CN114835818A - 一种基因编辑融合蛋白、其构建的腺嘌呤碱基编辑器及其应用 - Google Patents

一种基因编辑融合蛋白、其构建的腺嘌呤碱基编辑器及其应用 Download PDF

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CN114835818A
CN114835818A CN202210265179.8A CN202210265179A CN114835818A CN 114835818 A CN114835818 A CN 114835818A CN 202210265179 A CN202210265179 A CN 202210265179A CN 114835818 A CN114835818 A CN 114835818A
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刘龙
陈坚
吕雪芹
堵国成
李江华
刘延峰
武耀康
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Abstract

本发明涉及一种基因编辑融合蛋白、其构建的腺嘌呤碱基编辑器及其应用。基因编辑融合蛋白的氨基酸序列为SEQ ID NO.2‑4所示的序列之一，基于该基因编辑融合蛋白构建的腺嘌呤碱基编辑器还包括crRNA阵列***区，将基因组多位点腺嘌呤转化为鸟嘌呤。本发明构建的腺嘌呤碱基编辑器解决了现有的腺嘌呤碱基编辑器在基因组上的可操作范围受限，而且在多位点的碱基编辑过程中效率低且操作复杂的问题，提供了一种编辑位点多、编辑窗口大、编辑效率和编辑活性高的腺嘌呤碱基编辑器。

Description

一种基因编辑融合蛋白、其构建的腺嘌呤碱基编辑器及其 应用
技术领域
本发明涉及生物技术领域，尤其涉及一种基因编辑融合蛋白、其构建的腺嘌呤碱基编辑器及其应用。
背景技术
基因编辑是通过在DNA上特定位点引入序列改变，达到基因的敲除、外源DNA片段***或者DNA碱基突变的技术手段。CRISPR/Cas***在基因编辑中的应用非常广泛，目前已经发现了三种CRISPR/Cas***，Type I，Type II和Type III，其中Type II***是目前改造最成功的人工核酸酶，只需一个Cas9蛋白就能发挥CRISPR***的免疫作用。在该***中，crRNA(CRISPR-derived RNA)通过碱基配对与tracrRNA(trans-activating crRNA)结合形成tracrRNA/crRNA复合物，此复合物引导核酸酶Cas9蛋白在与crRNA配对的序列靶位点剪切双链DNA。作用过程包括三个阶段：(1)获得CRISPR的间隔区：细菌通过入侵的噬菌体或质粒的一小段DNA序列整合到自身基因组的重复序列之间，即CRISPR 5′端的两个重复序列之间，从而获得高度可变的间隔区域；(2)CRISPR基因座的表达与crRNA的成熟：CRISPR基因座首先被转录成前体CRISPR RNA(pre-crRNA)，然后在Cas9和核酸酶的作用下被剪切成成熟的crRNA；(3)CRISPR/Cas***对外源遗传物质的切割：成熟的tracrRNA、crRNA和Cas9形成核糖核蛋白复合物，crRNA识别外源匹配的DNA序列并与之结合，从而介导核糖核蛋白复合物对外源遗传物质的切割。
为了简化操作过程并提高编辑效率，常常将crRNA和tracrRNA构建成一个嵌合体sgRNA(small guide RNA)进行表达，即只需要表达sgRNA和Cas9蛋白便可进行基因编辑。目前除了CRISPR/Cas9***以外，CRISPR/Cpf1***也常被应用于基因编辑，与CRISPR/Cas9***不同的是，CRISPR/Cpf1只需要crRNA便可发挥作用，且Cpf1自身就具有RNA酶的活力，可以对未成熟的包含多个crRNA的mRNA序列进行切割加工，因此可以设计一段包含多个crRNA的crRNA阵列，可产生多个具有独立功能的crRNA并引导Cpf1靶向基因组的对应靶点，实现多个位点的同时编辑。
很多物种都缺少NHEJ途径(即使有活性也较弱)，而HDR途径需要同源模板的参与才能发挥作用，且这两种修复机制之间还会存在竞争关系，这都导致基于上述方式的基因编辑过程致死率较高且编辑效率较低。将Cas9的DNA酶进行失活后，可以得到只能切割一条DNA链的nCas9和无法切割DNA的dCas9。nCas9与dCas9仍然可以在sgRNA的引导下结合到基因组的特定位点，但却不会产生致死的双链断裂。大肠杆菌的tRNA腺苷脱氨酶TadA突变体TadA7.10可以将腺嘌呤(A)脱氨形成肌苷(I)，在DNA水平肌苷会被当作鸟嘌呤(G)进行读码和复制，最终实现A→G的转换。因此，可以将TadA7.10融合到nCas9或dCas9的N末端后获得的融合蛋白(dCas9-ABE或nCas9-ABE)可以在sgRNA的引导下实现基因组上特定位点的A→G转化，被称为腺嘌呤碱基编辑***。
中国专利CN201811613264.9公开了一种CRISPR/Cas9介导的腺嘌呤碱基编辑***，实现了单碱基定点替换(A>G)，并将其用于改良水稻稻瘟病广谱抗性，但该***用于多位点碱基编辑时需要构建多个对应的sgRNA表达框，这不仅增加了构建过程的复杂性，还会由于启动子等元件的重复使用而降低DNA序列的稳定性，且Cas9所识别的位点需要具有NGG(N＝A,T,C,G)的PAM序列，这限制了基于Cas9的胞嘧啶碱基编辑器在基因组上的可操作范围；中国专利CN201811563073.6公开了一种植物碱基编辑方法，包括由核酸酶失活的CRISPR效应蛋白和DNA依赖的腺嘌呤脱氨酶形成的碱基编辑融合蛋白，所述CRISPR效应蛋白是Cas9核酸酶或Cpf1核酸酶，DNA依赖的腺嘌呤脱氨酶是大肠杆菌tRNA腺嘌呤脱氨酶TadA(ecTadA)的变体，所述变体相对于野生型ecTadA包含一或多组选自以下的突变：1)A106V和D108N；2)D147Y和E155V；3)L84F、H123Y和I156F；4)A142N；5)H36L、R51L、S146C和K157N；6)P48S/T/A；7)A142N；8)W23L/R；9)R152H/P；中国专利201811578853.8公开了一种基于CPF1蛋白的碱基编辑***，其中，碱基编辑融合蛋白包含DNA切割活性缺失的Cpf1(D917A突变)和脱氨酶，脱氨酶的突变体同上，能够实现靶序列中一或多个C至T或者A至G的取代。上述***都只使用了目前应用较为广泛的ecTadA的ABE7.9(TadA7.9)和ABE7.10(TadA7.10)突变体，碱基编辑器的性能还需进一步提高。因此，本发明旨在寻找一种编辑位点多、编辑窗口大、编辑效率和编辑活性高的腺嘌呤碱基编辑器。
发明内容
为解决上述技术问题，本发明提供了一种基于CRISPR/Cpf1的腺嘌呤碱基编辑器，克服了现有的A>G腺嘌呤碱基编辑器在基因组上的可操作范围受限，而且在多位点的碱基编辑过程中效率低且操作复杂的问题。
本发明的第一个目的是提供一种基因编辑融合蛋白，该基因编辑融合蛋白的氨基酸序列为SEQ ID NO.2-4所示的序列之一。其中，SEQ ID NO.2为含有TadA8.20的融合蛋白序列，SEQ ID NO.3为含有TadA8e的融合蛋白序列，SEQ ID NO.4为含有TadA9的融合蛋白序列。
本发明提供了一种基因编辑融合蛋白，该基因编辑融合蛋白可识别TTV作为PAM，其中，V＝A,C,G，该融合蛋白用于基因编辑时，编辑窗口在crRNA的第9个碱基、第7个碱基到第9个碱基之间或第4个碱基到第23个碱基之间，将腺嘌呤碱基编辑为鸟嘌呤。
本发明的第二个目的是提供一种腺嘌呤碱基编辑器，该腺嘌呤碱基编辑器包括上述基因编辑融合蛋白和crRNA阵列***区。crRNA阵列***区***的crRNA与DNA酶失活的Cpf1突变体dCpf1相匹配，引导DNA酶失活的Cpf1突变体dCpf1剪切编辑特定靶点。
进一步地，crRNA阵列***区的核苷酸序列如SEQ ID NO.5所示。该crRNA阵列***区两端包含两个正向重复的crRNA把手序列，并在中间***了两个反向的Eco31I酶切位点，通过酶切连接便可将所需的识别序列或crRNA阵列放置到两个把手序列之间。
进一步地，上述基因编辑融合蛋白通过诱导型启动子Pgrac100调控表达。
进一步地，crRNA阵列***区通过组成型启动子Pveg调控表达。
进一步地，上述腺嘌呤碱基编辑器是将编码SEQ ID NO.2-4之一融合蛋白的基因整合到表达载体以及将crRNA阵列***区***到表达载体上得到。
进一步地，腺嘌呤碱基编辑器用于基因编辑时，将上述表达载体导入真核生物或原核生物中。
进一步地，腺嘌呤碱基编辑器包括：含有上述腺嘌呤碱基编辑器以及上述crRNA阵列***区的质粒。
进一步地，上述质粒的核苷酸序列为SEQ ID NO.9-11所示之一。其中，SEQ IDNO.9为含有TadA8.20的质粒序列，SEQ ID NO.10为含有TadA8e的质粒序列，SEQ ID NO.11为含有TadA9的质粒序列。
本发明的第三个目的是提供上述腺嘌呤碱基编辑器在基因编辑中的应用。
进一步地，上述腺嘌呤碱基编辑器用于将枯草芽孢杆菌基因组上多位点腺嘌呤转化为鸟嘌呤。
进一步地，上述腺嘌呤碱基编辑器用于获取突变体或用于胞外蛋白酶失活。
进一步地，枯草芽孢杆菌为枯草芽孢杆菌Bacillus subtilis 168。
借由上述方案，本发明至少具有以下优点：
本发明设计构建了基于CRISPR/Cpf1的腺嘌呤碱基编辑器(dCpf1-ABE)，利用一个crRNA阵列便可以同时在基因组的5个位点进行碱基(A→G)，从而产生极为丰富的突变体组合，并且拓宽了腺嘌呤碱基编辑器在基因组上的可操作范围，部分位点的编辑效率达到100％，编辑活性高，编辑窗口大。
上述说明仅是本发明技术方案的概述，为了能够更清楚了解本发明的技术手段，并可依照说明书的内容予以实施，以下以本发明的较佳实施例并配合详细附图说明如后。
附图说明
为了使本发明的内容更容易被清楚的理解，下面根据本发明的具体实施例并结合附图，对本发明作进一步详细的说明。
图1为基于CRISPR/Cpf1的腺嘌呤碱基编辑器示意图；
图2为枯草芽孢杆菌腺嘌呤碱基编辑***质粒图谱；
图3为腺苷脱氨酶选择TadA9时构建的腺嘌呤碱基编辑器介导的多位点碱基编辑；
图4为多位点腺嘌呤碱基编辑***处理后产生的突变体的具体组成；
图5为使用多位点腺嘌呤碱基编辑***对胞外蛋白酶进行失活后的蛋白水解情况。
具体实施方式
下面结合附图和具体实施例对本发明作进一步说明，以使本领域的技术人员可以更好地理解本发明并能予以实施，但所举实施例不作为对本发明的限定。
实验涉及的材料及试剂：
DNA聚合酶购自Takara公司，限制性内切酶与T4连接酶购买自NEB公司，质粒提取试剂盒购买自生工生物工程(上海)股份有限公司，PCR产物核酸纯化试剂盒购买自ThermoScientific公司。
细胞均使用LB培养基培养，其中含有：胰蛋白胨10g/L，酵母粉5g/L，NaCl 10g/L。培养基中卡那霉素的终浓度为50μg/mL，IPTG的终浓度为1mM。
实施例1基于CRISPR/Cpf1的腺嘌呤碱基编辑器的设计构建
如图1A所示，构建了基于CRISPR/Cpf1的腺嘌呤编辑***，该***由以下两个基本元件组成：
(1)由大肠杆菌的tRNA腺苷脱氨酶TadA突变体与Cpf1的DNA酶失活突变体(D917A)dCpf1构成的融合蛋白，即腺嘌呤碱基编辑器dCpf1-ABE，该融合蛋白可以在dCpf1的帮助下结合到基因组的特定靶点，并在腺嘌呤脱氨酶的作用下将该处的腺嘌呤(A)转化为鸟嘌呤(G)；
具体地，分别将tRNA腺苷脱氨酶TadA7.10的突变体TadA8.17(TadA7.10 V82S、Q154R)、TadA8.20(TadA7.10 I76Y、V82S、Y123H、Y147R、Q154R)、TadA8e(TadA7.10 A109S、T111R、D119N、H122N、Y147D、F149Y、T166I、D167N)和TadA9(TadA7.10 V82S、A109S、T111R、D119N、H122N、Y147D、F149Y、Q154R、T166I、D167N)通过短肽linker(SGGSSGGSSGSETPGTSESATPESSGGSSGGS)融合到dCpf1的N末端，得到的融合氨基酸序列分别如序列1-4所示。此外，还设计了DNA序列如5所示的crRNA阵列***区，该区域两端包含两个正向重复(directrepeat)的crRNA把手序列，并在中间***了两个反向的Eco31I酶切位点，通过酶切连接便可将所需的识别序列或crRNA阵列放置到两个把手序列之间。
(2)与Cpf1相匹配的crRNA，该crRNA包含固定的把手序列和与目标靶点互补的识别序列，在dCpf1与crRNA把手序列间的相互作用下dCpf1-ABE会与crRNA结合形成复合物，并在互补序列的作用下识别和靶向基因组的特异性靶点，进而实现碱基编辑(A→G)。由于dCpf1仍然具备RNA酶活力，多个crRNA可通过阵列的形式进行表达，crRNA阵列被dCpf1切割处理以后，便可以将dCpf1-ABE靶向多个位点实现碱基编辑(A→G)过程(图1B)。
实施例2基于CRISPR/Cpf1的腺嘌呤碱基编辑器在多位点碱基编辑中的应用
在枯草芽孢杆菌中进行了基于CRISPR/Cpf1的腺嘌呤碱基编辑***的验证与应用。如图2所示，使用受IPTG诱导的Pgrac100启动子表达不同构造的腺嘌呤碱基编辑器dCpf1-ABE，并将crRNA阵列***区放置到组成型启动子Pveg之后以实现crRNA阵列的表达，将上述两个表达框都放置到了含有温敏型复制子pE194的质粒上用于枯草芽孢杆菌中的碱基编辑(A→G)。
质粒构建具体包括以下步骤：所使用的质粒骨架来自于pJOE8999(Altenbuchner,J.,2016.Editing of the Bacillus subtilis genome by the CRISPR-Cas9system.Applied and Environmental Microbiology 82,5421–5427)，该质粒在大肠杆菌与枯草芽孢杆菌中均为卡那霉素抗性(KanR)，且具有多拷贝复制子pBR322可在大肠杆菌中进行质粒的构建与保存，而在枯草芽孢杆菌中则带有温敏型复制子pE194(30℃下可以稳定复制，50℃则会被消除)；所使用Pgrac100启动子及其阻遏蛋白基因lacI和dCpf1蛋白均来自于质粒pLCg6-dCpf1(Wu,Y.,Liu,Y.,Lv,X.,Li,J.,Du,G.,Liu,L.,2020.CAMERS-B:CRISPR/Cpf1 assisted multiple-genes editing and regulation system for Bacillussubtilis.Biotechnology and Bioengineering 117,1817–1825)；crRNA阵列***区及启动子Pveg来自于质粒pcra2(Wu,Y.,Liu,Y.,Lv,X.,Li,J.,Du,G.,Liu,L.,2020.CAMERS-B:CRISPR/Cpf1 assisted multiple-genes editing and regulation system forBacillus subtilis.Biotechnology and Bioengineering 117,1817–1825)；几种腺嘌呤脱氨酶的突变体都是通过基因合成的方式获得。上述片段通过PCR扩增之后，使用无缝连接试剂盒(碧云天D7010M)进行连接，并转化到大肠杆菌中进行测序和保存。包含有TadA7.10的突变体TadA8.17(TadA7.10 V82S、Q154R)、TadA8.20(TadA7.10 I76Y、V82S、Y123H、Y147R、Q154R)、TadA8e(TadA7.10 A109S、T111R、D119N、H122N、Y147D、F149Y、T166I、D167N)和TadA9(TadA7.10V82S、A109S、T111R、D119N、H122N、Y147D、F149Y、Q154R、T166I、D167N)的质粒序列分别如SEQ ID NO.8-11所示，序列9-11所示质粒具体见https://benchling.com/s/seq-8cOWQiJn0nzb9DwUWoQo？m＝slm-JrgZjL6OUp4Dod4GvNOX，https://benchling.com/s/seq-7MMzFTpA45YTqhfwW7KV？m＝slm-PWCavDFpL7MmrQywyplb,https://benchling.com/s/seq-jtNb6AEo2Q5lPsP42jhH？m＝slm-HJP8s8AVmOcAoKe0UnJv。
以枯草芽孢杆菌aprE基因和nprE基因中的5个位点作为靶点，进行多位点的碱基编辑，共设计了如表1中所示的5个crRNA，通过SOMACA(Synthetic Oligos MediatedAssembly of crRNA Array)方法(参考文献：Wu,Y.,Liu,Y.,Lv,X.,Li,J.,Du,G.,Liu,L.,2020.CAMERS-B:CRISPR/Cpf1assisted multiple-genes editing and regulationsystem for Bacillus subtilis.Biotechnology and Bioengineering 117,1817–1825.)组装到了crRNA阵列***区中，得到可以完成以上5个位点碱基编辑(A→G)的编辑质粒。
具体地，当表达单个crRNA时，crRNA可以直接通过带有重叠区的一对引物退火得到(引物浓度为10uM，20uL体系中上下游引物各10uL；反应条件为：98℃2min，0.1℃/S降温至4℃后保温。)，退产物稀释10倍后取1uL与使用Eco31I酶切后的载体连接即可。当设计多个crRNA形成crRNA阵列时，首先将多对具有重叠区的引物进行PCR(引物浓度为10uM，20uL体系中含DNA聚合酶10uL以及上下游引物各5uL；使用标准PCR程序，延伸时间5秒，10个循环)，结束后稀释10倍便可得到两端具有Eco31I的双链DNA，取上述双链DNA各1μL与质粒进行golden gate组装即可。将上述产物转化到大肠杆菌中之后，在质粒骨架和crRNA上各选一个引物进行菌落PCR，并挑取有条带的单菌落进行测序筛选含有所需crRNA的阳性克隆。
表1用于引导多位点碱基编辑(A→G)的crRNA
Figure BDA0003552340720000081
多位点碱基编辑(A→G)与分析的操作步骤如下：首先，将编辑质粒转化到枯草芽孢杆菌Bacillus subtilis 168中，涂布含有卡那霉素的平板，30℃培养12h；然后，挑取任意1个单菌落接种到装有2mL LB培养基(含有卡那霉素)的14mL摇菌管中，30℃振荡培养12h；随后，取5μL菌液转接到含装有2mL LB培养基(含有卡那霉素和IPTG)的14mL摇菌管中，30℃振荡培养12h诱导碱基编辑***表达进行多位点碱基编辑(A→G)；最后，取1μL诱导后的菌液到50μL PCR反应体系中，扩增aprE和nprE基因并进行sanger测序分析。aprE和nprE扩增使用的引物分别如表2所示，上述DNA片段经过纯化后使用表2中所示引物进行sanger测序，并使用BEAT软件(https://hanlab.cc/beat/)对测序结果进行分析。由于所使用的质粒是温敏的，编辑完成之后，可以将菌液划线到不含抗生素的LB平板上，50℃培养过夜即可将质粒消除。
表2引物序列
Figure BDA0003552340720000091
含有不同腺嘌呤脱氨酶TadA突变体的腺嘌呤碱基编辑器(dCpf1-ABE)介导的多位点碱基编辑情况如表3所示：其中将TadA9与dCpf1融合时碱基编辑效率最高，5个位点都有A转化为G，且编辑窗口为crRNA的第4个碱基到第23个碱基之间，而且部分位点的编辑效率达到100％；融合TadA8e时碱基编辑效率略低于TadA9，只在位点2、位点3和位点4个中存在A转化为G，且编辑窗口也较窄，为crRNA的第7个碱基到第9个碱基之间；融合TadA8.20时碱基编辑效率低于TadA9与TadA8e，只在位点2和位点3中存在A转化为G，且编辑窗口也较窄，仅在crRNA的第9个碱基处发生编辑；而融合TadA8.17时在5个位点都无法实现碱基编辑。
表3腺嘌呤碱基编辑器介导的多位点碱基编辑(A→G)及其效率
Figure BDA0003552340720000101
实施例3诱导时间对多位点碱基编辑效率的影响
由于含有TadA7.10突变体TadA9的腺嘌呤碱基编辑器效率最高，可以实现五个位点的同时编辑。因此，又考察了IPTG的诱导时间对该碱基编辑器的碱基编辑效率的影响。
如表4所示，随着诱导时间的增加，各个位点的编辑效率都有一定的提高，但是幅度有限，而且编辑的窗口也并未随着诱导时间的延长而发生变化。上述结果说明诱导12h已经足够实现较为完全的编辑。其中，诱导36h之后的编辑测序结果如图3所示。
表4诱导时间对多位点碱基编辑效率的影响
Figure BDA0003552340720000102
实施例4多位点腺嘌呤碱基编辑***产生突变体情况分析
为了分析使用腺嘌呤碱基编辑处理后获得的突变体的具体组成，将含有腺嘌呤脱氨酶TadA7.10突变体TadA9的腺嘌呤碱基编辑器诱导处理36h后的菌液划线到了不含抗生素LB平板上，并置于37℃过夜培养。然后，挑取了8个单菌落分别扩增aprE和nprE位点进行测序分析。
如图4所示，只需要设计和构建一个特定的crRNA阵列，便可以同时产生丰富的具有不同突变组合的突变体，这对于蛋白进化及菌株改造都是非常有价值的；此外，我们也观察到了使用混合模板测序没有测到的突变位点(如位点2中的A10G，位点3中的A11、A12G和A15G)，说明某些位点可能存在效率很低的突变，这些突变无法通过实施例2中的方法被检测到，这也说明该碱基编辑器具有更大的编辑窗口。此外，由于挑取的菌落较少，表4中某些低频突变位点也可能在上述突变体中未被检测到。
实施例5多位点腺嘌呤碱基编辑器在胞外蛋白酶失活中的应用
枯草芽孢杆菌中共存在6个主要的蛋白酶，这些蛋白酶会将分泌到胞外的蛋白质水解，因此对于目的蛋白的高效分泌表达是极为不利的。因此，我们使用表5中所示的6个crRNA对表中加粗的6个起始密码子(ATG、TTG或GTG)互补链中的A进行了突变(A→G)，如此上述起始密码子中的T会突变为C而无法起始表达，从而造成上述蛋白酶的失活。通过测序分析挑选到了6个蛋白酶全部失活的突变菌株之后，并使用脱脂乳平板对胞外蛋白酶活进行了检测，结果显示与野生菌相比突变株不再具有水解圈，说明胞外蛋白酶活已经被成功消除(图5)。
表5用于引导胞外蛋白酶失活的crRNA
Figure BDA0003552340720000111
对比例1其他腺嘌呤脱氨酶TadA突变体介导的碱基编辑情况
基于CRISPR/Cas9的腺嘌呤碱基编辑***中最早应用的腺嘌呤脱氨酶TadA突变体为TadA7.10，而且常常将一个原始TadA和一个突变体TadA7.10同时融合到nCas9的N末端。中国专利CN201811563073.6与中国专利201811578853.8也都使用了此突变体构建了腺嘌呤碱基编辑***并在水稻中进行了应用。因此，在此进一步尝试了将TadA与TadA7.10融合到dCpf1的N末端构建如序列6所示的基于CRISPR/Cpf1的腺嘌呤碱基编辑***。
然而验证发现，与TadA8.17一样，TadA7.10与dCpf1融合后也不具备任何碱基编辑(A→G)的能力，说明dCpf1与上述TadA突变体不匹配，无法完成碱基编辑(A→G)；或者是上述腺嘌呤碱基编辑器所产生的突变效率过低，因而无法通过实施例2中的分析方法被检测到。
对比例2不同DNA酶失活的Cpf1突变体介导的碱基编辑情况
dCpf1除了常用的D917A以外，其双位点失活的突变体ddCpf1(D917A、E1006A)同样也只是将其DNase活性进行了失活，而保留了其RNase活性，因此我们构建了序列7所示有TadA9和ddCpf1构成的腺嘌呤碱基编辑器，并使用实施例2中的5个crRNA进行了验证。
如表6所示，使用TadA9与ddCpf1构成的腺嘌呤碱基编辑器同样可以完成5个位点的同时突变，但是突变效率较dCpf1有降低。
表6不同DNA酶失活的Cpf1突变体介导的碱基编辑情况
Figure BDA0003552340720000121
对比例3连接方式对构建的碱基编辑***的影响
如实施例1中所述，本发明中的腺嘌呤脱氨酶突变体与dCpf1均使用了短肽linker(SGGSSGGSSGSETPGTSESATPESSGGSSGGS)进行连接，为了验证连接对碱基编辑***的影响，我们尝试将TadA9与dCpf1上的短肽去除，从而使两个蛋白直接相连。然而，去除连接肽后的融合蛋白，在使用实施例3中的5个crRNA进行验证时，已不再具有碱基编辑的活性，说明连接肽对于碱基编辑器的正常工作是必须的。
显然，上述实施例仅仅是为清楚地说明所作的举例，并非对实施方式的限定。对于所属领域的普通技术人员来说，在上述说明的基础上还可以做出其它不同形式变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引申出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
序列表
<110> 江南大学
<120> 一种基因编辑融合蛋白、其构建的腺嘌呤碱基编辑器及其应用
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Asn Asn Gln Glu Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu
1475 1480 1485
Tyr Phe Glu Phe Val Gln Asn Arg Asn Asn Ser Gly Gly Ser Pro Lys
1490 1495 1500
Lys Lys Arg Lys Val
1505
<210> 2
<211> 1509
<212> PRT
<213> （人工序列）
<400> 2
Met Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu
1 5 10 15
Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala
20 25 30
Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala
35 40 45
Ile Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg
50 55 60
Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Tyr Asp Ala Thr Leu
65 70 75 80
Tyr Ser Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His
85 90 95
Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ala Lys Thr Gly
100 105 110
Ala Ala Gly Ser Leu Met Asp Val Leu His His Pro Gly Met Asn His
115 120 125
Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu
130 135 140
Leu Cys Arg Phe Phe Arg Met Pro Arg Arg Val Phe Asn Ala Gln Lys
145 150 155 160
Lys Ala Gln Ser Ser Thr Asp Ser Gly Gly Ser Ser Gly Gly Ser Ser
165 170 175
Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser
180 185 190
Gly Gly Ser Ser Gly Gly Ser Ser Ile Tyr Gln Glu Phe Val Asn Lys
195 200 205
Tyr Ser Leu Ser Lys Thr Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys
210 215 220
Thr Leu Glu Asn Ile Lys Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys
225 230 235 240
Arg Ala Lys Asp Tyr Lys Lys Ala Lys Gln Ile Ile Asp Lys Tyr His
245 250 255
Gln Phe Phe Ile Glu Glu Ile Leu Ser Ser Val Cys Ile Ser Glu Asp
260 265 270
Leu Leu Gln Asn Tyr Ser Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp
275 280 285
Asp Asp Asn Leu Gln Lys Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys
290 295 300
Lys Gln Ile Ser Glu Tyr Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu
305 310 315 320
Phe Asn Gln Asn Leu Ile Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu
325 330 335
Ile Leu Trp Leu Lys Gln Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys
340 345 350
Ala Asn Ser Asp Ile Thr Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys
355 360 365
Ser Phe Lys Gly Trp Thr Thr Tyr Phe Lys Gly Phe His Glu Asn Arg
370 375 380
Lys Asn Val Tyr Ser Ser Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg
385 390 395 400
Ile Val Asp Asp Asn Leu Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr
405 410 415
Glu Ser Leu Lys Asp Lys Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile
420 425 430
Lys Lys Asp Leu Ala Glu Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr
435 440 445
Ser Glu Val Asn Gln Arg Val Phe Ser Leu Asp Glu Val Phe Glu Ile
450 455 460
Ala Asn Phe Asn Asn Tyr Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn
465 470 475 480
Thr Ile Ile Gly Gly Lys Phe Val Asn Gly Glu Asn Thr Lys Arg Lys
485 490 495
Gly Ile Asn Glu Tyr Ile Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys
500 505 510
Thr Leu Lys Lys Tyr Lys Met Ser Val Leu Phe Lys Gln Ile Leu Ser
515 520 525
Asp Thr Glu Ser Lys Ser Phe Val Ile Asp Lys Leu Glu Asp Asp Ser
530 535 540
Asp Val Val Thr Thr Met Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe
545 550 555 560
Lys Thr Val Glu Glu Lys Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe
565 570 575
Asp Asp Leu Lys Ala Gln Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys
580 585 590
Asn Asp Lys Ser Leu Thr Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr
595 600 605
Ser Val Ile Gly Thr Ala Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala
610 615 620
Pro Lys Asn Leu Asp Asn Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala
625 630 635 640
Lys Lys Thr Glu Lys Ala Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu
645 650 655
Ala Leu Glu Glu Phe Asn Lys His Arg Asp Ile Asp Lys Gln Cys Arg
660 665 670
Phe Glu Glu Ile Leu Ala Asn Phe Ala Ala Ile Pro Met Ile Phe Asp
675 680 685
Glu Ile Ala Gln Asn Lys Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr
690 695 700
Gln Asn Gln Gly Lys Lys Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp
705 710 715 720
Val Lys Ala Ile Lys Asp Leu Leu Asp Gln Thr Asn Asn Leu Leu His
725 730 735
Lys Leu Lys Ile Phe His Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile
740 745 750
Leu Asp Lys Asp Glu His Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe
755 760 765
Glu Leu Ala Asn Ile Val Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile
770 775 780
Thr Gln Lys Pro Tyr Ser Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn
785 790 795 800
Ser Thr Leu Ala Asn Gly Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr
805 810 815
Ala Ile Leu Phe Ile Lys Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn
820 825 830
Lys Lys Asn Asn Lys Ile Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys
835 840 845
Gly Glu Gly Tyr Lys Lys Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn
850 855 860
Lys Met Leu Pro Lys Val Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr
865 870 875 880
Asn Pro Ser Glu Asp Ile Leu Arg Ile Arg Asn His Ser Thr His Thr
885 890 895
Lys Asn Gly Ser Pro Gln Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile
900 905 910
Glu Asp Cys Arg Lys Phe Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys
915 920 925
His Pro Glu Trp Lys Asp Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg
930 935 940
Tyr Asn Ser Ile Asp Glu Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr
945 950 955 960
Lys Leu Thr Phe Glu Asn Ile Ser Glu Ser Tyr Ile Asp Ser Val Val
965 970 975
Asn Gln Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser
980 985 990
Ala Tyr Ser Lys Gly Arg Pro Asn Leu His Thr Leu Tyr Trp Lys Ala
995 1000 1005
Leu Phe Asp Glu Arg Asn Leu Gln Asp Val Val Tyr Lys Leu Asn Gly
1010 1015 1020
Glu Ala Glu Leu Phe Tyr Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr
1025 1030 1035 1040
His Pro Ala Lys Glu Ala Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys
1045 1050 1055
Lys Glu Ser Val Phe Glu Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr
1060 1065 1070
Glu Asp Lys Phe Phe Phe His Cys Pro Ile Thr Ile Asn Phe Lys Ser
1075 1080 1085
Ser Gly Ala Asn Lys Phe Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu
1090 1095 1100
Lys Ala Asn Asp Val His Ile Leu Ser Ile Ala Arg Gly Glu Arg His
1105 1110 1115 1120
Leu Ala Tyr Tyr Thr Leu Val Asp Gly Lys Gly Asn Ile Ile Lys Gln
1125 1130 1135
Asp Thr Phe Asn Ile Ile Gly Asn Asp Arg Met Lys Thr Asn Tyr His
1140 1145 1150
Asp Lys Leu Ala Ala Ile Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp
1155 1160 1165
Trp Lys Lys Ile Asn Asn Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser
1170 1175 1180
Gln Val Val His Glu Ile Ala Lys Leu Val Ile Glu Tyr Asn Ala Ile
1185 1190 1195 1200
Val Val Phe Glu Asp Leu Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys
1205 1210 1215
Val Glu Lys Gln Val Tyr Gln Lys Leu Glu Lys Met Leu Ile Glu Lys
1220 1225 1230
Leu Asn Tyr Leu Val Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly
1235 1240 1245
Val Leu Arg Ala Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys
1250 1255 1260
Met Gly Lys Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr
1265 1270 1275 1280
Ser Lys Ile Cys Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys
1285 1290 1295
Tyr Glu Ser Val Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp Lys
1300 1305 1310
Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe Asp Tyr
1315 1320 1325
Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly Lys Trp Thr Ile Ala Ser
1330 1335 1340
Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn Ser Asp Lys Asn His Asn
1345 1350 1355 1360
Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys Glu Leu Glu Lys Leu Leu
1365 1370 1375
Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly Glu Cys Ile Lys Ala Ala
1380 1385 1390
Ile Cys Gly Glu Ser Asp Lys Lys Phe Phe Ala Lys Leu Thr Ser Val
1395 1400 1405
Leu Asn Thr Ile Leu Gln Met Arg Asn Ser Lys Thr Gly Thr Glu Leu
1410 1415 1420
Asp Tyr Leu Ile Ser Pro Val Ala Asp Val Asn Gly Asn Phe Phe Asp
1425 1430 1435 1440
Ser Arg Gln Ala Pro Lys Asn Met Pro Gln Asp Ala Asp Ala Asn Gly
1445 1450 1455
Ala Tyr His Ile Gly Leu Lys Gly Leu Met Leu Leu Gly Arg Ile Lys
1460 1465 1470
Asn Asn Gln Glu Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu
1475 1480 1485
Tyr Phe Glu Phe Val Gln Asn Arg Asn Asn Ser Gly Gly Ser Pro Lys
1490 1495 1500
Lys Lys Arg Lys Val
1505
<210> 3
<211> 1509
<212> PRT
<213> （人工序列）
<400> 3
Met Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu
1 5 10 15
Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala
20 25 30
Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala
35 40 45
Ile Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg
50 55 60
Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu
65 70 75 80
Tyr Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His
85 90 95
Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly
100 105 110
Ala Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His
115 120 125
Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu
130 135 140
Leu Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys
145 150 155 160
Lys Ala Gln Ser Ser Ile Asn Ser Gly Gly Ser Ser Gly Gly Ser Ser
165 170 175
Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser
180 185 190
Gly Gly Ser Ser Gly Gly Ser Ser Ile Tyr Gln Glu Phe Val Asn Lys
195 200 205
Tyr Ser Leu Ser Lys Thr Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys
210 215 220
Thr Leu Glu Asn Ile Lys Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys
225 230 235 240
Arg Ala Lys Asp Tyr Lys Lys Ala Lys Gln Ile Ile Asp Lys Tyr His
245 250 255
Gln Phe Phe Ile Glu Glu Ile Leu Ser Ser Val Cys Ile Ser Glu Asp
260 265 270
Leu Leu Gln Asn Tyr Ser Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp
275 280 285
Asp Asp Asn Leu Gln Lys Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys
290 295 300
Lys Gln Ile Ser Glu Tyr Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu
305 310 315 320
Phe Asn Gln Asn Leu Ile Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu
325 330 335
Ile Leu Trp Leu Lys Gln Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys
340 345 350
Ala Asn Ser Asp Ile Thr Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys
355 360 365
Ser Phe Lys Gly Trp Thr Thr Tyr Phe Lys Gly Phe His Glu Asn Arg
370 375 380
Lys Asn Val Tyr Ser Ser Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg
385 390 395 400
Ile Val Asp Asp Asn Leu Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr
405 410 415
Glu Ser Leu Lys Asp Lys Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile
420 425 430
Lys Lys Asp Leu Ala Glu Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr
435 440 445
Ser Glu Val Asn Gln Arg Val Phe Ser Leu Asp Glu Val Phe Glu Ile
450 455 460
Ala Asn Phe Asn Asn Tyr Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn
465 470 475 480
Thr Ile Ile Gly Gly Lys Phe Val Asn Gly Glu Asn Thr Lys Arg Lys
485 490 495
Gly Ile Asn Glu Tyr Ile Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys
500 505 510
Thr Leu Lys Lys Tyr Lys Met Ser Val Leu Phe Lys Gln Ile Leu Ser
515 520 525
Asp Thr Glu Ser Lys Ser Phe Val Ile Asp Lys Leu Glu Asp Asp Ser
530 535 540
Asp Val Val Thr Thr Met Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe
545 550 555 560
Lys Thr Val Glu Glu Lys Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe
565 570 575
Asp Asp Leu Lys Ala Gln Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys
580 585 590
Asn Asp Lys Ser Leu Thr Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr
595 600 605
Ser Val Ile Gly Thr Ala Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala
610 615 620
Pro Lys Asn Leu Asp Asn Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala
625 630 635 640
Lys Lys Thr Glu Lys Ala Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu
645 650 655
Ala Leu Glu Glu Phe Asn Lys His Arg Asp Ile Asp Lys Gln Cys Arg
660 665 670
Phe Glu Glu Ile Leu Ala Asn Phe Ala Ala Ile Pro Met Ile Phe Asp
675 680 685
Glu Ile Ala Gln Asn Lys Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr
690 695 700
Gln Asn Gln Gly Lys Lys Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp
705 710 715 720
Val Lys Ala Ile Lys Asp Leu Leu Asp Gln Thr Asn Asn Leu Leu His
725 730 735
Lys Leu Lys Ile Phe His Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile
740 745 750
Leu Asp Lys Asp Glu His Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe
755 760 765
Glu Leu Ala Asn Ile Val Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile
770 775 780
Thr Gln Lys Pro Tyr Ser Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn
785 790 795 800
Ser Thr Leu Ala Asn Gly Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr
805 810 815
Ala Ile Leu Phe Ile Lys Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn
820 825 830
Lys Lys Asn Asn Lys Ile Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys
835 840 845
Gly Glu Gly Tyr Lys Lys Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn
850 855 860
Lys Met Leu Pro Lys Val Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr
865 870 875 880
Asn Pro Ser Glu Asp Ile Leu Arg Ile Arg Asn His Ser Thr His Thr
885 890 895
Lys Asn Gly Ser Pro Gln Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile
900 905 910
Glu Asp Cys Arg Lys Phe Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys
915 920 925
His Pro Glu Trp Lys Asp Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg
930 935 940
Tyr Asn Ser Ile Asp Glu Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr
945 950 955 960
Lys Leu Thr Phe Glu Asn Ile Ser Glu Ser Tyr Ile Asp Ser Val Val
965 970 975
Asn Gln Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser
980 985 990
Ala Tyr Ser Lys Gly Arg Pro Asn Leu His Thr Leu Tyr Trp Lys Ala
995 1000 1005
Leu Phe Asp Glu Arg Asn Leu Gln Asp Val Val Tyr Lys Leu Asn Gly
1010 1015 1020
Glu Ala Glu Leu Phe Tyr Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr
1025 1030 1035 1040
His Pro Ala Lys Glu Ala Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys
1045 1050 1055
Lys Glu Ser Val Phe Glu Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr
1060 1065 1070
Glu Asp Lys Phe Phe Phe His Cys Pro Ile Thr Ile Asn Phe Lys Ser
1075 1080 1085
Ser Gly Ala Asn Lys Phe Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu
1090 1095 1100
Lys Ala Asn Asp Val His Ile Leu Ser Ile Ala Arg Gly Glu Arg His
1105 1110 1115 1120
Leu Ala Tyr Tyr Thr Leu Val Asp Gly Lys Gly Asn Ile Ile Lys Gln
1125 1130 1135
Asp Thr Phe Asn Ile Ile Gly Asn Asp Arg Met Lys Thr Asn Tyr His
1140 1145 1150
Asp Lys Leu Ala Ala Ile Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp
1155 1160 1165
Trp Lys Lys Ile Asn Asn Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser
1170 1175 1180
Gln Val Val His Glu Ile Ala Lys Leu Val Ile Glu Tyr Asn Ala Ile
1185 1190 1195 1200
Val Val Phe Glu Asp Leu Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys
1205 1210 1215
Val Glu Lys Gln Val Tyr Gln Lys Leu Glu Lys Met Leu Ile Glu Lys
1220 1225 1230
Leu Asn Tyr Leu Val Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly
1235 1240 1245
Val Leu Arg Ala Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys
1250 1255 1260
Met Gly Lys Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr
1265 1270 1275 1280
Ser Lys Ile Cys Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys
1285 1290 1295
Tyr Glu Ser Val Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp Lys
1300 1305 1310
Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe Asp Tyr
1315 1320 1325
Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly Lys Trp Thr Ile Ala Ser
1330 1335 1340
Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn Ser Asp Lys Asn His Asn
1345 1350 1355 1360
Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys Glu Leu Glu Lys Leu Leu
1365 1370 1375
Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly Glu Cys Ile Lys Ala Ala
1380 1385 1390
Ile Cys Gly Glu Ser Asp Lys Lys Phe Phe Ala Lys Leu Thr Ser Val
1395 1400 1405
Leu Asn Thr Ile Leu Gln Met Arg Asn Ser Lys Thr Gly Thr Glu Leu
1410 1415 1420
Asp Tyr Leu Ile Ser Pro Val Ala Asp Val Asn Gly Asn Phe Phe Asp
1425 1430 1435 1440
Ser Arg Gln Ala Pro Lys Asn Met Pro Gln Asp Ala Asp Ala Asn Gly
1445 1450 1455
Ala Tyr His Ile Gly Leu Lys Gly Leu Met Leu Leu Gly Arg Ile Lys
1460 1465 1470
Asn Asn Gln Glu Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu
1475 1480 1485
Tyr Phe Glu Phe Val Gln Asn Arg Asn Asn Ser Gly Gly Ser Pro Lys
1490 1495 1500
Lys Lys Arg Lys Val
1505
<210> 4
<211> 1509
<212> PRT
<213> （人工序列）
<400> 4
Met Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu
1 5 10 15
Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala
20 25 30
Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala
35 40 45
Ile Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg
50 55 60
Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu
65 70 75 80
Tyr Ser Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His
85 90 95
Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly
100 105 110
Ala Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His
115 120 125
Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu
130 135 140
Leu Cys Asp Phe Tyr Arg Met Pro Arg Arg Val Phe Asn Ala Gln Lys
145 150 155 160
Lys Ala Gln Ser Ser Ile Asn Ser Gly Gly Ser Ser Gly Gly Ser Ser
165 170 175
Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser
180 185 190
Gly Gly Ser Ser Gly Gly Ser Ser Ile Tyr Gln Glu Phe Val Asn Lys
195 200 205
Tyr Ser Leu Ser Lys Thr Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys
210 215 220
Thr Leu Glu Asn Ile Lys Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys
225 230 235 240
Arg Ala Lys Asp Tyr Lys Lys Ala Lys Gln Ile Ile Asp Lys Tyr His
245 250 255
Gln Phe Phe Ile Glu Glu Ile Leu Ser Ser Val Cys Ile Ser Glu Asp
260 265 270
Leu Leu Gln Asn Tyr Ser Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp
275 280 285
Asp Asp Asn Leu Gln Lys Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys
290 295 300
Lys Gln Ile Ser Glu Tyr Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu
305 310 315 320
Phe Asn Gln Asn Leu Ile Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu
325 330 335
Ile Leu Trp Leu Lys Gln Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys
340 345 350
Ala Asn Ser Asp Ile Thr Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys
355 360 365
Ser Phe Lys Gly Trp Thr Thr Tyr Phe Lys Gly Phe His Glu Asn Arg
370 375 380
Lys Asn Val Tyr Ser Ser Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg
385 390 395 400
Ile Val Asp Asp Asn Leu Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr
405 410 415
Glu Ser Leu Lys Asp Lys Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile
420 425 430
Lys Lys Asp Leu Ala Glu Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr
435 440 445
Ser Glu Val Asn Gln Arg Val Phe Ser Leu Asp Glu Val Phe Glu Ile
450 455 460
Ala Asn Phe Asn Asn Tyr Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn
465 470 475 480
Thr Ile Ile Gly Gly Lys Phe Val Asn Gly Glu Asn Thr Lys Arg Lys
485 490 495
Gly Ile Asn Glu Tyr Ile Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys
500 505 510
Thr Leu Lys Lys Tyr Lys Met Ser Val Leu Phe Lys Gln Ile Leu Ser
515 520 525
Asp Thr Glu Ser Lys Ser Phe Val Ile Asp Lys Leu Glu Asp Asp Ser
530 535 540
Asp Val Val Thr Thr Met Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe
545 550 555 560
Lys Thr Val Glu Glu Lys Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe
565 570 575
Asp Asp Leu Lys Ala Gln Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys
580 585 590
Asn Asp Lys Ser Leu Thr Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr
595 600 605
Ser Val Ile Gly Thr Ala Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala
610 615 620
Pro Lys Asn Leu Asp Asn Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala
625 630 635 640
Lys Lys Thr Glu Lys Ala Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu
645 650 655
Ala Leu Glu Glu Phe Asn Lys His Arg Asp Ile Asp Lys Gln Cys Arg
660 665 670
Phe Glu Glu Ile Leu Ala Asn Phe Ala Ala Ile Pro Met Ile Phe Asp
675 680 685
Glu Ile Ala Gln Asn Lys Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr
690 695 700
Gln Asn Gln Gly Lys Lys Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp
705 710 715 720
Val Lys Ala Ile Lys Asp Leu Leu Asp Gln Thr Asn Asn Leu Leu His
725 730 735
Lys Leu Lys Ile Phe His Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile
740 745 750
Leu Asp Lys Asp Glu His Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe
755 760 765
Glu Leu Ala Asn Ile Val Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile
770 775 780
Thr Gln Lys Pro Tyr Ser Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn
785 790 795 800
Ser Thr Leu Ala Asn Gly Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr
805 810 815
Ala Ile Leu Phe Ile Lys Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn
820 825 830
Lys Lys Asn Asn Lys Ile Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys
835 840 845
Gly Glu Gly Tyr Lys Lys Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn
850 855 860
Lys Met Leu Pro Lys Val Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr
865 870 875 880
Asn Pro Ser Glu Asp Ile Leu Arg Ile Arg Asn His Ser Thr His Thr
885 890 895
Lys Asn Gly Ser Pro Gln Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile
900 905 910
Glu Asp Cys Arg Lys Phe Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys
915 920 925
His Pro Glu Trp Lys Asp Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg
930 935 940
Tyr Asn Ser Ile Asp Glu Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr
945 950 955 960
Lys Leu Thr Phe Glu Asn Ile Ser Glu Ser Tyr Ile Asp Ser Val Val
965 970 975
Asn Gln Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser
980 985 990
Ala Tyr Ser Lys Gly Arg Pro Asn Leu His Thr Leu Tyr Trp Lys Ala
995 1000 1005
Leu Phe Asp Glu Arg Asn Leu Gln Asp Val Val Tyr Lys Leu Asn Gly
1010 1015 1020
Glu Ala Glu Leu Phe Tyr Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr
1025 1030 1035 1040
His Pro Ala Lys Glu Ala Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys
1045 1050 1055
Lys Glu Ser Val Phe Glu Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr
1060 1065 1070
Glu Asp Lys Phe Phe Phe His Cys Pro Ile Thr Ile Asn Phe Lys Ser
1075 1080 1085
Ser Gly Ala Asn Lys Phe Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu
1090 1095 1100
Lys Ala Asn Asp Val His Ile Leu Ser Ile Ala Arg Gly Glu Arg His
1105 1110 1115 1120
Leu Ala Tyr Tyr Thr Leu Val Asp Gly Lys Gly Asn Ile Ile Lys Gln
1125 1130 1135
Asp Thr Phe Asn Ile Ile Gly Asn Asp Arg Met Lys Thr Asn Tyr His
1140 1145 1150
Asp Lys Leu Ala Ala Ile Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp
1155 1160 1165
Trp Lys Lys Ile Asn Asn Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser
1170 1175 1180
Gln Val Val His Glu Ile Ala Lys Leu Val Ile Glu Tyr Asn Ala Ile
1185 1190 1195 1200
Val Val Phe Glu Asp Leu Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys
1205 1210 1215
Val Glu Lys Gln Val Tyr Gln Lys Leu Glu Lys Met Leu Ile Glu Lys
1220 1225 1230
Leu Asn Tyr Leu Val Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly
1235 1240 1245
Val Leu Arg Ala Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys
1250 1255 1260
Met Gly Lys Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr
1265 1270 1275 1280
Ser Lys Ile Cys Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys
1285 1290 1295
Tyr Glu Ser Val Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp Lys
1300 1305 1310
Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe Asp Tyr
1315 1320 1325
Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly Lys Trp Thr Ile Ala Ser
1330 1335 1340
Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn Ser Asp Lys Asn His Asn
1345 1350 1355 1360
Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys Glu Leu Glu Lys Leu Leu
1365 1370 1375
Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly Glu Cys Ile Lys Ala Ala
1380 1385 1390
Ile Cys Gly Glu Ser Asp Lys Lys Phe Phe Ala Lys Leu Thr Ser Val
1395 1400 1405
Leu Asn Thr Ile Leu Gln Met Arg Asn Ser Lys Thr Gly Thr Glu Leu
1410 1415 1420
Asp Tyr Leu Ile Ser Pro Val Ala Asp Val Asn Gly Asn Phe Phe Asp
1425 1430 1435 1440
Ser Arg Gln Ala Pro Lys Asn Met Pro Gln Asp Ala Asp Ala Asn Gly
1445 1450 1455
Ala Tyr His Ile Gly Leu Lys Gly Leu Met Leu Leu Gly Arg Ile Lys
1460 1465 1470
Asn Asn Gln Glu Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu
1475 1480 1485
Tyr Phe Glu Phe Val Gln Asn Arg Asn Asn Ser Gly Gly Ser Pro Lys
1490 1495 1500
Lys Lys Arg Lys Val
1505
<210> 5
<211> 82
<212> DNA
<213> （人工序列）
<400> 5
gtctaagaac tttaaataat ttctactgtt gtagatagag accgtgaagt taataaggtc 60
tcaaatttct actgttgtag at 82
<210> 6
<211> 1707
<212> PRT
<213> （人工序列）
<400> 6
Met Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu
1 5 10 15
Thr Leu Ala Lys Arg Ala Trp Asp Glu Arg Glu Val Pro Val Gly Ala
20 25 30
Val Leu Val His Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Pro
35 40 45
Ile Gly Arg His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg
50 55 60
Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu
65 70 75 80
Tyr Val Thr Leu Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His
85 90 95
Ser Arg Ile Gly Arg Val Val Phe Gly Ala Arg Asp Ala Lys Thr Gly
100 105 110
Ala Ala Gly Ser Leu Met Asp Val Leu His His Pro Gly Met Asn His
115 120 125
Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu
130 135 140
Leu Ser Asp Phe Phe Arg Met Arg Arg Gln Glu Ile Lys Ala Gln Lys
145 150 155 160
Lys Ala Gln Ser Ser Thr Asp Ser Gly Gly Ser Ser Gly Gly Ser Ser
165 170 175
Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser
180 185 190
Gly Gly Ser Ser Gly Gly Ser Ser Glu Val Glu Phe Ser His Glu Tyr
195 200 205
Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg
210 215 220
Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg Val Ile Gly
225 230 235 240
Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr Ala His Ala
245 250 255
Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln Asn Tyr Arg
260 265 270
Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys Val Met Cys
275 280 285
Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val Phe Gly Val
290 295 300
Arg Asn Ala Lys Thr Gly Ala Ala Gly Ser Leu Met Asp Val Leu His
305 310 315 320
Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly Ile Leu Ala
325 330 335
Asp Glu Cys Ala Ala Leu Leu Cys Tyr Phe Phe Arg Met Pro Arg Gln
340 345 350
Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Thr Asp Ser Gly Gly
355 360 365
Ser Ser Gly Gly Ser Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser
370 375 380
Ala Thr Pro Glu Ser Ser Gly Gly Ser Ser Gly Gly Ser Ser Ile Tyr
385 390 395 400
Gln Glu Phe Val Asn Lys Tyr Ser Leu Ser Lys Thr Leu Arg Phe Glu
405 410 415
Leu Ile Pro Gln Gly Lys Thr Leu Glu Asn Ile Lys Ala Arg Gly Leu
420 425 430
Ile Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys Lys Ala Lys Gln
435 440 445
Ile Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu Ile Leu Ser Ser
450 455 460
Val Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser Asp Val Tyr Phe
465 470 475 480
Lys Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys Asp Phe Lys Ser
485 490 495
Ala Lys Asp Thr Ile Lys Lys Gln Ile Ser Glu Tyr Ile Lys Asp Ser
500 505 510
Glu Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile Asp Ala Lys Lys
515 520 525
Gly Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln Ser Lys Asp Asn
530 535 540
Gly Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr Asp Ile Asp Glu
545 550 555 560
Ala Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr Thr Tyr Phe Lys
565 570 575
Gly Phe His Glu Asn Arg Lys Asn Val Tyr Ser Ser Asn Asp Ile Pro
580 585 590
Thr Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu Pro Lys Phe Leu
595 600 605
Glu Asn Lys Ala Lys Tyr Glu Ser Leu Lys Asp Lys Ala Pro Glu Ala
610 615 620
Ile Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu Glu Leu Thr Phe
625 630 635 640
Asp Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg Val Phe Ser Leu
645 650 655
Asp Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr Leu Asn Gln Ser
660 665 670
Gly Ile Thr Lys Phe Asn Thr Ile Ile Gly Gly Lys Phe Val Asn Gly
675 680 685
Glu Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile Asn Leu Tyr Ser
690 695 700
Gln Gln Ile Asn Asp Lys Thr Leu Lys Lys Tyr Lys Met Ser Val Leu
705 710 715 720
Phe Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser Phe Val Ile Asp
725 730 735
Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met Gln Ser Phe Tyr
740 745 750
Glu Gln Ile Ala Ala Phe Lys Thr Val Glu Glu Lys Ser Ile Lys Glu
755 760 765
Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln Lys Leu Asp Leu
770 775 780
Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr Asp Leu Ser Gln
785 790 795 800
Gln Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala Val Leu Glu Tyr
805 810 815
Ile Thr Gln Gln Ile Ala Pro Lys Asn Leu Asp Asn Pro Ser Lys Lys
820 825 830
Glu Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala Lys Tyr Leu Ser
835 840 845
Leu Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn Lys His Arg Asp
850 855 860
Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Leu Ala Asn Phe Ala Ala
865 870 875 880
Ile Pro Met Ile Phe Asp Glu Ile Ala Gln Asn Lys Asp Asn Leu Ala
885 890 895
Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys Asp Leu Leu Gln
900 905 910
Ala Ser Ala Glu Asp Asp Val Lys Ala Ile Lys Asp Leu Leu Asp Gln
915 920 925
Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His Ile Ser Gln Ser
930 935 940
Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His Phe Tyr Leu Val
945 950 955 960
Phe Glu Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val Pro Leu Tyr Asn
965 970 975
Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser Asp Glu Lys Phe
980 985 990
Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly Trp Asp Lys Asn
995 1000 1005
Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys Asp Asp Lys Tyr
1010 1015 1020
Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile Phe Asp Asp Lys
1025 1030 1035 1040
Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys Ile Val Tyr Lys
1045 1050 1055
Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val Phe Phe Ser Ala
1060 1065 1070
Lys Ser Ile Lys Phe Tyr Asn Pro Ser Glu Asp Ile Leu Arg Ile Arg
1075 1080 1085
Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln Lys Gly Tyr Glu
1090 1095 1100
Lys Phe Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe Ile Asp Phe Tyr
1105 1110 1115 1120
Lys Gln Ser Ile Ser Lys His Pro Glu Trp Lys Asp Phe Gly Phe Arg
1125 1130 1135
Phe Ser Asp Thr Gln Arg Tyr Asn Ser Ile Asp Glu Phe Tyr Arg Glu
1140 1145 1150
Val Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn Ile Ser Glu Ser
1155 1160 1165
Tyr Ile Asp Ser Val Val Asn Gln Gly Lys Leu Tyr Leu Phe Gln Ile
1170 1175 1180
Tyr Asn Lys Asp Phe Ser Ala Tyr Ser Lys Gly Arg Pro Asn Leu His
1185 1190 1195 1200
Thr Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn Leu Gln Asp Val
1205 1210 1215
Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr Arg Lys Gln Ser
1220 1225 1230
Ile Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala Ile Ala Asn Lys
1235 1240 1245
Asn Lys Asp Asn Pro Lys Lys Glu Ser Val Phe Glu Tyr Asp Leu Ile
1250 1255 1260
Lys Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe His Cys Pro Ile
1265 1270 1275 1280
Thr Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe Asn Asp Glu Ile
1285 1290 1295
Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His Ile Leu Ser Ile
1300 1305 1310
Ala Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu Val Asp Gly Lys
1315 1320 1325
Gly Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile Gly Asn Asp Arg
1330 1335 1340
Met Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile Glu Lys Asp Arg
1345 1350 1355 1360
Asp Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn Ile Lys Glu Met
1365 1370 1375
Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile Ala Lys Leu Val
1380 1385 1390
Ile Glu Tyr Asn Ala Ile Val Val Phe Glu Asp Leu Asn Phe Gly Phe
1395 1400 1405
Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Val Tyr Gln Lys Leu Glu
1410 1415 1420
Lys Met Leu Ile Glu Lys Leu Asn Tyr Leu Val Phe Lys Asp Asn Glu
1425 1430 1435 1440
Phe Asp Lys Thr Gly Gly Val Leu Arg Ala Tyr Gln Leu Thr Ala Pro
1445 1450 1455
Phe Glu Thr Phe Lys Lys Met Gly Lys Gln Thr Gly Ile Ile Tyr Tyr
1460 1465 1470
Val Pro Ala Gly Phe Thr Ser Lys Ile Cys Pro Val Thr Gly Phe Val
1475 1480 1485
Asn Gln Leu Tyr Pro Lys Tyr Glu Ser Val Ser Lys Ser Gln Glu Phe
1490 1495 1500
Phe Ser Lys Phe Asp Lys Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe
1505 1510 1515 1520
Glu Phe Ser Phe Asp Tyr Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly
1525 1530 1535
Lys Trp Thr Ile Ala Ser Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn
1540 1545 1550
Ser Asp Lys Asn His Asn Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys
1555 1560 1565
Glu Leu Glu Lys Leu Leu Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly
1570 1575 1580
Glu Cys Ile Lys Ala Ala Ile Cys Gly Glu Ser Asp Lys Lys Phe Phe
1585 1590 1595 1600
Ala Lys Leu Thr Ser Val Leu Asn Thr Ile Leu Gln Met Arg Asn Ser
1605 1610 1615
Lys Thr Gly Thr Glu Leu Asp Tyr Leu Ile Ser Pro Val Ala Asp Val
1620 1625 1630
Asn Gly Asn Phe Phe Asp Ser Arg Gln Ala Pro Lys Asn Met Pro Gln
1635 1640 1645
Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Gly Leu Lys Gly Leu Met
1650 1655 1660
Leu Leu Gly Arg Ile Lys Asn Asn Gln Glu Gly Lys Lys Leu Asn Leu
1665 1670 1675 1680
Val Ile Lys Asn Glu Glu Tyr Phe Glu Phe Val Gln Asn Arg Asn Asn
1685 1690 1695
Ser Gly Gly Ser Pro Lys Lys Lys Arg Lys Val
1700 1705
<210> 7
<211> 1509
<212> PRT
<213> （人工序列）
<400> 7
Met Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu
1 5 10 15
Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala
20 25 30
Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala
35 40 45
Ile Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg
50 55 60
Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu
65 70 75 80
Tyr Ser Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His
85 90 95
Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly
100 105 110
Ala Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His
115 120 125
Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu
130 135 140
Leu Cys Asp Phe Tyr Arg Met Pro Arg Arg Val Phe Asn Ala Gln Lys
145 150 155 160
Lys Ala Gln Ser Ser Ile Asn Ser Gly Gly Ser Ser Gly Gly Ser Ser
165 170 175
Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser
180 185 190
Gly Gly Ser Ser Gly Gly Ser Ser Ile Tyr Gln Glu Phe Val Asn Lys
195 200 205
Tyr Ser Leu Ser Lys Thr Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys
210 215 220
Thr Leu Glu Asn Ile Lys Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys
225 230 235 240
Arg Ala Lys Asp Tyr Lys Lys Ala Lys Gln Ile Ile Asp Lys Tyr His
245 250 255
Gln Phe Phe Ile Glu Glu Ile Leu Ser Ser Val Cys Ile Ser Glu Asp
260 265 270
Leu Leu Gln Asn Tyr Ser Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp
275 280 285
Asp Asp Asn Leu Gln Lys Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys
290 295 300
Lys Gln Ile Ser Glu Tyr Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu
305 310 315 320
Phe Asn Gln Asn Leu Ile Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu
325 330 335
Ile Leu Trp Leu Lys Gln Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys
340 345 350
Ala Asn Ser Asp Ile Thr Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys
355 360 365
Ser Phe Lys Gly Trp Thr Thr Tyr Phe Lys Gly Phe His Glu Asn Arg
370 375 380
Lys Asn Val Tyr Ser Ser Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg
385 390 395 400
Ile Val Asp Asp Asn Leu Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr
405 410 415
Glu Ser Leu Lys Asp Lys Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile
420 425 430
Lys Lys Asp Leu Ala Glu Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr
435 440 445
Ser Glu Val Asn Gln Arg Val Phe Ser Leu Asp Glu Val Phe Glu Ile
450 455 460
Ala Asn Phe Asn Asn Tyr Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn
465 470 475 480
Thr Ile Ile Gly Gly Lys Phe Val Asn Gly Glu Asn Thr Lys Arg Lys
485 490 495
Gly Ile Asn Glu Tyr Ile Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys
500 505 510
Thr Leu Lys Lys Tyr Lys Met Ser Val Leu Phe Lys Gln Ile Leu Ser
515 520 525
Asp Thr Glu Ser Lys Ser Phe Val Ile Asp Lys Leu Glu Asp Asp Ser
530 535 540
Asp Val Val Thr Thr Met Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe
545 550 555 560
Lys Thr Val Glu Glu Lys Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe
565 570 575
Asp Asp Leu Lys Ala Gln Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys
580 585 590
Asn Asp Lys Ser Leu Thr Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr
595 600 605
Ser Val Ile Gly Thr Ala Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala
610 615 620
Pro Lys Asn Leu Asp Asn Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala
625 630 635 640
Lys Lys Thr Glu Lys Ala Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu
645 650 655
Ala Leu Glu Glu Phe Asn Lys His Arg Asp Ile Asp Lys Gln Cys Arg
660 665 670
Phe Glu Glu Ile Leu Ala Asn Phe Ala Ala Ile Pro Met Ile Phe Asp
675 680 685
Glu Ile Ala Gln Asn Lys Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr
690 695 700
Gln Asn Gln Gly Lys Lys Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp
705 710 715 720
Val Lys Ala Ile Lys Asp Leu Leu Asp Gln Thr Asn Asn Leu Leu His
725 730 735
Lys Leu Lys Ile Phe His Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile
740 745 750
Leu Asp Lys Asp Glu His Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe
755 760 765
Glu Leu Ala Asn Ile Val Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile
770 775 780
Thr Gln Lys Pro Tyr Ser Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn
785 790 795 800
Ser Thr Leu Ala Asn Gly Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr
805 810 815
Ala Ile Leu Phe Ile Lys Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn
820 825 830
Lys Lys Asn Asn Lys Ile Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys
835 840 845
Gly Glu Gly Tyr Lys Lys Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn
850 855 860
Lys Met Leu Pro Lys Val Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr
865 870 875 880
Asn Pro Ser Glu Asp Ile Leu Arg Ile Arg Asn His Ser Thr His Thr
885 890 895
Lys Asn Gly Ser Pro Gln Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile
900 905 910
Glu Asp Cys Arg Lys Phe Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys
915 920 925
His Pro Glu Trp Lys Asp Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg
930 935 940
Tyr Asn Ser Ile Asp Glu Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr
945 950 955 960
Lys Leu Thr Phe Glu Asn Ile Ser Glu Ser Tyr Ile Asp Ser Val Val
965 970 975
Asn Gln Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser
980 985 990
Ala Tyr Ser Lys Gly Arg Pro Asn Leu His Thr Leu Tyr Trp Lys Ala
995 1000 1005
Leu Phe Asp Glu Arg Asn Leu Gln Asp Val Val Tyr Lys Leu Asn Gly
1010 1015 1020
Glu Ala Glu Leu Phe Tyr Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr
1025 1030 1035 1040
His Pro Ala Lys Glu Ala Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys
1045 1050 1055
Lys Glu Ser Val Phe Glu Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr
1060 1065 1070
Glu Asp Lys Phe Phe Phe His Cys Pro Ile Thr Ile Asn Phe Lys Ser
1075 1080 1085
Ser Gly Ala Asn Lys Phe Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu
1090 1095 1100
Lys Ala Asn Asp Val His Ile Leu Ser Ile Ala Arg Gly Glu Arg His
1105 1110 1115 1120
Leu Ala Tyr Tyr Thr Leu Val Asp Gly Lys Gly Asn Ile Ile Lys Gln
1125 1130 1135
Asp Thr Phe Asn Ile Ile Gly Asn Asp Arg Met Lys Thr Asn Tyr His
1140 1145 1150
Asp Lys Leu Ala Ala Ile Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp
1155 1160 1165
Trp Lys Lys Ile Asn Asn Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser
1170 1175 1180
Gln Val Val His Glu Ile Ala Lys Leu Val Ile Glu Tyr Asn Ala Ile
1185 1190 1195 1200
Val Val Phe Ala Asp Leu Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys
1205 1210 1215
Val Glu Lys Gln Val Tyr Gln Lys Leu Glu Lys Met Leu Ile Glu Lys
1220 1225 1230
Leu Asn Tyr Leu Val Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly
1235 1240 1245
Val Leu Arg Ala Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys
1250 1255 1260
Met Gly Lys Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr
1265 1270 1275 1280
Ser Lys Ile Cys Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys
1285 1290 1295
Tyr Glu Ser Val Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp Lys
1300 1305 1310
Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe Asp Tyr
1315 1320 1325
Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly Lys Trp Thr Ile Ala Ser
1330 1335 1340
Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn Ser Asp Lys Asn His Asn
1345 1350 1355 1360
Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys Glu Leu Glu Lys Leu Leu
1365 1370 1375
Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly Glu Cys Ile Lys Ala Ala
1380 1385 1390
Ile Cys Gly Glu Ser Asp Lys Lys Phe Phe Ala Lys Leu Thr Ser Val
1395 1400 1405
Leu Asn Thr Ile Leu Gln Met Arg Asn Ser Lys Thr Gly Thr Glu Leu
1410 1415 1420
Asp Tyr Leu Ile Ser Pro Val Ala Asp Val Asn Gly Asn Phe Phe Asp
1425 1430 1435 1440
Ser Arg Gln Ala Pro Lys Asn Met Pro Gln Asp Ala Asp Ala Asn Gly
1445 1450 1455
Ala Tyr His Ile Gly Leu Lys Gly Leu Met Leu Leu Gly Arg Ile Lys
1460 1465 1470
Asn Asn Gln Glu Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu
1475 1480 1485
Tyr Phe Glu Phe Val Gln Asn Arg Asn Asn Ser Gly Gly Ser Pro Lys
1490 1495 1500
Lys Lys Arg Lys Val
1505
<210> 8
<211> 9445
<212> DNA
<213> （人工序列）
<400> 8
atgtcatgac attggtgtac agaaatggcg cagcaatggc aagaacgtcc cgggcggagc 60
tcaggcctta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 120
tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 180
ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc 240
gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa 300
tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat 360
cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg 420
cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc 480
atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc 540
ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag 600
acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc 660
tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg 720
tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca 780
tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg 840
tgcaccgccg ttttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg 900
gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg 960
gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc 1020
acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc 1080
gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca 1140
tactctgcga catcgtataa cgttactggt ttcatcaaaa tcgtctccct ccgtttgaat 1200
atttgattga tcgtaaccag atgaagcact ctttccacta tccctacagt gttatggctt 1260
gaacaatcac gaaacaataa ttggtacgta cgatctttca gccgactcaa acatcaaatc 1320
ttacaaatgt agtctttgaa agtattacat atgtaagatt taaatgcaac cgttttttcg 1380
gaaggaaatg atgacctcgt ttccaccgga attagcttgg taccagctat tgtaacataa 1440
tcggtacggg ggtgaaaaag ctaacggaaa agggagcgga aaagaatgat gtaagcgtga 1500
aaaatttttt aaaaaatctc ttgacattgg aagggagata tgttattata agaattgcgg 1560
aattgtgagc ggataacaat tcataattgt gagcggataa caattcaacc ccaaaggagg 1620
tgggatccat gagcgaagtt gaattcagcc acgagtactg gatgcgccat gcccttacac 1680
ttgccaaacg cgctcgcgat gaacgcgaag tcccagttgg cgccgttctt gttcttaaca 1740
accgcgtcat cggagaaggc tggaaccgcg ctatcggact tcatgatccg acagctcatg 1800
ccgaaattat ggctctgcgc caaggcggac ttgtcatgca gaattacaga cttattgatg 1860
ccacgcttta ctcaacgttc gaaccgtgcg tcatgtgcgc cggcgccatg attcatagcc 1920
gcatcggcag agttgtcttc ggcgtccgca atgctaaaac gggcgccgcc ggctccctta 1980
tggacgtcct tcattacccg ggcatgaatc accgcgttga aatcacagaa ggcattctgg 2040
ccgatgagtg cgccgctctg ctgtgctact tcttcagaat gccgagaaga gtcttcaacg 2100
cccagaagaa agcccaaagc agcacagact ctggaggatc atccggaggc agctctggaa 2160
gtgaaacacc aggaacaagc gaatcagcta caccagagtc ctctggaggc tcatctggag 2220
gaagctcaat ttatcaagaa tttgttaata aatatagttt aagtaaaact ctaagatttg 2280
agttaatccc acagggtaaa acacttgaaa acataaaagc aagaggtttg attttagatg 2340
atgagaaaag agctaaagac tacaaaaagg ctaaacaaat aattgataaa tatcatcagt 2400
tttttataga ggagatatta agttcggttt gtattagcga agatttatta caaaactatt 2460
ctgatgttta ttttaaactt aaaaagagtg atgatgataa tctacaaaaa gattttaaaa 2520
gtgcaaaaga tacgataaag aaacaaatat ctgaatatat aaaggactca gagaaattta 2580
agaatttgtt taatcaaaac cttatcgatg ctaaaaaagg gcaagagtca gatttaattc 2640
tatggctaaa gcaatctaag gataatggta tagaactatt taaagccaat agtgatatca 2700
cagatataga tgaggcgtta gaaataatca aatcttttaa aggttggaca acttatttta 2760
agggttttca tgaaaataga aaaaatgttt atagtagcaa tgatattcct acatctatta 2820
tttataggat agtagatgat aatttgccta aatttctaga aaataaagct aagtatgaga 2880
gtttaaaaga caaagctcca gaagctataa actatgaaca aattaaaaaa gatttggcag 2940
aagagctaac ctttgatatt gactacaaaa catctgaagt taatcaaaga gttttttcac 3000
ttgatgaagt ttttgagata gcaaacttta ataattatct aaatcaaagt ggtattacta 3060
aatttaatac tattattggt ggtaaatttg taaatggtga aaatacaaag agaaaaggta 3120
taaatgaata tataaatcta tactcacagc aaataaatga taaaacactc aaaaaatata 3180
aaatgagtgt tttatttaag caaattttaa gtgatacaga atctaaatct tttgtaattg 3240
ataagttaga agatgatagt gatgtagtta caacgatgca aagtttttat gagcaaatag 3300
cagcttttaa aacagtagaa gaaaaatcta ttaaagaaac actatcttta ttatttgatg 3360
atttaaaagc tcaaaaactt gatttgagta aaatttattt taaaaatgat aaatctctta 3420
ctgatctatc acaacaagtt tttgatgatt atagtgttat tggtacagcg gtactagaat 3480
atataactca acaaatagca cctaaaaatc ttgataaccc tagtaagaaa gagcaagaat 3540
taatagccaa aaaaactgaa aaagcaaaat acttatctct agaaactata aagcttgcct 3600
tagaagaatt taataagcat agagatatag ataaacagtg taggtttgaa gaaatacttg 3660
caaactttgc ggctattccg atgatatttg atgaaatagc tcaaaacaaa gacaatttgg 3720
cacagatatc tatcaaatat caaaatcaag gtaaaaaaga cctacttcaa gctagtgcgg 3780
aagatgatgt taaagctatc aaggatcttt tagatcaaac taataatctc ttacataaac 3840
taaaaatatt tcatattagt cagtcagaag ataaggcaaa tattttagac aaggatgagc 3900
atttttatct agtatttgag gagtgctact ttgagctagc gaatatagtg cctctttata 3960
acaaaattag aaactatata actcaaaagc catatagtga tgagaaattt aagctcaatt 4020
ttgagaactc gactttggct aatggttggg ataaaaataa agagcctgac aatacggcaa 4080
ttttatttat caaagatgat aaatattatc tgggtgtgat gaataagaaa aataacaaaa 4140
tatttgatga taaagctatc aaagaaaata aaggcgaggg ttataaaaaa attgtttata 4200
aacttttacc tggcgcaaat aaaatgttac ctaaggtttt cttttctgct aaatctataa 4260
aattttataa tcctagtgaa gatatactta gaataagaaa tcattccaca catacaaaaa 4320
atggtagtcc tcaaaaagga tatgaaaaat ttgagtttaa tattgaagat tgccgaaaat 4380
ttatagattt ttataaacag tctataagta agcatccgga gtggaaagat tttggattta 4440
gattttctga tactcaaaga tataattcta tagatgaatt ttatagagaa gttgaaaatc 4500
aaggctacaa actaactttt gaaaatatat cagagagcta tattgatagc gtagttaatc 4560
agggtaaatt gtacctattc caaatctata ataaagattt ttcagcttat agcaaagggc 4620
gaccaaatct acatacttta tattggaaag cgctgtttga tgagagaaat cttcaagatg 4680
tggtttataa gctaaatggt gaggcagagc ttttttatcg taaacaatca atacctaaaa 4740
aaatcactca cccagctaaa gaggcaatag ctaataaaaa caaagataat cctaaaaaag 4800
agagtgtttt tgaatatgat ttaatcaaag ataaacgctt tactgaagat aagtttttct 4860
ttcactgtcc tattacaatc aattttaaat ctagtggagc taataagttt aatgatgaaa 4920
tcaatttatt gctaaaagaa aaagcaaatg atgttcatat attaagtata gctagaggtg 4980
aaagacattt agcttactat actttggtag atggtaaagg caatatcatc aaacaagata 5040
ctttcaacat cattggtaat gatagaatga aaacaaacta ccatgataag cttgctgcaa 5100
tagagaaaga tagggattca gctaggaaag actggaaaaa gataaataac atcaaagaga 5160
tgaaagaggg ctatctatct caggtagttc atgaaatagc taagctagtt atagagtata 5220
atgctattgt ggtttttgag gatttaaatt ttggatttaa aagagggcgt ttcaaggtag 5280
agaagcaggt ctatcaaaag ttagaaaaaa tgctaattga gaaactaaac tatctagttt 5340
tcaaagataa tgagtttgat aaaactgggg gagtgcttag agcttatcag ctaacagcac 5400
cttttgagac ttttaaaaag atgggtaaac aaacaggtat tatctactat gtaccagctg 5460
gttttacttc aaaaatttgt cctgtaactg gttttgtaaa tcagttatat cctaagtatg 5520
aaagtgtcag caaatctcaa gagttcttta gtaagtttga caagatttgt tataaccttg 5580
ataagggcta ttttgagttt agttttgatt ataaaaactt tggtgacaag gctgccaaag 5640
gcaagtggac tatagctagc tttgggagta gattgattaa ctttagaaat tcagataaaa 5700
atcataattg ggatactcga gaagtttatc caactaaaga gttggagaaa ttgctaaaag 5760
attattctat cgaatatggg catggcgaat gtatcaaagc agctatttgc ggtgagagcg 5820
acaaaaagtt ttttgctaag ctaactagtg tcctaaatac tatcttacaa atgcgtaact 5880
caaaaacagg tactgagtta gattatctaa tttcaccagt agcagatgta aatggcaatt 5940
tctttgattc gcgacaggcg ccaaaaaata tgcctcaaga tgctgatgcc aatggtgctt 6000
atcatattgg gctaaaaggt ctgatgctac taggtaggat caaaaataat caagagggca 6060
aaaaactcaa tttggttatc aaaaatgaag agtattttga gttcgtgcag aataggaata 6120
actctggtgg ttctcccaag aagaagagga aagtctaact gcagtataat cagaaacagc 6180
ccgcggatgt tgatctgcgg gctgtttttt attgatcgaa tggccatgac caaaatccct 6240
taacgtgaag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 6300
ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 6360
agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 6420
cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 6480
caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 6540
tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 6600
ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 6660
ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 6720
gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 6780
gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 6840
tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 6900
cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 6960
gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg agacaggtca 7020
ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caactcaaaa tggtatgcgt 7080
tttgacacat ccactatata tccgtgtcgt tctgtccact cctgaatccc attccagaaa 7140
ttctctagcg attccagaag tttctcagag tcggaaagtt gaccagacat tacgaactgg 7200
cacagatggt cataacctga aggaagatct gattgcttaa ctgcttcagt taagaccgaa 7260
gcgctcgtcg tataacagat gcgatgatgc agaccaatca acatggcacc tgccattgct 7320
acctgcacag tcaaggatgg tagaaatgtt gtcggtcctt gcacacgaat attacgccat 7380
ttgcctgcat attcaaacag ctcttctacg ataagggcac aaatcgcatc gtggaacgtt 7440
tgggcttcta ccgatttagc agtttgatac actttctcta agtatccacc tgaatcataa 7500
atcggcaaaa tagagaaaaa ttgaccatgt gtaagcggcc aatctgattc cacctgagat 7560
gcataatcta gtagaatctc ttcgctatca aaattcactt ccaccttcca ctcaccggtt 7620
gtccattcat ggctgaactc tgcttcctct gttgacatga cacacatcat ctcaatatcc 7680
gaatagggcc catcagtctg acgaccaaga gagccataaa caccaatagc cttaacatca 7740
tccccatatt tatccaatat tcgttcctta atttcatgaa caatcttcat tctttcttct 7800
ctagtcatta ttattggtcc attcactatt ctcattcccc tttcagataa ttttagattt 7860
gcttttctaa ataagaatat ttggagagca ccgttcttat tcagctatta aacccattat 7920
atcgggtttt tgaggggatt tcaactgcag acacctaaat tcaaaatcta tcggtcagat 7980
ttataccgat ttgattttat atattcttga ataacatacg ccgagttatc acataaaagc 8040
gggaaccaat catcaaattt aaacttcatt gcataatcca ttaaactctt aaattctacg 8100
attccttgtt catcaataaa ctcaatcatt tctttaatta atttatatct atctgttgtt 8160
gttttcttta ataattcatc aacatctaca ccgccataaa ctatcatatc ttctttttga 8220
tatttaaatt tattaggatc gtccatgtga agcatatatc tcacaagacc tttcacactt 8280
cctgcaatct gcggaatagt cgcattcaat tcttctgtaa ttatttttat ctgttcataa 8340
gatttattac cctcatacat cactagaata tgataatgct cttttttcat cctatcttct 8400
gtatcagtat ccctatcatg taatggagac actacaaatt gaatgtgtaa ctcttttaaa 8460
tactctaacc actcggcttt tgctgattct ggatataaaa caaatgtcca attacgtcct 8520
cttgaatttt tcttgttttc agtttctttt attacatttt cgctcatgat ataataacgg 8580
tgctaataca tttaacaaaa tttagtcata gataggcagc atgccagtgc tgtctatctt 8640
tttttgttta aaatgcaccg tattcctcct ttgcatattt ttttattaga ataccggttg 8700
catctgattt gctaatatta tatttttctt tgattctatt taatatctca ttttcttctg 8760
ttgtaagtct taaagtaaca gcaacttttt tctcttcttt tctatctaca accatcactg 8820
tacctcccaa catctgtttt tttcacttta acataaaaaa caacctttta acattaaaaa 8880
cccaatattt atttatttgt ttggacaatg gacaatggac acctaggggg gaggtcgtag 8940
taccccccta tgttttctcc cctaaataac cccaaaaatc taagaaaaaa agacctcaaa 9000
aaggtcttta attaacatct caaatttcgc atttattcca atttcctttt tgcgtgtgat 9060
gcgttattaa cgttgatata atttaaattt tatttgacaa aaatgggctc gtgttgtaca 9120
ataaatgtag aggtagagac gcgaggtcta agaactttaa ataatttcta ctgttgtaga 9180
tagagaccgt gaagttaata aggtctcaaa tttctactgt tgtagatcgt ctctgaactg 9240
attcaagcaa gcttaaaccc agctcaatga gctgggtttt ttgtttgttt tttcaaactt 9300
agttagcttg gccagtgcct ctagagtcaa gtaaagagtc gacctgttac gaacggcaga 9360
tcagaatttt gtaataaaaa aagagcctgc tcattacact gcgggctctt tttcatggtc 9420
agaagacggg taaccaagat aacaa 9445
<210> 9
<211> 9445
<212> DNA
<213> （人工序列）
<400> 9
atgtcatgac attggtgtac agaaatggcg cagcaatggc aagaacgtcc cgggcggagc 60
tcaggcctta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 120
tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 180
ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc 240
gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa 300
tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat 360
cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg 420
cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc 480
atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc 540
ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag 600
acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc 660
tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg 720
tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca 780
tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg 840
tgcaccgccg ttttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg 900
gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg 960
gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc 1020
acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc 1080
gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca 1140
tactctgcga catcgtataa cgttactggt ttcatcaaaa tcgtctccct ccgtttgaat 1200
atttgattga tcgtaaccag atgaagcact ctttccacta tccctacagt gttatggctt 1260
gaacaatcac gaaacaataa ttggtacgta cgatctttca gccgactcaa acatcaaatc 1320
ttacaaatgt agtctttgaa agtattacat atgtaagatt taaatgcaac cgttttttcg 1380
gaaggaaatg atgacctcgt ttccaccgga attagcttgg taccagctat tgtaacataa 1440
tcggtacggg ggtgaaaaag ctaacggaaa agggagcgga aaagaatgat gtaagcgtga 1500
aaaatttttt aaaaaatctc ttgacattgg aagggagata tgttattata agaattgcgg 1560
aattgtgagc ggataacaat tcataattgt gagcggataa caattcaacc ccaaaggagg 1620
tgggatccat gagcgaagtc gagttctccc acgaatactg gatgcgccat gcccttacac 1680
ttgccaaacg cgctcgcgat gaacgcgaag tcccagttgg cgccgttctt gttcttaaca 1740
accgcgtcat cggagaaggc tggaaccgcg ctatcggact tcatgatccg acagctcatg 1800
ccgaaattat ggctctgcgc caaggcggac ttgtcatgca gaattacaga ctttatgatg 1860
ccacgcttta ctcaacgttc gaaccgtgcg tcatgtgcgc cggcgccatg attcatagcc 1920
gcatcggcag agttgtcttc ggcgtccgca atgctaaaac gggcgccgcc ggctccctta 1980
tggacgtcct tcatcacccg ggcatgaatc accgcgttga aatcacagaa ggcattctgg 2040
ccgatgagtg cgccgctctg ctgtgcagat tcttcagaat gccgagaaga gtcttcaacg 2100
cccagaagaa agcccaaagc agcacagact ctggaggatc atccggaggc agctctggaa 2160
gtgaaacacc aggaacaagc gaatcagcta caccagagtc ctctggaggc tcatctggag 2220
gaagctcaat ttatcaagaa tttgttaata aatatagttt aagtaaaact ctaagatttg 2280
agttaatccc acagggtaaa acacttgaaa acataaaagc aagaggtttg attttagatg 2340
atgagaaaag agctaaagac tacaaaaagg ctaaacaaat aattgataaa tatcatcagt 2400
tttttataga ggagatatta agttcggttt gtattagcga agatttatta caaaactatt 2460
ctgatgttta ttttaaactt aaaaagagtg atgatgataa tctacaaaaa gattttaaaa 2520
gtgcaaaaga tacgataaag aaacaaatat ctgaatatat aaaggactca gagaaattta 2580
agaatttgtt taatcaaaac cttatcgatg ctaaaaaagg gcaagagtca gatttaattc 2640
tatggctaaa gcaatctaag gataatggta tagaactatt taaagccaat agtgatatca 2700
cagatataga tgaggcgtta gaaataatca aatcttttaa aggttggaca acttatttta 2760
agggttttca tgaaaataga aaaaatgttt atagtagcaa tgatattcct acatctatta 2820
tttataggat agtagatgat aatttgccta aatttctaga aaataaagct aagtatgaga 2880
gtttaaaaga caaagctcca gaagctataa actatgaaca aattaaaaaa gatttggcag 2940
aagagctaac ctttgatatt gactacaaaa catctgaagt taatcaaaga gttttttcac 3000
ttgatgaagt ttttgagata gcaaacttta ataattatct aaatcaaagt ggtattacta 3060
aatttaatac tattattggt ggtaaatttg taaatggtga aaatacaaag agaaaaggta 3120
taaatgaata tataaatcta tactcacagc aaataaatga taaaacactc aaaaaatata 3180
aaatgagtgt tttatttaag caaattttaa gtgatacaga atctaaatct tttgtaattg 3240
ataagttaga agatgatagt gatgtagtta caacgatgca aagtttttat gagcaaatag 3300
cagcttttaa aacagtagaa gaaaaatcta ttaaagaaac actatcttta ttatttgatg 3360
atttaaaagc tcaaaaactt gatttgagta aaatttattt taaaaatgat aaatctctta 3420
ctgatctatc acaacaagtt tttgatgatt atagtgttat tggtacagcg gtactagaat 3480
atataactca acaaatagca cctaaaaatc ttgataaccc tagtaagaaa gagcaagaat 3540
taatagccaa aaaaactgaa aaagcaaaat acttatctct agaaactata aagcttgcct 3600
tagaagaatt taataagcat agagatatag ataaacagtg taggtttgaa gaaatacttg 3660
caaactttgc ggctattccg atgatatttg atgaaatagc tcaaaacaaa gacaatttgg 3720
cacagatatc tatcaaatat caaaatcaag gtaaaaaaga cctacttcaa gctagtgcgg 3780
aagatgatgt taaagctatc aaggatcttt tagatcaaac taataatctc ttacataaac 3840
taaaaatatt tcatattagt cagtcagaag ataaggcaaa tattttagac aaggatgagc 3900
atttttatct agtatttgag gagtgctact ttgagctagc gaatatagtg cctctttata 3960
acaaaattag aaactatata actcaaaagc catatagtga tgagaaattt aagctcaatt 4020
ttgagaactc gactttggct aatggttggg ataaaaataa agagcctgac aatacggcaa 4080
ttttatttat caaagatgat aaatattatc tgggtgtgat gaataagaaa aataacaaaa 4140
tatttgatga taaagctatc aaagaaaata aaggcgaggg ttataaaaaa attgtttata 4200
aacttttacc tggcgcaaat aaaatgttac ctaaggtttt cttttctgct aaatctataa 4260
aattttataa tcctagtgaa gatatactta gaataagaaa tcattccaca catacaaaaa 4320
atggtagtcc tcaaaaagga tatgaaaaat ttgagtttaa tattgaagat tgccgaaaat 4380
ttatagattt ttataaacag tctataagta agcatccgga gtggaaagat tttggattta 4440
gattttctga tactcaaaga tataattcta tagatgaatt ttatagagaa gttgaaaatc 4500
aaggctacaa actaactttt gaaaatatat cagagagcta tattgatagc gtagttaatc 4560
agggtaaatt gtacctattc caaatctata ataaagattt ttcagcttat agcaaagggc 4620
gaccaaatct acatacttta tattggaaag cgctgtttga tgagagaaat cttcaagatg 4680
tggtttataa gctaaatggt gaggcagagc ttttttatcg taaacaatca atacctaaaa 4740
aaatcactca cccagctaaa gaggcaatag ctaataaaaa caaagataat cctaaaaaag 4800
agagtgtttt tgaatatgat ttaatcaaag ataaacgctt tactgaagat aagtttttct 4860
ttcactgtcc tattacaatc aattttaaat ctagtggagc taataagttt aatgatgaaa 4920
tcaatttatt gctaaaagaa aaagcaaatg atgttcatat attaagtata gctagaggtg 4980
aaagacattt agcttactat actttggtag atggtaaagg caatatcatc aaacaagata 5040
ctttcaacat cattggtaat gatagaatga aaacaaacta ccatgataag cttgctgcaa 5100
tagagaaaga tagggattca gctaggaaag actggaaaaa gataaataac atcaaagaga 5160
tgaaagaggg ctatctatct caggtagttc atgaaatagc taagctagtt atagagtata 5220
atgctattgt ggtttttgag gatttaaatt ttggatttaa aagagggcgt ttcaaggtag 5280
agaagcaggt ctatcaaaag ttagaaaaaa tgctaattga gaaactaaac tatctagttt 5340
tcaaagataa tgagtttgat aaaactgggg gagtgcttag agcttatcag ctaacagcac 5400
cttttgagac ttttaaaaag atgggtaaac aaacaggtat tatctactat gtaccagctg 5460
gttttacttc aaaaatttgt cctgtaactg gttttgtaaa tcagttatat cctaagtatg 5520
aaagtgtcag caaatctcaa gagttcttta gtaagtttga caagatttgt tataaccttg 5580
ataagggcta ttttgagttt agttttgatt ataaaaactt tggtgacaag gctgccaaag 5640
gcaagtggac tatagctagc tttgggagta gattgattaa ctttagaaat tcagataaaa 5700
atcataattg ggatactcga gaagtttatc caactaaaga gttggagaaa ttgctaaaag 5760
attattctat cgaatatggg catggcgaat gtatcaaagc agctatttgc ggtgagagcg 5820
acaaaaagtt ttttgctaag ctaactagtg tcctaaatac tatcttacaa atgcgtaact 5880
caaaaacagg tactgagtta gattatctaa tttcaccagt agcagatgta aatggcaatt 5940
tctttgattc gcgacaggcg ccaaaaaata tgcctcaaga tgctgatgcc aatggtgctt 6000
atcatattgg gctaaaaggt ctgatgctac taggtaggat caaaaataat caagagggca 6060
aaaaactcaa tttggttatc aaaaatgaag agtattttga gttcgtgcag aataggaata 6120
actctggtgg ttctcccaag aagaagagga aagtctaact gcagtataat cagaaacagc 6180
ccgcggatgt tgatctgcgg gctgtttttt attgatcgaa tggccatgac caaaatccct 6240
taacgtgaag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 6300
ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 6360
agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 6420
cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 6480
caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 6540
tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 6600
ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 6660
ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 6720
gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 6780
gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 6840
tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 6900
cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 6960
gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg agacaggtca 7020
ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caactcaaaa tggtatgcgt 7080
tttgacacat ccactatata tccgtgtcgt tctgtccact cctgaatccc attccagaaa 7140
ttctctagcg attccagaag tttctcagag tcggaaagtt gaccagacat tacgaactgg 7200
cacagatggt cataacctga aggaagatct gattgcttaa ctgcttcagt taagaccgaa 7260
gcgctcgtcg tataacagat gcgatgatgc agaccaatca acatggcacc tgccattgct 7320
acctgcacag tcaaggatgg tagaaatgtt gtcggtcctt gcacacgaat attacgccat 7380
ttgcctgcat attcaaacag ctcttctacg ataagggcac aaatcgcatc gtggaacgtt 7440
tgggcttcta ccgatttagc agtttgatac actttctcta agtatccacc tgaatcataa 7500
atcggcaaaa tagagaaaaa ttgaccatgt gtaagcggcc aatctgattc cacctgagat 7560
gcataatcta gtagaatctc ttcgctatca aaattcactt ccaccttcca ctcaccggtt 7620
gtccattcat ggctgaactc tgcttcctct gttgacatga cacacatcat ctcaatatcc 7680
gaatagggcc catcagtctg acgaccaaga gagccataaa caccaatagc cttaacatca 7740
tccccatatt tatccaatat tcgttcctta atttcatgaa caatcttcat tctttcttct 7800
ctagtcatta ttattggtcc attcactatt ctcattcccc tttcagataa ttttagattt 7860
gcttttctaa ataagaatat ttggagagca ccgttcttat tcagctatta aacccattat 7920
atcgggtttt tgaggggatt tcaactgcag acacctaaat tcaaaatcta tcggtcagat 7980
ttataccgat ttgattttat atattcttga ataacatacg ccgagttatc acataaaagc 8040
gggaaccaat catcaaattt aaacttcatt gcataatcca ttaaactctt aaattctacg 8100
attccttgtt catcaataaa ctcaatcatt tctttaatta atttatatct atctgttgtt 8160
gttttcttta ataattcatc aacatctaca ccgccataaa ctatcatatc ttctttttga 8220
tatttaaatt tattaggatc gtccatgtga agcatatatc tcacaagacc tttcacactt 8280
cctgcaatct gcggaatagt cgcattcaat tcttctgtaa ttatttttat ctgttcataa 8340
gatttattac cctcatacat cactagaata tgataatgct cttttttcat cctatcttct 8400
gtatcagtat ccctatcatg taatggagac actacaaatt gaatgtgtaa ctcttttaaa 8460
tactctaacc actcggcttt tgctgattct ggatataaaa caaatgtcca attacgtcct 8520
cttgaatttt tcttgttttc agtttctttt attacatttt cgctcatgat ataataacgg 8580
tgctaataca tttaacaaaa tttagtcata gataggcagc atgccagtgc tgtctatctt 8640
tttttgttta aaatgcaccg tattcctcct ttgcatattt ttttattaga ataccggttg 8700
catctgattt gctaatatta tatttttctt tgattctatt taatatctca ttttcttctg 8760
ttgtaagtct taaagtaaca gcaacttttt tctcttcttt tctatctaca accatcactg 8820
tacctcccaa catctgtttt tttcacttta acataaaaaa caacctttta acattaaaaa 8880
cccaatattt atttatttgt ttggacaatg gacaatggac acctaggggg gaggtcgtag 8940
taccccccta tgttttctcc cctaaataac cccaaaaatc taagaaaaaa agacctcaaa 9000
aaggtcttta attaacatct caaatttcgc atttattcca atttcctttt tgcgtgtgat 9060
gcgttattaa cgttgatata atttaaattt tatttgacaa aaatgggctc gtgttgtaca 9120
ataaatgtag aggtagagac gcgaggtcta agaactttaa ataatttcta ctgttgtaga 9180
tagagaccgt gaagttaata aggtctcaaa tttctactgt tgtagatcgt ctctgaactg 9240
attcaagcaa gcttaaaccc agctcaatga gctgggtttt ttgtttgttt tttcaaactt 9300
agttagcttg gccagtgcct ctagagtcaa gtaaagagtc gacctgttac gaacggcaga 9360
tcagaatttt gtaataaaaa aagagcctgc tcattacact gcgggctctt tttcatggtc 9420
agaagacggg taaccaagat aacaa 9445
<210> 10
<211> 9445
<212> DNA
<213> （人工序列）
<400> 10
atgtcatgac attggtgtac agaaatggcg cagcaatggc aagaacgtcc cgggcggagc 60
tcaggcctta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 120
tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 180
ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc 240
gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa 300
tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat 360
cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg 420
cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc 480
atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc 540
ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag 600
acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc 660
tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg 720
tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca 780
tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg 840
tgcaccgccg ttttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg 900
gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg 960
gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc 1020
acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc 1080
gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca 1140
tactctgcga catcgtataa cgttactggt ttcatcaaaa tcgtctccct ccgtttgaat 1200
atttgattga tcgtaaccag atgaagcact ctttccacta tccctacagt gttatggctt 1260
gaacaatcac gaaacaataa ttggtacgta cgatctttca gccgactcaa acatcaaatc 1320
ttacaaatgt agtctttgaa agtattacat atgtaagatt taaatgcaac cgttttttcg 1380
gaaggaaatg atgacctcgt ttccaccgga attagcttgg taccagctat tgtaacataa 1440
tcggtacggg ggtgaaaaag ctaacggaaa agggagcgga aaagaatgat gtaagcgtga 1500
aaaatttttt aaaaaatctc ttgacattgg aagggagata tgttattata agaattgcgg 1560
aattgtgagc ggataacaat tcataattgt gagcggataa caattcaacc ccaaaggagg 1620
tgggatccat gagcgaagtc gagttctccc acgaatactg gatgcgccat gcccttacac 1680
ttgccaaacg cgctcgcgat gaacgcgaag tcccagttgg cgccgttctt gttcttaaca 1740
accgcgtcat cggagaaggc tggaaccgcg ctatcggact tcatgatccg acagctcatg 1800
ccgaaattat ggctctgcgc caaggcggac ttgtcatgca gaattacaga cttattgatg 1860
ccacgcttta cgtcacgttc gaaccgtgcg tcatgtgcgc cggcgccatg attcatagcc 1920
gcatcggcag agttgtcttc ggcgtccgca attcaaaaag aggcgccgcc ggctccctta 1980
tgaacgtcct taactacccg ggcatgaatc accgcgttga aatcacagaa ggcattctgg 2040
ccgatgagtg cgccgctctg ctgtgcgact tctatagaat gccgagacaa gtcttcaacg 2100
cccagaagaa agcccaaagc agcattaact ctggaggatc atccggaggc agctctggaa 2160
gtgaaacacc aggaacaagc gaatcagcta caccagagtc ctctggaggc tcatctggag 2220
gaagctcaat ttatcaagaa tttgttaata aatatagttt aagtaaaact ctaagatttg 2280
agttaatccc acagggtaaa acacttgaaa acataaaagc aagaggtttg attttagatg 2340
atgagaaaag agctaaagac tacaaaaagg ctaaacaaat aattgataaa tatcatcagt 2400
tttttataga ggagatatta agttcggttt gtattagcga agatttatta caaaactatt 2460
ctgatgttta ttttaaactt aaaaagagtg atgatgataa tctacaaaaa gattttaaaa 2520
gtgcaaaaga tacgataaag aaacaaatat ctgaatatat aaaggactca gagaaattta 2580
agaatttgtt taatcaaaac cttatcgatg ctaaaaaagg gcaagagtca gatttaattc 2640
tatggctaaa gcaatctaag gataatggta tagaactatt taaagccaat agtgatatca 2700
cagatataga tgaggcgtta gaaataatca aatcttttaa aggttggaca acttatttta 2760
agggttttca tgaaaataga aaaaatgttt atagtagcaa tgatattcct acatctatta 2820
tttataggat agtagatgat aatttgccta aatttctaga aaataaagct aagtatgaga 2880
gtttaaaaga caaagctcca gaagctataa actatgaaca aattaaaaaa gatttggcag 2940
aagagctaac ctttgatatt gactacaaaa catctgaagt taatcaaaga gttttttcac 3000
ttgatgaagt ttttgagata gcaaacttta ataattatct aaatcaaagt ggtattacta 3060
aatttaatac tattattggt ggtaaatttg taaatggtga aaatacaaag agaaaaggta 3120
taaatgaata tataaatcta tactcacagc aaataaatga taaaacactc aaaaaatata 3180
aaatgagtgt tttatttaag caaattttaa gtgatacaga atctaaatct tttgtaattg 3240
ataagttaga agatgatagt gatgtagtta caacgatgca aagtttttat gagcaaatag 3300
cagcttttaa aacagtagaa gaaaaatcta ttaaagaaac actatcttta ttatttgatg 3360
atttaaaagc tcaaaaactt gatttgagta aaatttattt taaaaatgat aaatctctta 3420
ctgatctatc acaacaagtt tttgatgatt atagtgttat tggtacagcg gtactagaat 3480
atataactca acaaatagca cctaaaaatc ttgataaccc tagtaagaaa gagcaagaat 3540
taatagccaa aaaaactgaa aaagcaaaat acttatctct agaaactata aagcttgcct 3600
tagaagaatt taataagcat agagatatag ataaacagtg taggtttgaa gaaatacttg 3660
caaactttgc ggctattccg atgatatttg atgaaatagc tcaaaacaaa gacaatttgg 3720
cacagatatc tatcaaatat caaaatcaag gtaaaaaaga cctacttcaa gctagtgcgg 3780
aagatgatgt taaagctatc aaggatcttt tagatcaaac taataatctc ttacataaac 3840
taaaaatatt tcatattagt cagtcagaag ataaggcaaa tattttagac aaggatgagc 3900
atttttatct agtatttgag gagtgctact ttgagctagc gaatatagtg cctctttata 3960
acaaaattag aaactatata actcaaaagc catatagtga tgagaaattt aagctcaatt 4020
ttgagaactc gactttggct aatggttggg ataaaaataa agagcctgac aatacggcaa 4080
ttttatttat caaagatgat aaatattatc tgggtgtgat gaataagaaa aataacaaaa 4140
tatttgatga taaagctatc aaagaaaata aaggcgaggg ttataaaaaa attgtttata 4200
aacttttacc tggcgcaaat aaaatgttac ctaaggtttt cttttctgct aaatctataa 4260
aattttataa tcctagtgaa gatatactta gaataagaaa tcattccaca catacaaaaa 4320
atggtagtcc tcaaaaagga tatgaaaaat ttgagtttaa tattgaagat tgccgaaaat 4380
ttatagattt ttataaacag tctataagta agcatccgga gtggaaagat tttggattta 4440
gattttctga tactcaaaga tataattcta tagatgaatt ttatagagaa gttgaaaatc 4500
aaggctacaa actaactttt gaaaatatat cagagagcta tattgatagc gtagttaatc 4560
agggtaaatt gtacctattc caaatctata ataaagattt ttcagcttat agcaaagggc 4620
gaccaaatct acatacttta tattggaaag cgctgtttga tgagagaaat cttcaagatg 4680
tggtttataa gctaaatggt gaggcagagc ttttttatcg taaacaatca atacctaaaa 4740
aaatcactca cccagctaaa gaggcaatag ctaataaaaa caaagataat cctaaaaaag 4800
agagtgtttt tgaatatgat ttaatcaaag ataaacgctt tactgaagat aagtttttct 4860
ttcactgtcc tattacaatc aattttaaat ctagtggagc taataagttt aatgatgaaa 4920
tcaatttatt gctaaaagaa aaagcaaatg atgttcatat attaagtata gctagaggtg 4980
aaagacattt agcttactat actttggtag atggtaaagg caatatcatc aaacaagata 5040
ctttcaacat cattggtaat gatagaatga aaacaaacta ccatgataag cttgctgcaa 5100
tagagaaaga tagggattca gctaggaaag actggaaaaa gataaataac atcaaagaga 5160
tgaaagaggg ctatctatct caggtagttc atgaaatagc taagctagtt atagagtata 5220
atgctattgt ggtttttgag gatttaaatt ttggatttaa aagagggcgt ttcaaggtag 5280
agaagcaggt ctatcaaaag ttagaaaaaa tgctaattga gaaactaaac tatctagttt 5340
tcaaagataa tgagtttgat aaaactgggg gagtgcttag agcttatcag ctaacagcac 5400
cttttgagac ttttaaaaag atgggtaaac aaacaggtat tatctactat gtaccagctg 5460
gttttacttc aaaaatttgt cctgtaactg gttttgtaaa tcagttatat cctaagtatg 5520
aaagtgtcag caaatctcaa gagttcttta gtaagtttga caagatttgt tataaccttg 5580
ataagggcta ttttgagttt agttttgatt ataaaaactt tggtgacaag gctgccaaag 5640
gcaagtggac tatagctagc tttgggagta gattgattaa ctttagaaat tcagataaaa 5700
atcataattg ggatactcga gaagtttatc caactaaaga gttggagaaa ttgctaaaag 5760
attattctat cgaatatggg catggcgaat gtatcaaagc agctatttgc ggtgagagcg 5820
acaaaaagtt ttttgctaag ctaactagtg tcctaaatac tatcttacaa atgcgtaact 5880
caaaaacagg tactgagtta gattatctaa tttcaccagt agcagatgta aatggcaatt 5940
tctttgattc gcgacaggcg ccaaaaaata tgcctcaaga tgctgatgcc aatggtgctt 6000
atcatattgg gctaaaaggt ctgatgctac taggtaggat caaaaataat caagagggca 6060
aaaaactcaa tttggttatc aaaaatgaag agtattttga gttcgtgcag aataggaata 6120
actctggtgg ttctcccaag aagaagagga aagtctaact gcagtataat cagaaacagc 6180
ccgcggatgt tgatctgcgg gctgtttttt attgatcgaa tggccatgac caaaatccct 6240
taacgtgaag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 6300
ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 6360
agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 6420
cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 6480
caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 6540
tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 6600
ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 6660
ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 6720
gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 6780
gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 6840
tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 6900
cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 6960
gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg agacaggtca 7020
ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caactcaaaa tggtatgcgt 7080
tttgacacat ccactatata tccgtgtcgt tctgtccact cctgaatccc attccagaaa 7140
ttctctagcg attccagaag tttctcagag tcggaaagtt gaccagacat tacgaactgg 7200
cacagatggt cataacctga aggaagatct gattgcttaa ctgcttcagt taagaccgaa 7260
gcgctcgtcg tataacagat gcgatgatgc agaccaatca acatggcacc tgccattgct 7320
acctgcacag tcaaggatgg tagaaatgtt gtcggtcctt gcacacgaat attacgccat 7380
ttgcctgcat attcaaacag ctcttctacg ataagggcac aaatcgcatc gtggaacgtt 7440
tgggcttcta ccgatttagc agtttgatac actttctcta agtatccacc tgaatcataa 7500
atcggcaaaa tagagaaaaa ttgaccatgt gtaagcggcc aatctgattc cacctgagat 7560
gcataatcta gtagaatctc ttcgctatca aaattcactt ccaccttcca ctcaccggtt 7620
gtccattcat ggctgaactc tgcttcctct gttgacatga cacacatcat ctcaatatcc 7680
gaatagggcc catcagtctg acgaccaaga gagccataaa caccaatagc cttaacatca 7740
tccccatatt tatccaatat tcgttcctta atttcatgaa caatcttcat tctttcttct 7800
ctagtcatta ttattggtcc attcactatt ctcattcccc tttcagataa ttttagattt 7860
gcttttctaa ataagaatat ttggagagca ccgttcttat tcagctatta aacccattat 7920
atcgggtttt tgaggggatt tcaactgcag acacctaaat tcaaaatcta tcggtcagat 7980
ttataccgat ttgattttat atattcttga ataacatacg ccgagttatc acataaaagc 8040
gggaaccaat catcaaattt aaacttcatt gcataatcca ttaaactctt aaattctacg 8100
attccttgtt catcaataaa ctcaatcatt tctttaatta atttatatct atctgttgtt 8160
gttttcttta ataattcatc aacatctaca ccgccataaa ctatcatatc ttctttttga 8220
tatttaaatt tattaggatc gtccatgtga agcatatatc tcacaagacc tttcacactt 8280
cctgcaatct gcggaatagt cgcattcaat tcttctgtaa ttatttttat ctgttcataa 8340
gatttattac cctcatacat cactagaata tgataatgct cttttttcat cctatcttct 8400
gtatcagtat ccctatcatg taatggagac actacaaatt gaatgtgtaa ctcttttaaa 8460
tactctaacc actcggcttt tgctgattct ggatataaaa caaatgtcca attacgtcct 8520
cttgaatttt tcttgttttc agtttctttt attacatttt cgctcatgat ataataacgg 8580
tgctaataca tttaacaaaa tttagtcata gataggcagc atgccagtgc tgtctatctt 8640
tttttgttta aaatgcaccg tattcctcct ttgcatattt ttttattaga ataccggttg 8700
catctgattt gctaatatta tatttttctt tgattctatt taatatctca ttttcttctg 8760
ttgtaagtct taaagtaaca gcaacttttt tctcttcttt tctatctaca accatcactg 8820
tacctcccaa catctgtttt tttcacttta acataaaaaa caacctttta acattaaaaa 8880
cccaatattt atttatttgt ttggacaatg gacaatggac acctaggggg gaggtcgtag 8940
taccccccta tgttttctcc cctaaataac cccaaaaatc taagaaaaaa agacctcaaa 9000
aaggtcttta attaacatct caaatttcgc atttattcca atttcctttt tgcgtgtgat 9060
gcgttattaa cgttgatata atttaaattt tatttgacaa aaatgggctc gtgttgtaca 9120
ataaatgtag aggtagagac gcgaggtcta agaactttaa ataatttcta ctgttgtaga 9180
tagagaccgt gaagttaata aggtctcaaa tttctactgt tgtagatcgt ctctgaactg 9240
attcaagcaa gcttaaaccc agctcaatga gctgggtttt ttgtttgttt tttcaaactt 9300
agttagcttg gccagtgcct ctagagtcaa gtaaagagtc gacctgttac gaacggcaga 9360
tcagaatttt gtaataaaaa aagagcctgc tcattacact gcgggctctt tttcatggtc 9420
agaagacggg taaccaagat aacaa 9445
<210> 11
<211> 9445
<212> DNA
<213> （人工序列）
<400> 11
atgtcatgac attggtgtac agaaatggcg cagcaatggc aagaacgtcc cgggcggagc 60
tcaggcctta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 120
tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 180
ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc 240
gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa 300
tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat 360
cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg 420
cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc 480
atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc 540
ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag 600
acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc 660
tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg 720
tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca 780
tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg 840
tgcaccgccg ttttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg 900
gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg 960
gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc 1020
acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc 1080
gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca 1140
tactctgcga catcgtataa cgttactggt ttcatcaaaa tcgtctccct ccgtttgaat 1200
atttgattga tcgtaaccag atgaagcact ctttccacta tccctacagt gttatggctt 1260
gaacaatcac gaaacaataa ttggtacgta cgatctttca gccgactcaa acatcaaatc 1320
ttacaaatgt agtctttgaa agtattacat atgtaagatt taaatgcaac cgttttttcg 1380
gaaggaaatg atgacctcgt ttccaccgga attagcttgg taccagctat tgtaacataa 1440
tcggtacggg ggtgaaaaag ctaacggaaa agggagcgga aaagaatgat gtaagcgtga 1500
aaaatttttt aaaaaatctc ttgacattgg aagggagata tgttattata agaattgcgg 1560
aattgtgagc ggataacaat tcataattgt gagcggataa caattcaacc ccaaaggagg 1620
tgggatccat gagcgaagtc gagttctccc acgaatactg gatgcgccat gcccttacac 1680
ttgccaaacg cgctcgcgat gaacgcgaag tcccagttgg cgccgttctt gttcttaaca 1740
accgcgtcat cggagaaggc tggaaccgcg ctatcggact tcatgatccg acagctcatg 1800
ccgaaattat ggctctgcgc caaggcggac ttgtcatgca gaattacaga cttattgatg 1860
ccacgcttta ctcaacgttc gaaccgtgcg tcatgtgcgc cggcgccatg attcatagcc 1920
gcatcggcag agttgtcttc ggcgtccgca attcaaaaag aggcgccgcc ggctccctta 1980
tgaacgtcct taactacccg ggcatgaatc accgcgttga aatcacagaa ggcattctgg 2040
ccgatgagtg cgccgctctg ctgtgcgact tctatagaat gccgagaaga gtcttcaacg 2100
cccagaagaa agcccaaagc agcattaact ctggaggatc atccggaggc agctctggaa 2160
gtgaaacacc aggaacaagc gaatcagcta caccagagtc ctctggaggc tcatctggag 2220
gaagctcaat ttatcaagaa tttgttaata aatatagttt aagtaaaact ctaagatttg 2280
agttaatccc acagggtaaa acacttgaaa acataaaagc aagaggtttg attttagatg 2340
atgagaaaag agctaaagac tacaaaaagg ctaaacaaat aattgataaa tatcatcagt 2400
tttttataga ggagatatta agttcggttt gtattagcga agatttatta caaaactatt 2460
ctgatgttta ttttaaactt aaaaagagtg atgatgataa tctacaaaaa gattttaaaa 2520
gtgcaaaaga tacgataaag aaacaaatat ctgaatatat aaaggactca gagaaattta 2580
agaatttgtt taatcaaaac cttatcgatg ctaaaaaagg gcaagagtca gatttaattc 2640
tatggctaaa gcaatctaag gataatggta tagaactatt taaagccaat agtgatatca 2700
cagatataga tgaggcgtta gaaataatca aatcttttaa aggttggaca acttatttta 2760
agggttttca tgaaaataga aaaaatgttt atagtagcaa tgatattcct acatctatta 2820
tttataggat agtagatgat aatttgccta aatttctaga aaataaagct aagtatgaga 2880
gtttaaaaga caaagctcca gaagctataa actatgaaca aattaaaaaa gatttggcag 2940
aagagctaac ctttgatatt gactacaaaa catctgaagt taatcaaaga gttttttcac 3000
ttgatgaagt ttttgagata gcaaacttta ataattatct aaatcaaagt ggtattacta 3060
aatttaatac tattattggt ggtaaatttg taaatggtga aaatacaaag agaaaaggta 3120
taaatgaata tataaatcta tactcacagc aaataaatga taaaacactc aaaaaatata 3180
aaatgagtgt tttatttaag caaattttaa gtgatacaga atctaaatct tttgtaattg 3240
ataagttaga agatgatagt gatgtagtta caacgatgca aagtttttat gagcaaatag 3300
cagcttttaa aacagtagaa gaaaaatcta ttaaagaaac actatcttta ttatttgatg 3360
atttaaaagc tcaaaaactt gatttgagta aaatttattt taaaaatgat aaatctctta 3420
ctgatctatc acaacaagtt tttgatgatt atagtgttat tggtacagcg gtactagaat 3480
atataactca acaaatagca cctaaaaatc ttgataaccc tagtaagaaa gagcaagaat 3540
taatagccaa aaaaactgaa aaagcaaaat acttatctct agaaactata aagcttgcct 3600
tagaagaatt taataagcat agagatatag ataaacagtg taggtttgaa gaaatacttg 3660
caaactttgc ggctattccg atgatatttg atgaaatagc tcaaaacaaa gacaatttgg 3720
cacagatatc tatcaaatat caaaatcaag gtaaaaaaga cctacttcaa gctagtgcgg 3780
aagatgatgt taaagctatc aaggatcttt tagatcaaac taataatctc ttacataaac 3840
taaaaatatt tcatattagt cagtcagaag ataaggcaaa tattttagac aaggatgagc 3900
atttttatct agtatttgag gagtgctact ttgagctagc gaatatagtg cctctttata 3960
acaaaattag aaactatata actcaaaagc catatagtga tgagaaattt aagctcaatt 4020
ttgagaactc gactttggct aatggttggg ataaaaataa agagcctgac aatacggcaa 4080
ttttatttat caaagatgat aaatattatc tgggtgtgat gaataagaaa aataacaaaa 4140
tatttgatga taaagctatc aaagaaaata aaggcgaggg ttataaaaaa attgtttata 4200
aacttttacc tggcgcaaat aaaatgttac ctaaggtttt cttttctgct aaatctataa 4260
aattttataa tcctagtgaa gatatactta gaataagaaa tcattccaca catacaaaaa 4320
atggtagtcc tcaaaaagga tatgaaaaat ttgagtttaa tattgaagat tgccgaaaat 4380
ttatagattt ttataaacag tctataagta agcatccgga gtggaaagat tttggattta 4440
gattttctga tactcaaaga tataattcta tagatgaatt ttatagagaa gttgaaaatc 4500
aaggctacaa actaactttt gaaaatatat cagagagcta tattgatagc gtagttaatc 4560
agggtaaatt gtacctattc caaatctata ataaagattt ttcagcttat agcaaagggc 4620
gaccaaatct acatacttta tattggaaag cgctgtttga tgagagaaat cttcaagatg 4680
tggtttataa gctaaatggt gaggcagagc ttttttatcg taaacaatca atacctaaaa 4740
aaatcactca cccagctaaa gaggcaatag ctaataaaaa caaagataat cctaaaaaag 4800
agagtgtttt tgaatatgat ttaatcaaag ataaacgctt tactgaagat aagtttttct 4860
ttcactgtcc tattacaatc aattttaaat ctagtggagc taataagttt aatgatgaaa 4920
tcaatttatt gctaaaagaa aaagcaaatg atgttcatat attaagtata gctagaggtg 4980
aaagacattt agcttactat actttggtag atggtaaagg caatatcatc aaacaagata 5040
ctttcaacat cattggtaat gatagaatga aaacaaacta ccatgataag cttgctgcaa 5100
tagagaaaga tagggattca gctaggaaag actggaaaaa gataaataac atcaaagaga 5160
tgaaagaggg ctatctatct caggtagttc atgaaatagc taagctagtt atagagtata 5220
atgctattgt ggtttttgag gatttaaatt ttggatttaa aagagggcgt ttcaaggtag 5280
agaagcaggt ctatcaaaag ttagaaaaaa tgctaattga gaaactaaac tatctagttt 5340
tcaaagataa tgagtttgat aaaactgggg gagtgcttag agcttatcag ctaacagcac 5400
cttttgagac ttttaaaaag atgggtaaac aaacaggtat tatctactat gtaccagctg 5460
gttttacttc aaaaatttgt cctgtaactg gttttgtaaa tcagttatat cctaagtatg 5520
aaagtgtcag caaatctcaa gagttcttta gtaagtttga caagatttgt tataaccttg 5580
ataagggcta ttttgagttt agttttgatt ataaaaactt tggtgacaag gctgccaaag 5640
gcaagtggac tatagctagc tttgggagta gattgattaa ctttagaaat tcagataaaa 5700
atcataattg ggatactcga gaagtttatc caactaaaga gttggagaaa ttgctaaaag 5760
attattctat cgaatatggg catggcgaat gtatcaaagc agctatttgc ggtgagagcg 5820
acaaaaagtt ttttgctaag ctaactagtg tcctaaatac tatcttacaa atgcgtaact 5880
caaaaacagg tactgagtta gattatctaa tttcaccagt agcagatgta aatggcaatt 5940
tctttgattc gcgacaggcg ccaaaaaata tgcctcaaga tgctgatgcc aatggtgctt 6000
atcatattgg gctaaaaggt ctgatgctac taggtaggat caaaaataat caagagggca 6060
aaaaactcaa tttggttatc aaaaatgaag agtattttga gttcgtgcag aataggaata 6120
actctggtgg ttctcccaag aagaagagga aagtctaact gcagtataat cagaaacagc 6180
ccgcggatgt tgatctgcgg gctgtttttt attgatcgaa tggccatgac caaaatccct 6240
taacgtgaag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 6300
ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 6360
agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 6420
cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 6480
caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 6540
tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 6600
ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 6660
ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 6720
gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 6780
gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 6840
tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 6900
cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 6960
gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg agacaggtca 7020
ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caactcaaaa tggtatgcgt 7080
tttgacacat ccactatata tccgtgtcgt tctgtccact cctgaatccc attccagaaa 7140
ttctctagcg attccagaag tttctcagag tcggaaagtt gaccagacat tacgaactgg 7200
cacagatggt cataacctga aggaagatct gattgcttaa ctgcttcagt taagaccgaa 7260
gcgctcgtcg tataacagat gcgatgatgc agaccaatca acatggcacc tgccattgct 7320
acctgcacag tcaaggatgg tagaaatgtt gtcggtcctt gcacacgaat attacgccat 7380
ttgcctgcat attcaaacag ctcttctacg ataagggcac aaatcgcatc gtggaacgtt 7440
tgggcttcta ccgatttagc agtttgatac actttctcta agtatccacc tgaatcataa 7500
atcggcaaaa tagagaaaaa ttgaccatgt gtaagcggcc aatctgattc cacctgagat 7560
gcataatcta gtagaatctc ttcgctatca aaattcactt ccaccttcca ctcaccggtt 7620
gtccattcat ggctgaactc tgcttcctct gttgacatga cacacatcat ctcaatatcc 7680
gaatagggcc catcagtctg acgaccaaga gagccataaa caccaatagc cttaacatca 7740
tccccatatt tatccaatat tcgttcctta atttcatgaa caatcttcat tctttcttct 7800
ctagtcatta ttattggtcc attcactatt ctcattcccc tttcagataa ttttagattt 7860
gcttttctaa ataagaatat ttggagagca ccgttcttat tcagctatta aacccattat 7920
atcgggtttt tgaggggatt tcaactgcag acacctaaat tcaaaatcta tcggtcagat 7980
ttataccgat ttgattttat atattcttga ataacatacg ccgagttatc acataaaagc 8040
gggaaccaat catcaaattt aaacttcatt gcataatcca ttaaactctt aaattctacg 8100
attccttgtt catcaataaa ctcaatcatt tctttaatta atttatatct atctgttgtt 8160
gttttcttta ataattcatc aacatctaca ccgccataaa ctatcatatc ttctttttga 8220
tatttaaatt tattaggatc gtccatgtga agcatatatc tcacaagacc tttcacactt 8280
cctgcaatct gcggaatagt cgcattcaat tcttctgtaa ttatttttat ctgttcataa 8340
gatttattac cctcatacat cactagaata tgataatgct cttttttcat cctatcttct 8400
gtatcagtat ccctatcatg taatggagac actacaaatt gaatgtgtaa ctcttttaaa 8460
tactctaacc actcggcttt tgctgattct ggatataaaa caaatgtcca attacgtcct 8520
cttgaatttt tcttgttttc agtttctttt attacatttt cgctcatgat ataataacgg 8580
tgctaataca tttaacaaaa tttagtcata gataggcagc atgccagtgc tgtctatctt 8640
tttttgttta aaatgcaccg tattcctcct ttgcatattt ttttattaga ataccggttg 8700
catctgattt gctaatatta tatttttctt tgattctatt taatatctca ttttcttctg 8760
ttgtaagtct taaagtaaca gcaacttttt tctcttcttt tctatctaca accatcactg 8820
tacctcccaa catctgtttt tttcacttta acataaaaaa caacctttta acattaaaaa 8880
cccaatattt atttatttgt ttggacaatg gacaatggac acctaggggg gaggtcgtag 8940
taccccccta tgttttctcc cctaaataac cccaaaaatc taagaaaaaa agacctcaaa 9000
aaggtcttta attaacatct caaatttcgc atttattcca atttcctttt tgcgtgtgat 9060
gcgttattaa cgttgatata atttaaattt tatttgacaa aaatgggctc gtgttgtaca 9120
ataaatgtag aggtagagac gcgaggtcta agaactttaa ataatttcta ctgttgtaga 9180
tagagaccgt gaagttaata aggtctcaaa tttctactgt tgtagatcgt ctctgaactg 9240
attcaagcaa gcttaaaccc agctcaatga gctgggtttt ttgtttgttt tttcaaactt 9300
agttagcttg gccagtgcct ctagagtcaa gtaaagagtc gacctgttac gaacggcaga 9360
tcagaatttt gtaataaaaa aagagcctgc tcattacact gcgggctctt tttcatggtc 9420
agaagacggg taaccaagat aacaa 9445

Claims (10)

1.一种基因编辑融合蛋白，其特征在于，所述基因编辑融合蛋白的氨基酸序列为SEQID NO.2-4所示的序列之一。
2.一种腺嘌呤碱基编辑器，其特征在于：所述腺嘌呤碱基编辑器包括权利要求1所述的基因编辑融合蛋白和crRNA阵列***区。
3.根据权利要求2所述的腺嘌呤碱基编辑器，其特征在于：所述crRNA阵列***区的核苷酸序列如SEQ ID NO.5所示。
4.根据权利要求2所述的腺嘌呤碱基编辑器，其特征在于：所述基因编辑融合蛋白通过诱导型启动子Pgrac100调控表达。
5.根据权利要求2所述的腺嘌呤碱基编辑器，其特征在于：crRNA阵列***区通过组成型启动子Pveg调控表达。
6.一种枯草芽孢杆菌腺嘌呤碱基编辑器，其特征在于，所述枯草芽孢杆菌腺嘌呤碱基编辑器包括：含有编码权利要求1所述基因编辑融合蛋白的基因以及权利要求2所述crRNA阵列***区的质粒。
7.根据权利要求6所述的枯草芽孢杆菌腺嘌呤碱基编辑器，其特征在于：所述质粒的核苷酸序列为SEQ ID NO.9-11所示之一。
8.权利要求1所述的基因编辑融合蛋白、权利要求2-5任一项所述的腺嘌呤碱基编辑器或权利要求6或7所述的枯草芽孢杆菌腺嘌呤碱基编辑器在基因编辑中的应用。
9.根据权利要求8所述的应用，其特征在于：所述的基因编辑为将基因组上多位点腺嘌呤转化为鸟嘌呤。
10.根据权利要求8所述的应用，其特征在于：所述的基因编辑融合蛋白、腺嘌呤碱基编辑器或枯草芽孢杆菌腺嘌呤碱基编辑器用于突变体的获取或胞外蛋白酶的失活。
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