taaatgagtg ccgctttctt cgtttttcat catcttaatc 7620
gcttcctgca ccgattcgta agattcgctt gaaggtgcaa ggaaccggcg cccgtctgag 7680
ctttcatata aatagtagca tttttgcgtc tggtgcataa tcgccatatc ggcgagcatt 7740
ccgaatgttt caaattcaac acccgattta tcattggaaa taaacagaac agaatcatac 7800
gatccccatt taaaggtttc gttgatcaca tttttcagcc gttcgaaatc ttcgactgat 7860
agctccggta ttttctcatc aacttgaatc ttcagttttt tattgttttt ctgctctttg 7920
aacttcacct tatcaaggta agctgtgtca aatgatgtaa actggtccac tccaagccgg 7980
ctgtaagcgt gaagcgcatc ttcaagattt gtcatgccag tgcttttctc gaggcttacc 8040
gggacaacga cagacttgga ctcgtcaagg aaagcgaagg tgatatagtc gtctttttga 8100
ttctgtgaga cgacaaacgt atttgcaggt tcagacttgg cagcatcagc ctccgtctgc 8160
accaattttc cgtcagaaga aatgttggcg tcggcgctgt tttgagatct gatctgttcg 8220
attaactggg gagtgatcag catcagaaga aagagaacca aaactgtagc agcaaatgcg 8280
ccgacccgtt ttttcggtga tttacgcttt ggtgcgagaa tcagcttttg atagatctga 8340
tttgccgaac gattatcctt aaccgttgga agttggctta gtaacgcctt cagccgttct 8400
tcgttccatt ctgacttctt cattctttgg atcctccttc aaaagctcca tctgtttacg 8460
aagcactttc agaccgcggt gctgagtggt ttttaccttg ctttcggaaa aattcaaggc 8520
ttttgctgtt tcactgatcg aatatccttg aataaaacgc aagacgataa ctgatctttg 8580
gtcaagcgta cacttgtcta gggcctcgaa aatttccttt aggttttcat tttgcatcac 8640
gatgtcctca ggcagaggct tgcggtcttt tacatcttgt ttctcccagt caaacgtccc 8700
caaaatccgc tggcggatcg tctgctgctt tctgaaccag tcgatcgcaa cgtgccgcgc 8760
aatcgaaaga agccaggttt tttcgctgct cctgccttca aatgtttcgt aagaatgcag 8820
gacgcggatg tatacttcct gaactaagtc ttccgcctga tttttgtctt ttaccatata 8880
aaataaaaac tgaaataaat cctgatgata ctgatcatat attttctgaa aggtttcttc 8940
cacctgaaac ccctccgttc aatttattgt cgtttgtcaa tcttaaaagg ttacattaca 9000
actattacaa ctatattacg aacatatgaa aatggaaagg gggttttgcg aaagttaagc 9060
ttaattttaa cttaacaagc acaaaagcac ccgttctaaa tgaacaggtg ccaaggttat 9120
aggagcccac attttcacta agctgtgccc ttacaaggct ttcgttctcc tgaccggagc 9180
gttgcggatc cgctgaaatg aactaatttc aatccgttta tgactttaag tccaattgtt 9240
ggcgaagctt tttggaaatc tccattctct tttcgtcagt cactaggtga taccataagc 9300
cgtcaacttt tttgtcttca ccttcaattt ctacctgctt aatatcttta gcagcttctt 9360
tataagaact ttgaatatcg atcatcttgt ccagtgttaa gtttgttttg atgttttttt 9420
ctaatgctgc caaaatctct tgatagtttg ttaatgattt taattgtgca ccttcatgaa 9480
taatttcgtt gataatctca cgctgacgct cttgac 9516
<210> 149
<211> 1452
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 149
gcagcgacaa acggaacaat gatgcagtat ttcgagtggt atgtacctaa cgacggccag 60
caatggaaca gactgagaac agatgcccct tacttgtcat ctgttggtat tacagcagta 120
tggacaccgc cggcttataa gggcacgtct caagcagatg tggggtacgg cccgtacgat 180
ctgtatgatt taggcgagtt taatcaaaaa ggtacagtca gaacgaagta tggcacaaaa 240
ggagaactta aatctgctgt caacacgctg cattcaaatg gaatccaagt gtatggtgat 300
gtcgtgatga atcataaagc aggtgctgat tatacagaaa acgtaacggc ggtggaggtg 360
aatccgtcta atagaaatca ggaaacgagc ggcgaatata atattcaggc atggacaggc 420
ttcaactttc cgggcagagg aacaacgtat tctaacttca aatggcagtg gttccatttt 480
gatggaacgg attgggacca gagcagaagc ctctctagaa tcttcaaatt cacgggaaag 540
gcgtgggact ggccggtttc ttcagaaaac ggaaattatg actatctgat gtacgcggac 600
tacgattatg accatccgga tgtcgtgaat gaaatgaaaa agtggggcgt ctggtatgcc 660
aacgaagttg ggttagatgg atacagactt gacgcggtca aacatattaa atttagcttt 720
ctcaaagact gggtggataa cgcaagagca gcgacgggaa aagaaatgtt tacggttggc 780
gaatattggc aaaatgattt aggggccctg aataactacc tggcaaaggt aaattacaac 840
caatctcttt ttgatgcgcc gttgcattac aacttttacg ctgcctcaac agggggtgga 900
tattacgata tgagaaatat tcttaataac acgttagtcg caagcaatcc gacaaaggct 960
gttacgttag ttgagaatca tgacacacag cctggacaat cactggaatc aacagtccaa 1020
ccgtggttta aaccgttagc ctacgcgttt attctcacga gaagcggagg ctatccttct 1080
gtattttatg gagatatgta cggtacaaaa ggaacgacaa caagagagat ccctgctctt 1140
aaatctaaaa tcgaaccttt gcttaaggct agaaaagact atgcttatgg aacacagaga 1200
gactatattg ataacccgga tgtcattggc tggacgagag aaggggactc aacgaaagcc 1260
aagagcggtc tggccacagt gattacagat gggccgggcg gttcaaaaag aatgtatgtt 1320
ggcacgagca atgcgggtga aatctggtat gatttgacag ggaataactc aacaaaaatc 1380
acgattggaa gcgatggcta tgcaacattt cctgtcaata aaggctcagt ttcagtatgg 1440
gtgcagcaat ag 1452
<210> 150
<211> 9215
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 150
catcggacag ctcttgcttg atatcttcaa aatgacgccg gctcatgtca tgtcaacttt 60
tgtcgtatct ggagcgatcc ttgacggatt cggcatttac gaccgtttta tcgaatttgc 120
cggtgccggg gctacagtcc cgattgtcag cttcggccac tctcttttgc acggcgcgat 180
gcaccaggct gagaaacatg gctttatcgg aatcggcatg gggatatttg aactgacatc 240
tgccggtata tctgccgcta tcttgttcgc ttttcttgtt gccgtgattt ttaaaccgaa 300
aggataaagg aaaatgccag caaaacgcaa ggtcattttg gtcacagacg gcgatatata 360
cgctgcaaaa gcaatcgaat atgcagcaag aaaaacgggt ggccgctgca tttcccaatc 420
ggcggggaat ccgagcgtta aaacaggacc ggagcttgta accatgatcc tgcaaacccc 480
tcatgatcct gtattcgtca tgtttgatga ttccggactt caaggtgaag gcccgggaga 540
gacagctatg aaatatgtag cgatgcatcc cgatatcgag gtgctcggag tcatcgccgt 600
cgcttcaaaa actcattatg cagagtggac gagagtcgat gtatcaatcg atgcagaagg 660
cgaactgaca gagtacggcg tcgataaaca cggggtcaaa gagttcgatg tcaaacgaat 720
gaatggtgat acagtctatt gccttgacca gctggatgtt ccgatcattg tcggaatcgg 780
tgatatcggt aagatgaaca gaaaagacga tgtggaaaaa ggttcgccga ttacaatgaa 840
agcggtcgag ctcattttag aaaggagcgg gtatcatgag tgctcaaaag caagagaaga 900
cgaacgtatt ccttgatcct tctaagaatg aagcgtattt caagaagcgg gtcggcatgg 960
gagaaagctt tgaccttggc gtacggaagg tctttattct cggacatgaa gttcagcttt 1020
attatgtcaa cggattgtgc gacacacaat acatcattca cctgttaaga gaactggtgc 1080
atctgaatga taaagaaaaa gaatcgggcg aggtcgaaga catcgtcgaa aacaggcttt 1140
tgaaccagca ggtttcaaaa gcggaaacgc ttgatgaagc tgtcgaccaa gtgttgtcag 1200
gactggttgc catcatcgtc gaagatgcgg gctttgcttt tatcatcgat gtcagaagct 1260
acccgggcag aacgccggaa gaacctgata cagaaaaagt cgtacgcggt gcaagggacg 1320
gactcgtcga gaacatcatc gtcaacacag ccctgattag acgccggatc agagatgagc 1380
gcttgcgcta caaaatgctt catatcggtg aacgctctaa aacagacatc tgcctctgct 1440
atttggaaga cgttgcagat cccgatcttg ttgaagtatt aaaaaaagaa attgaagatg 1500
tgaagatcga cgggctgccg atgtcggata aatcggtaga ggaattcctg gtcggccaag 1560
gctacaatcc gtttccgctt gtcaggttta cggaaagggc agacgtagcc gcaagccata 1620
ttttagaggg gcatgtcatc gtgatcgtcg atacgtcgcc aagcgtcatc atcacaccga 1680
ccactttgtt tcaccatgtt cagcatgctg aggaatacag acagacgccg gctgttggga 1740
cgtttttaag gtgggtgcgg tttttcggta ttttggcctc cacctttttg ctgccgcttt 1800
ggctgctgtt tgtcattcat ccgtcgctct tgcctgataa tttatcgttt atcgggttga 1860
ataaagacac ccatattccg attatcatgc agattttcct ggcggatctc ggcgtcgaat 1920
ttttaagaat ggccgccatt catacgccga cggcgctttc gactgcaatg ggcctgatcg 1980
ccgctgtatt gatcggcgat atcgcgatca atgtcggctt gttttctccc gaagtcattt 2040
tatacgtttc cctctcggca atcggagcct acacgacacc aagctacgag ctgagcctgg 2100
cgaataaaat ggtgaagctg tttatgctga tattggtggc gctttttaaa gtggagggat 2160
ttgtcatcgg attaacgatc ttaactatag tgatgacttc gatcaggtca ttgcgaacgc 2220
cttacttatg gcctctcctc ccgttcaatg gaaaagcgtt ttggcatgtt ctcgtgcgca 2280
cgtccgttcc agggggaaaa gtcaggccga gcatcgttca tccgagaaac cgctccagac 2340
agccgtgaag ccggcattcg aagaggcttt tccccgggga aaagcctctt tttcaataat 2400
cgaattccgg tctttgagta ccgatgcctc tgtattcatt ggcagagatc gcgactgccc 2460
ggaggctgca gatgttgttc tgtcttctga tcggatagac gacatacagc atttcgcggc 2520
cgtacgggtc aatcgttgac gaatgaagga aaacctcagt tcctctccgc caaaatctcg 2580
tattcgccgg agctgtaata atctgccctt cataaggctc ataaattctc tgttcataat 2640
gcgcagccgg ctgataaggg gcgtatacat cttcaggtgc atagccggga gcgggggtgt 2700
agggatagcg atttggatac atatgataac ctctttccca cttcgttttt tggttttcat 2760
ctttaagatt atattcaggt aaatgcctat ttgtatgggc gaaaatctca gcttttcggc 2820
tcttttttta ttgaatggac gttgtgtatg cctatttcta tcaagcgctg ttttctgtta 2880
ttctataatc aatagaatgg attagttgtt tagggaatca tttcctttat aaatcaagaa 2940
aatttggaca aatggtggtt tagtttttaa aacgaaatgt tataatacaa cataagaatc 3000
gcactatcat gaagccggaa gatgcatcgg gcagcaaccg gagcgcccct tgcacctttg 3060
tcgatagaga aagagggaat gacaattgtt tttacacggt actagcagac aaaatgaaag 3120
agggcacctc gaaatcggcg gtgtcgatgt tctatcattg gcagaaagat acggaacacc 3180
tctttatgta tacgatgtcg cgctgattag agagcgcgcc cgaaaattcc agaaggcatt 3240
caaggaagcc ggtttaaaag cgcaggtagc gtatgcaagc aaggcgtttt catcggttgc 3300
catgattcag cttgccgaac aagaggggct gtctctggat gtggtatcgg gaggagagct 3360
tttcactgcg atcaaagcag ggttcccagc tgagcggatt cattttcacg gaaacaataa 3420
gagccctgaa gaactagcca tggcgctgga gcatcaaatc ggctgcatcg tgctcgataa 3480
ctttcacgag atcgccatta cagaagatct ttgcaagcga tcaggacaaa ctgtagacgt 3540
tttgctcaga atcactccgg gagttgaagc gcacacgcac gattatatta cgacggggca 3600
ggaagattcc aaattcggtt ttgatctgca taatggacag gtcgaacaag ccatcgaaca 3660
agtcctccgc tcgtctgcgt ttaagctcct cggcgtgcac tgccacatcg gttcgcaaat 3720
ttttgatacg gcaggatttg tccttgcagc agacaagatt ttcgagaagc ttgcggaatg 3780
gcgggagact tactctttca ttccggaagt gctcaatctt ggcgggggct tcggcatccg 3840
ctatacaaaa gacgacgagc cgcttgcagc tgatgtttat gttgaaaaaa tcatcgaggc 3900
ggtcaaagca aatgccgagc atttcggctt tgacatccct gagatttgga tcgaaccagg 3960
ccggtctctc gtcggtgatg cggggactac gctgtacacg atcggttctc aaaaagaggt 4020
gccgggcatt cgcaaatatg tagccatcga cggcggcatg agcgataata tcaggccggc 4080
gctttatgag gcaaaatatg aagcagccgt cgccaacagg atgaacgatg cttgtcatga 4140
taccgcatca atcgcaggaa aatgctgcga aagcggagat atgctgattt gggatttgga 4200
aatccccgaa gttcgcgacg gagatgtgct cgccgttttc tgcaccggtg cgtacggcta 4260
cagcatggcc aacaactaca accgcattcc gcgcccggcc gtcgtctttg tcgaggacgg 4320
ggaagcgcag ctcgtcattc agagagagac gtatgaggat atcgtcaagc tggatctgcc 4380
gctgaaatcg aaagtcaaac aataaaaaaa tggagattcc ctaagagggg ggtctccatt 4440
tttaattcaa gctgataaac agctgacatc aactaaaagt ttcattaaat actttgaaaa 4500
aagttgttga cttaaaagaa gctaaatgtt atagtaattg tacagaatag tcttttaagt 4560
aagtctactc tgaatttttt taaaaggaga gggtaaagaa tgaaacaaca aaaacggctt 4620
tacgcccgat tgctgacgct gttatttgcg ctcatcttct tgctgcctca ttctgcagct 4680
agcgcagcag cgacaaacgg aacaatgatg cagtatttcg agtggtatgt acctaacgac 4740
ggccagcaat ggaacagact gagaacagat gccccttact tgtcatctgt tggtattaca 4800
gcagtatgga caccgccggc ttataagggc acgtctcaag cagatgtggg gtacggcccg 4860
tacgatctgt atgatttagg cgagtttaat caaaaaggta cagtcagaac gaagtatggc 4920
acaaaaggag aacttaaatc tgctgtcaac acgctgcatt caaatggaat ccaagtgtat 4980
ggtgatgtcg tgatgaatca taaagcaggt gctgattata cagaaaacgt aacggcggtg 5040
gaggtgaatc cgtctaatag aaatcaggaa acgagcggcg aatataatat tcaggcatgg 5100
acaggcttca actttccggg cagaggaaca acgtattcta acttcaaatg gcagtggttc 5160
cattttgatg gaacggattg ggaccagagc agaagcctct ctagaatctt caaattcacg 5220
ggaaaggcgt gggactggcc ggtttcttca gaaaacggaa attatgacta tctgatgtac 5280
gcggactacg attatgacca tccggatgtc gtgaatgaaa tgaaaaagtg gggcgtctgg 5340
tatgccaacg aagttgggtt agatggatac agacttgacg cggtcaaaca tattaaattt 5400
agctttctca aagactgggt ggataacgca agagcagcga cgggaaaaga aatgtttacg 5460
gttggcgaat attggcaaaa tgatttaggg gccctgaata actacctggc aaaggtaaat 5520
tacaaccaat ctctttttga tgcgccgttg cattacaact tttacgctgc ctcaacaggg 5580
ggtggatatt acgatatgag aaatattctt aataacacgt tagtcgcaag caatccgaca 5640
aaggctgtta cgttagttga gaatcatgac acacagcctg gacaatcact ggaatcaaca 5700
gtccaaccgt ggtttaaacc gttagcctac gcgtttattc tcacgagaag cggaggctat 5760
ccttctgtat tttatggaga tatgtacggt acaaaaggaa cgacaacaag agagatccct 5820
gctcttaaat ctaaaatcga acctttgctt aaggctagaa aagactatgc ttatggaaca 5880
cagagagact atattgataa cccggatgtc attggctgga cgagagaagg ggactcaacg 5940
aaagccaaga gcggtctggc cacagtgatt acagatgggc cgggcggttc aaaaagaatg 6000
tatgttggca cgagcaatgc gggtgaaatc tggtatgatt tgacagggaa taactcaaca 6060
aaaatcacga ttggaagcga tggctatgca acatttcctg tcaataaagg ctcagtttca 6120
gtatgggtgc agcaatagaa gcttctcgag gttaacagag gacggatttc ctgaaggaaa 6180
tccgtttttt tattttcaag cacgaaaaac acttcccggt gatcgggagg tgttttttgt 6240
taaaaagatc atgacatgca tagaacagcg accgggctaa ttgtatataa tattgtgaat 6300
ttaacaaaaa atttacaaag gagatgataa aggcaatgac cagggtgaaa aggatgagat 6360
ttgctgattt gttggattta gaggcggagt agatgaaacc ggccaaagta tccctactcc 6420
accgattgct ccagtgcctg aagcaatgtg ttgattgtaa cacagtaaat cgttttacag 6480
caataaacat ttttgtgaat attttattga tttcggctgt gatctcattc ccatattctg 6540
ctgcggccca tggcgcaaca cagtccggcg atcaatattc aagctttgaa gaattggagc 6600
ggaatgaaga tccagcttct taccgaatta cggagaagaa cgcaagagtg ccgatgctca 6660
tcatggccat ccatggaggc ggcatcgaac ccggaacgag cgaaatcgcc aatgaagtgt 6720
ccaaaaacta ttccctgtac ttgtttgaag ggctgaaatc atcaggcaat acggaccttc 6780
acattacaag cacgcgtttt gacgagccag cggcgctcgc aattactgca agccaccagt 6840
atgtcatgtc gctccacggc tattacagtg aagaccgcga tattaaagta ggcggcacag 6900
accgcgctaa aatcagaata ttggttgatg agctgaaccg ctcggggttt gccgctgaaa 6960
tgctggggac agatgacaag tatgccggaa cccatccgaa taacatcgcc aacaagtcgc 7020
tttccgggct gagcattcag cttgaaatga gcacgggttt ccgcaaatct ttattcgacc 7080
ggtttacact aaaagacagg gcggcgacgc aaaacgaaac gttttaccga tttacaaagc 7140
tgctgacaga ttttattcat gaaaactatg aagaagacgg aggggatttc ccctctgcaa 7200
aaataaaaca cccccttcaa gtgaaaaagg aggtgtttcg gcggttgtgt taaccgttgg 7260
actctgaggt gccgccgccg gtgaatacgg aaacgatggc gttccacaga gacacaaaga 7320
agtcgatcag tttttgaaga aagttttgtc cttcttcaga atccaagaat ttcgtgattt 7380
tatcctttgc tttgtcaagc tggtctccaa cctggttcca gtcgatatta atatttttca 7440
tgttattaaa taaagatata agagagtttt tctgatcttc tgtgagtgtc acgccaagtt 7500
cggaagcagc cgaatcaatc gttttctcca attcctcttt tgactcggga actccgtttt 7560
tcgagatttc ttccttgact ttggccatca gcgctgacgc gttttcactg ccgattttct 7620
cgccaagctc tgaagtggtg acaagctctt cattcgcgac ctttttcaca tcttcggaaa 7680
ttttttcgcc cgaagtcgtt tcatacgctt tcatcaatcc ggttaaagcg gctgtgcctg 7740
acacttcaaa cggagcggtg acatagactt tggcgtcttt tacaccggcc gtcatcagcg 7800
cgttcaaata catctcatct gtaattctgc tgatattgtg tgtctgaact tccaaaccgg 7860
tgcctttttt cgctacggta attgaagaag aagaaatcgc tcttgttccg atttgtgctt 7920
tcggtatata atcccctaaa tatttatgct cctcatcatt tgtcacctcg atgatggtcg 7980
cattttcagg cgcattcatt tcttttaata ctttttgtct gtcctggctt gacaagtctt 8040
tccccagcgt gacgatgaca tcacccactg cggcgtcagc gaagctgacc tgcgggaaaa 8100
tgagcagaca caatgctgta aagattccta gtatcgattt tttcaagctc aatgccctcc 8160
ttaaaaatgc aggcttcagg cagaattgct gtacttttaa agaagcctgc cggaacggaa 8220
ataatgcgtt ccgaaatata gacggatgaa agatgagtga ggtttcaaag aaaaaaagag 8280
agaattttct cttcaagtca aatgccctcc cggcatcgta tctcgccgct cttttatcat 8340
tcatgatttt cacaggcgat tcaacctttt tttaaaattt tttacaaaaa cgatacaaga 8400
gcggcgttta tttcggtcga ttggctctct gcttcttcaa tatgatataa tgacccttgt 8460
gaaatgaaag gagagaatca agatggctaa aaaaggatac atacaactga caaacggcaa 8520
aaaaatcgag tttgaactat atccggatgc ggcgccggga actgtcgcca actttgaaaa 8580
acttgcaaac gaagggttct atgacgggct gaagttccac cgcgtcatcc cgggcttcgt 8640
cagccaggga ggctgcccgc acggcaccgg aacaggcgga cctggatata cgattaaatg 8700
cgagacagaa gggaatccgc acaaacacga agccggttct ctctcaatgg ctcacgcagg 8760
aaaagatacc ggaggcagcc aattttttat cgtccatgag cctcagccgc acttgaacgg 8820
cgttcacacc gttttcggaa aggtcacatc aggccttgat gccgtcactt caatggagca 8880
gggacaaggc atggaaaaag tcgaagtatt tgatgcataa tcagagagcg caaaaaacag 8940
cccgcttagc cgggctgttt ttttgtctgt aacggtgttt attttccagg tgcaacagga 9000
cttgaggccg attcttcgtc cacatcctga taggaaataa cgatgctaat aaataaaata 9060
attgtgaaaa aatgaccctt tatgtaaaat atattcaagt gaagagctag atagagaacg 9120
caatctgtaa aaaaggaagg ggcgtaaggg gtgagcgtaa aaatcccatc gacggcagtc 9180
ggcgtaaaaa ttaatgactg gtataacgcg atacg 9215
<210> 151
<211> 485
<212> PRT
<213> 人工序列
<220>
<223> 合成的
<400> 151
His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr
1 5 10 15
Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Asn Ser Asp Ala Ser
20 25 30
Asn Leu Lys Ser Lys Gly Ile Thr Ala Val Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Ala Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly
65 70 75 80
Thr Arg Ser Gln Leu Gln Ala Ala Val Thr Ser Leu Lys Asn Asn Gly
85 90 95
Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Ala Thr Glu Met Val Arg Ala Val Glu Val Asn Pro Asn Asn Arg Asn
115 120 125
Gln Glu Val Thr Gly Glu Tyr Thr Ile Glu Ala Trp Thr Arg Phe Asp
130 135 140
Phe Pro Gly Arg Gly Asn Thr His Ser Ser Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Arg Leu Asn Asn Arg
165 170 175
Ile Tyr Lys Phe Arg Gly His Gly Lys Ala Trp Asp Trp Glu Val Asp
180 185 190
Thr Glu Asn Gly Asn Tyr Asp Tyr Leu Leu Tyr Ala Asp Ile Asp Met
195 200 205
Asp His Pro Glu Val Val Asn Glu Leu Arg Asn Trp Gly Val Trp Tyr
210 215 220
Thr Asn Thr Leu Gly Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His
225 230 235 240
Ile Lys Tyr Ser Phe Thr Arg Asp Trp Ile Asn His Val Arg Ser Ala
245 250 255
Thr Gly Lys Asn Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu
260 265 270
Gly Ala Ile Glu Asn Tyr Leu Gln Lys Thr Asn Trp Asn His Ser Val
275 280 285
Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Lys Ser Gly
290 295 300
Gly Asn Tyr Asp Met Arg Asn Ile Phe Asn Gly Thr Val Val Gln Arg
305 310 315 320
His Pro Ser His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro
325 330 335
Glu Glu Ala Leu Glu Ser Phe Val Glu Glu Trp Phe Lys Pro Leu Ala
340 345 350
Tyr Ala Leu Thr Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr
355 360 365
Gly Asp Tyr Tyr Gly Ile Pro Thr His Gly Val Pro Ala Met Arg Ser
370 375 380
Lys Ile Asp Pro Ile Leu Glu Ala Arg Gln Lys Tyr Ala Tyr Gly Lys
385 390 395 400
Gln Asn Asp Tyr Leu Asp His His Asn Ile Ile Gly Trp Thr Arg Glu
405 410 415
Gly Asn Thr Ala His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp
420 425 430
Gly Ala Gly Gly Ser Lys Trp Met Phe Val Gly Arg Asn Lys Ala Gly
435 440 445
Gln Val Trp Ser Asp Ile Thr Gly Asn Arg Thr Gly Thr Val Thr Ile
450 455 460
Asn Ala Asp Gly Trp Gly Asn Phe Ser Val Asn Gly Gly Ser Val Ser
465 470 475 480
Ile Trp Val Asn Lys
485
<210> 152
<211> 9522
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 152
ctgggagctg atagcacagc gatttcccgc ctcctgtcgg aagcatcgcc accgtgtccc 60
gtccctccag cacgcttttg atgatcgttt cctggccttt tttaaaagca tgataaccaa 120
aatacctgcg gagcgcctga tgaagtttat ccattcctga tcaccgtttt cgacagggcc 180
aggcggattt taaaataact gtatcggccg ccgagtccct ctttaatttg tctgatttta 240
ttggtttggt tcgctatcgc ataatcggca ataattttct gttcctcttt cgacacgtac 300
tgatcgatgg aaaaagacgg atcatgaatg gcgatttcca caatgtgatc ctcgatcgtc 360
gccaatttca gctttctaat ttttgcgatc cggtctattg tatacccttc ttttataaga 420
aagagcgttt ttctcgtcga ttgtgtcaga ccatcattca aaggaatgtc atgaatgagc 480
gtttgaaaaa gcggactttc accattttgg gcggattgaa taaaatggtg cagaacatcc 540
caaaacagtg catatatgta ccactcatca aggttcattt tttcggaaag ctgtctgaat 600
gtatagcccg cttttgttct ggatgtcagg gagtgaacaa aaatagcggc ctgttcatca 660
tgatttaaaa cagacagctt ctctttcagc tcttggtgaa actgggccgc tgtttcagca 720
gcattccggt ttcttaaata ctgtttaacc cagttctgaa tctgataatc ttttacgatc 780
ggcaaataca cgcgttcgcg gtaaagcttg ttggagagca cttggatcaa aagcgacatc 840
cttgcccaca tcactttggc tgccgcctga taatagccgc cgtgaaaatg gcggggccac 900
ggataaaggg caaaaaagcc ggcaagttcc gcttctcctt tttcggtgac ggtgtaagcg 960
ccgctttccg ctttttctct gaccaaagac tcctgcttca atcgttgaac gctggctgcc 1020
acctgctctc tcgacagagc cgaacaaaag ccgaagtatt ttgaaacggc aaataaaccg 1080
gcgtcctgta tcgtctgtga cgaccttttt ccttttaata aatgatagac cgcgcttgga 1140
gaacgctcac ccttcatgga tgacagaatg tcaagcacaa tcgcgtcaaa aaaatgaacc 1200
ggcatatcat cacctgcaat cttccggcaa cattcgatca tttcttcctt ttattttaac 1260
agattttgcg gagaaatcga cgtttaaact catataaaag gggtatgtta gcagtagaac 1320
ccttgtgtga taagcattct caatattttt gagttgaaat gtaagattaa caccattaca 1380
ataaggaatg ggaataggtt tcatatcgga tagatagagg gttaaaccat ttgttccaac 1440
gaagaacaat ctgggaggtt ttttattcat gccaaaatat acaattgtag acaaagatac 1500
gtgcatcgca tgcggagctt gtggtgctgc ggctcctgat atttatgatt acgacgatga 1560
gggaatcgca tttgtcaccc ttgacgacaa tcagggtgtc gtcgaagtcc ctgacgtctt 1620
agaagaagac atgatggacg cgtttgaagg ctgtcctaca gattcgatca aagttgcgga 1680
tgagccgttc gaaggcgacc cgcttaaaca cgaataaagc caaaaaacat ccggtgcaca 1740
aagtgccgga tgttttttta tgagataagc acggctttac caacaagcaa aaagaagccg 1800
gctaaagaca tccggcttct tctgcagctg acaatatccg ggaacatgca cccgatattg 1860
tcatgtttat ttatttggcc atgcggacgt tttccttcag ccgcggtttc agcgaaagga 1920
aaatcggcgt ggacacgagg gccacagcga tgcctttaat gaaattaaaa ggcaggattc 1980
cggccagaac tgttgtcttg agcgcctctc cagtcagcgc tggagcattt aaaaaccaag 2040
tgtaggcagg cagaaacagc agataattta aaatgctcat cgaaacggcc atcacaagcg 2100
tccctgcgaa aagagctgtg acaaaccctt tggcagaact tgattttttc agcagtacag 2160
ctgccggcag gataaacaat gttccggcaa tgaagttagc cgcctgatca atcggaacgc 2220
ccgaggcgct tcctgcaata aagtaattca gcacgttttt gatcgcttca acggcaatcc 2280
cggctcccgg accgtacaaa ataacagcga gcaatgccgg gatatcactg aaatcgattt 2340
ttaaatacgg gaatgccccc aggatcggaa agctcagcat cattaaaata aatgcgatgc 2400
tgctcagcat gctgatagag acgagacgtc tcaccttgtt gtgtttcatt ttgtcactct 2460
ctccttttcg atcacatctc acgaaaagag gaatggttct ttcccctgtc ctaaacaaaa 2520
aacccgcttt attgaaaaag cggggctgtt ttacagacag gtcaaataaa cgtttgaaaa 2580
tgttcatttc aaaacgcgcg gaacctccat cttctcccat ccagactata ctgtcggctt 2640
cggaatcgca ccgaatcctg cccataaaaa ggctcgcggg cttagagcgc ttgctcatca 2700
ccgccggtag ggaatttcac cctgccccga agattgatct tatttatttt taatactgat 2760
attattataa attaattgtg aaaaaatgta caggtgcaaa gcttattgcg ctgttttggg 2820
acatcctgca cgatatttcg gtaaactcac tttttccgca tactaaaaac cgcacattca 2880
cagttatttc atttttaatt ttcgtctttc cgcgtgaaac tcattgacac tctttatgga 2940
atatggtaaa ttatcagata tttatgacgc ttatttagga ggaaatctta catgtttcga 3000
gtattggtct cagataaaat gtccagcgac ggcctcaaac cattaatgga agcagatttt 3060
attgaaattg tagaaaagaa tgttgcggaa gcggaagacg agcttcatac gtttgacgcg 3120
ctcttggtgc ggagcgccac gaaggtaacc gaagagctgt ttaaaaagat gacttcgctg 3180
aaaatcgtcg ccagagcagg tgtcggcgtc gacaatatcg atattgacga ggcgacaaaa 3240
cacggtgtta tcgtcgtaaa cgcgccaaac gggaatacaa tttcaaccgc tgaacatacc 3300
tttgcaatgt tttcagcgtt aatgagacat attccgcagg caaacatctc cgtgaaatca 3360
agggagtgga atcgttcggc ttacgtcggt tcagagcttt acggaaaaac gctcggcatc 3420
atcggaatgg gccgcatcgg aagcgaaatc gcgagccgcg caaaagcatt cggtatgacc 3480
gttcatgtat ttgacccgtt cctgacccaa gaaagggcaa gcaagctcgg cgttaacgcg 3540
aacagctttg aagaagttct ggcatgcgcc gacatcatta cggttcatac cccgctcacg 3600
aaagaaacga agggactttt gaacaaagaa accatcgcaa aaacgaaaaa aggcgttcgt 3660
ctcgttaact gtgcaagagg cggcatcatc gatgaagcag cgcttttgga agctctggaa 3720
agcggacatg tcgctggcgc tgccttggat gtattcgaag tcgagcctcc ggtcgattca 3780
aaactgatcg atcatccgct tgtagtcgcg actcctcact tgggcgcctc aacaaaagaa 3840
gcccagctga atgtcgctgc acaagtgtcc gaagaagtcc ttcagtatgc gcaaggaaac 3900
cctgtgatgt ccgcgatcaa ccttccggcc atgacaaagg attcattcga aaaaatccag 3960
ccttatcatc agtttgccaa tacgatcgga aaccttgtgt ctcagtgcat gaatgagcct 4020
gttcaagatg tagccatcca atatgaaggc tccatcgcca aacttgaaac gtcatttatt 4080
acgaaaagcc ttttggccgg atttctgaag ccgagggtcg cggctaccgt taacgaagtg 4140
aatgccggca ccgttgcgaa agagcgcggc atcagcttca gcgaaaaaat ttcttccaat 4200
gagtcaggct atgaaaactg catctctgtg actgtcacgg gagatgtaac aacattctct 4260
ttaagagcga cgtacattcc gcacttcggc ggacgcatcg ttgccttaaa cggctttgat 4320
attgattttt atccggctgg acaccttgtc tacattcacc accaggataa accaggggct 4380
atcggccatg tcggacgaat tttaggagac catgacatca atatcgccac tatgcaggta 4440
ggccgaaaag aaaaaggcgg agaagcgatc atgatgcttt cctttgaccg ccaccttgag 4500
gacgatattt tagctgagct gaaaaacatc ccggatatcg tgtctgttaa agccatcgac 4560
cttccttaag tcgctgataa acagctgaca tcaatatcct attttttcaa aaaatatttt 4620
aaaaagttgt tgacttaaaa gaagctaaat gttatagtaa taaaacagaa tagtctttta 4680
agtaagtcta ctctgaattt ttttaaaagg agagggtaaa gaatgaaaca acaaaaacgg 4740
ctttacgccc gattgctgac gctgttattt gcgctcatct tcttgctgcc tcattctgca 4800
gctagcgcac atcataacgg tacgaacggg acaatgatgc aatactttga atggtatcta 4860
cctaatgacg gaaatcattg gaatcgatta aactctgatg cgagtaacct taaaagcaaa 4920
gggattacag cggtgtggat tcctccagca tggaagggcg cttctcaaaa tgacgtagga 4980
tacggagcct atgacctgta tgatctggga gaatttaatc aaaaaggtac cgtccgtaca 5040
aaatatggaa cacgtagtca gttacaagct gcggtaacct ccttaaaaaa taatggaatt 5100
caagtatatg gtgacgttgt tatgaatcac aaaggtggcg cagacgctac tgaaatggta 5160
agggccgttg aagtgaatcc caataaccgt aaccaagaag tgactggtga atataccatt 5220
gaagcttgga ctagatttga ttttccaggg cgaggaaata ctcattctag ctttaaatgg 5280
agatggtatc attttgatgg tgtggattgg gatcagtcac gtagactgaa caatcgcatc 5340
tataaattta gaggtcatgg caaagcttgg gattgggaag ttgatacgga aaatggtaat 5400
tatgattatt tactatacgc tgatattgat atggatcacc cagaagtagt aaatgaatta 5460
agaaattggg gtgtttggta cacaaacaca ttaggactcg atggatttag aatagatgcg 5520
gttaaacata taaagtatag ctttacgcgc gattggatta atcacgttag aagtgcaaca 5580
ggtaaaaata tgtttgcggt tgctgagttt tggaagaatg atttaggtgc aattgaaaac 5640
tatctgcaga aaacaaactg gaaccattca gtctttgatg tgccgttaca ttataatctt 5700
tataatgcat caaaaagcgg agggaactat gatatgcgaa acatatttaa tggaacggtt 5760
gttcaacgac atccaagtca tgctgtaaca tttgttgata atcatgattc gcagcctgaa 5820
gaagcattag aatcttttgt tgaagaatgg tttaaaccat tagcgtatgc gcttacatta 5880
acgcgtgaac aaggataccc ttctgtattt tacggagatt attatgggat tccaacacat 5940
ggagtgccag caatgagatc aaaaatcgat ccgattttag aagcacgtca aaagtatgca 6000
tacggaaaac aaaatgatta cttagaccat cataatatca ttggttggac gcgtgaaggg 6060
aatacagcac accccaattc aggtctagct accatcatgt ctgatggagc gggtggaagt 6120
aagtggatgt ttgttgggcg taataaggct ggtcaagtat ggagtgatat tacaggaaac 6180
cgtacaggta cggttacaat caatgcagac ggttggggca atttctctgt gaatggaggg 6240
tcagtttcta tttgggtcaa caaataaaag cttctcgagg ttaacagagg acggatttcc 6300
tgaaggaaat ccgttttttt attttacaga agctgcggaa cctgaaaaga attcctttca 6360
ggttccgttt tttttaggaa ttctccctga tctcaagcat ctggcgggga taaatccgct 6420
ctcctttcaa atcgttccat tctttgaggc gctgtacagt tacgcccatt ttttcggcga 6480
tatgatgaag cgtatcccct ttccgcacta catatgtacc ggtcttcgat tcatcgtcat 6540
gaaggcggag tgtttggccg gccttgagat ttgaatgttt caacccgttt attctcatga 6600
tctcctcgat ggatataccg ctatccttgc tgattctcca gagcgtgtcc cctttttgaa 6660
cggtcaccgc accgctcatt gtcccggcgt tttgataaac gtggatagaa ttttgccgga 6720
acgcctcctc acgaagcacc gtcagcggat tgattgcata tcttttatct tcagtccatg 6780
aaccgtgatg catttcaaaa tgcaggtggg ttccggtcga tattcccgta ttgccgatga 6840
ttccgatttg ctcgcctttt ttcacccgct ccttttcctt tttcaggcgt ttgcttaagt 6900
gggcataaac ggtttcatat ccgttgtcat gtttaataaa tatcacttgg ccgtaggagt 6960
cggattgata cgatttgctt atcgttccgt ctgcggctgc cgctactgct tccccttcgg 7020
gagcagcgat gtcaagcccc ttatgctttc cgcctctcgt accgaattga tctgtgatct 7080
ctcctttaat cggttcaatc cactctgagg cttccgcccc cggggcattg acgaaaagcg 7140
ccaatcccga aagccatgcg atcgcgaaca ggaagttttg atgtctgagt ttcttcaagg 7200
ttttccatat cctcctatta catgcatctt cggtaaaatt gccccctatt cggagacagc 7260
ttagtatact tccaaatcaa tacaatttat acattaaaaa aagactccgc acagggagtc 7320
ttttagtttt ctatcgtcat cggattcggt gcgtacggaa cctgtacaga tttcgacagg 7380
tcataggcgc cgaccttggt tatggatgcg tttttaaatt tcacttttgt gaagccgaaa 7440
tctttcgcgg tcaatagaag gccttccacc atcaagacat cttcgggttt attttcaata 7500
ttcgcggagg aagaaaattg aatgatcagt tcttttccat tcttttgaat atcttcaatc 7560
ggcgtatcat cggataaaat gggttttaaa tgagtgccgc tttcttcgtt tttcatcatc 7620
ttaatcgctt cctgcaccga ttcgtaagat tcgcttgaag gtgcaaggaa ccggcgcccg 7680
tctgagcttt catataaata gtagcatttt tgcgtctggt gcataatcgc catatcggcg 7740
agcattccga atgtttcaaa ttcaacaccc gatttatcat tggaaataaa cagaacagaa 7800
tcatacgatc cccatttaaa ggtttcgttg atcacatttt tcagccgttc gaaatcttcg 7860
actgatagct ccggtatttt ctcatcaact tgaatcttca gttttttatt gtttttctgc 7920
tctttgaact tcaccttatc aaggtaagct gtgtcaaatg atgtaaactg gtccactcca 7980
agccggctgt aagcgtgaag cgcatcttca agatttgtca tgccagtgct tttctcgagg 8040
cttaccggga caacgacaga cttggactcg tcaaggaaag cgaaggtgat atagtcgtct 8100
ttttgattct gtgagacgac aaacgtattt gcaggttcag acttggcagc atcagcctcc 8160
gtctgcacca attttccgtc agaagaaatg ttggcgtcgg cgctgttttg agatctgatc 8220
tgttcgatta actggggagt gatcagcatc agaagaaaga gaaccaaaac tgtagcagca 8280
aatgcgccga cccgtttttt cggtgattta cgctttggtg cgagaatcag cttttgatag 8340
atctgatttg ccgaacgatt atccttaacc gttggaagtt ggcttagtaa cgccttcagc 8400
cgttcttcgt tccattctga cttcttcatt ctttggatcc tccttcaaaa gctccatctg 8460
tttacgaagc actttcagac cgcggtgctg agtggttttt accttgcttt cggaaaaatt 8520
caaggctttt gctgtttcac tgatcgaata tccttgaata aaacgcaaga cgataactga 8580
tctttggtca agcgtacact tgtctagggc ctcgaaaatt tcctttaggt tttcattttg 8640
catcacgatg tcctcaggca gaggcttgcg gtcttttaca tcttgtttct cccagtcaaa 8700
cgtccccaaa atccgctggc ggatcgtctg ctgctttctg aaccagtcga tcgcaacgtg 8760
ccgcgcaatc gaaagaagcc aggttttttc gctgctcctg ccttcaaatg tttcgtaaga 8820
atgcaggacg cggatgtata cttcctgaac taagtcttcc gcctgatttt tgtcttttac 8880
catataaaat aaaaactgaa ataaatcctg atgatactga tcatatattt tctgaaaggt 8940
ttcttccacc tgaaacccct ccgttcaatt tattgtcgtt tgtcaatctt aaaaggttac 9000
attacaacta ttacaactat attacgaaca tatgaaaatg gaaagggggt tttgcgaaag 9060
ttaagcttaa ttttaactta acaagcacaa aagcacccgt tctaaatgaa caggtgccaa 9120
ggttatagga gcccacattt tcactaagct gtgcccttac aaggctttcg ttctcctgac 9180
cggagcgttg cggatccgct gaaatgaact aatttcaatc cgtttatgac tttaagtcca 9240
attgttggcg aagctttttg gaaatctcca ttctcttttc gtcagtcact aggtgatacc 9300
ataagccgtc aacttttttg tcttcacctt caatttctac ctgcttaata tctttagcag 9360
cttctttata agaactttga atatcgatca tcttgtccag tgttaagttt gttttgatgt 9420
ttttttctaa tgctgccaaa atctcttgat agtttgttaa tgattttaat tgtgcacctt 9480
catgaataat ttcgttgata atctcacgct gacgctcttg ac 9522
<210> 153
<211> 1458
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 153
catcataacg gtacgaacgg gacaatgatg caatactttg aatggtatct acctaatgac 60
ggaaatcatt ggaatcgatt aaactctgat gcgagtaacc ttaaaagcaa agggattaca 120
gcggtgtgga ttcctccagc atggaagggc gcttctcaaa atgacgtagg atacggagcc 180
tatgacctgt atgatctggg agaatttaat caaaaaggta ccgtccgtac aaaatatgga 240
acacgtagtc agttacaagc tgcggtaacc tccttaaaaa ataatggaat tcaagtatat 300
ggtgacgttg ttatgaatca caaaggtggc gcagacgcta ctgaaatggt aagggccgtt 360
gaagtgaatc ccaataaccg taaccaagaa gtgactggtg aatataccat tgaagcttgg 420
actagatttg attttccagg gcgaggaaat actcattcta gctttaaatg gagatggtat 480
cattttgatg gtgtggattg ggatcagtca cgtagactga acaatcgcat ctataaattt 540
agaggtcatg gcaaagcttg ggattgggaa gttgatacgg aaaatggtaa ttatgattat 600
ttactatacg ctgatattga tatggatcac ccagaagtag taaatgaatt aagaaattgg 660
ggtgtttggt acacaaacac attaggactc gatggattta gaatagatgc ggttaaacat 720
ataaagtata gctttacgcg cgattggatt aatcacgtta gaagtgcaac aggtaaaaat 780
atgtttgcgg ttgctgagtt ttggaagaat gatttaggtg caattgaaaa ctatctgcag 840
aaaacaaact ggaaccattc agtctttgat gtgccgttac attataatct ttataatgca 900
tcaaaaagcg gagggaacta tgatatgcga aacatattta atggaacggt tgttcaacga 960
catccaagtc atgctgtaac atttgttgat aatcatgatt cgcagcctga agaagcatta 1020
gaatcttttg ttgaagaatg gtttaaacca ttagcgtatg cgcttacatt aacgcgtgaa 1080
caaggatacc cttctgtatt ttacggagat tattatggga ttccaacaca tggagtgcca 1140
gcaatgagat caaaaatcga tccgatttta gaagcacgtc aaaagtatgc atacggaaaa 1200
caaaatgatt acttagacca tcataatatc attggttgga cgcgtgaagg gaatacagca 1260
caccccaatt caggtctagc taccatcatg tctgatggag cgggtggaag taagtggatg 1320
tttgttgggc gtaataaggc tggtcaagta tggagtgata ttacaggaaa ccgtacaggt 1380
acggttacaa tcaatgcaga cggttggggc aatttctctg tgaatggagg gtcagtttct 1440
atttgggtca acaaataa 1458
<210> 154
<211> 9219
<212> DNA
<213> 人工序列
<220>
<223> 合成的
<400> 154
catcggacag ctcttgcttg atatcttcaa aatgacgccg gctcatgtca tgtcaacttt 60
tgtcgtatct ggagcgatcc ttgacggatt cggcatttac gaccgtttta tcgaatttgc 120
cggtgccggg gctacagtcc cgattgtcag cttcggccac tctcttttgc acggcgcgat 180
gcaccaggct gagaaacatg gctttatcgg aatcggcatg gggatatttg aactgacatc 240
tgccggtata tctgccgcta tcttgttcgc ttttcttgtt gccgtgattt ttaaaccgaa 300
aggataaagg aaaatgccag caaaacgcaa ggtcattttg gtcacagacg gcgatatata 360
cgctgcaaaa gcaatcgaat atgcagcaag aaaaacgggt ggccgctgca tttcccaatc 420
ggcggggaat ccgagcgtta aaacaggacc ggagcttgta accatgatcc tgcaaacccc 480
tcatgatcct gtattcgtca tgtttgatga ttccggactt caaggtgaag gcccgggaga 540
gacagctatg aaatatgtag cgatgcatcc cgatatcgag gtgctcggag tcatcgccgt 600
cgcttcaaaa actcattatg cagagtggac gagagtcgat gtatcaatcg atgcagaagg 660
cgaactgaca gagtacggcg tcgataaaca cggggtcaaa gagttcgatg tcaaacgaat 720
gaatggtgat acagtctatt gccttgacca gctggatgtt ccgatcattg tcggaatcgg 780
tgatatcggt aagatgaaca gaaaagacga tgtggaaaaa ggttcgccga ttacaatgaa 840
agcggtcgag ctcattttag aaaggagcgg gtatcatgag tgctcaaaag caagagaaga 900
cgaacgtatt ccttgatcct tctaagaatg aagcgtattt caagaagcgg gtcggcatgg 960
gagaaagctt tgaccttggc gtacggaagg tctttattct cggacatgaa gttcagcttt 1020
attatgtcaa cggattgtgc gacacacaat acatcattca cctgttaaga gaactggtgc 1080
atctgaatga taaagaaaaa gaatcgggcg aggtcgaaga catcgtcgaa aacaggcttt 1140
tgaaccagca ggtttcaaaa gcggaaacgc ttgatgaagc tgtcgaccaa gtgttgtcag 1200
gactggttgc catcatcgtc gaagatgcgg gctttgcttt tatcatcgat gtcagaagct 1260
acccgggcag aacgccggaa gaacctgata cagaaaaagt cgtacgcggt gcaagggacg 1320
gactcgtcga gaacatcatc gtcaacacag ccctgattag acgccggatc agagatgagc 1380
gcttgcgcta caaaatgctt catatcggtg aacgctctaa aacagacatc tgcctctgct 1440
atttggaaga cgttgcagat cccgatcttg ttgaagtatt aaaaaaagaa attgaagatg 1500
tgaagatcga cgggctgccg atgtcggata aatcggtaga ggaattcctg gtcggccaag 1560
gctacaatcc gtttccgctt gtcaggttta cggaaagggc agacgtagcc gcaagccata 1620
ttttagaggg gcatgtcatc gtgatcgtcg atacgtcgcc aagcgtcatc atcacaccga 1680
ccactttgtt tcaccatgtt cagcatgctg aggaatacag acagacgccg gctgttggga 1740
cgtttttaag gtgggtgcgg tttttcggta ttttggcctc cacctttttg ctgccgcttt 1800
ggctgctgtt tgtcattcat ccgtcgctct tgcctgataa tttatcgttt atcgggttga 1860
ataaagacac ccatattccg attatcatgc agattttcct ggcggatctc ggcgtcgaat 1920
ttttaagaat ggccgccatt catacgccga cggcgctttc gactgcaatg ggcctgatcg 1980
ccgctgtatt gatcggcgat atcgcgatca atgtcggctt gttttctccc gaagtcattt 2040
tatacgtttc cctctcggca atcggagcct acacgacacc aagctacgag ctgagcctgg 2100
cgaataaaat ggtgaagctg tttatgctga tattggtggc gctttttaaa gtggagggat 2160
ttgtcatcgg attaacgatc ttaactatag tgatgacttc gatcaggtca ttgcgaacgc 2220
cttacttatg gcctctcctc ccgttcaatg gaaaagcgtt ttggcatgtt ctcgtgcgca 2280
cgtccgttcc agggggaaaa gtcaggccga gcatcgttca tccgagaaac cgctccagac 2340
agccgtgaag ccggcattcg aagaggcttt tccccgggga aaagcctctt tttcaataat 2400
cgaattccgg tctttgagta ccgatgcctc tgtattcatt ggcagagatc gcgactgccc 2460
ggaggctgca gatgttgttc tgtcttctga tcggatagac gacatacagc atttcgcggc 2520
cgtacgggtc aatcgttgac gaatgaagga aaacctcagt tcctctccgc caaaatctcg 2580
tattcgccgg agctgtaata atctgccctt cataaggctc ataaattctc tgttcataat 2640
gcgcagccgg ctgataaggg gcgtatacat cttcaggtgc atagccggga gcgggggtgt 2700
agggatagcg atttggatac atatgataac ctctttccca cttcgttttt tggttttcat 2760
ctttaagatt atattcaggt aaatgcctat ttgtatgggc gaaaatctca gcttttcggc 2820
tcttttttta ttgaatggac gttgtgtatg cctatttcta tcaagcgctg ttttctgtta 2880
ttctataatc aatagaatgg attagttgtt tagggaatca tttcctttat aaatcaagaa 2940
aatttggaca aatggtggtt tagtttttaa aacgaaatgt tataatacaa cataagaatc 3000
gcactatcat gaagccggaa gatgcatcgg gcagcaaccg gagcgcccct tgcacctttg 3060
tcgatagaga aagagggaat gacaattgtt tttacacggt actagcagac aaaatgaaag 3120
agggcacctc gaaatcggcg gtgtcgatgt tctatcattg gcagaaagat acggaacacc 3180
tctttatgta tacgatgtcg cgctgattag agagcgcgcc cgaaaattcc agaaggcatt 3240
caaggaagcc ggtttaaaag cgcaggtagc gtatgcaagc aaggcgtttt catcggttgc 3300
catgattcag cttgccgaac aagaggggct gtctctggat gtggtatcgg gaggagagct 3360
tttcactgcg atcaaagcag ggttcccagc tgagcggatt cattttcacg gaaacaataa 3420
gagccctgaa gaactagcca tggcgctgga gcatcaaatc ggctgcatcg tgctcgataa 3480
ctttcacgag atcgccatta cagaagatct ttgcaagcga tcaggacaaa ctgtagacgt 3540
tttgctcaga atcactccgg gagttgaagc gcacacgcac gattatatta cgacggggca 3600
ggaagattcc aaattcggtt ttgatctgca taatggacag gtcgaacaag ccatcgaaca 3660
agtcctccgc tcgtctgcgt ttaagctcct cggcgtgcac tgccacatcg gttcgcaaat 3720
ttttgatacg gcaggatttg tccttgcagc agacaagatt ttcgagaagc ttgcggaatg 3780
gcgggagact tactctttca ttccggaagt gctcaatctt ggcgggggct tcggcatccg 3840
ctatacaaaa gacgacgagc cgcttgcagc tgatgtttat gttgaaaaaa tcatcgaggc 3900
ggtcaaagca aatgccgagc atttcggctt tgacatccct gagatttgga tcgaaccagg 3960
ccggtctctc gtcggtgatg cggggactac gctgtacacg atcggttctc aaaaagaggt 4020
gccgggcatt cgcaaatatg tagccatcga cggcggcatg agcgataata tcaggccggc 4080
gctttatgag gcaaaatatg aagcagccgt cgccaacagg atgaacgatg cttgtcatga 4140
taccgcatca atcgcaggaa aatgctgcga aagcggagat atgctgattt gggatttgga 4200
aatccccgaa gttcgcgacg gagatgtgct cgccgttttc tgcaccggtg cgtacggcta 4260
cagcatggcc aacaactaca accgcattcc gcgcccggcc gtcgtctttg tcgaggacgg 4320
ggaagcgcag ctcgtcattc agagagagac gtatgaggat atcgtcaagc tggatctgcc 4380
gctgaaatcg aaagtcaaac aataaaaaaa tggagattcc ctaagagggg ggtctccatt 4440
tttaattcaa gctgataaac agctgacatc aactaaaagt ttcattaaat actttgaaaa 4500
aagttgttga cttaaaagaa gctaaatgtt atagtaattg tacagaatag tcttttaagt 4560
aagtctactc tgaatttttt taaaaggaga gggtaaagaa tgaaacaaca aaaacggctt 4620
tacgcccgat tgctgacgct gttatttgcg ctcatcttct tgctgcctca ttctgcagct 4680
agcgcacatc ataacggtac gaacgggaca atgatgcaat actttgaatg gtatctacct 4740
aatgacggaa atcattggaa tcgattaaac tctgatgcga gtaaccttaa aagcaaaggg 4800
attacagcgg tgtggattcc tccagcatgg aagggcgctt ctcaaaatga cgtaggatac 4860
ggagcctatg acctgtatga tctgggagaa tttaatcaaa aaggtaccgt ccgtacaaaa 4920
tatggaacac gtagtcagtt acaagctgcg gtaacctcct taaaaaataa tggaattcaa 4980
gtatatggtg acgttgttat gaatcacaaa ggtggcgcag acgctactga aatggtaagg 5040
gccgttgaag tgaatcccaa taaccgtaac caagaagtga ctggtgaata taccattgaa 5100
gcttggacta gatttgattt tccagggcga ggaaatactc attctagctt taaatggaga 5160
tggtatcatt ttgatggtgt ggattgggat cagtcacgta gactgaacaa tcgcatctat 5220
aaatttagag gtcatggcaa agcttgggat tgggaagttg atacggaaaa tggtaattat 5280
gattatttac tatacgctga tattgatatg gatcacccag aagtagtaaa tgaattaaga 5340
aattggggtg tttggtacac aaacacatta ggactcgatg gatttagaat agatgcggtt 5400
aaacatataa agtatagctt tacgcgcgat tggattaatc acgttagaag tgcaacaggt 5460
aaaaatatgt ttgcggttgc tgagttttgg aagaatgatt taggtgcaat tgaaaactat 5520
ctgcagaaaa caaactggaa ccattcagtc tttgatgtgc cgttacatta taatctttat 5580
aatgcatcaa aaagcggagg gaactatgat atgcgaaaca tatttaatgg aacggttgtt 5640
caacgacatc caagtcatgc tgtaacattt gttgataatc atgattcgca gcctgaagaa 5700
gcattagaat cttttgttga agaatggttt aaaccattag cgtatgcgct tacattaacg 5760
cgtgaacaag gatacccttc tgtattttac ggagattatt atgggattcc aacacatgga 5820
gtgccagcaa tgagatcaaa aatcgatccg attttagaag cacgtcaaaa gtatgcatac 5880
ggaaaacaaa atgattactt agaccatcat aatatcattg gttggacgcg tgaagggaat 5940
acagcacacc ccaattcagg tctagctacc atcatgtctg atggagcggg tggaagtaag 6000
tggatgtttg ttgggcgtaa taaggctggt caagtatgga gtgatattac aggaaaccgt 6060
acaggtacgg ttacaatcaa tgcagacggt tggggcaatt tctctgtgaa tggagggtca 6120
gtttctattt gggtcaacaa ataaaagctt ctcgaggtta acagaggacg gatttcctga 6180
aggaaatccg tttttttatt ttagcacgaa aaacacttcc cggtgatcgg gaggtgtttt 6240
ttgttaaaaa gatcatgaca tgcatagaac agcgaccggg ctaattgtat ataatattgt 6300
gaatttaaca aaaaatttac aaaggagatg ataaaggcaa tgaccagggt gaaaaggatg 6360
agatttgctg atttgttgga tttagaggcg gagtagatga aaccggccaa agtatcccta 6420
ctccaccgat tgctccagtg cctgaagcaa tgtgttgatt gtaacacagt aaatcgtttt 6480
acagcaataa acatttttgt gaatatttta ttgatttcgg ctgtgatctc attcccatat 6540
tctgctgcgg cccatggcgc aacacagtcc ggcgatcaat attcaagctt tgaagaattg 6600
gagcggaatg aagatccagc ttcttaccga attacggaga agaacgcaag agtgccgatg 6660
ctcatcatgg ccatccatgg aggcggcatc gaacccggaa cgagcgaaat cgccaatgaa 6720
gtgtccaaaa actattccct gtacttgttt gaagggctga aatcatcagg caatacggac 6780
cttcacatta caagcacgcg ttttgacgag ccagcggcgc tcgcaattac tgcaagccac 6840
cagtatgtca tgtcgctcca cggctattac agtgaagacc gcgatattaa agtaggcggc 6900
acagaccgcg ctaaaatcag aatattggtt gatgagctga accgctcggg gtttgccgct 6960
gaaatgctgg ggacagatga caagtatgcc ggaacccatc cgaataacat cgccaacaag 7020
tcgctttccg ggctgagcat tcagcttgaa atgagcacgg gtttccgcaa atctttattc 7080
gaccggttta cactaaaaga cagggcggcg acgcaaaacg aaacgtttta ccgatttaca 7140
aagctgctga cagattttat tcatgaaaac tatgaagaag acggagggga tttcccctct 7200
gcaaaaataa aacaccccct tcaagtgaaa aaggaggtgt ttcggcggtt gtgttaaccg 7260
ttggactctg aggtgccgcc gccggtgaat acggaaacga tggcgttcca cagagacaca 7320
aagaagtcga tcagtttttg aagaaagttt tgtccttctt cagaatccaa gaatttcgtg 7380
attttatcct ttgctttgtc aagctggtct ccaacctggt tccagtcgat attaatattt 7440
ttcatgttat taaataaaga tataagagag tttttctgat cttctgtgag tgtcacgcca 7500
agttcggaag cagccgaatc aatcgttttc tccaattcct cttttgactc gggaactccg 7560
tttttcgaga tttcttcctt gactttggcc atcagcgctg acgcgttttc actgccgatt 7620
ttctcgccaa gctctgaagt ggtgacaagc tcttcattcg cgaccttttt cacatcttcg 7680
gaaatttttt cgcccgaagt cgtttcatac gctttcatca atccggttaa agcggctgtg 7740
cctgacactt caaacggagc ggtgacatag actttggcgt cttttacacc ggccgtcatc 7800
agcgcgttca aatacatctc atctgtaatt ctgctgatat tgtgtgtctg aacttccaaa 7860
ccggtgcctt ttttcgctac ggtaattgaa gaagaagaaa tcgctcttgt tccgatttgt 7920
gctttcggta tataatcccc taaatattta tgctcctcat catttgtcac ctcgatgatg 7980
gtcgcatttt caggcgcatt catttctttt aatacttttt gtctgtcctg gcttgacaag 8040
tctttcccca gcgtgacgat gacatcaccc actgcggcgt cagcgaagct gacctgcggg 8100
aaaatgagca gacacaatgc tgtaaagatt cctagtatcg attttttcaa gctcaatgcc 8160
ctccttaaaa atgcaggctt caggcagaat tgctgtactt ttaaagaagc ctgccggaac 8220
ggaaataatg cgttccgaaa tatagacgga tgaaagatga gtgaggtttc aaagaaaaaa 8280
agagagaatt ttctcttcaa gtcaaatgcc ctcccggcat cgtatctcgc cgctctttta 8340
tcattcatga ttttcacagg cgattcaacc tttttttaaa attttttaca aaaacgatac 8400
aagagcggcg tttatttcgg tcgattggct ctctgcttct tcaatatgat ataatgaccc 8460
ttgtgaaatg aaaggagaga atcaagatgg ctaaaaaagg atacatacaa ctgacaaacg 8520
gcaaaaaaat cgagtttgaa ctatatccgg atgcggcgcc gggaactgtc gccaactttg 8580
aaaaacttgc aaacgaaggg ttctatgacg ggctgaagtt ccaccgcgtc atcccgggct 8640
tcgtcagcca gggaggctgc ccgcacggca ccggaacagg cggacctgga tatacgatta 8700
aatgcgagac agaagggaat ccgcacaaac acgaagccgg ttctctctca atggctcacg 8760
caggaaaaga taccggaggc agccaatttt ttatcgtcca tgagcctcag ccgcacttga 8820
acggcgttca caccgttttc ggaaaggtca catcaggcct tgatgccgtc acttcaatgg 8880
agcagggaca aggcatggaa aaagtcgaag tatttgatgc ataatcagag agcgcaaaaa 8940
acagcccgct tagccgggct gtttttttgt ctgtaacggt gtttattttc caggtgcaac 9000
aggacttgag gccgattctt cgtccacatc ctgataggaa ataacgatgc taataaataa 9060
aataattgtg aaaaaatgac cctttatgta aaatatattc aagtgaagag ctagatagag 9120
aacgcaatct gtaaaaaagg aaggggcgta aggggtgagc gtaaaaatcc catcgacggc 9180
agtcggcgta aaaattaatg actggtataa cgcgatacg 9219
<210> 155
<211> 286
<212> PRT
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 155
Met Lys Lys Ile Ala Ile Ala Ala Ile Thr Ala Thr Ser Val Leu Ala
1 5 10 15
Leu Ser Ala Cys Ser Gly Gly Asp Ser Glu Val Val Ala Glu Thr Lys
20 25 30
Ala Gly Asn Ile Thr Lys Glu Asp Leu Tyr Gln Thr Leu Lys Asp Asn
35 40 45
Ala Gly Ala Asp Ala Leu Asn Met Leu Val Gln Gln Lys Val Leu Asp
50 55 60
Asp Lys Tyr Asp Val Ser Asp Lys Glu Ile Asp Lys Lys Leu Asn Glu
65 70 75 80
Tyr Lys Lys Ser Met Gly Asp Gln Leu Asn Gln Leu Ile Asp Gln Lys
85 90 95
Gly Glu Asp Phe Val Lys Glu Gln Ile Lys Tyr Glu Leu Leu Met Gln
100 105 110
Lys Ala Ala Lys Asp Asn Ile Lys Val Thr Asp Asp Asp Val Lys Glu
115 120 125
Tyr Tyr Asp Gly Leu Lys Gly Lys Ile His Leu Ser His Ile Leu Val
130 135 140
Lys Glu Lys Lys Thr Ala Glu Glu Val Glu Lys Lys Leu Lys Lys Gly
145 150 155 160
Glu Lys Phe Glu Asp Leu Ala Lys Glu Tyr Ser Thr Asp Gly Thr Ala
165 170 175
Glu Lys Gly Gly Asp Leu Gly Trp Val Gly Lys Asp Asp Asn Met Asp
180 185 190
Lys Asp Phe Val Lys Ala Ala Phe Ala Leu Lys Thr Gly Glu Ile Ser
195 200 205
Gly Pro Val Lys Ser Gln Phe Gly Tyr His Ile Ile Lys Lys Asp Glu
210 215 220
Glu Arg Gly Lys Tyr Glu Asp Met Lys Lys Glu Leu Lys Lys Glu Val
225 230 235 240
Gln Glu Gln Lys Gln Asn Asp Gln Thr Glu Leu Gln Ser Val Ile Asp
245 250 255
Lys Leu Val Lys Asp Ala Asp Leu Lys Val Lys Asp Lys Glu Leu Lys
260 265 270
Lys Gln Val Asp Gln Arg Gln Ala Gln Thr Ser Ser Ser Ser
275 280 285
<210> 156
<211> 134
<212> PRT
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 156
Met Ala Met Thr Arg Phe Gly Glu Arg Leu Lys Glu Leu Arg Glu Gln
1 5 10 15
Arg Ser Leu Ser Val Asn Gln Leu Ala Met Tyr Ala Gly Val Ser Ala
20 25 30
Ala Ala Ile Ser Arg Ile Glu Asn Gly His Arg Gly Val Pro Lys Pro
35 40 45
Ala Thr Ile Arg Lys Leu Ala Glu Ala Leu Lys Met Pro Tyr Glu Gln
50 55 60
Leu Met Asp Ile Ala Gly Tyr Met Arg Ala Asp Glu Ile Arg Glu Gln
65 70 75 80
Pro Arg Gly Tyr Val Thr Met Gln Glu Ile Ala Ala Lys His Gly Val
85 90 95
Glu Asp Leu Trp Leu Phe Lys Pro Glu Lys Trp Asp Cys Leu Ser Arg
100 105 110
Glu Asp Leu Leu Asn Leu Glu Gln Tyr Phe His Phe Leu Val Asn Glu
115 120 125
Ala Lys Lys Arg Gln Ser
130
<210> 157
<211> 140
<212> PRT
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 157
Met Ala Met Thr Arg Phe Gly Glu Arg Leu Lys Glu Leu Arg Glu Gln
1 5 10 15
Arg Ser Leu Ser Val Asn Gln Leu Ala Met Tyr Ala Gly Val Ser Ala
20 25 30
Ala Ala Ile Ser Arg Ala Ala Ala Ile Ser Arg Ile Glu Asn Gly His
35 40 45
Arg Gly Val Pro Lys Pro Ala Thr Ile Arg Lys Leu Ala Glu Ala Leu
50 55 60
Lys Met Pro Tyr Glu Gln Leu Met Asp Ile Ala Gly Tyr Met Arg Ala
65 70 75 80
Asp Glu Ile Arg Glu Gln Pro Arg Gly Tyr Val Thr Met Gln Glu Ile
85 90 95
Ala Ala Lys His Gly Val Glu Asp Leu Trp Leu Phe Lys Pro Glu Lys
100 105 110
Trp Asp Cys Leu Ser Arg Glu Asp Leu Leu Asn Leu Glu Gln Tyr Phe
115 120 125
His Phe Leu Val Asn Glu Ala Lys Lys Arg Gln Ser
130 135 140
<210> 158
<211> 423
<212> DNA
<213> 地衣芽孢杆菌(Bacillus licheniformis)
<400> 158
atggcgatga caaggttcgg cgagcggctc aaagagctga gggaacaaag aagcctgtcg 60
gttaatcagc ttgccatgta tgccggtgtg agcgccgcag ccatttccag agccgcagcc 120
atttccagaa tcgaaaacgg ccaccgcggc gttcccaagc ccgcgacgat cagaaaattg 180
gccgaggctc tgaaaatgcc gtacgagcag ctcatggata ttgccggtta tatgagagct 240
gacgagattc gcgaacagcc gcgcggctat gtcacgatgc aggagatcgc ggccaagcac 300
ggcgtcgaag acctgtggct gtttaaaccc gagaaatggg actgtttgtc ccgcgaagac 360
ctgctcaacc tcgaacagta ttttcatttt ttggttaatg aagcgaagaa gcgccaatca 420
taa 423

Claims (16)

1.一种用于在经修饰的地衣芽孢杆菌细胞中产生增加量的目的蛋白(POI)的方法,所述方法包括:
(a)通过在表达POI的亲本地衣芽孢杆菌细胞中引入包含与天然prsA可读框(ORF)可操作地连接的天然prsA启动子的多核苷酸来修饰所述表达POI的亲本地衣芽孢杆菌细胞,以及
(b)在适于产生所述POI的条件下发酵所述经修饰的细胞,
其中当在相同条件下发酵时,所述经修饰的细胞相对于所述亲本细胞产生增加量的所述POI。
2.一种用于在经修饰的地衣芽孢杆菌细胞中产生增加量的目的蛋白(POI)的方法,所述方法包括:
(a)向亲本地衣芽孢杆菌细胞中引入(i)编码POI的表达盒,和(ii)
包含可操作地连接到天然prsA可读框(ORF)的天然prsA启动子的多核苷酸,以及
(b)在适于产生所述POI的条件下发酵步骤(a)的经修饰的细胞,
其中当在相同条件下发酵时,所述经修饰的细胞相对于所述亲本细胞产生增加量的所述POI。
3.如权利要求1或权利要求2所述的方法,其中所述引入的多核苷酸包含与SEQ ID NO:100具有至少95%序列同一性的天然prsA启动子序列。
4.如权利要求1或权利要求2所述的方法,其中所述引入的多核苷酸包含与SEQ ID NO:101具有至少90%序列同一性的天然prsA ORF序列。
5.如权利要求1或权利要求2所述的方法,其中所述亲本细胞包含编码天然prsA蛋白的内源prsA基因。
6.如权利要求1或权利要求2所述的方法,其中使所述引入的多核苷酸整合到所述经修饰的细胞的基因组中。
7.如权利要求1或权利要求2所述的方法,其中所述经修饰的细胞进一步包含与SEQ IDNO:122具有至少90%序列同一性的dltA基因的缺失或破坏和/或与SEQ ID NO:121或SEQID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。
8.如权利要求1或权利要求2所述的方法,其中所述POI是酶。
9.一种衍生自亲本地衣芽孢杆菌细胞的经修饰的地衣芽孢杆菌细胞,其中所述经修饰的细胞包含含有可操作地连接到天然prsA可读框(ORF)的天然prsA启动子的引入的多核苷酸。
10.如权利要求9所述的经修饰的细胞,其中所述引入的多核苷酸包含与SEQ ID NO:100具有至少95%序列同一性的天然prsA启动子。
11.如权利要求9所述的经修饰的细胞,其中所述引入的多核苷酸包含与SEQ ID NO:101具有至少90%序列同一性的天然prsA ORF。
12.如权利要求9所述的经修饰的细胞,其中所述引入的多核苷酸编码与SEQ ID NO:155具有约90%序列同一性的天然prsA蛋白。
13.如权利要求9所述的经修饰的细胞,其包含与SEQ ID NO:122具有至少90%序列同一性的dltA基因的缺失或破坏和/或与SEQ ID NO:121或SEQ ID NO:158具有至少90%序列同一性的rghR2基因的缺失或破坏。
14.如权利要求9所述的经修饰的细胞,其包含引入的表达构建体,所述引入的表达构建体编码异源目的蛋白(POI)。
15.如权利要求14所述的经修饰的细胞,其中所述POI是酶。
16.一种目的蛋白,所述目的蛋白由如权利要求14所述的经修饰的细胞产生。
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Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4387982A2 (en) * 2021-08-20 2024-06-26 Danisco US Inc. Methods and compositions for enhanced protein production in bacillus cells
WO2023091878A1 (en) 2021-11-16 2023-05-25 Danisco Us Inc. Compositions and methods for enhanced protein production in bacillus cells
CA3238467A1 (en) * 2021-11-18 2023-05-25 Veli Alkan High performance alpha-amylases for starch liquefaction
WO2024050343A1 (en) 2022-09-02 2024-03-07 Danisco Us Inc. Subtilisin variants and methods related thereto

Family Cites Families (41)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5310675A (en) 1983-06-24 1994-05-10 Genencor, Inc. Procaryotic carbonyl hydrolases
US4760025A (en) 1984-05-29 1988-07-26 Genencor, Inc. Modified enzymes and methods for making same
US5801038A (en) 1984-05-29 1998-09-01 Genencor International Inc. Modified subtilisins having amino acid alterations
EP0254735B2 (en) 1986-01-15 1998-06-17 Amgen Inc. METHOD FOR PRODUCTION OF THERMALLY STABLE AND pH STABLE SUBTILISIN ANALOGS
US4980288A (en) 1986-02-12 1990-12-25 Genex Corporation Subtilisin with increased thermal stability
US4914031A (en) 1987-04-10 1990-04-03 Amgen, Inc. Subtilisin analogs
DK6488D0 (da) 1988-01-07 1988-01-07 Novo Industri As Enzymer
CN1056187C (zh) 1988-02-11 2000-09-06 金克克国际有限公司 新的蛋白水解酶及其在洗涤剂中的应用
DE3909096A1 (de) 1989-03-20 1990-09-27 Garabed Antranikian Alpha-amylase
US5665587A (en) 1989-06-26 1997-09-09 Novo Nordisk A/S Modified subtilisins and detergent compositions containing same
DK97190D0 (da) 1990-04-19 1990-04-19 Novo Nordisk As Oxidationsstabile detergentenzymer
US5482849A (en) 1990-12-21 1996-01-09 Novo Nordisk A/S Subtilisin mutants
ES2121014T3 (es) 1991-05-01 1998-11-16 Novo Nordisk As Enzimas estabilizadas y composiciones detergentes.
CA2155831C (en) 1993-02-11 2009-11-10 Richard L. Antrim Oxidatively stable alpha-amylase
DE4411223A1 (de) 1994-03-31 1995-10-05 Solvay Enzymes Gmbh & Co Kg Verwendung alkalischer Proteasen in gewerblichen Textilwaschverfahren
KR20010015754A (ko) 1997-10-13 2001-02-26 한센 핀 베네드, 안네 제헤르, 웨이콥 마리안느 α-아밀라제 변이체
AR016969A1 (es) 1997-10-23 2001-08-01 Procter & Gamble VARIANTE DE PROTEASA, ADN, VECTOR DE EXPRESIoN, MICROORGANISMO HUESPED, COMPOSICIoN DE LIMPIEZA, ALIMENTO PARA ANIMALES Y COMPOSICIoN PARA TRATAR UN TEXTIL
CN103352033B (zh) 1998-02-27 2016-05-11 诺维信公司 麦芽α淀粉酶变体
US6835550B1 (en) 1998-04-15 2004-12-28 Genencor International, Inc. Mutant proteins having lower allergenic response in humans and methods for constructing, identifying and producing such proteins
US6197565B1 (en) 1998-11-16 2001-03-06 Novo-Nordisk A/S α-Amylase variants
DK1818396T3 (da) 1999-03-30 2014-08-11 Novozymes As Alpha-amylase varianter
US6509185B1 (en) 2000-01-07 2003-01-21 Genencor International, Inc. Mutant aprE promotor
CA2418317A1 (en) 2000-08-11 2002-02-21 Genencor International, Inc. Bacillus transformation, transformants and mutant libraries
EP2341070A3 (en) 2002-03-29 2011-10-19 Genencor International, Inc. Ehanced protein expression in bacillus
CA2483006C (en) 2002-04-22 2013-10-29 Genencor International, Inc. Methods of creating modified promoters resulting in varying levels of gene expression
DE102004047777B4 (de) 2004-10-01 2018-05-09 Basf Se Alpha-Amylase-Varianten mit erhöhter Lösungsmittelstabilität, Verfahren zu deren Herstellung sowie deren Verwendung
DE102004047776B4 (de) 2004-10-01 2018-05-09 Basf Se Gegen Di- und/oder Multimerisierung stabilisierte Alpha-Amylase-Varianten, Verfahren zu deren Herstellung sowie deren Verwendung
WO2006089107A1 (en) 2005-02-18 2006-08-24 Genencor International, Inc. Polypeptides having alpha-amylase and granular starch hydrolyzing activity
JP5226958B2 (ja) * 2007-02-22 2013-07-03 花王株式会社 組換え微生物
BRPI0808513A2 (pt) 2007-03-09 2014-08-19 Danisco Us Inc Genencor Div Variantes de alfa-amilase de espécies de bacillus alcalifílico, composições compreendendo variantes de alfa-amilase e métodos de uso
CN101815783A (zh) 2007-05-30 2010-08-25 丹尼斯科美国公司 地衣芽孢杆菌α-淀粉酶的改良变体
CN101679960B (zh) 2007-05-30 2013-01-16 丹尼斯科美国公司 在发酵过程中具有改良生产水平的α-淀粉酶的变体
JP5687566B2 (ja) 2011-06-01 2015-03-18 日東電工株式会社 防水通音部材
US20140329309A1 (en) 2011-12-09 2014-11-06 Danisco Us Inc. Ribosomal Promoters for Production in Microorganisms
FR2992915B1 (fr) 2012-07-03 2015-08-07 Renault Sa "ensemble de verrouillage pour une ceinture de securite de vehicule automobile"
CN110777132B (zh) 2013-03-11 2024-03-22 丹尼斯科美国公司 α-淀粉酶组合变体
JP7231228B2 (ja) 2017-02-24 2023-03-01 ダニスコ・ユーエス・インク バチルス・リケニフォルミスにおける増加したタンパク質産生のための組成物及び方法
US20210095268A1 (en) 2017-03-31 2021-04-01 Danisco Us Inc Alpha-amylase combinatorial variants
US11879127B2 (en) 2017-08-23 2024-01-23 Danisco Us Inc. Methods and compositions for efficient genetic modifications of Bacillus licheniformis strains
JP7218985B2 (ja) 2017-09-13 2023-02-07 ダニスコ・ユーエス・インク バチルス属(Bacillus)におけるタンパク質産生の増加のための改変5’-非翻訳領域(UTR)配列
WO2019118463A1 (en) 2017-12-15 2019-06-20 Danisco Us Inc Cas9 variants and methods of use

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