CN115074369A - 表达全长abca4基因的腺相关病毒载体及应用 - Google Patents

表达全长abca4基因的腺相关病毒载体及应用 Download PDF

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CN115074369A
CN115074369A CN202210721461.2A CN202210721461A CN115074369A CN 115074369 A CN115074369 A CN 115074369A CN 202210721461 A CN202210721461 A CN 202210721461A CN 115074369 A CN115074369 A CN 115074369A
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杨阳
魏于全
王青楠
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Chengdu Jinweike Biological Technology Co ltd
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Abstract

本发明公开了表达全长ABCA4基因的腺相关病毒载体及应用。ABCA4基因片段，所述的基因片段编码ABCA4全长蛋白以p1150Cys为***位点断裂得到的任意一个片段。一种重组腺相关病毒，由含有含内含肽片段的人ABCA4‑N或ABCA4‑C蛋白片段的编码基因的载体、AAV的Rep及Cap蛋白表达质粒、辅助质粒共转染HEK293细胞包装制备得到。利用本发明所重组腺相关病毒可在STGDI疾病模型中高效的转导视网膜光感受器细胞，并在高效地表达全长的ABCA4蛋白，实现疾病治疗的目标。

Description

表达全长ABCA4基因的腺相关病毒载体及应用
技术领域
本发明属于生物医药领域，涉及表达全长ABCA4基因的腺相关病毒载体及应用。
背景技术
STARGARDT病(STGDI)称眼底黄色斑点症，是青少年最常见的黄斑营养不良性疾病，可导致严重的、不可逆的双侧视力损害，患病率约为1/10000～1/8000。其临床特点是青少年时期发病(10～20岁)，进行性双眼中心视力下降伴不同程度的黄斑区及周边视网膜色素上皮层(retinal pigment epithelium，RPE)和神经上皮层萎缩，眼底检查后极部大面积的脂褐质沉积于黄斑区，呈典型的“金箔样”反光，早期可仅表现为“淤青样”或黄白斑点限局于黄斑中心凹及旁中心凹，晚期脂褐质遍布RPE层，荧光素眼底血管造影(fluoresceinfundus angiography，FFA)呈“脉络膜淹没征”，相干光断层扫描(OCT)示视网膜脉络膜萎缩薄变伴高反光颗粒物沉积。STGDI的遗传方式为常染色体隐性遗传，其唯一致病基因是ABCA4(ATP-Binding Cassette,subfamily A,member 4)基因，该基因位于染色体1p22.1，由50个外显子组成，全长约8kb，编码2235个氨基酸，称为Rim跨膜蛋白，该蛋白属于ATP结合盒式转运超家族基因亚科，主要位于光感受器外节，是视觉循环必需的运输蛋白。现已发现900余种ABCA4基因的致病突变(The Human Gene Mutation Database，HGDM；http://www.hgmd.cf.ac.uk/ac/index.php)，除STGDI，其突变还可导致常染色体隐性锥杆细胞营养不良和视网膜色素变性。在STGDI患者中，ABCA4基因突变会阻碍类视黄醇的运输和再循环，引起类视黄醇积聚蓄积，最终导致形成有毒的类视黄醇二聚体(N-视黄基-N-视黄醇胺，A2E)，从而使得视网膜色素上皮和感光细胞变性，患者视力丧失。
对于正常基因功能缺失疾病的治疗，最直接的方法就是利用载体将正常基因导入患者体内实现治疗作用。近年来，针对基因治疗的临床研究不断涌现，并已在一些遗传病及恶性肿瘤领域取得了重大突破，为一些传统疗法无效、容易复发或难治疾病患者带来了新的希望。2008年，Kong等将全长ABCA4编码序列包装到马传染性贫血病毒(Equineinfectious anaemia virus，EIAV)改造的慢病毒载体中，并通过视网膜下注射递送至出生后第5天ABCA4基因敲除小鼠[1]。ABCA4蛋白表达持续超过12个月，减少了A2E的积累。2011年，英国Oxford Biomedica公司开展了基于EIAV的慢病毒载体基因治疗I/II期临床试验(NCT01367444)，该试验于2019年终止，治疗效果不理想。尽管慢病毒很强的基因包装能力，但其大小和病毒结构限制了慢病毒载体在视网膜中的转导能力。由于缺乏临床试验的疗效数据，目前业界很少鼓励继续使用慢病毒载体进行STGDI基因治疗。在基因治疗领域，腺相关病毒载体(Adeno associated virus，AAV)已广泛用于治疗性转基因传递和视网膜疾病的临床试验，显示出良好的安全性，且不良事件发生较少。2017年，Spark Therapeutics公司的Luxturna(voretigene neparvovec)成为FDA批准的首款AAV基因疗法，用于治疗由RPE65基因突变导致的遗传性视网膜疾病病变。然而ABCA4基因编码序列约为6.8kb，使用AAV载体递送十分困难，因为AAV的最佳包装能力约为4.7kb。在早期的STGDI基因治疗研究中，Allocca等尝试构建包含完整6.8kb ABCA4编码序列的单一“超大”AAV。研究发现“超大”的AAV确实能在体内和体外产生全长的ABCA4蛋白[2]。然而，由于无法可靠地制备含有确定转基因的均一AAV制剂，这种方法的临床应用十分困难。2014年，Trapani等开发出基于反式剪切(trans-splicing)、重叠(overlapping)以及混合(hybrid)等双AAV载体基因治疗策略，用于递送大基因片段[3]。在Trapani等研究中发现，“Overlapping”双AAV载体策略在光感受器细胞中不能有效的表达全长的目的基因。因此，在后续的研究中，研究人员更加关注反式剪切(trans-splicing)以及混合(hybrid)的双AAV载体递送技术。2019年，Dyka等发现在ABCA4基因敲除小鼠中，注射反式剪切(trans-splicing)双AAV载体后，18％眼睛表达全长ABCA4蛋白；而注射混合(hybrid)双AAV载体后，50％的眼睛表达全长ABCA4蛋白。此外，与未治疗的眼睛相比，反式剪切(trans-splicing)和混合(hybrid)双AAV载体治疗组的眼睛显示出脂褐质颗粒的显著减少[4]。由于双AAV载体有效转导所需的各种限制步骤，包括：适当的DNA多联体形成、异质mRNA的稳定性和跨载体连接的剪接效率，在光感受器中使用双AAV载体实现的转基因表达效率远低于使用单一AAV载体实现的转基因表达效率[5]。
断裂内含肽(Split inteins)是存在于古菌、蓝细菌等单细胞生物体中的一种遗传元件，它们在宿主蛋白的末端表达为两个独立的多肽并催化它们反式剪接形成一个较大的蛋白，催化后并不会在最终蛋白产物中产生遗留[6,7]。断裂内含肽被广泛用于蛋白质纯化、蛋白标记步骤以及CRISPR/Cas9基因编辑核酸酶的重组等。2019年，Tornabene等研究发现，断裂内含肽成功将两截绿色荧光蛋白片段拼接成完整的绿色荧光蛋白，且拼接效率高，跟单个AAV载体表达的蛋白水平无显著差异。这些实验结果表明，通过断裂内含肽的剪接作用，可以成功的将多AAV载体递送多肽片段拼接成完整且具有功能的蛋白质[5]。
由于AAV介导的基因治疗现已成为遗传性视网膜变性的批准治疗方法，因此增加AAV转基因大小的能力将大大拓宽治疗适应症的范围。STGDI的遗传方式为常染色体隐性遗传，其唯一致病基因是ABCA4基因，编码序列约为6.8kb，超出了单个AAV的最佳包装容量(<4.7kb)。断裂内含肽双AAV载体策略最关键的是内含肽序列和***位点，因为这将极大的影响目的蛋白的剪切效率，以及目的蛋白质的三级结构和翻译后修饰，进而影响蛋白功能。此外，由于断裂内含肽双AAV载体分别表达ABCA4蛋白的N端和C端。因此，未有效进行拼接的N端和C端副产物具有潜在的安全性风险。
发明内容
本发明的目的是针对现有技术的上述不足，提供一种ABCA4-N或ABCA4-C端蛋白表达框。
本发明的第二个目的是提供含有所述表达框的载体及载体***。
本发明的第三个目的是提供表达ABCA4蛋白的重组腺相关病毒及其组合物。
本发明的第四个目的是提供所述的蛋白表达框、载体、重组腺相关病毒的应用。
本发明的目的可通过以下技术方案实现：
ABCA4基因片段，所述的基因片段编码ABCA4全长蛋白以p1150Cys为***位点断裂得到的任意一个片段。
作为本发明的一种优选，所述的ABCA4基因片段的一端含有内含肽片段；内含肽选择来自于Synechocystis sp.strain PCC6803 DnaB内含肽(Ssp intein，SEQ ID NO.2)，Rhodothermus marinus DnaB内含肽(Rma intein，SEQ ID NO.3)，Nostoc punctiformePCC73102 DnaE内含肽(Npu intein，SEQ ID NO.4)和Consensus DnaE内含肽(Cfa intein，SEQ ID NO.5)。
所述的内含肽选择SEQ ID NO.3所示的Rma intein或SEQ ID NO.4所示的Npuintein。
作为本发明的一种优选，所述的ABCA4基因片段选自以下任意一个：
(1)如SEQ ID NO.8所示的含内含肽片段的人ABCA4-N蛋白片段的编码基因，
(2)如SEQ ID NO.9所示的含内含肽片段的人ABCA4-C蛋白片段的编码基因。
一种ABCA4-N或ABCA4-C端蛋白表达框，由增强子-启动子-内含子-目的基因序列-polyA信号组成，所述的增强子选自人214bp interphotoreceptor retinoid-bindingprotein(IRBP)enhancer增强子序列(PMID:9723991或US20140275231)，启动子选自视网膜感光细胞特异的人RS1(Retinoschisin 1)启动子，人Rho(Rhodopsin)启动子、人RK(rhodopsin kinase)启动子或小鼠CAR(Cone arrestin)启动子；内含子选自SV40内含子；目的基因选自含内含肽片段的人ABCA4-N或ABCA4-C蛋白片段的编码基因；polyA信号序列选自SV40 PolyA，bGH polyA，hGH polyA或rBG polyA序列；所述的增强子、启动子、内含子、目的基因序列和polyA之间通过键或核苷酸连接序列连接；所述的人ABCA4-N和ABCA4-C蛋白片段为ABCA4全长蛋白以p1150Cys为***位点断裂所得。
作为本发明的一种优选，所述的内含肽选择SEQ ID NO.3所示的Rma intein或SEQID NO.4所示的Npu intein；优选SEQ ID NO.3所示的Rma intein。
作为本发明的进一步优选，所述的含内含肽片段的人ABCA4-N端氨基酸序列如SEQID NO.6所示，所述的含内含肽片段的人ABCA4-C端氨基酸序列如SEQ ID NO.7所示。所述的含内含肽片段的人ABCA4-N蛋白片段的编码基因核苷酸序列如SEQ ID NO.8所示，含内含肽片段的人ABCA4-C蛋白片段的编码基因核苷酸序列如SEQ ID NO.9所示。
作为本发明的一种优选，所述的启动子选自RK启动子，序列如SEQ ID NO.10所示；内含子序列选自SV40内含子，序列如GenBank登录号MK225672.1公开的核苷酸序列的第4863bp-4959bp所示；polyA选自bGH ployA，序列如GenBank登录号MT267334.1公开的核苷酸序列的第957bp-1181bp所示。
作为本发明的更进一步优选，所述的ABCA4-N端蛋白表达框的核苷酸序列如SEQID NO.11；所述的ABCA4-C端蛋白表达框的核苷酸序列如SEQ ID NO.12。
一种载体，含有本发明所述的ABCA4基因片段、或本发明所述的ABCA4-N或ABCA4-C端蛋白表达框。
作为本发明的一种优选，所述载体选自以下任意一种重组腺相关病毒载体血清型：AAV1、AAV2、AAV3B、AAV4、AAV5、AAV6、AAV7、AAV8、AAV9、AAVrh10、AAV-LK03或AAVAnc80d，优选AAV2、AAV5、AAV8或AAV9。
一种质粒组合物，包含本发明所述的含有所述的ABCA4-N端蛋白表达框的载体和含有所述的ABCA4-C端蛋白表达框的载体；优选包含所述的载体、AAV的Rep及Cap蛋白表达质粒、辅助质粒；进一步优选包含所述的载体、AAV的Rep及Cap蛋白表达质粒pAAV2/8以及辅助质粒pAdΔF6。
一种重组腺相关病毒，由本发明所述的载体、AAV的Rep及Cap蛋白表达质粒、辅助质粒共转染HEK293细胞包装制备得到。
作为本发明的一种优选，所述的重组腺相关病毒由本发明所述的含有ABCA4-N蛋白表达框的载体、AAV的Rep及Cap蛋白表达质粒、辅助质粒共转染HEK293细胞包装制备得到。
作为本发明的一种优选，所述的重组腺相关病毒，由本发明所述的含有ABCA4-C蛋白表达框的载体、AAV的Rep及Cap蛋白表达质粒、辅助质粒共转染HEK293细胞包装制备得到。
一种重组腺相关病毒组合物，包含由本发明所述的含有ABCA4-N蛋白表达框的载体、AAV的Rep及Cap蛋白表达质粒，以及辅助质粒共转染HEK293细胞包装制备得到重组腺相关病毒；以及由本发明所述的含有ABCA4-C蛋白表达框的载体、AAV的Rep及Cap蛋白表达质粒、以及辅助质粒共转染HEK293细胞包装制备得到重组腺相关病毒。
本发明所述的ABCA4基因片段、所述的ABCA4-N或ABCA4-C端蛋白表达框、所述的载体、所述的质粒组合物、所述的重组腺相关病毒、所述的重组腺相关病毒组合物在制备治疗STGDI疾病的药物中的应用。
有益效果：
本发明提供了一种新设计高效且组织特异性表达ABCA4蛋白的重组腺相关病毒及其治疗STGDI的应用。本发明首先对ABCA4基因进行密码优化，提高了ABCA4蛋白表达水平。现有技术中未见对目标蛋白的断裂位点和内含肽序列进行***筛选。而对于断裂内含肽介导的双AAV载体技术，确定目标蛋白的最佳序列和***点将是至关重要的，因为这将极大的影响其在细胞内的蛋白剪切效率，以及目的蛋白质的三级结构和翻译后修饰，进而影响蛋白功能。在本发明中，我们基于蛋白结构和优化的ABCA4基因序列，对内含肽对目的蛋白的断裂位点和内含肽序列进行了***的筛选。在本发明中，我们还在体内外发现使用Rma内含肽能够使ABCA4蛋白的p1150Cys位点***形成的两段多肽反式剪接形成全长蛋白，副产物几乎检测不到，显示出非常好安全性。最后，构建高效组织特异表达全长ABCA4的AAV8intein病毒载体，制备了AAV8.RK.ABCA4-N与AAV8.RK.ABCA4-C病毒，用于STGDI疾病的基因治疗研究。在STGDI(ABCA4-KO)疾病模型中，将低剂量(1×108GC/μl的AAV8.RK.ABCA4-N与AAV8.RK.ABCA4-C病毒等体积混合，每只眼注射1μl)，中剂量(1×109GC/μl的AAV8.RK.ABCA4-N与AAV8.RK.ABCA4-C病毒等体积混合，每只眼注射1μl)和高剂量(1×1010GC/μl的AAV8.RK.ABCA4-N与AAV8.RK.ABCA4-C病毒等体积混合，每只眼注射1μl)重组腺相关病毒组合分别通过视网膜下腔，注射至疾病小鼠眼底后评估基因治疗疗效。在给药后第3个月和第6个月，低剂量、中剂量和高剂量治疗组的眼底自发荧光强度均明显低于未治疗组的STGDI疾病小鼠(ABCA4-KO)，与野生型小鼠相似；给药后六个月，通过视网膜电图(Electroretinogram，ERG)检测发现，治疗组小鼠b波波幅均高于未治疗组，视网膜功能得到显著改善；高效液相色谱(HPLC)检测发现AAV8 intein基因治疗后3个月，有毒的类视黄醇二聚体A2E降低至正常水平，且全长的ABCA4蛋白在视网膜感光细胞中的表达恢复至正常水平的22％(低剂量治疗组)、24％(中剂量治疗组)和30％(高剂量治疗组)。以上结果显示，通过新设计优化的ABCA4蛋白编码序列、对内含肽断裂位点和内含肽序列的设计、以及对ABCA4蛋白AAV8 intein表达框的设计，可在STGDI疾病模型中高效的转导视网膜光感受器细胞，并在高效地表达全长的ABCA4蛋白，实现疾病治疗的目标。
附图说明
图1 pAAV.CBh.ABCA4wt.bGH(A图)和pAAV.CBh.ABCA4co.bGH(B图)表达载体的质粒谱图
图2 ABCA4蛋白表达载体体外表达水平验证
图3 ABCA4目的基因序列内含肽断裂位点的设计、载体构建及验证
A为ABCA4目的基因序列内含肽断裂位点的设计
B为四种断裂内含肽***示意图
C为实施例3中蛋白免疫印迹法对12个实验组以及对照组的载体全长ABCA4蛋白及副产物表达水平进行比较
D为C的统计图
图4 pAAV.RK.ABCA4.RmaintN.bGH和pAAV.RK.RmaintC.ABCA4.bGH两个载体的质粒图谱
图5重组腺相关病毒AAV8.RK.ABCA4-N和重组腺相关病毒AAV8.RK.ABCA4-C结构示意图
图6在病毒给药后一个月(A图)和六个月(B图)，低剂量治疗组、中剂量治疗组、高剂量治疗组、未治疗组、野生型小鼠b波波幅统计图
图7 AAV8 intein基因治疗后视网膜细胞中ABCA4蛋白表达情况
A为ABCA4蛋白免疫荧光检测。基因治疗后3月通过免疫荧光检测所有组别小鼠眼球组织中ABCA4蛋白的表达，标尺＝50μm；
B为体内ABCA4蛋白表达水平检测。WB检测基因治疗后3月小鼠视网膜组织中ABCA4和GAPDH；
C为体内ABCA4蛋白表达水平WB检测灰度值统计。用image J对(A)图进行分析统计相对灰度值。
图8各组小鼠眼底脂褐质的积累结果
图9用海德堡激光眼科诊断仪FAF模块检测各组小鼠眼底自发荧光强弱结果
具体实施方式
实施例1：ABCA4蛋白目的基因序列优化及表达载体构建
ABCA4蛋白氨基酸序列如Genbank数据库NP_000341.2所示，全基因合成野生型ABCA4基因(ABCA4wt，Genbank数据库NM_000350.3(104bp-6925bp))和密码子优化的ABCA4基因(ABCA4co，SEQ ID NO.1)，通过酶切连接分别构建pAAV.CBh.ABCA4wt.bGH(图1A，SEQID NO.15)和pAAV.CBh.ABCA4co.bGH(图1B，SEQ ID NO.16)表达载体。
实施例2：ABCA4蛋白表达载体体外表达水平验证
将实施例1中的pAAV.CBh.ABCA4wt.bGH和pAAV.CBh.ABCA4wt.bGH以相同的质粒量(250ng),通过PEImax转染试剂转染培养于24孔板中的HEK293细胞，72h后提取蛋白，利用蛋白免疫印迹法(Western blot)对ABCA4蛋白的表达水平进行比较。结果显示密码子优化可以显著提高蛋白的表达水平(图2A)，通过对蛋白条带灰度值进行对比，密码子优化后ABCA4蛋白表达水平提高了4.7倍(图2B)。
实施例3：ABCA4目的基因序列内含肽断裂位点的设计、载体构建及验证
使用内含肽分别在ABCA4蛋白的p1150Cys、p1140cys、p1177cys位点进行反式剪接，从1号位到1150号位半胱氨酸、从1号位到1140号位半胱氨酸或从1号位到1177号位半胱氨酸作为氮端序列(ABCA4-N)，从1151号位半胱氨酸到2235号位、或从1141号位半胱氨酸到2235号位、或从1178号位半胱氨酸到2235号位作为碳端序列(ABCA4-C)(图3A)。分别选择Ssp内含肽(Sspintein，SEQ ID NO.2)、Npu内含肽(Npuintein，SEQ ID NO.3)、Rma内含肽(Rmaintein，SEQ ID NO.4)和Cfa内含肽(Cfaintein，SEQ ID NO.5)四种断裂内含肽***(图3B)，构建断裂内含肽ABCA4表达载体，共转染HEK293细胞，检测全长ABCA4蛋白的表达效率。
按照Sspintein、Npuintein、Rmaintein、Cfaintein四种断裂内含肽***和ABCA4蛋白的p1150cys、p1140cys、p1177cys三种内含肽断裂位点设置12组实验组，用全长ABCA4目的基因和eGFP作为对照组，转染培养于24孔板中的HEK293细胞，72h后提取蛋白，利用蛋白免疫印迹法对各组载体全长ABCA4蛋白表达水平进行比较。结果显示使用Rmaintein在ABCA4蛋白的p1150Cys位点进行反式剪接，副产物几乎检测不到，全长ABCA4蛋白的表达效率最高(图3C，D)。
实施例4：基因治疗候选载体设计构建
为了进一步评估构建Rmaintein在ABCA4蛋白的p1150Cys位点进行反式剪接后在体内的表达，基因治疗疗效和安全性。我们全基因合成了视网膜感光细胞特异RK启动子(SEQID NO.10)，通过T4连接亚克隆构建了病毒包装顺式质粒载体pAAV.RK.ABCA4.RmaintN.bGH(图4A，SEQ ID NO.17)和pAAV.RK.RmaintC.ABCA4.bGH两个载体(图4B，SEQ ID NO.18)，分别包含本发明所述的ABCA4-N端蛋白表达框(SEQ ID NO.11)和ABCA4-C端蛋白表达框(SEQID NO.12)。
实施例5：AAV病毒制备与纯化
参考Martin Lock等报道包装和纯化重组AAV病毒的方法[8]，采用PEI将AAV的Rep及Cap蛋白表达质粒(pAAV2/8)、辅助质粒(pAdΔF6)和AAV包装顺式质粒(pAAV.RK.RmaintC.ABCA4.bGH，SEQ ID NO.18；pAAV.RK.ABCA4.RmaintN.bGH，SEQ IDNO.17)分别共转染HEK293细胞包装制备病毒AAV8.RK.ABCA4-N(图5A，SEQ ID NO.13)和AAV8.RK.ABCA4-C病毒(图5B，SEQ ID NO.14)，转染48h后，收获细胞和培养上清，使用碘克沙醇超速梯度离心纯化AAV病毒，用数字定量PCR(ddPCR)法测定病毒滴度AAV8.RK.ABCA4-N病毒滴度为7.38×1013GC/mL，AAV8.RK.ABCA4-C病毒滴度为2.77×1013GC/mL。
实施例6：AAV8 intein基因治疗改善STGDI疾病小鼠ERG功能
使用实施例5中制备的AAV8.RK.ABCA4-N和AAV8.RK.ABCA4-C病毒等比例混合(简称AAV8 intein)，通过视网膜下腔共同注射STGDI疾病小鼠(ABCA4-KO，4周龄，n＝10)。根据病毒注射剂量分为低剂量治疗组(1×108GC AAV8 intein)，中剂量治疗组(1×109GC AAV8intein)和高剂量治疗组(1×1010GC AAV8 intein)。病毒给药后一个月和六个月，分别通过ERG检测不同刺激强度下各治疗组、未治疗组、野生型小鼠b波振幅。结果显示，在病毒给药后一个月，低剂量治疗组、中剂量治疗组、高剂量治疗组、未治疗组、野生型小鼠b波波幅均未出现统计学差异(图6A)；在病毒给药后六个月，治疗组小鼠b波波幅均高于未治疗组，且其中低剂量治疗组、中剂量治疗组与未治疗组的统计学差异最为显著(图6B，*p<0.05，**p<0.01，***p<0.001)，表明AAV8 intein治疗后可以显著改善STGDI疾病小鼠的眼睛视网膜电生理功能。
实施例7：AAV8 intein基因治疗后恢复视网膜细胞中ABCA4蛋白表达
在注射后3个月，我们收集小鼠的眼球组织，通过冰冻切片和免疫荧光染色，检测了视网膜组织中的ABCA4蛋白表达情况。结果显示，相比与未治疗组(n＝6)，全部治疗组(低剂量组n＝4，中剂量组n＝4，高剂量组n＝4)和野生型小鼠(n＝4)小鼠眼部组织中ABCA4表达恢复至正常水平，主要分布在视网膜感光细胞层(图7A)。同时，我们还收集眼球的视网膜组织，进行蛋白裂解，通过蛋白免疫印迹法进一步检测分析了ABCA4全长蛋白的表达情况。结果显示，相比与未治疗组(n＝6)，治疗组(低剂量组n＝6，中剂量组n＝6，高剂量组n＝6)和野生型(n＝6)小鼠眼部组织中均高效表达ABCA4蛋白(图7B)。低剂量治疗组(1×108GCAAV8 intein)，中剂量治疗组(1×109GC AAV8 intein)和高剂量治疗组(1×1010GC AAV8intein)的ABCA4蛋白水平分别达到了正常小鼠的22％，24％和30％。并且在所有WB检测的治疗组样本中，都检测到了全长的ABCA4蛋白且均未检测到截短的副产物蛋白。
实施例8：AAV8 intein基因治疗降低STGDI小鼠有毒的类视黄醇二聚体(N-视黄基-N-视黄醇胺，A2E)累积
在注射后3个月，我们收集小鼠的眼球组织，进行组织内色素提取与高效液相色谱检测，检测小鼠眼球组织中有毒的类视黄醇二聚体(N-视黄基-N-视黄醇胺，A2E)累积情况。结果显示，注射后3个月的野生型、低剂量治疗组、中剂量治疗组小鼠中A2E的含量均为0，在相同年龄的未治疗组小鼠中含量为7.03×10-6mol/l，高剂量治疗组小鼠中为6.21×10- 6mol/l，表示低剂量治疗组和中剂量治疗组中眼底脂褐质的积累均有明显减少(图8)。
实施例9：AAV8 intein基因治疗降低STGDI小鼠模型的眼底自发荧光
在注射后3个月、6个月，用海德堡激光眼科诊断仪FAF模块检测小鼠眼底自发荧光强弱。结果显示，治疗组小鼠自发荧光强度均更接近野生型小鼠，明显弱于未治疗组自发荧光强度,表明AAV8 intein治疗后可以显著改善疾病小鼠的眼睛色素沉积(图9)。
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8.Lock M,Alvira M,Vandenberghe LH,Samanta A,Toelen J,Debyser Z,WilsonJM:Rapid,simple,and versatile manufacturing of recombinant adeno-associatedviral vectors at scale.Human gene therapy 2010,21(10):1259-1271.
序列表
<110> 成都金唯科生物科技有限公司
<120> 表达全长ABCA4基因的腺相关病毒载体及应用
<160> 18
<170> SIPOSequenceListing 1.0
<210> 1
<211> 6822
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
atgggctttg tgcgacagat tcagctgctg ctgtggaaga actggaccct gcggaagcgg 60
cagaaaatca gattcgtggt ggaactcgtg tggcccctga gcctgtttct ggtgctgatc 120
tggctgcgga acgccaatcc tctgtacagc caccacgagt gtcacttccc caacaaggcc 180
atgccttctg ccggaatgct gccttggctg cagggcatct tctgcaacgt gaacaacccc 240
tgctttcaga gccccacacc tggcgaaagc cctggcatcg tgtccaacta caacaacagc 300
atcctggcca gagtgtaccg ggacttccaa gagctgctga tgaacgcccc tgagtctcag 360
cacctgggca gaatctggac cgagctgcac atcctgagcc agttcatgga caccctgaga 420
acacaccccg agagaatcgc cggcaggggc atcagaatcc gggacatcct gaaggacgag 480
gaaaccctga cactgttcct catcaagaac atcggcctga gcgacagcgt ggtgtacctg 540
ctgatcaaca gccaagtgcg gcccgagcag tttgctcatg gcgtgccaga tctcgccctg 600
aaggatatcg cctgttctga ggccctgctg gaacggttca tcatcttcag ccagcggaga 660
ggcgccaaga ccgtcagata tgccctgtgc agtctgagcc agggaaccct gcagtggatc 720
gaggataccc tgtacgccaa cgtggacttc ttcaagctgt tccgggtgct gcccacactg 780
ctggattcta gatcccaggg catcaacctg agaagctggg gcggcatcct gtccgacatg 840
agcccaagaa tccaagagtt catccaccgg cctagcatgc aggacctgct gtgggttacc 900
agacctctga tgcagaacgg cggacccgag acattcacca agctgatggg aattctgagc 960
gatctgctgt gcggctaccc tgaaggcgga ggatctagag tgctgagctt caattggtac 1020
gaggacaaca actacaaggc cttcctgggc atcgactcca ccagaaagga ccccatctac 1080
agctacgacc ggcggacaac cagcttctgc aatgccctga tccagagcct ggaaagcaac 1140
cctctgacca agatcgcttg gagggccgcc aaacctctgc tgatgggaaa gatcctgtac 1200
acccctgaca gccctgccgc cagaagaatc ctgaagaacg ccaacagcac cttcgaggaa 1260
ctggaacacg tgcgcaagct ggtcaaggcc tgggaagaag tgggacctca gatctggtac 1320
ttcttcgaca atagcaccca gatgaacatg atcagagaca ccctgggcaa ccctaccgtg 1380
aaggacttcc tgaacagaca gctgggcgaa gagggcatta ccgccgaggc catcctgaac 1440
tttctgtaca agggccccag agagtcccag gccgacgaca tggccaactt cgattggcgg 1500
gacatcttca acatcaccga cagaaccctg cggctggtca accagtacct ggaatgcctg 1560
gtgctggaca agttcgagag ctacaacgac gagacacagc tgacccagag agccctgtct 1620
ctgctggaag agaatatgtt ctgggctggc gtggtgttcc ccgacatgta cccttggaca 1680
agcagcctgc ctcctcacgt gaagtacaag atccggatgg acatcgacgt ggtcgaaaag 1740
accaacaaga tcaaggatcg gtactgggac agcggcccta gagctgatcc cgtggaagat 1800
tttcggtaca tctggggcgg attcgcatac ctgcaggaca tggtggaaca gggaatcaca 1860
cggtcccagg tgcaggctga agctcctgtg ggaatctacc tgcagcagat gccttatcct 1920
tgcttcgtgg acgacagctt catgatcatc ctgaatcggt gcttccccat cttcatggtg 1980
ctggcctgga tctactccgt gtctatgacc gtgaagtcca tcgtgctgga aaaagagctg 2040
cggctgaaag agacactgaa gaaccagggc gtgtccaatg ccgtgatctg gtgcacctgg 2100
tttctggaca gcttctccat tatgagcatg agcatctttc tgctgacgat cttcatcatg 2160
cacggccgga tcctgcacta cagcgacccc tttatcctct tcctgttcct gctggccttc 2220
agcaccgcta caatcatgct gtgttttctg ctgtccacct tcttcagcaa ggcctctctg 2280
gccgctgctt gtagcggcgt gatctacttc accctgtacc tgcctcacat cctgtgcttc 2340
gcatggcagg acagaatgac cgccgagctg aagaaagctg tgtccctgct gagccctgtg 2400
gcctttggct ttggcaccga gtacctcgtc agatttgagg aacaaggact gggactgcag 2460
tggtccaaca tcggcaatag ccctacagag ggcgacgagt tcagcttcct gctgtctatg 2520
cagatgatgc tgctggacgc cgccgtgtat ggactgctgg cttggtatct ggaccaggtg 2580
ttcccaggcg attacggcac tcctctgcct tggtatttcc tgctgcaaga gagctactgg 2640
ctcggcggcg agggatgtag caccagagaa gaaagagccc tggaaaagac cgagcctctg 2700
accgaggaaa cagaggaccc tgaacaccca gagggcatcc acgatagctt tttcgagaga 2760
gaacaccccg gctgggtgcc aggcgtgtgt gtgaagaatc tggtcaagat cttcgagccc 2820
tgcggcagac ctgccgtgga cagactgaac atcaccttct acgagaacca gattaccgcc 2880
tttctgggcc acaacggcgc tggcaagaca accacactga gcatcctcac aggcctgctg 2940
cctccaacaa gcggcacagt tctcgttggc ggcagagaca tcgagacaag cctggatgcc 3000
gtcagacagt ccctgggcat gtgccctcag cacaacatcc tgtttcacca cctgaccgtg 3060
gccgagcaca tgctgtttta tgcccagctg aagggcaaga gccaagaaga ggctcagctg 3120
gaaatggaag ccatgctcga ggacaccggc ctgcaccaca agagaaatga ggaagcccag 3180
gatctgagcg gcggcatgca gagaaaactg agcgtggcca ttgccttcgt gggcgacgcc 3240
aaggttgtga tcctggatga gcctacaagc ggcgtggacc cttacagcag aagatccatc 3300
tgggatctgc tgctgaagta cagaagcggc cggaccatca tcatgagcac ccaccacatg 3360
gacgaggccg atctgctcgg agacagaatc gccatcattg ctcagggcag actgtactgc 3420
agcggcaccc cactgtttct gaagaactgt tttggcaccg gcctgtacct gacactcgtg 3480
cggaagatga agaacatcca gagccagcgg aagggcagcg agggcacatg tagctgtagc 3540
agcaagggct tcagcaccac ctgtcctgct cacgtggacg atctgacccc tgaacaggtg 3600
ctggatggcg acgtgaacga gctgatggac gtggtgctgc accacgtgcc tgaggccaag 3660
ctggtggaat gcatcggcca agagctgatc ttcctgctgc ctaacaagaa cttcaagcac 3720
cgggcctacg ccagcctgtt cagagagctg gaagagacac tggccgatct gggcctgagc 3780
agctttggca tcagcgacac ccctctggaa gaaatcttcc tgaaagtgac cgaggacagc 3840
gacagcggcc cactttttgc tggcggagca cagcagaaac gcgagaacgt gaaccctaga 3900
cacccctgtc tgggccctag agagaaagcc ggacagaccc ctcaggacag caatgtgtgc 3960
tctcctggtg ctcctgccgc tcaccctgaa ggacaacctc cacctgagcc tgagtgtcct 4020
ggacctcagc tgaataccgg cacacagctg gttctgcagc acgtgcaagc cctgctggtc 4080
aagagattcc agcacaccat cagaagccac aaggactttc tggcccagat tgtgctgccc 4140
gccaccttcg tttttctggc tctgatgctg agcatcgtga tccctccatt cggcgagtac 4200
cccgctctga cactgcaccc ttggatctac ggccagcagt acaccttctt cagcatggac 4260
gagcctggca gcgagcagtt tacagtgctg gctgatgtgc tgctgaacaa gcccggcttc 4320
ggcaaccgct gtctgaaaga aggatggctg cctgagtacc cctgcggcaa tagcacccct 4380
tggaaaaccc ctagcgtgtc ccctaacatc acccagctgt tccagaaaca gaaatggacc 4440
caagtgaacc cctctccatc ctgccggtgc tccaccagag aaaagctgac catgctgccc 4500
gagtgtccag aaggtgcagg cggacttcct ccacctcaga gaacccagag aagcaccgag 4560
attctgcagg acctgaccga ccggaatatc agcgacttcc tggttaagac ataccccgca 4620
ctgatccggt ccagcctgaa gtccaagttc tgggtcaacg agcagagata cggcggcatc 4680
tctatcggcg gaaagctgcc tgtggtgcct atcacaggcg aagccctcgt gggctttctg 4740
tctgacctgg gcagaatcat gaacgtgtcc ggcggaccca tcaccagaga ggccagcaaa 4800
gagatccccg atttcctgaa gcacctggaa accgaggata acatcaaagt gtggttcaac 4860
aacaaaggct ggcacgccct ggtgtccttt ctgaacgtgg cccacaacgc catcctgaga 4920
gccagcctgc ctaaggacag aagccctgag gaatacggca tcaccgtgat ctcccagcca 4980
ctgaacctga ccaaagagca gctgagcgag atcaccgtgc tgaccacatc tgtggatgcc 5040
gtggtggcca tctgcgtgat cttctccatg agcttcgtgc ctgcctcctt cgtgctgtat 5100
ctgatccaag agagagtgaa caagagcaag cacctccagt ttatcagcgg agtgtcccca 5160
accacctact gggtcaccaa ctttctgtgg gacatcatga actactccgt gtctgccggc 5220
ctggtcgtgg gcatcttcat cggctttcag aagaaggcct acacaagccc cgagaacctg 5280
cctgctctgg ttgctctgct gctgctgtat ggctgggccg tgattcccat gatgtacccc 5340
gccagcttcc tgttcgatgt gcctagcaca gcctacgtgg ccctgagctg cgccaatctg 5400
ttcatcggca tcaacagcag cgccatcacc ttcatcctgg aactgttcga gaacaacaga 5460
accctgctgc ggttcaacgc cgtgctgaga aagctgctga tcgtgttccc tcacttctgc 5520
ctcggcagag gcctgattga tctggccctg tctcaggccg tgaccgacgt gtacgctaga 5580
tttggcgagg aacacagcgc caatcctttc cactgggacc tgatcggcaa gaacctgttc 5640
gccatggtgg tggaaggcgt ggtgtatttc ctgctgactc tgctggtgca gcggcacttc 5700
tttctgtccc aatggatcgc cgagcctacc aaagaaccca tcgtggacga ggacgacgac 5760
gtggccgaag agagacagag aatcatcacc ggcggcaaca agaccgacat cctgaggctg 5820
cacgagctga ccaagatcta ccctggcaca tctagccctg ccgtggacag actgtgtgtg 5880
ggagttagac ctggcgagtg ctttggcctg ctgggagtta atggcgccgg aaagaccacc 5940
accttcaaga tgctgaccgg cgacaccaca gtgaccagcg gagatgctac tgtggccggc 6000
aagagcatcc tgaccaacat cagcgaggtg caccagaaca tgggctactg ccctcagttc 6060
gacgccatcg acgaactgct gacaggcaga gagcacctgt atctgtacgc cagactgaga 6120
ggcgtgcccg ccgaagaaat tgagaaggtg gccaactggt ccatcaagag cctgggactg 6180
accgtgtacg ccgattgtct ggccggcaca tatagcggcg gaaacaagcg gaagctgagc 6240
accgccattg ctctgatcgg atgccctcct ctggtcctgc tggatgagcc tacaaccggc 6300
atggatcccc aggctagacg gatgctgtgg aacgtgatcg tgtccatcat ccgcgaaggc 6360
agagccgtgg tcctgacaag ccacagcatg gaagagtgcg aggccctgtg taccagactg 6420
gccatcatgg tcaagggcgc cttcagatgc atgggcacca ttcagcacct gaaaagcaag 6480
ttcggcgacg gctacatcgt gaccatgaag atcaagtccc caaaggacga cctgctgcca 6540
gatctgaacc ccgtggaaca gttcttccag ggcaacttcc ctggctccgt gcagcgggaa 6600
agacactaca acatgctgca gtttcaggtg tccagcagct ccctggccag aatctttcag 6660
ctgctgctct cccacaagga tagcctgctg attgaagagt acagcgtgac ccagaccaca 6720
ctggaccagg tgttcgtgaa cttcgccaag cagcagaccg agagccacga cttgcctctg 6780
caccctagag ctgccggcgc ttctagacag gcccaggatt aa 6822
<210> 2
<211> 154
<212> PRT
<213> 集胞藻(Synechocystis sp. strain PCC6803)
<400> 2
Cys Ile Ser Gly Asp Ser Leu Ile Ser Leu Ala Ser Thr Gly Lys Arg
1 5 10 15
Val Pro Ile Lys Asp Leu Leu Gly Glu Lys Asp Phe Glu Ile Trp Ala
20 25 30
Ile Asn Glu Gln Thr Met Lys Leu Glu Ser Ala Lys Val Ser Arg Val
35 40 45
Phe Cys Thr Gly Lys Lys Leu Val Tyr Thr Leu Lys Thr Arg Leu Gly
50 55 60
Arg Thr Ile Lys Ala Thr Ala Asn His Arg Phe Leu Thr Ile Asp Gly
65 70 75 80
Trp Lys Arg Leu Asp Glu Leu Ser Leu Lys Glu His Ile Ala Leu Pro
85 90 95
Arg Lys Leu Glu Ser Ser Ser Leu Gln Leu Ala Pro Glu Ile Glu Lys
100 105 110
Leu Pro Gln Ser Asp Ile Tyr Trp Asp Pro Ile Val Ser Ile Thr Glu
115 120 125
Thr Gly Val Glu Glu Val Phe Asp Leu Thr Val Pro Gly Leu Arg Asn
130 135 140
Phe Val Ala Asn Asp Ile Ile Val His Asn
145 150
<210> 3
<211> 153
<212> PRT
<213> 海洋红嗜热盐菌(Rhodothermus marinus)
<400> 3
Cys Leu Ala Gly Asp Thr Leu Ile Thr Leu Ala Asp Gly Arg Arg Val
1 5 10 15
Pro Ile Arg Glu Leu Val Ser Gln Gln Asn Phe Ser Val Trp Ala Leu
20 25 30
Asn Pro Gln Thr Tyr Arg Leu Glu Arg Ala Arg Val Ser Arg Ala Phe
35 40 45
Cys Thr Gly Ile Lys Pro Val Tyr Arg Leu Thr Thr Arg Leu Gly Arg
50 55 60
Ser Ile Arg Ala Thr Ala Asn His Arg Phe Leu Thr Pro Gln Gly Trp
65 70 75 80
Lys Arg Val Asp Glu Leu Gln Pro Gly Asp Tyr Leu Ala Leu Pro Arg
85 90 95
Arg Ile Pro Thr Ala Ser Ala Ala Ala Cys Pro Glu Leu Arg Gln Leu
100 105 110
Ala Gln Ser Asp Val Tyr Trp Asp Pro Ile Val Ser Ile Glu Pro Asp
115 120 125
Gly Val Glu Glu Val Phe Asp Leu Thr Val Pro Gly Pro His Asn Phe
130 135 140
Val Ala Asn Asp Ile Ile Ala His Asn
145 150
<210> 4
<211> 137
<212> PRT
<213> 念珠藻(Nostoc punctiforme)
<400> 4
Cys Leu Ser Tyr Glu Thr Glu Ile Leu Thr Val Glu Tyr Gly Leu Leu
1 5 10 15
Pro Ile Gly Lys Ile Val Glu Lys Arg Ile Glu Cys Thr Val Tyr Ser
20 25 30
Val Asp Asn Asn Gly Asn Ile Tyr Thr Gln Pro Val Ala Gln Trp His
35 40 45
Asp Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser
50 55 60
Leu Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Val Asp Gly Gln
65 70 75 80
Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Glu Leu Asp Leu Met Arg
85 90 95
Val Asp Asn Leu Pro Asn Ile Lys Ile Ala Thr Arg Lys Tyr Leu Gly
100 105 110
Lys Gln Asn Val Tyr Asp Ile Gly Val Glu Arg Asp His Asn Phe Ala
115 120 125
Leu Lys Asn Gly Phe Ile Ala Ser Asn
130 135
<210> 5
<211> 136
<212> PRT
<213> Consensus
<400> 5
Cys Leu Ser Tyr Asp Thr Glu Ile Leu Thr Val Glu Tyr Gly Phe Leu
1 5 10 15
Pro Ile Gly Lys Ile Val Glu Glu Arg Ile Glu Cys Thr Val Tyr Thr
20 25 30
Val Asp Lys Asn Gly Phe Val Tyr Thr Gln Pro Ile Ala Gln Trp His
35 40 45
Asn Arg Gly Glu Gln Glu Val Phe Glu Tyr Cys Leu Glu Asp Gly Ser
50 55 60
Ile Ile Arg Ala Thr Lys Asp His Lys Phe Met Thr Thr Asp Gly Gln
65 70 75 80
Met Leu Pro Ile Asp Glu Ile Phe Glu Arg Gly Leu Asp Leu Lys Gln
85 90 95
Val Asp Gly Leu Pro Val Lys Ile Ile Ser Arg Lys Ser Leu Gly Thr
100 105 110
Gln Asn Val Tyr Asp Ile Gly Val Glu Lys Asp His Asn Phe Leu Leu
115 120 125
Lys Asn Gly Leu Val Ala Ser Asn
130 135
<210> 6
<211> 1251
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 6
Met Gly Phe Val Arg Gln Ile Gln Leu Leu Leu Trp Lys Asn Trp Thr
1 5 10 15
Leu Arg Lys Arg Gln Lys Ile Arg Phe Val Val Glu Leu Val Trp Pro
20 25 30
Leu Ser Leu Phe Leu Val Leu Ile Trp Leu Arg Asn Ala Asn Pro Leu
35 40 45
Tyr Ser His His Glu Cys His Phe Pro Asn Lys Ala Met Pro Ser Ala
50 55 60
Gly Met Leu Pro Trp Leu Gln Gly Ile Phe Cys Asn Val Asn Asn Pro
65 70 75 80
Cys Phe Gln Ser Pro Thr Pro Gly Glu Ser Pro Gly Ile Val Ser Asn
85 90 95
Tyr Asn Asn Ser Ile Leu Ala Arg Val Tyr Arg Asp Phe Gln Glu Leu
100 105 110
Leu Met Asn Ala Pro Glu Ser Gln His Leu Gly Arg Ile Trp Thr Glu
115 120 125
Leu His Ile Leu Ser Gln Phe Met Asp Thr Leu Arg Thr His Pro Glu
130 135 140
Arg Ile Ala Gly Arg Gly Ile Arg Ile Arg Asp Ile Leu Lys Asp Glu
145 150 155 160
Glu Thr Leu Thr Leu Phe Leu Ile Lys Asn Ile Gly Leu Ser Asp Ser
165 170 175
Val Val Tyr Leu Leu Ile Asn Ser Gln Val Arg Pro Glu Gln Phe Ala
180 185 190
His Gly Val Pro Asp Leu Ala Leu Lys Asp Ile Ala Cys Ser Glu Ala
195 200 205
Leu Leu Glu Arg Phe Ile Ile Phe Ser Gln Arg Arg Gly Ala Lys Thr
210 215 220
Val Arg Tyr Ala Leu Cys Ser Leu Ser Gln Gly Thr Leu Gln Trp Ile
225 230 235 240
Glu Asp Thr Leu Tyr Ala Asn Val Asp Phe Phe Lys Leu Phe Arg Val
245 250 255
Leu Pro Thr Leu Leu Asp Ser Arg Ser Gln Gly Ile Asn Leu Arg Ser
260 265 270
Trp Gly Gly Ile Leu Ser Asp Met Ser Pro Arg Ile Gln Glu Phe Ile
275 280 285
His Arg Pro Ser Met Gln Asp Leu Leu Trp Val Thr Arg Pro Leu Met
290 295 300
Gln Asn Gly Gly Pro Glu Thr Phe Thr Lys Leu Met Gly Ile Leu Ser
305 310 315 320
Asp Leu Leu Cys Gly Tyr Pro Glu Gly Gly Gly Ser Arg Val Leu Ser
325 330 335
Phe Asn Trp Tyr Glu Asp Asn Asn Tyr Lys Ala Phe Leu Gly Ile Asp
340 345 350
Ser Thr Arg Lys Asp Pro Ile Tyr Ser Tyr Asp Arg Arg Thr Thr Ser
355 360 365
Phe Cys Asn Ala Leu Ile Gln Ser Leu Glu Ser Asn Pro Leu Thr Lys
370 375 380
Ile Ala Trp Arg Ala Ala Lys Pro Leu Leu Met Gly Lys Ile Leu Tyr
385 390 395 400
Thr Pro Asp Ser Pro Ala Ala Arg Arg Ile Leu Lys Asn Ala Asn Ser
405 410 415
Thr Phe Glu Glu Leu Glu His Val Arg Lys Leu Val Lys Ala Trp Glu
420 425 430
Glu Val Gly Pro Gln Ile Trp Tyr Phe Phe Asp Asn Ser Thr Gln Met
435 440 445
Asn Met Ile Arg Asp Thr Leu Gly Asn Pro Thr Val Lys Asp Phe Leu
450 455 460
Asn Arg Gln Leu Gly Glu Glu Gly Ile Thr Ala Glu Ala Ile Leu Asn
465 470 475 480
Phe Leu Tyr Lys Gly Pro Arg Glu Ser Gln Ala Asp Asp Met Ala Asn
485 490 495
Phe Asp Trp Arg Asp Ile Phe Asn Ile Thr Asp Arg Thr Leu Arg Leu
500 505 510
Val Asn Gln Tyr Leu Glu Cys Leu Val Leu Asp Lys Phe Glu Ser Tyr
515 520 525
Asn Asp Glu Thr Gln Leu Thr Gln Arg Ala Leu Ser Leu Leu Glu Glu
530 535 540
Asn Met Phe Trp Ala Gly Val Val Phe Pro Asp Met Tyr Pro Trp Thr
545 550 555 560
Ser Ser Leu Pro Pro His Val Lys Tyr Lys Ile Arg Met Asp Ile Asp
565 570 575
Val Val Glu Lys Thr Asn Lys Ile Lys Asp Arg Tyr Trp Asp Ser Gly
580 585 590
Pro Arg Ala Asp Pro Val Glu Asp Phe Arg Tyr Ile Trp Gly Gly Phe
595 600 605
Ala Tyr Leu Gln Asp Met Val Glu Gln Gly Ile Thr Arg Ser Gln Val
610 615 620
Gln Ala Glu Ala Pro Val Gly Ile Tyr Leu Gln Gln Met Pro Tyr Pro
625 630 635 640
Cys Phe Val Asp Asp Ser Phe Met Ile Ile Leu Asn Arg Cys Phe Pro
645 650 655
Ile Phe Met Val Leu Ala Trp Ile Tyr Ser Val Ser Met Thr Val Lys
660 665 670
Ser Ile Val Leu Glu Lys Glu Leu Arg Leu Lys Glu Thr Leu Lys Asn
675 680 685
Gln Gly Val Ser Asn Ala Val Ile Trp Cys Thr Trp Phe Leu Asp Ser
690 695 700
Phe Ser Ile Met Ser Met Ser Ile Phe Leu Leu Thr Ile Phe Ile Met
705 710 715 720
His Gly Arg Ile Leu His Tyr Ser Asp Pro Phe Ile Leu Phe Leu Phe
725 730 735
Leu Leu Ala Phe Ser Thr Ala Thr Ile Met Leu Cys Phe Leu Leu Ser
740 745 750
Thr Phe Phe Ser Lys Ala Ser Leu Ala Ala Ala Cys Ser Gly Val Ile
755 760 765
Tyr Phe Thr Leu Tyr Leu Pro His Ile Leu Cys Phe Ala Trp Gln Asp
770 775 780
Arg Met Thr Ala Glu Leu Lys Lys Ala Val Ser Leu Leu Ser Pro Val
785 790 795 800
Ala Phe Gly Phe Gly Thr Glu Tyr Leu Val Arg Phe Glu Glu Gln Gly
805 810 815
Leu Gly Leu Gln Trp Ser Asn Ile Gly Asn Ser Pro Thr Glu Gly Asp
820 825 830
Glu Phe Ser Phe Leu Leu Ser Met Gln Met Met Leu Leu Asp Ala Ala
835 840 845
Val Tyr Gly Leu Leu Ala Trp Tyr Leu Asp Gln Val Phe Pro Gly Asp
850 855 860
Tyr Gly Thr Pro Leu Pro Trp Tyr Phe Leu Leu Gln Glu Ser Tyr Trp
865 870 875 880
Leu Gly Gly Glu Gly Cys Ser Thr Arg Glu Glu Arg Ala Leu Glu Lys
885 890 895
Thr Glu Pro Leu Thr Glu Glu Thr Glu Asp Pro Glu His Pro Glu Gly
900 905 910
Ile His Asp Ser Phe Phe Glu Arg Glu His Pro Gly Trp Val Pro Gly
915 920 925
Val Cys Val Lys Asn Leu Val Lys Ile Phe Glu Pro Cys Gly Arg Pro
930 935 940
Ala Val Asp Arg Leu Asn Ile Thr Phe Tyr Glu Asn Gln Ile Thr Ala
945 950 955 960
Phe Leu Gly His Asn Gly Ala Gly Lys Thr Thr Thr Leu Ser Ile Leu
965 970 975
Thr Gly Leu Leu Pro Pro Thr Ser Gly Thr Val Leu Val Gly Gly Arg
980 985 990
Asp Ile Glu Thr Ser Leu Asp Ala Val Arg Gln Ser Leu Gly Met Cys
995 1000 1005
Pro Gln His Asn Ile Leu Phe His His Leu Thr Val Ala Glu His Met
1010 1015 1020
Leu Phe Tyr Ala Gln Leu Lys Gly Lys Ser Gln Glu Glu Ala Gln Leu
1025 1030 1035 1040
Glu Met Glu Ala Met Leu Glu Asp Thr Gly Leu His His Lys Arg Asn
1045 1050 1055
Glu Glu Ala Gln Asp Leu Ser Gly Gly Met Gln Arg Lys Leu Ser Val
1060 1065 1070
Ala Ile Ala Phe Val Gly Asp Ala Lys Val Val Ile Leu Asp Glu Pro
1075 1080 1085
Thr Ser Gly Val Asp Pro Tyr Ser Arg Arg Ser Ile Trp Asp Leu Leu
1090 1095 1100
Leu Lys Tyr Arg Ser Gly Arg Thr Ile Ile Met Ser Thr His His Met
1105 1110 1115 1120
Asp Glu Ala Asp Leu Leu Gly Asp Arg Ile Ala Ile Ile Ala Gln Gly
1125 1130 1135
Arg Leu Tyr Cys Ser Gly Thr Pro Leu Phe Leu Lys Asn Cys Leu Ala
1140 1145 1150
Gly Asp Thr Leu Ile Thr Leu Ala Asp Gly Arg Arg Val Pro Ile Arg
1155 1160 1165
Glu Leu Val Ser Gln Gln Asn Phe Ser Val Trp Ala Leu Asn Pro Gln
1170 1175 1180
Thr Tyr Arg Leu Glu Arg Ala Arg Val Ser Arg Ala Phe Cys Thr Gly
1185 1190 1195 1200
Ile Lys Pro Val Tyr Arg Leu Thr Thr Arg Leu Gly Arg Ser Ile Arg
1205 1210 1215
Ala Thr Ala Asn His Arg Phe Leu Thr Pro Gln Gly Trp Lys Arg Val
1220 1225 1230
Asp Glu Leu Gln Pro Gly Asp Tyr Leu Ala Leu Pro Arg Arg Ile Pro
1235 1240 1245
Thr Ala Ser
1250
<210> 7
<211> 1176
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 7
Met Ala Ala Ala Cys Pro Glu Leu Arg Gln Leu Ala Gln Ser Asp Val
1 5 10 15
Tyr Trp Asp Pro Ile Val Ser Ile Glu Pro Asp Gly Val Glu Glu Val
20 25 30
Phe Asp Leu Thr Val Pro Gly Pro His Asn Phe Val Ala Asn Asp Ile
35 40 45
Ile Ala His Asn Cys Phe Gly Thr Gly Leu Tyr Leu Thr Leu Val Arg
50 55 60
Lys Met Lys Asn Ile Gln Ser Gln Arg Lys Gly Ser Glu Gly Thr Cys
65 70 75 80
Ser Cys Ser Ser Lys Gly Phe Ser Thr Thr Cys Pro Ala His Val Asp
85 90 95
Asp Leu Thr Pro Glu Gln Val Leu Asp Gly Asp Val Asn Glu Leu Met
100 105 110
Asp Val Val Leu His His Val Pro Glu Ala Lys Leu Val Glu Cys Ile
115 120 125
Gly Gln Glu Leu Ile Phe Leu Leu Pro Asn Lys Asn Phe Lys His Arg
130 135 140
Ala Tyr Ala Ser Leu Phe Arg Glu Leu Glu Glu Thr Leu Ala Asp Leu
145 150 155 160
Gly Leu Ser Ser Phe Gly Ile Ser Asp Thr Pro Leu Glu Glu Ile Phe
165 170 175
Leu Lys Val Thr Glu Asp Ser Asp Ser Gly Pro Leu Phe Ala Gly Gly
180 185 190
Ala Gln Gln Lys Arg Glu Asn Val Asn Pro Arg His Pro Cys Leu Gly
195 200 205
Pro Arg Glu Lys Ala Gly Gln Thr Pro Gln Asp Ser Asn Val Cys Ser
210 215 220
Pro Gly Ala Pro Ala Ala His Pro Glu Gly Gln Pro Pro Pro Glu Pro
225 230 235 240
Glu Cys Pro Gly Pro Gln Leu Asn Thr Gly Thr Gln Leu Val Leu Gln
245 250 255
His Val Gln Ala Leu Leu Val Lys Arg Phe Gln His Thr Ile Arg Ser
260 265 270
His Lys Asp Phe Leu Ala Gln Ile Val Leu Pro Ala Thr Phe Val Phe
275 280 285
Leu Ala Leu Met Leu Ser Ile Val Ile Pro Pro Phe Gly Glu Tyr Pro
290 295 300
Ala Leu Thr Leu His Pro Trp Ile Tyr Gly Gln Gln Tyr Thr Phe Phe
305 310 315 320
Ser Met Asp Glu Pro Gly Ser Glu Gln Phe Thr Val Leu Ala Asp Val
325 330 335
Leu Leu Asn Lys Pro Gly Phe Gly Asn Arg Cys Leu Lys Glu Gly Trp
340 345 350
Leu Pro Glu Tyr Pro Cys Gly Asn Ser Thr Pro Trp Lys Thr Pro Ser
355 360 365
Val Ser Pro Asn Ile Thr Gln Leu Phe Gln Lys Gln Lys Trp Thr Gln
370 375 380
Val Asn Pro Ser Pro Ser Cys Arg Cys Ser Thr Arg Glu Lys Leu Thr
385 390 395 400
Met Leu Pro Glu Cys Pro Glu Gly Ala Gly Gly Leu Pro Pro Pro Gln
405 410 415
Arg Thr Gln Arg Ser Thr Glu Ile Leu Gln Asp Leu Thr Asp Arg Asn
420 425 430
Ile Ser Asp Phe Leu Val Lys Thr Tyr Pro Ala Leu Ile Arg Ser Ser
435 440 445
Leu Lys Ser Lys Phe Trp Val Asn Glu Gln Arg Tyr Gly Gly Ile Ser
450 455 460
Ile Gly Gly Lys Leu Pro Val Val Pro Ile Thr Gly Glu Ala Leu Val
465 470 475 480
Gly Phe Leu Ser Asp Leu Gly Arg Ile Met Asn Val Ser Gly Gly Pro
485 490 495
Ile Thr Arg Glu Ala Ser Lys Glu Ile Pro Asp Phe Leu Lys His Leu
500 505 510
Glu Thr Glu Asp Asn Ile Lys Val Trp Phe Asn Asn Lys Gly Trp His
515 520 525
Ala Leu Val Ser Phe Leu Asn Val Ala His Asn Ala Ile Leu Arg Ala
530 535 540
Ser Leu Pro Lys Asp Arg Ser Pro Glu Glu Tyr Gly Ile Thr Val Ile
545 550 555 560
Ser Gln Pro Leu Asn Leu Thr Lys Glu Gln Leu Ser Glu Ile Thr Val
565 570 575
Leu Thr Thr Ser Val Asp Ala Val Val Ala Ile Cys Val Ile Phe Ser
580 585 590
Met Ser Phe Val Pro Ala Ser Phe Val Leu Tyr Leu Ile Gln Glu Arg
595 600 605
Val Asn Lys Ser Lys His Leu Gln Phe Ile Ser Gly Val Ser Pro Thr
610 615 620
Thr Tyr Trp Val Thr Asn Phe Leu Trp Asp Ile Met Asn Tyr Ser Val
625 630 635 640
Ser Ala Gly Leu Val Val Gly Ile Phe Ile Gly Phe Gln Lys Lys Ala
645 650 655
Tyr Thr Ser Pro Glu Asn Leu Pro Ala Leu Val Ala Leu Leu Leu Leu
660 665 670
Tyr Gly Trp Ala Val Ile Pro Met Met Tyr Pro Ala Ser Phe Leu Phe
675 680 685
Asp Val Pro Ser Thr Ala Tyr Val Ala Leu Ser Cys Ala Asn Leu Phe
690 695 700
Ile Gly Ile Asn Ser Ser Ala Ile Thr Phe Ile Leu Glu Leu Phe Glu
705 710 715 720
Asn Asn Arg Thr Leu Leu Arg Phe Asn Ala Val Leu Arg Lys Leu Leu
725 730 735
Ile Val Phe Pro His Phe Cys Leu Gly Arg Gly Leu Ile Asp Leu Ala
740 745 750
Leu Ser Gln Ala Val Thr Asp Val Tyr Ala Arg Phe Gly Glu Glu His
755 760 765
Ser Ala Asn Pro Phe His Trp Asp Leu Ile Gly Lys Asn Leu Phe Ala
770 775 780
Met Val Val Glu Gly Val Val Tyr Phe Leu Leu Thr Leu Leu Val Gln
785 790 795 800
Arg His Phe Phe Leu Ser Gln Trp Ile Ala Glu Pro Thr Lys Glu Pro
805 810 815
Ile Val Asp Glu Asp Asp Asp Val Ala Glu Glu Arg Gln Arg Ile Ile
820 825 830
Thr Gly Gly Asn Lys Thr Asp Ile Leu Arg Leu His Glu Leu Thr Lys
835 840 845
Ile Tyr Pro Gly Thr Ser Ser Pro Ala Val Asp Arg Leu Cys Val Gly
850 855 860
Val Arg Pro Gly Glu Cys Phe Gly Leu Leu Gly Val Asn Gly Ala Gly
865 870 875 880
Lys Thr Thr Thr Phe Lys Met Leu Thr Gly Asp Thr Thr Val Thr Ser
885 890 895
Gly Asp Ala Thr Val Ala Gly Lys Ser Ile Leu Thr Asn Ile Ser Glu
900 905 910
Val His Gln Asn Met Gly Tyr Cys Pro Gln Phe Asp Ala Ile Asp Glu
915 920 925
Leu Leu Thr Gly Arg Glu His Leu Tyr Leu Tyr Ala Arg Leu Arg Gly
930 935 940
Val Pro Ala Glu Glu Ile Glu Lys Val Ala Asn Trp Ser Ile Lys Ser
945 950 955 960
Leu Gly Leu Thr Val Tyr Ala Asp Cys Leu Ala Gly Thr Tyr Ser Gly
965 970 975
Gly Asn Lys Arg Lys Leu Ser Thr Ala Ile Ala Leu Ile Gly Cys Pro
980 985 990
Pro Leu Val Leu Leu Asp Glu Pro Thr Thr Gly Met Asp Pro Gln Ala
995 1000 1005
Arg Arg Met Leu Trp Asn Val Ile Val Ser Ile Ile Arg Glu Gly Arg
1010 1015 1020
Ala Val Val Leu Thr Ser His Ser Met Glu Glu Cys Glu Ala Leu Cys
1025 1030 1035 1040
Thr Arg Leu Ala Ile Met Val Lys Gly Ala Phe Arg Cys Met Gly Thr
1045 1050 1055
Ile Gln His Leu Lys Ser Lys Phe Gly Asp Gly Tyr Ile Val Thr Met
1060 1065 1070
Lys Ile Lys Ser Pro Lys Asp Asp Leu Leu Pro Asp Leu Asn Pro Val
1075 1080 1085
Glu Gln Phe Phe Gln Gly Asn Phe Pro Gly Ser Val Gln Arg Glu Arg
1090 1095 1100
His Tyr Asn Met Leu Gln Phe Gln Val Ser Ser Ser Ser Leu Ala Arg
1105 1110 1115 1120
Ile Phe Gln Leu Leu Leu Ser His Lys Asp Ser Leu Leu Ile Glu Glu
1125 1130 1135
Tyr Ser Val Thr Gln Thr Thr Leu Asp Gln Val Phe Val Asn Phe Ala
1140 1145 1150
Lys Gln Gln Thr Glu Ser His Asp Leu Pro Leu His Pro Arg Ala Ala
1155 1160 1165
Gly Ala Ser Arg Gln Ala Gln Asp
1170 1175
<210> 8
<211> 3756
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
atgggctttg tgcgacagat tcagctgctg ctgtggaaga actggaccct gcggaagcgg 60
cagaaaatca gattcgtggt ggaactcgtg tggcccctga gcctgtttct ggtgctgatc 120
tggctgcgga acgccaatcc tctgtacagc caccacgagt gtcacttccc caacaaggcc 180
atgccttctg ccggaatgct gccttggctg cagggcatct tctgcaacgt gaacaacccc 240
tgctttcaga gccccacacc tggcgaaagc cctggcatcg tgtccaacta caacaacagc 300
atcctggcca gagtgtaccg ggacttccaa gagctgctga tgaacgcccc tgagtctcag 360
cacctgggca gaatctggac cgagctgcac atcctgagcc agttcatgga caccctgaga 420
acacaccccg agagaatcgc cggcaggggc atcagaatcc gggacatcct gaaggacgag 480
gaaaccctga cactgttcct catcaagaac atcggcctga gcgacagcgt ggtgtacctg 540
ctgatcaaca gccaagtgcg gcccgagcag tttgctcatg gcgtgccaga tctcgccctg 600
aaggatatcg cctgttctga ggccctgctg gaacggttca tcatcttcag ccagcggaga 660
ggcgccaaga ccgtcagata tgccctgtgc agtctgagcc agggaaccct gcagtggatc 720
gaggataccc tgtacgccaa cgtggacttc ttcaagctgt tccgggtgct gcccacactg 780
ctggattcta gatcccaggg catcaacctg agaagctggg gcggcatcct gtccgacatg 840
agcccaagaa tccaagagtt catccaccgg cctagcatgc aggacctgct gtgggttacc 900
agacctctga tgcagaacgg cggacccgag acattcacca agctgatggg aattctgagc 960
gatctgctgt gcggctaccc tgaaggcgga ggatctagag tgctgagctt caattggtac 1020
gaggacaaca actacaaggc cttcctgggc atcgactcca ccagaaagga ccccatctac 1080
agctacgacc ggcggacaac cagcttctgc aatgccctga tccagagcct ggaaagcaac 1140
cctctgacca agatcgcttg gagggccgcc aaacctctgc tgatgggaaa gatcctgtac 1200
acccctgaca gccctgccgc cagaagaatc ctgaagaacg ccaacagcac cttcgaggaa 1260
ctggaacacg tgcgcaagct ggtcaaggcc tgggaagaag tgggacctca gatctggtac 1320
ttcttcgaca atagcaccca gatgaacatg atcagagaca ccctgggcaa ccctaccgtg 1380
aaggacttcc tgaacagaca gctgggcgaa gagggcatta ccgccgaggc catcctgaac 1440
tttctgtaca agggccccag agagtcccag gccgacgaca tggccaactt cgattggcgg 1500
gacatcttca acatcaccga cagaaccctg cggctggtca accagtacct ggaatgcctg 1560
gtgctggaca agttcgagag ctacaacgac gagacacagc tgacccagag agccctgtct 1620
ctgctggaag agaatatgtt ctgggctggc gtggtgttcc ccgacatgta cccttggaca 1680
agcagcctgc ctcctcacgt gaagtacaag atccggatgg acatcgacgt ggtcgaaaag 1740
accaacaaga tcaaggatcg gtactgggac agcggcccta gagctgatcc cgtggaagat 1800
tttcggtaca tctggggcgg attcgcatac ctgcaggaca tggtggaaca gggaatcaca 1860
cggtcccagg tgcaggctga agctcctgtg ggaatctacc tgcagcagat gccttatcct 1920
tgcttcgtgg acgacagctt catgatcatc ctgaatcggt gcttccccat cttcatggtg 1980
ctggcctgga tctactccgt gtctatgacc gtgaagtcca tcgtgctgga aaaagagctg 2040
cggctgaaag agacactgaa gaaccagggc gtgtccaatg ccgtgatctg gtgcacctgg 2100
tttctggaca gcttctccat tatgagcatg agcatctttc tgctgacgat cttcatcatg 2160
cacggccgga tcctgcacta cagcgacccc tttatcctct tcctgttcct gctggccttc 2220
agcaccgcta caatcatgct gtgttttctg ctgtccacct tcttcagcaa ggcctctctg 2280
gccgctgctt gtagcggcgt gatctacttc accctgtacc tgcctcacat cctgtgcttc 2340
gcatggcagg acagaatgac cgccgagctg aagaaagctg tgtccctgct gagccctgtg 2400
gcctttggct ttggcaccga gtacctcgtc agatttgagg aacaaggact gggactgcag 2460
tggtccaaca tcggcaatag ccctacagag ggcgacgagt tcagcttcct gctgtctatg 2520
cagatgatgc tgctggacgc cgccgtgtat ggactgctgg cttggtatct ggaccaggtg 2580
ttcccaggcg attacggcac tcctctgcct tggtatttcc tgctgcaaga gagctactgg 2640
ctcggcggcg agggatgtag caccagagaa gaaagagccc tggaaaagac cgagcctctg 2700
accgaggaaa cagaggaccc tgaacaccca gagggcatcc acgatagctt tttcgagaga 2760
gaacaccccg gctgggtgcc aggcgtgtgt gtgaagaatc tggtcaagat cttcgagccc 2820
tgcggcagac ctgccgtgga cagactgaac atcaccttct acgagaacca gattaccgcc 2880
tttctgggcc acaacggcgc tggcaagaca accacactga gcatcctcac aggcctgctg 2940
cctccaacaa gcggcacagt tctcgttggc ggcagagaca tcgagacaag cctggatgcc 3000
gtcagacagt ccctgggcat gtgccctcag cacaacatcc tgtttcacca cctgaccgtg 3060
gccgagcaca tgctgtttta tgcccagctg aagggcaaga gccaagaaga ggctcagctg 3120
gaaatggaag ccatgctcga ggacaccggc ctgcaccaca agagaaatga ggaagcccag 3180
gatctgagcg gcggcatgca gagaaaactg agcgtggcca ttgccttcgt gggcgacgcc 3240
aaggttgtga tcctggatga gcctacaagc ggcgtggacc cttacagcag aagatccatc 3300
tgggatctgc tgctgaagta cagaagcggc cggaccatca tcatgagcac ccaccacatg 3360
gacgaggccg atctgctcgg agacagaatc gccatcattg ctcagggcag actgtactgc 3420
agcggcaccc cactgtttct gaagaactgt ctggctggcg atactctcat taccctggcc 3480
gatggacgac gagtgcctat tagagaactg gtgtcacagc agaatttttc cgtgtgggct 3540
ctgaatcctc agacttaccg cctggagagg gctagagtga gtagagcttt ctgtaccggc 3600
atcaaacctg tgtaccgcct caccactaga ctggggagat ccattagggc cactgccaac 3660
caccgatttc tcacacctca gggctggaaa cgagtcgatg aactccagcc tggagattac 3720
ctggctctgc ctaggagaat ccctactgcc tcctaa 3756
<210> 9
<211> 3531
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
atggcggcgg cgtgcccgga actgcgtcag ctggcgcaga gcgatgtgta ttgggatccg 60
attgtgagca ttgaaccgga tggcgtggaa gaagtgtttg atctgaccgt gccgggcccg 120
cataactttg tggcgaacga tattattgcg cataactgtt ttggcaccgg cctgtacctg 180
acactcgtgc ggaagatgaa gaacatccag agccagcgga agggcagcga gggcacatgt 240
agctgtagca gcaagggctt cagcaccacc tgtcctgctc acgtggacga tctgacccct 300
gaacaggtgc tggatggcga cgtgaacgag ctgatggacg tggtgctgca ccacgtgcct 360
gaggccaagc tggtggaatg catcggccaa gagctgatct tcctgctgcc taacaagaac 420
ttcaagcacc gggcctacgc cagcctgttc agagagctgg aagagacact ggccgatctg 480
ggcctgagca gctttggcat cagcgacacc cctctggaag aaatcttcct gaaagtgacc 540
gaggacagcg acagcggccc actttttgct ggcggagcac agcagaaacg cgagaacgtg 600
aaccctagac acccctgtct gggccctaga gagaaagccg gacagacccc tcaggacagc 660
aatgtgtgct ctcctggtgc tcctgccgct caccctgaag gacaacctcc acctgagcct 720
gagtgtcctg gacctcagct gaataccggc acacagctgg ttctgcagca cgtgcaagcc 780
ctgctggtca agagattcca gcacaccatc agaagccaca aggactttct ggcccagatt 840
gtgctgcccg ccaccttcgt ttttctggct ctgatgctga gcatcgtgat ccctccattc 900
ggcgagtacc ccgctctgac actgcaccct tggatctacg gccagcagta caccttcttc 960
agcatggacg agcctggcag cgagcagttt acagtgctgg ctgatgtgct gctgaacaag 1020
cccggcttcg gcaaccgctg tctgaaagaa ggatggctgc ctgagtaccc ctgcggcaat 1080
agcacccctt ggaaaacccc tagcgtgtcc cctaacatca cccagctgtt ccagaaacag 1140
aaatggaccc aagtgaaccc ctctccatcc tgccggtgct ccaccagaga aaagctgacc 1200
atgctgcccg agtgtccaga aggtgcaggc ggacttcctc cacctcagag aacccagaga 1260
agcaccgaga ttctgcagga cctgaccgac cggaatatca gcgacttcct ggttaagaca 1320
taccccgcac tgatccggtc cagcctgaag tccaagttct gggtcaacga gcagagatac 1380
ggcggcatct ctatcggcgg aaagctgcct gtggtgccta tcacaggcga agccctcgtg 1440
ggctttctgt ctgacctggg cagaatcatg aacgtgtccg gcggacccat caccagagag 1500
gccagcaaag agatccccga tttcctgaag cacctggaaa ccgaggataa catcaaagtg 1560
tggttcaaca acaaaggctg gcacgccctg gtgtcctttc tgaacgtggc ccacaacgcc 1620
atcctgagag ccagcctgcc taaggacaga agccctgagg aatacggcat caccgtgatc 1680
tcccagccac tgaacctgac caaagagcag ctgagcgaga tcaccgtgct gaccacatct 1740
gtggatgccg tggtggccat ctgcgtgatc ttctccatga gcttcgtgcc tgcctccttc 1800
gtgctgtatc tgatccaaga gagagtgaac aagagcaagc acctccagtt tatcagcgga 1860
gtgtccccaa ccacctactg ggtcaccaac tttctgtggg acatcatgaa ctactccgtg 1920
tctgccggcc tggtcgtggg catcttcatc ggctttcaga agaaggccta cacaagcccc 1980
gagaacctgc ctgctctggt tgctctgctg ctgctgtatg gctgggccgt gattcccatg 2040
atgtaccccg ccagcttcct gttcgatgtg cctagcacag cctacgtggc cctgagctgc 2100
gccaatctgt tcatcggcat caacagcagc gccatcacct tcatcctgga actgttcgag 2160
aacaacagaa ccctgctgcg gttcaacgcc gtgctgagaa agctgctgat cgtgttccct 2220
cacttctgcc tcggcagagg cctgattgat ctggccctgt ctcaggccgt gaccgacgtg 2280
tacgctagat ttggcgagga acacagcgcc aatcctttcc actgggacct gatcggcaag 2340
aacctgttcg ccatggtggt ggaaggcgtg gtgtatttcc tgctgactct gctggtgcag 2400
cggcacttct ttctgtccca atggatcgcc gagcctacca aagaacccat cgtggacgag 2460
gacgacgacg tggccgaaga gagacagaga atcatcaccg gcggcaacaa gaccgacatc 2520
ctgaggctgc acgagctgac caagatctac cctggcacat ctagccctgc cgtggacaga 2580
ctgtgtgtgg gagttagacc tggcgagtgc tttggcctgc tgggagttaa tggcgccgga 2640
aagaccacca ccttcaagat gctgaccggc gacaccacag tgaccagcgg agatgctact 2700
gtggccggca agagcatcct gaccaacatc agcgaggtgc accagaacat gggctactgc 2760
cctcagttcg acgccatcga cgaactgctg acaggcagag agcacctgta tctgtacgcc 2820
agactgagag gcgtgcccgc cgaagaaatt gagaaggtgg ccaactggtc catcaagagc 2880
ctgggactga ccgtgtacgc cgattgtctg gccggcacat atagcggcgg aaacaagcgg 2940
aagctgagca ccgccattgc tctgatcgga tgccctcctc tggtcctgct ggatgagcct 3000
acaaccggca tggatcccca ggctagacgg atgctgtgga acgtgatcgt gtccatcatc 3060
cgcgaaggca gagccgtggt cctgacaagc cacagcatgg aagagtgcga ggccctgtgt 3120
accagactgg ccatcatggt caagggcgcc ttcagatgca tgggcaccat tcagcacctg 3180
aaaagcaagt tcggcgacgg ctacatcgtg accatgaaga tcaagtcccc aaaggacgac 3240
ctgctgccag atctgaaccc cgtggaacag ttcttccagg gcaacttccc tggctccgtg 3300
cagcgggaaa gacactacaa catgctgcag tttcaggtgt ccagcagctc cctggccaga 3360
atctttcagc tgctgctctc ccacaaggat agcctgctga ttgaagagta cagcgtgacc 3420
cagaccacac tggaccaggt gttcgtgaac ttcgccaagc agcagaccga gagccacgac 3480
ttgcctctgc accctagagc tgccggcgct tctagacagg cccaggatta a 3531
<210> 10
<211> 296
<212> DNA
<213> 人类(Homo sapiens)
<400> 10
ggccccagaa gcctggtggt tgtttgtcct tctcagggga aaagtgaggc ggccccttgg 60
aggaaggggc cgggcagaat gatctaatcg gattccaagc agctcagggg attgtctttt 120
tctagcacct tcttgccact cctaagcgtc ctccgtgacc ccggctggga tttagcctgg 180
tgctgtgtca gccccggtct cccaggggct tcccagtggt ccccaggaac cctcgacagg 240
gcccggtctc tctcgtccag caagggcagg gacgggccac aggccaaggg cggtac 296
<210> 11
<211> 4495
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
ggccccagaa gcctggtggt tgtttgtcct tctcagggga aaagtgaggc ggccccttgg 60
aggaaggggc cgggcagaat gatctaatcg gattccaagc agctcagggg attgtctttt 120
tctagcacct tcttgccact cctaagcgtc ctccgtgacc ccggctggga tttagcctgg 180
tgctgtgtca gccccggtct cccaggggct tcccagtggt ccccaggaac cctcgacagg 240
gcccggtctc tctcgtccag caagggcagg gacgggccac aggccaaggg cggtactcta 300
gaggatccgg tactcgagga actgaaaaac cagaaagtta actggtaagt ttagtctttt 360
tgtcttttat ttcaggtccc ggatccggtg gtggtgcaaa tcaaagaact gctcctcagt 420
ggatgttgcc tttacttcta ggcctgtacg gaagtgttac ttctgctcta aaagctgcgg 480
aattgtaccc gcggccgcca ccatgggctt tgtgcgacag attcagctgc tgctgtggaa 540
gaactggacc ctgcggaagc ggcagaaaat cagattcgtg gtggaactcg tgtggcccct 600
gagcctgttt ctggtgctga tctggctgcg gaacgccaat cctctgtaca gccaccacga 660
gtgtcacttc cccaacaagg ccatgccttc tgccggaatg ctgccttggc tgcagggcat 720
cttctgcaac gtgaacaacc cctgctttca gagccccaca cctggcgaaa gccctggcat 780
cgtgtccaac tacaacaaca gcatcctggc cagagtgtac cgggacttcc aagagctgct 840
gatgaacgcc cctgagtctc agcacctggg cagaatctgg accgagctgc acatcctgag 900
ccagttcatg gacaccctga gaacacaccc cgagagaatc gccggcaggg gcatcagaat 960
ccgggacatc ctgaaggacg aggaaaccct gacactgttc ctcatcaaga acatcggcct 1020
gagcgacagc gtggtgtacc tgctgatcaa cagccaagtg cggcccgagc agtttgctca 1080
tggcgtgcca gatctcgccc tgaaggatat cgcctgttct gaggccctgc tggaacggtt 1140
catcatcttc agccagcgga gaggcgccaa gaccgtcaga tatgccctgt gcagtctgag 1200
ccagggaacc ctgcagtgga tcgaggatac cctgtacgcc aacgtggact tcttcaagct 1260
gttccgggtg ctgcccacac tgctggattc tagatcccag ggcatcaacc tgagaagctg 1320
gggcggcatc ctgtccgaca tgagcccaag aatccaagag ttcatccacc ggcctagcat 1380
gcaggacctg ctgtgggtta ccagacctct gatgcagaac ggcggacccg agacattcac 1440
caagctgatg ggaattctga gcgatctgct gtgcggctac cctgaaggcg gaggatctag 1500
agtgctgagc ttcaattggt acgaggacaa caactacaag gccttcctgg gcatcgactc 1560
caccagaaag gaccccatct acagctacga ccggcggaca accagcttct gcaatgccct 1620
gatccagagc ctggaaagca accctctgac caagatcgct tggagggccg ccaaacctct 1680
gctgatggga aagatcctgt acacccctga cagccctgcc gccagaagaa tcctgaagaa 1740
cgccaacagc accttcgagg aactggaaca cgtgcgcaag ctggtcaagg cctgggaaga 1800
agtgggacct cagatctggt acttcttcga caatagcacc cagatgaaca tgatcagaga 1860
caccctgggc aaccctaccg tgaaggactt cctgaacaga cagctgggcg aagagggcat 1920
taccgccgag gccatcctga actttctgta caagggcccc agagagtccc aggccgacga 1980
catggccaac ttcgattggc gggacatctt caacatcacc gacagaaccc tgcggctggt 2040
caaccagtac ctggaatgcc tggtgctgga caagttcgag agctacaacg acgagacaca 2100
gctgacccag agagccctgt ctctgctgga agagaatatg ttctgggctg gcgtggtgtt 2160
ccccgacatg tacccttgga caagcagcct gcctcctcac gtgaagtaca agatccggat 2220
ggacatcgac gtggtcgaaa agaccaacaa gatcaaggat cggtactggg acagcggccc 2280
tagagctgat cccgtggaag attttcggta catctggggc ggattcgcat acctgcagga 2340
catggtggaa cagggaatca cacggtccca ggtgcaggct gaagctcctg tgggaatcta 2400
cctgcagcag atgccttatc cttgcttcgt ggacgacagc ttcatgatca tcctgaatcg 2460
gtgcttcccc atcttcatgg tgctggcctg gatctactcc gtgtctatga ccgtgaagtc 2520
catcgtgctg gaaaaagagc tgcggctgaa agagacactg aagaaccagg gcgtgtccaa 2580
tgccgtgatc tggtgcacct ggtttctgga cagcttctcc attatgagca tgagcatctt 2640
tctgctgacg atcttcatca tgcacggccg gatcctgcac tacagcgacc cctttatcct 2700
cttcctgttc ctgctggcct tcagcaccgc tacaatcatg ctgtgttttc tgctgtccac 2760
cttcttcagc aaggcctctc tggccgctgc ttgtagcggc gtgatctact tcaccctgta 2820
cctgcctcac atcctgtgct tcgcatggca ggacagaatg accgccgagc tgaagaaagc 2880
tgtgtccctg ctgagccctg tggcctttgg ctttggcacc gagtacctcg tcagatttga 2940
ggaacaagga ctgggactgc agtggtccaa catcggcaat agccctacag agggcgacga 3000
gttcagcttc ctgctgtcta tgcagatgat gctgctggac gccgccgtgt atggactgct 3060
ggcttggtat ctggaccagg tgttcccagg cgattacggc actcctctgc cttggtattt 3120
cctgctgcaa gagagctact ggctcggcgg cgagggatgt agcaccagag aagaaagagc 3180
cctggaaaag accgagcctc tgaccgagga aacagaggac cctgaacacc cagagggcat 3240
ccacgatagc tttttcgaga gagaacaccc cggctgggtg ccaggcgtgt gtgtgaagaa 3300
tctggtcaag atcttcgagc cctgcggcag acctgccgtg gacagactga acatcacctt 3360
ctacgagaac cagattaccg cctttctggg ccacaacggc gctggcaaga caaccacact 3420
gagcatcctc acaggcctgc tgcctccaac aagcggcaca gttctcgttg gcggcagaga 3480
catcgagaca agcctggatg ccgtcagaca gtccctgggc atgtgccctc agcacaacat 3540
cctgtttcac cacctgaccg tggccgagca catgctgttt tatgcccagc tgaagggcaa 3600
gagccaagaa gaggctcagc tggaaatgga agccatgctc gaggacaccg gcctgcacca 3660
caagagaaat gaggaagccc aggatctgag cggcggcatg cagagaaaac tgagcgtggc 3720
cattgccttc gtgggcgacg ccaaggttgt gatcctggat gagcctacaa gcggcgtgga 3780
cccttacagc agaagatcca tctgggatct gctgctgaag tacagaagcg gccggaccat 3840
catcatgagc acccaccaca tggacgaggc cgatctgctc ggagacagaa tcgccatcat 3900
tgctcagggc agactgtact gcagcggcac cccactgttt ctgaagaact gtctggctgg 3960
cgatactctc attaccctgg ccgatggacg acgagtgcct attagagaac tggtgtcaca 4020
gcagaatttt tccgtgtggg ctctgaatcc tcagacttac cgcctggaga gggctagagt 4080
gagtagagct ttctgtaccg gcatcaaacc tgtgtaccgc ctcaccacta gactggggag 4140
atccattagg gccactgcca accaccgatt tctcacacct cagggctgga aacgagtcga 4200
tgaactccag cctggagatt acctggctct gcctaggaga atccctactg cctcctaaac 4260
gcgtcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 4320
tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 4380
tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 4440
ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatgg 4495
<210> 12
<211> 4273
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
ggccccagaa gcctggtggt tgtttgtcct tctcagggga aaagtgaggc ggccccttgg 60
aggaaggggc cgggcagaat gatctaatcg gattccaagc agctcagggg attgtctttt 120
tctagcacct tcttgccact cctaagcgtc ctccgtgacc ccggctggga tttagcctgg 180
tgctgtgtca gccccggtct cccaggggct tcccagtggt ccccaggaac cctcgacagg 240
gcccggtctc tctcgtccag caagggcagg gacgggccac aggccaaggg cggtactcta 300
gaggatccgg tactcgagga actgaaaaac cagaaagtta actggtaagt ttagtctttt 360
tgtcttttat ttcaggtccc ggatccggtg gtggtgcaaa tcaaagaact gctcctcagt 420
ggatgttgcc tttacttcta ggcctgtacg gaagtgttac ttctgctcta aaagctgcgg 480
aattgtaccc gcggccgcca ccatggcggc ggcgtgcccg gaactgcgtc agctggcgca 540
gagcgatgtg tattgggatc cgattgtgag cattgaaccg gatggcgtgg aagaagtgtt 600
tgatctgacc gtgccgggcc cgcataactt tgtggcgaac gatattattg cgcataactg 660
ttttggcacc ggcctgtacc tgacactcgt gcggaagatg aagaacatcc agagccagcg 720
gaagggcagc gagggcacat gtagctgtag cagcaagggc ttcagcacca cctgtcctgc 780
tcacgtggac gatctgaccc ctgaacaggt gctggatggc gacgtgaacg agctgatgga 840
cgtggtgctg caccacgtgc ctgaggccaa gctggtggaa tgcatcggcc aagagctgat 900
cttcctgctg cctaacaaga acttcaagca ccgggcctac gccagcctgt tcagagagct 960
ggaagagaca ctggccgatc tgggcctgag cagctttggc atcagcgaca cccctctgga 1020
agaaatcttc ctgaaagtga ccgaggacag cgacagcggc ccactttttg ctggcggagc 1080
acagcagaaa cgcgagaacg tgaaccctag acacccctgt ctgggcccta gagagaaagc 1140
cggacagacc cctcaggaca gcaatgtgtg ctctcctggt gctcctgccg ctcaccctga 1200
aggacaacct ccacctgagc ctgagtgtcc tggacctcag ctgaataccg gcacacagct 1260
ggttctgcag cacgtgcaag ccctgctggt caagagattc cagcacacca tcagaagcca 1320
caaggacttt ctggcccaga ttgtgctgcc cgccaccttc gtttttctgg ctctgatgct 1380
gagcatcgtg atccctccat tcggcgagta ccccgctctg acactgcacc cttggatcta 1440
cggccagcag tacaccttct tcagcatgga cgagcctggc agcgagcagt ttacagtgct 1500
ggctgatgtg ctgctgaaca agcccggctt cggcaaccgc tgtctgaaag aaggatggct 1560
gcctgagtac ccctgcggca atagcacccc ttggaaaacc cctagcgtgt cccctaacat 1620
cacccagctg ttccagaaac agaaatggac ccaagtgaac ccctctccat cctgccggtg 1680
ctccaccaga gaaaagctga ccatgctgcc cgagtgtcca gaaggtgcag gcggacttcc 1740
tccacctcag agaacccaga gaagcaccga gattctgcag gacctgaccg accggaatat 1800
cagcgacttc ctggttaaga cataccccgc actgatccgg tccagcctga agtccaagtt 1860
ctgggtcaac gagcagagat acggcggcat ctctatcggc ggaaagctgc ctgtggtgcc 1920
tatcacaggc gaagccctcg tgggctttct gtctgacctg ggcagaatca tgaacgtgtc 1980
cggcggaccc atcaccagag aggccagcaa agagatcccc gatttcctga agcacctgga 2040
aaccgaggat aacatcaaag tgtggttcaa caacaaaggc tggcacgccc tggtgtcctt 2100
tctgaacgtg gcccacaacg ccatcctgag agccagcctg cctaaggaca gaagccctga 2160
ggaatacggc atcaccgtga tctcccagcc actgaacctg accaaagagc agctgagcga 2220
gatcaccgtg ctgaccacat ctgtggatgc cgtggtggcc atctgcgtga tcttctccat 2280
gagcttcgtg cctgcctcct tcgtgctgta tctgatccaa gagagagtga acaagagcaa 2340
gcacctccag tttatcagcg gagtgtcccc aaccacctac tgggtcacca actttctgtg 2400
ggacatcatg aactactccg tgtctgccgg cctggtcgtg ggcatcttca tcggctttca 2460
gaagaaggcc tacacaagcc ccgagaacct gcctgctctg gttgctctgc tgctgctgta 2520
tggctgggcc gtgattccca tgatgtaccc cgccagcttc ctgttcgatg tgcctagcac 2580
agcctacgtg gccctgagct gcgccaatct gttcatcggc atcaacagca gcgccatcac 2640
cttcatcctg gaactgttcg agaacaacag aaccctgctg cggttcaacg ccgtgctgag 2700
aaagctgctg atcgtgttcc ctcacttctg cctcggcaga ggcctgattg atctggccct 2760
gtctcaggcc gtgaccgacg tgtacgctag atttggcgag gaacacagcg ccaatccttt 2820
ccactgggac ctgatcggca agaacctgtt cgccatggtg gtggaaggcg tggtgtattt 2880
cctgctgact ctgctggtgc agcggcactt ctttctgtcc caatggatcg ccgagcctac 2940
caaagaaccc atcgtggacg aggacgacga cgtggccgaa gagagacaga gaatcatcac 3000
cggcggcaac aagaccgaca tcctgaggct gcacgagctg accaagatct accctggcac 3060
atctagccct gccgtggaca gactgtgtgt gggagttaga cctggcgagt gctttggcct 3120
gctgggagtt aatggcgccg gaaagaccac caccttcaag atgctgaccg gcgacaccac 3180
agtgaccagc ggagatgcta ctgtggccgg caagagcatc ctgaccaaca tcagcgaggt 3240
gcaccagaac atgggctact gccctcagtt cgacgccatc gacgaactgc tgacaggcag 3300
agagcacctg tatctgtacg ccagactgag aggcgtgccc gccgaagaaa ttgagaaggt 3360
ggccaactgg tccatcaaga gcctgggact gaccgtgtac gccgattgtc tggccggcac 3420
atatagcggc ggaaacaagc ggaagctgag caccgccatt gctctgatcg gatgccctcc 3480
tctggtcctg ctggatgagc ctacaaccgg catggatccc caggctagac ggatgctgtg 3540
gaacgtgatc gtgtccatca tccgcgaagg cagagccgtg gtcctgacaa gccacagcat 3600
ggaagagtgc gaggccctgt gtaccagact ggccatcatg gtcaagggcg ccttcagatg 3660
catgggcacc attcagcacc tgaaaagcaa gttcggcgac ggctacatcg tgaccatgaa 3720
gatcaagtcc ccaaaggacg acctgctgcc agatctgaac cccgtggaac agttcttcca 3780
gggcaacttc cctggctccg tgcagcggga aagacactac aacatgctgc agtttcaggt 3840
gtccagcagc tccctggcca gaatctttca gctgctgctc tcccacaagg atagcctgct 3900
gattgaagag tacagcgtga cccagaccac actggaccag gtgttcgtga acttcgccaa 3960
gcagcagacc gagagccacg acttgcctct gcaccctaga gctgccggcg cttctagaca 4020
ggcccaggat taataaacgc gtcctcgact gtgccttcta gttgccagcc atctgttgtt 4080
tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 4140
taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 4200
gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 4260
gtgggctcta tgg 4273
<210> 13
<211> 4918
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 60
ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120
aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct 180
aggaagatcg gaattcgccc ttaagggccc cagaagcctg gtggttgttt gtccttctca 240
ggggaaaagt gaggcggccc cttggaggaa ggggccgggc agaatgatct aatcggattc 300
caagcagctc aggggattgt ctttttctag caccttcttg ccactcctaa gcgtcctccg 360
tgaccccggc tgggatttag cctggtgctg tgtcagcccc ggtctcccag gggcttccca 420
gtggtcccca ggaaccctcg acagggcccg gtctctctcg tccagcaagg gcagggacgg 480
gccacaggcc aagggcggta ctctagagga tccggtactc gaggaactga aaaaccagaa 540
agttaactgg taagtttagt ctttttgtct tttatttcag gtcccggatc cggtggtggt 600
gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct gtacggaagt 660
gttacttctg ctctaaaagc tgcggaattg tacccgcggc cgccaccatg ggctttgtgc 720
gacagattca gctgctgctg tggaagaact ggaccctgcg gaagcggcag aaaatcagat 780
tcgtggtgga actcgtgtgg cccctgagcc tgtttctggt gctgatctgg ctgcggaacg 840
ccaatcctct gtacagccac cacgagtgtc acttccccaa caaggccatg ccttctgccg 900
gaatgctgcc ttggctgcag ggcatcttct gcaacgtgaa caacccctgc tttcagagcc 960
ccacacctgg cgaaagccct ggcatcgtgt ccaactacaa caacagcatc ctggccagag 1020
tgtaccggga cttccaagag ctgctgatga acgcccctga gtctcagcac ctgggcagaa 1080
tctggaccga gctgcacatc ctgagccagt tcatggacac cctgagaaca caccccgaga 1140
gaatcgccgg caggggcatc agaatccggg acatcctgaa ggacgaggaa accctgacac 1200
tgttcctcat caagaacatc ggcctgagcg acagcgtggt gtacctgctg atcaacagcc 1260
aagtgcggcc cgagcagttt gctcatggcg tgccagatct cgccctgaag gatatcgcct 1320
gttctgaggc cctgctggaa cggttcatca tcttcagcca gcggagaggc gccaagaccg 1380
tcagatatgc cctgtgcagt ctgagccagg gaaccctgca gtggatcgag gataccctgt 1440
acgccaacgt ggacttcttc aagctgttcc gggtgctgcc cacactgctg gattctagat 1500
cccagggcat caacctgaga agctggggcg gcatcctgtc cgacatgagc ccaagaatcc 1560
aagagttcat ccaccggcct agcatgcagg acctgctgtg ggttaccaga cctctgatgc 1620
agaacggcgg acccgagaca ttcaccaagc tgatgggaat tctgagcgat ctgctgtgcg 1680
gctaccctga aggcggagga tctagagtgc tgagcttcaa ttggtacgag gacaacaact 1740
acaaggcctt cctgggcatc gactccacca gaaaggaccc catctacagc tacgaccggc 1800
ggacaaccag cttctgcaat gccctgatcc agagcctgga aagcaaccct ctgaccaaga 1860
tcgcttggag ggccgccaaa cctctgctga tgggaaagat cctgtacacc cctgacagcc 1920
ctgccgccag aagaatcctg aagaacgcca acagcacctt cgaggaactg gaacacgtgc 1980
gcaagctggt caaggcctgg gaagaagtgg gacctcagat ctggtacttc ttcgacaata 2040
gcacccagat gaacatgatc agagacaccc tgggcaaccc taccgtgaag gacttcctga 2100
acagacagct gggcgaagag ggcattaccg ccgaggccat cctgaacttt ctgtacaagg 2160
gccccagaga gtcccaggcc gacgacatgg ccaacttcga ttggcgggac atcttcaaca 2220
tcaccgacag aaccctgcgg ctggtcaacc agtacctgga atgcctggtg ctggacaagt 2280
tcgagagcta caacgacgag acacagctga cccagagagc cctgtctctg ctggaagaga 2340
atatgttctg ggctggcgtg gtgttccccg acatgtaccc ttggacaagc agcctgcctc 2400
ctcacgtgaa gtacaagatc cggatggaca tcgacgtggt cgaaaagacc aacaagatca 2460
aggatcggta ctgggacagc ggccctagag ctgatcccgt ggaagatttt cggtacatct 2520
ggggcggatt cgcatacctg caggacatgg tggaacaggg aatcacacgg tcccaggtgc 2580
aggctgaagc tcctgtggga atctacctgc agcagatgcc ttatccttgc ttcgtggacg 2640
acagcttcat gatcatcctg aatcggtgct tccccatctt catggtgctg gcctggatct 2700
actccgtgtc tatgaccgtg aagtccatcg tgctggaaaa agagctgcgg ctgaaagaga 2760
cactgaagaa ccagggcgtg tccaatgccg tgatctggtg cacctggttt ctggacagct 2820
tctccattat gagcatgagc atctttctgc tgacgatctt catcatgcac ggccggatcc 2880
tgcactacag cgaccccttt atcctcttcc tgttcctgct ggccttcagc accgctacaa 2940
tcatgctgtg ttttctgctg tccaccttct tcagcaaggc ctctctggcc gctgcttgta 3000
gcggcgtgat ctacttcacc ctgtacctgc ctcacatcct gtgcttcgca tggcaggaca 3060
gaatgaccgc cgagctgaag aaagctgtgt ccctgctgag ccctgtggcc tttggctttg 3120
gcaccgagta cctcgtcaga tttgaggaac aaggactggg actgcagtgg tccaacatcg 3180
gcaatagccc tacagagggc gacgagttca gcttcctgct gtctatgcag atgatgctgc 3240
tggacgccgc cgtgtatgga ctgctggctt ggtatctgga ccaggtgttc ccaggcgatt 3300
acggcactcc tctgccttgg tatttcctgc tgcaagagag ctactggctc ggcggcgagg 3360
gatgtagcac cagagaagaa agagccctgg aaaagaccga gcctctgacc gaggaaacag 3420
aggaccctga acacccagag ggcatccacg atagcttttt cgagagagaa caccccggct 3480
gggtgccagg cgtgtgtgtg aagaatctgg tcaagatctt cgagccctgc ggcagacctg 3540
ccgtggacag actgaacatc accttctacg agaaccagat taccgccttt ctgggccaca 3600
acggcgctgg caagacaacc acactgagca tcctcacagg cctgctgcct ccaacaagcg 3660
gcacagttct cgttggcggc agagacatcg agacaagcct ggatgccgtc agacagtccc 3720
tgggcatgtg ccctcagcac aacatcctgt ttcaccacct gaccgtggcc gagcacatgc 3780
tgttttatgc ccagctgaag ggcaagagcc aagaagaggc tcagctggaa atggaagcca 3840
tgctcgagga caccggcctg caccacaaga gaaatgagga agcccaggat ctgagcggcg 3900
gcatgcagag aaaactgagc gtggccattg ccttcgtggg cgacgccaag gttgtgatcc 3960
tggatgagcc tacaagcggc gtggaccctt acagcagaag atccatctgg gatctgctgc 4020
tgaagtacag aagcggccgg accatcatca tgagcaccca ccacatggac gaggccgatc 4080
tgctcggaga cagaatcgcc atcattgctc agggcagact gtactgcagc ggcaccccac 4140
tgtttctgaa gaactgtctg gctggcgata ctctcattac cctggccgat ggacgacgag 4200
tgcctattag agaactggtg tcacagcaga atttttccgt gtgggctctg aatcctcaga 4260
cttaccgcct ggagagggct agagtgagta gagctttctg taccggcatc aaacctgtgt 4320
accgcctcac cactagactg gggagatcca ttagggccac tgccaaccac cgatttctca 4380
cacctcaggg ctggaaacga gtcgatgaac tccagcctgg agattacctg gctctgccta 4440
ggagaatccc tactgcctcc taaacgcgtc ctcgactgtg ccttctagtt gccagccatc 4500
tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 4560
ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 4620
gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 4680
ggatgcggtg ggctctatgg cctcgagtta agggcgaatt cccgataagg atcttcctag 4740
agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag 4800
tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa 4860
aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcag 4918
<210> 14
<211> 4696
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 60
ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120
aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct 180
aggaagatcg gaattcgccc ttaagggccc cagaagcctg gtggttgttt gtccttctca 240
ggggaaaagt gaggcggccc cttggaggaa ggggccgggc agaatgatct aatcggattc 300
caagcagctc aggggattgt ctttttctag caccttcttg ccactcctaa gcgtcctccg 360
tgaccccggc tgggatttag cctggtgctg tgtcagcccc ggtctcccag gggcttccca 420
gtggtcccca ggaaccctcg acagggcccg gtctctctcg tccagcaagg gcagggacgg 480
gccacaggcc aagggcggta ctctagagga tccggtactc gaggaactga aaaaccagaa 540
agttaactgg taagtttagt ctttttgtct tttatttcag gtcccggatc cggtggtggt 600
gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct gtacggaagt 660
gttacttctg ctctaaaagc tgcggaattg tacccgcggc cgccaccatg gcggcggcgt 720
gcccggaact gcgtcagctg gcgcagagcg atgtgtattg ggatccgatt gtgagcattg 780
aaccggatgg cgtggaagaa gtgtttgatc tgaccgtgcc gggcccgcat aactttgtgg 840
cgaacgatat tattgcgcat aactgttttg gcaccggcct gtacctgaca ctcgtgcgga 900
agatgaagaa catccagagc cagcggaagg gcagcgaggg cacatgtagc tgtagcagca 960
agggcttcag caccacctgt cctgctcacg tggacgatct gacccctgaa caggtgctgg 1020
atggcgacgt gaacgagctg atggacgtgg tgctgcacca cgtgcctgag gccaagctgg 1080
tggaatgcat cggccaagag ctgatcttcc tgctgcctaa caagaacttc aagcaccggg 1140
cctacgccag cctgttcaga gagctggaag agacactggc cgatctgggc ctgagcagct 1200
ttggcatcag cgacacccct ctggaagaaa tcttcctgaa agtgaccgag gacagcgaca 1260
gcggcccact ttttgctggc ggagcacagc agaaacgcga gaacgtgaac cctagacacc 1320
cctgtctggg ccctagagag aaagccggac agacccctca ggacagcaat gtgtgctctc 1380
ctggtgctcc tgccgctcac cctgaaggac aacctccacc tgagcctgag tgtcctggac 1440
ctcagctgaa taccggcaca cagctggttc tgcagcacgt gcaagccctg ctggtcaaga 1500
gattccagca caccatcaga agccacaagg actttctggc ccagattgtg ctgcccgcca 1560
ccttcgtttt tctggctctg atgctgagca tcgtgatccc tccattcggc gagtaccccg 1620
ctctgacact gcacccttgg atctacggcc agcagtacac cttcttcagc atggacgagc 1680
ctggcagcga gcagtttaca gtgctggctg atgtgctgct gaacaagccc ggcttcggca 1740
accgctgtct gaaagaagga tggctgcctg agtacccctg cggcaatagc accccttgga 1800
aaacccctag cgtgtcccct aacatcaccc agctgttcca gaaacagaaa tggacccaag 1860
tgaacccctc tccatcctgc cggtgctcca ccagagaaaa gctgaccatg ctgcccgagt 1920
gtccagaagg tgcaggcgga cttcctccac ctcagagaac ccagagaagc accgagattc 1980
tgcaggacct gaccgaccgg aatatcagcg acttcctggt taagacatac cccgcactga 2040
tccggtccag cctgaagtcc aagttctggg tcaacgagca gagatacggc ggcatctcta 2100
tcggcggaaa gctgcctgtg gtgcctatca caggcgaagc cctcgtgggc tttctgtctg 2160
acctgggcag aatcatgaac gtgtccggcg gacccatcac cagagaggcc agcaaagaga 2220
tccccgattt cctgaagcac ctggaaaccg aggataacat caaagtgtgg ttcaacaaca 2280
aaggctggca cgccctggtg tcctttctga acgtggccca caacgccatc ctgagagcca 2340
gcctgcctaa ggacagaagc cctgaggaat acggcatcac cgtgatctcc cagccactga 2400
acctgaccaa agagcagctg agcgagatca ccgtgctgac cacatctgtg gatgccgtgg 2460
tggccatctg cgtgatcttc tccatgagct tcgtgcctgc ctccttcgtg ctgtatctga 2520
tccaagagag agtgaacaag agcaagcacc tccagtttat cagcggagtg tccccaacca 2580
cctactgggt caccaacttt ctgtgggaca tcatgaacta ctccgtgtct gccggcctgg 2640
tcgtgggcat cttcatcggc tttcagaaga aggcctacac aagccccgag aacctgcctg 2700
ctctggttgc tctgctgctg ctgtatggct gggccgtgat tcccatgatg taccccgcca 2760
gcttcctgtt cgatgtgcct agcacagcct acgtggccct gagctgcgcc aatctgttca 2820
tcggcatcaa cagcagcgcc atcaccttca tcctggaact gttcgagaac aacagaaccc 2880
tgctgcggtt caacgccgtg ctgagaaagc tgctgatcgt gttccctcac ttctgcctcg 2940
gcagaggcct gattgatctg gccctgtctc aggccgtgac cgacgtgtac gctagatttg 3000
gcgaggaaca cagcgccaat cctttccact gggacctgat cggcaagaac ctgttcgcca 3060
tggtggtgga aggcgtggtg tatttcctgc tgactctgct ggtgcagcgg cacttctttc 3120
tgtcccaatg gatcgccgag cctaccaaag aacccatcgt ggacgaggac gacgacgtgg 3180
ccgaagagag acagagaatc atcaccggcg gcaacaagac cgacatcctg aggctgcacg 3240
agctgaccaa gatctaccct ggcacatcta gccctgccgt ggacagactg tgtgtgggag 3300
ttagacctgg cgagtgcttt ggcctgctgg gagttaatgg cgccggaaag accaccacct 3360
tcaagatgct gaccggcgac accacagtga ccagcggaga tgctactgtg gccggcaaga 3420
gcatcctgac caacatcagc gaggtgcacc agaacatggg ctactgccct cagttcgacg 3480
ccatcgacga actgctgaca ggcagagagc acctgtatct gtacgccaga ctgagaggcg 3540
tgcccgccga agaaattgag aaggtggcca actggtccat caagagcctg ggactgaccg 3600
tgtacgccga ttgtctggcc ggcacatata gcggcggaaa caagcggaag ctgagcaccg 3660
ccattgctct gatcggatgc cctcctctgg tcctgctgga tgagcctaca accggcatgg 3720
atccccaggc tagacggatg ctgtggaacg tgatcgtgtc catcatccgc gaaggcagag 3780
ccgtggtcct gacaagccac agcatggaag agtgcgaggc cctgtgtacc agactggcca 3840
tcatggtcaa gggcgccttc agatgcatgg gcaccattca gcacctgaaa agcaagttcg 3900
gcgacggcta catcgtgacc atgaagatca agtccccaaa ggacgacctg ctgccagatc 3960
tgaaccccgt ggaacagttc ttccagggca acttccctgg ctccgtgcag cgggaaagac 4020
actacaacat gctgcagttt caggtgtcca gcagctccct ggccagaatc tttcagctgc 4080
tgctctccca caaggatagc ctgctgattg aagagtacag cgtgacccag accacactgg 4140
accaggtgtt cgtgaacttc gccaagcagc agaccgagag ccacgacttg cctctgcacc 4200
ctagagctgc cggcgcttct agacaggccc aggattaata aacgcgtcct cgactgtgcc 4260
ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 4320
tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 4380
gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 4440
caatagcagg catgctgggg atgcggtggg ctctatggcc tcgagttaag ggcgaattcc 4500
cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta 4560
actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca 4620
ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga 4680
gcgagcgagc gcgcag 4696
<210> 15
<211> 11250
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 60
ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120
aggggttcct tgtagttaat gattaacccg ccatgctact tatctactta agcgttacat 180
aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa 240
tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg 300
cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg 360
acggtaaatg gcccgcctgg cattgtgccc agtacatgac cttatgggac tttcctactt 420
ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct 480
gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt 540
aattattttg tgcagcgatg ggggcggggg gggggggggg gcggggcgag gggcggggcg 600
gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt 660
ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag 720
tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc 780
ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg 840
gctgtaatta gctgagcaag aggtaagggt ttaagggatg gttggttggt ggggtattaa 900
tgtttaatta cctggagcac ctgcctgaaa tcactttttt tcaggttgga ccggtgccac 960
catgggcttc gtgagacaga tacagctttt gctctggaag aactggaccc tgcggaaaag 1020
gcaaaagatt cgctttgtgg tggaactcgt gtggccttta tctttatttc tggtcttgat 1080
ctggttaagg aatgccaacc cactctacag ccatcatgaa tgccatttcc ccaacaaggc 1140
gatgccctca gcaggaatgc tgccgtggct ccaggggatc ttctgcaatg tgaacaatcc 1200
ctgttttcaa agccccaccc caggagaatc tcctggaatt gtgtcaaact ataacaactc 1260
catcttggca agggtatatc gagattttca agaactcctc atgaatgcac cagagagcca 1320
gcaccttggc cgtatttgga cagagctaca catcttgtcc caattcatgg acaccctccg 1380
gactcacccg gagagaattg caggaagagg aatacgaata agggatatct tgaaagatga 1440
agaaacactg acactatttc tcattaaaaa catcggcctg tctgactcag tggtctacct 1500
tctgatcaac tctcaagtcc gtccagagca gttcgctcat ggagtcccgg acctggcgct 1560
gaaggacatc gcctgcagcg aggccctcct ggagcgcttc atcatcttca gccagagacg 1620
cggggcaaag acggtgcgct atgccctgtg ctccctctcc cagggcaccc tacagtggat 1680
agaagacact ctgtatgcca acgtggactt cttcaagctc ttccgtgtgc ttcccacact 1740
cctagacagc cgttctcaag gtatcaatct gagatcttgg ggaggaatat tatctgatat 1800
gtcaccaaga attcaagagt ttatccatcg gccgagtatg caggacttgc tgtgggtgac 1860
caggcccctc atgcagaatg gtggtccaga gacctttaca aagctgatgg gcatcctgtc 1920
tgacctcctg tgtggctacc ccgagggagg tggctctcgg gtgctctcct tcaactggta 1980
tgaagacaat aactataagg cctttctggg gattgactcc acaaggaagg atcctatcta 2040
ttcttatgac agaagaacaa catccttttg taatgcattg atccagagcc tggagtcaaa 2100
tcctttaacc aaaatcgctt ggagggcggc aaagcctttg ctgatgggaa aaatcctgta 2160
cactcctgat tcacctgcag cacgaaggat actgaagaat gccaactcaa cttttgaaga 2220
actggaacac gttaggaagt tggtcaaagc ctgggaagaa gtagggcccc agatctggta 2280
cttctttgac aacagcacac agatgaacat gatcagagat accctgggga acccaacagt 2340
aaaagacttt ttgaataggc agcttggtga agaaggtatt actgctgaag ccatcctaaa 2400
cttcctctac aagggccctc gggaaagcca ggctgacgac atggccaact tcgactggag 2460
ggacatattt aacatcactg atcgcaccct ccgcctggtc aatcaatacc tggagtgctt 2520
ggtcctggat aagtttgaaa gctacaatga tgaaactcag ctcacccaac gtgccctctc 2580
tctactggag gaaaacatgt tctgggccgg agtggtattc cctgacatgt atccctggac 2640
cagctctcta ccaccccacg tgaagtataa gatccgaatg gacatagacg tggtggagaa 2700
aaccaataag attaaagaca ggtattggga ttctggtccc agagctgatc ccgtggaaga 2760
tttccggtac atctggggcg ggtttgccta tctgcaggac atggttgaac aggggatcac 2820
aaggagccag gtgcaggcgg aggctccagt tggaatctac ctccagcaga tgccctaccc 2880
ctgcttcgtg gacgattctt tcatgatcat cctgaaccgc tgtttcccta tcttcatggt 2940
gctggcatgg atctactctg tctccatgac tgtgaagagc atcgtcttgg agaaggagtt 3000
gcgactgaag gagaccttga aaaatcaggg tgtctccaat gcagtgattt ggtgtacctg 3060
gttcctggac agcttctcca tcatgtcgat gagcatcttc ctcctgacga tattcatcat 3120
gcatggaaga atcctacatt acagcgaccc attcatcctc ttcctgttct tgttggcttt 3180
ctccactgcc accatcatgc tgtgctttct gctcagcacc ttcttctcca aggccagtct 3240
ggcagcagcc tgtagtggtg tcatctattt caccctctac ctgccacaca tcctgtgctt 3300
cgcctggcag gaccgcatga ccgctgagct gaagaaggct gtgagcttac tgtctccggt 3360
ggcatttgga tttggcactg agtacctggt tcgctttgaa gagcaaggcc tggggctgca 3420
gtggagcaac atcgggaaca gtcccacgga aggggacgaa ttcagcttcc tgctgtccat 3480
gcagatgatg ctccttgatg ctgctgtcta tggcttactc gcttggtacc ttgatcaggt 3540
gtttccagga gactatggaa ccccacttcc ttggtacttt cttctacaag agtcgtattg 3600
gcttggcggt gaagggtgtt caaccagaga agaaagagcc ctggaaaaga ccgagcccct 3660
aacagaggaa acggaggatc cagagcaccc agaaggaata cacgactcct tctttgaacg 3720
tgagcatcca gggtgggttc ctggggtatg cgtgaagaat ctggtaaaga tttttgagcc 3780
ctgtggccgg ccagctgtgg accgtctgaa catcaccttc tacgagaacc agatcaccgc 3840
attcctgggc cacaatggag ctgggaaaac caccaccttg tccatcctga cgggtctgtt 3900
gccaccaacc tctgggactg tgctcgttgg gggaagggac attgaaacca gcctggatgc 3960
agtccggcag agccttggca tgtgtccaca gcacaacatc ctgttccacc acctcacggt 4020
ggctgagcac atgctgttct atgcccagct gaaaggaaag tcccaggagg aggcccagct 4080
ggagatggaa gccatgttgg aggacacagg cctccaccac aagcggaatg aagaggctca 4140
ggacctatca ggtggcatgc agagaaagct gtcggttgcc attgcctttg tgggagatgc 4200
caaggtggtg attctggacg aacccacctc tggggtggac ccttactcga gacgctcaat 4260
ctgggatctg ctcctgaagt atcgctcagg cagaaccatc atcatgtcca ctcaccacat 4320
ggacgaggcc gacctccttg gggaccgcat tgccatcatt gcccagggaa ggctctactg 4380
ctcaggcacc ccactcttcc tgaagaactg ctttggcaca ggcttgtact taaccttggt 4440
gcgcaagatg aaaaacatcc agagccaaag gaaaggcagt gaggggacct gcagctgctc 4500
gtctaagggt ttctccacca cgtgtccagc ccacgtcgat gacctaactc cagaacaagt 4560
cctggatggg gatgtaaatg agctgatgga tgtagttctc caccatgttc cagaggcaaa 4620
gctggtggag tgcattggtc aagaacttat cttccttctt ccaaataaga acttcaagca 4680
cagagcatat gccagccttt tcagagagct ggaggagacg ctggctgacc ttggtctcag 4740
cagttttgga atttctgaca ctcccctgga agagattttt ctgaaggtca cggaggattc 4800
tgattcagga cctctgtttg cgggtggcgc tcagcagaaa agagaaaacg tcaacccccg 4860
acacccctgc ttgggtccca gagagaaggc tggacagaca ccccaggact ccaatgtctg 4920
ctccccaggg gcgccggctg ctcacccaga gggccagcct cccccagagc cagagtgccc 4980
aggcccgcag ctcaacacgg ggacacagct ggtcctccag catgtgcagg cgctgctggt 5040
caagagattc caacacacca tccgcagcca caaggacttc ctggcgcaga tcgtgctccc 5100
ggctaccttt gtgtttttgg ctctgatgct ttctattgtt atccctcctt ttggcgaata 5160
ccccgctttg acccttcacc cctggatata tgggcagcag tacaccttct tcagcatgga 5220
tgaaccaggc agtgagcagt tcacggtact tgcagacgtc ctcctgaata agccaggctt 5280
tggcaaccgc tgcctgaagg aagggtggct tccggagtac ccctgtggca actcaacacc 5340
ctggaagact ccttctgtgt ccccaaacat cacccagctg ttccagaagc agaaatggac 5400
acaggtcaac ccttcaccat cctgcaggtg cagcaccagg gagaagctca ccatgctgcc 5460
agagtgcccc gagggtgccg ggggcctccc gcccccccag agaacacagc gcagcacgga 5520
aattctacaa gacctgacgg acaggaacat ctccgacttc ttggtaaaaa cgtatcctgc 5580
tcttataaga agcagcttaa agagcaaatt ctgggtcaat gaacagaggt atggaggaat 5640
ttccattgga ggaaagctcc cagtcgtccc catcacgggg gaagcacttg ttgggttttt 5700
aagcgacctt ggccggatca tgaatgtgag cgggggccct atcactagag aggcctctaa 5760
agaaatacct gatttcctta aacatctaga aactgaagac aacattaagg tgtggtttaa 5820
taacaaaggc tggcatgccc tggtcagctt tctcaatgtg gcccacaacg ccatcttacg 5880
ggccagcctg cctaaggaca ggagccccga ggagtatgga atcaccgtca ttagccaacc 5940
cctgaacctg accaaggagc agctctcaga gattacagtg ctgaccactt cagtggatgc 6000
tgtggttgcc atctgcgtga ttttctccat gtccttcgtc ccagccagct ttgtccttta 6060
tttgatccag gagcgggtga acaaatccaa gcacctccag tttatcagtg gagtgagccc 6120
caccacctac tgggtgacca acttcctctg ggacatcatg aattattccg tgagtgctgg 6180
gctggtggtg ggcatcttca tcgggtttca gaagaaagcc tacacttctc cagaaaacct 6240
tcctgccctt gtggcactgc tcctgctgta tggatgggcg gtcattccca tgatgtaccc 6300
agcatccttc ctgtttgatg tccccagcac agcctatgtg gctttatctt gtgctaatct 6360
gttcatcggc atcaacagca gtgctattac cttcatcttg gaattatttg agaataaccg 6420
gacgctgctc aggttcaacg ccgtgctgag gaagctgctc attgtcttcc cccacttctg 6480
cctgggccgg ggcctcattg accttgcact gagccaggct gtgacagatg tctatgcccg 6540
gtttggtgag gagcactctg caaatccgtt ccactgggac ctgattggga agaacctgtt 6600
tgccatggtg gtggaagggg tggtgtactt cctcctgacc ctgctggtcc agcgccactt 6660
cttcctctcc caatggattg ccgagcccac taaggagccc attgttgatg aagatgatga 6720
tgtggctgaa gaaagacaaa gaattattac tggtggaaat aaaactgaca tcttaaggct 6780
acatgaacta accaagattt atccaggcac ctccagccca gcagtggaca ggctgtgtgt 6840
cggagttcgc cctggagagt gctttggcct cctgggagtg aatggtgccg gcaaaacaac 6900
cacattcaag atgctcactg gggacaccac agtgacctca ggggatgcca ccgtagcagg 6960
caagagtatt ttaaccaata tttctgaagt ccatcaaaat atgggctact gtcctcagtt 7020
tgatgcaatt gatgagctgc tcacaggacg agaacatctt tacctttatg cccggcttcg 7080
aggtgtacca gcagaagaaa tcgaaaaggt tgcaaactgg agtattaaga gcctgggcct 7140
gactgtctac gccgactgcc tggctggcac gtacagtggg ggcaacaagc ggaaactctc 7200
cacagccatc gcactcattg gctgcccacc gctggtgctg ctggatgagc ccaccacagg 7260
gatggacccc caggcacgcc gcatgctgtg gaacgtcatc gtgagcatca tcagagaagg 7320
gagggctgtg gtcctcacat cccacagcat ggaagaatgt gaggcactgt gtacccggct 7380
ggccatcatg gtaaagggcg cctttcgatg tatgggcacc attcagcatc tcaagtccaa 7440
atttggagat ggctatatcg tcacaatgaa gatcaaatcc ccgaaggacg acctgcttcc 7500
tgacctgaac cctgtggagc agttcttcca ggggaacttc ccaggcagtg tgcagaggga 7560
gaggcactac aacatgctcc agttccaggt ctcctcctcc tccctggcga ggatcttcca 7620
gctcctcctc tcccacaagg acagcctgct catcgaggag tactcagtca cacagaccac 7680
actggaccag gtgtttgtaa attttgctaa acagcagact gaaagtcatg acctccctct 7740
gcaccctcga gctgctggag ccagtcgaca agcccaggac tgagcggccg cgataatcaa 7800
cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 7860
acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 7920
ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc 7980
gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 8040
cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga 8100
ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt 8160
gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg 8220
attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct cgagtagata 8280
agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc 8340
cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg 8400
gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ccttaattaa cctaattcac 8460
tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc 8520
ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc 8580
cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa 8640
gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 8700
ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 8760
ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 8820
aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 8880
gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 8940
cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 9000
attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 9060
cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 9120
tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 9180
ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt 9240
ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga 9300
tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa 9360
gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct 9420
gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat 9480
acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga 9540
tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc 9600
caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat 9660
gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa 9720
cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac 9780
tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa 9840
agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc 9900
tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc 9960
ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag 10020
acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta 10080
ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 10140
gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 10200
gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 10260
ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 10320
gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 10380
tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 10440
cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 10500
cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 10560
ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 10620
tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 10680
cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 10740
ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 10800
aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 10860
ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 10920
tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga 10980
gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg 11040
gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg 11100
caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct 11160
tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta 11220
tgaccatgat tacgccagat ttaattaagg 11250
<210> 16
<211> 11250
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 60
ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120
aggggttcct tgtagttaat gattaacccg ccatgctact tatctactta agcgttacat 180
aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa 240
tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg 300
cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg 360
acggtaaatg gcccgcctgg cattgtgccc agtacatgac cttatgggac tttcctactt 420
ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct 480
gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt 540
aattattttg tgcagcgatg ggggcggggg gggggggggg gcggggcgag gggcggggcg 600
gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt 660
ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag 720
tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc 780
ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg 840
gctgtaatta gctgagcaag aggtaagggt ttaagggatg gttggttggt ggggtattaa 900
tgtttaatta cctggagcac ctgcctgaaa tcactttttt tcaggttgga ccggtgccac 960
catgggcttt gtgcgacaga ttcagctgct gctgtggaag aactggaccc tgcggaagcg 1020
gcagaaaatc agattcgtgg tggaactcgt gtggcccctg agcctgtttc tggtgctgat 1080
ctggctgcgg aacgccaatc ctctgtacag ccaccacgag tgtcacttcc ccaacaaggc 1140
catgccttct gccggaatgc tgccttggct gcagggcatc ttctgcaacg tgaacaaccc 1200
ctgctttcag agccccacac ctggcgaaag ccctggcatc gtgtccaact acaacaacag 1260
catcctggcc agagtgtacc gggacttcca agagctgctg atgaacgccc ctgagtctca 1320
gcacctgggc agaatctgga ccgagctgca catcctgagc cagttcatgg acaccctgag 1380
aacacacccc gagagaatcg ccggcagggg catcagaatc cgggacatcc tgaaggacga 1440
ggaaaccctg acactgttcc tcatcaagaa catcggcctg agcgacagcg tggtgtacct 1500
gctgatcaac agccaagtgc ggcccgagca gtttgctcat ggcgtgccag atctcgccct 1560
gaaggatatc gcctgttctg aggccctgct ggaacggttc atcatcttca gccagcggag 1620
aggcgccaag accgtcagat atgccctgtg cagtctgagc cagggaaccc tgcagtggat 1680
cgaggatacc ctgtacgcca acgtggactt cttcaagctg ttccgggtgc tgcccacact 1740
gctggattct agatcccagg gcatcaacct gagaagctgg ggcggcatcc tgtccgacat 1800
gagcccaaga atccaagagt tcatccaccg gcctagcatg caggacctgc tgtgggttac 1860
cagacctctg atgcagaacg gcggacccga gacattcacc aagctgatgg gaattctgag 1920
cgatctgctg tgcggctacc ctgaaggcgg aggatctaga gtgctgagct tcaattggta 1980
cgaggacaac aactacaagg ccttcctggg catcgactcc accagaaagg accccatcta 2040
cagctacgac cggcggacaa ccagcttctg caatgccctg atccagagcc tggaaagcaa 2100
ccctctgacc aagatcgctt ggagggccgc caaacctctg ctgatgggaa agatcctgta 2160
cacccctgac agccctgccg ccagaagaat cctgaagaac gccaacagca ccttcgagga 2220
actggaacac gtgcgcaagc tggtcaaggc ctgggaagaa gtgggacctc agatctggta 2280
cttcttcgac aatagcaccc agatgaacat gatcagagac accctgggca accctaccgt 2340
gaaggacttc ctgaacagac agctgggcga agagggcatt accgccgagg ccatcctgaa 2400
ctttctgtac aagggcccca gagagtccca ggccgacgac atggccaact tcgattggcg 2460
ggacatcttc aacatcaccg acagaaccct gcggctggtc aaccagtacc tggaatgcct 2520
ggtgctggac aagttcgaga gctacaacga cgagacacag ctgacccaga gagccctgtc 2580
tctgctggaa gagaatatgt tctgggctgg cgtggtgttc cccgacatgt acccttggac 2640
aagcagcctg cctcctcacg tgaagtacaa gatccggatg gacatcgacg tggtcgaaaa 2700
gaccaacaag atcaaggatc ggtactggga cagcggccct agagctgatc ccgtggaaga 2760
ttttcggtac atctggggcg gattcgcata cctgcaggac atggtggaac agggaatcac 2820
acggtcccag gtgcaggctg aagctcctgt gggaatctac ctgcagcaga tgccttatcc 2880
ttgcttcgtg gacgacagct tcatgatcat cctgaatcgg tgcttcccca tcttcatggt 2940
gctggcctgg atctactccg tgtctatgac cgtgaagtcc atcgtgctgg aaaaagagct 3000
gcggctgaaa gagacactga agaaccaggg cgtgtccaat gccgtgatct ggtgcacctg 3060
gtttctggac agcttctcca ttatgagcat gagcatcttt ctgctgacga tcttcatcat 3120
gcacggccgg atcctgcact acagcgaccc ctttatcctc ttcctgttcc tgctggcctt 3180
cagcaccgct acaatcatgc tgtgttttct gctgtccacc ttcttcagca aggcctctct 3240
ggccgctgct tgtagcggcg tgatctactt caccctgtac ctgcctcaca tcctgtgctt 3300
cgcatggcag gacagaatga ccgccgagct gaagaaagct gtgtccctgc tgagccctgt 3360
ggcctttggc tttggcaccg agtacctcgt cagatttgag gaacaaggac tgggactgca 3420
gtggtccaac atcggcaata gccctacaga gggcgacgag ttcagcttcc tgctgtctat 3480
gcagatgatg ctgctggacg ccgccgtgta tggactgctg gcttggtatc tggaccaggt 3540
gttcccaggc gattacggca ctcctctgcc ttggtatttc ctgctgcaag agagctactg 3600
gctcggcggc gagggatgta gcaccagaga agaaagagcc ctggaaaaga ccgagcctct 3660
gaccgaggaa acagaggacc ctgaacaccc agagggcatc cacgatagct ttttcgagag 3720
agaacacccc ggctgggtgc caggcgtgtg tgtgaagaat ctggtcaaga tcttcgagcc 3780
ctgcggcaga cctgccgtgg acagactgaa catcaccttc tacgagaacc agattaccgc 3840
ctttctgggc cacaacggcg ctggcaagac aaccacactg agcatcctca caggcctgct 3900
gcctccaaca agcggcacag ttctcgttgg cggcagagac atcgagacaa gcctggatgc 3960
cgtcagacag tccctgggca tgtgccctca gcacaacatc ctgtttcacc acctgaccgt 4020
ggccgagcac atgctgtttt atgcccagct gaagggcaag agccaagaag aggctcagct 4080
ggaaatggaa gccatgctcg aggacaccgg cctgcaccac aagagaaatg aggaagccca 4140
ggatctgagc ggcggcatgc agagaaaact gagcgtggcc attgccttcg tgggcgacgc 4200
caaggttgtg atcctggatg agcctacaag cggcgtggac ccttacagca gaagatccat 4260
ctgggatctg ctgctgaagt acagaagcgg ccggaccatc atcatgagca cccaccacat 4320
ggacgaggcc gatctgctcg gagacagaat cgccatcatt gctcagggca gactgtactg 4380
cagcggcacc ccactgtttc tgaagaactg ttttggcacc ggcctgtacc tgacactcgt 4440
gcggaagatg aagaacatcc agagccagcg gaagggcagc gagggcacat gtagctgtag 4500
cagcaagggc ttcagcacca cctgtcctgc tcacgtggac gatctgaccc ctgaacaggt 4560
gctggatggc gacgtgaacg agctgatgga cgtggtgctg caccacgtgc ctgaggccaa 4620
gctggtggaa tgcatcggcc aagagctgat cttcctgctg cctaacaaga acttcaagca 4680
ccgggcctac gccagcctgt tcagagagct ggaagagaca ctggccgatc tgggcctgag 4740
cagctttggc atcagcgaca cccctctgga agaaatcttc ctgaaagtga ccgaggacag 4800
cgacagcggc ccactttttg ctggcggagc acagcagaaa cgcgagaacg tgaaccctag 4860
acacccctgt ctgggcccta gagagaaagc cggacagacc cctcaggaca gcaatgtgtg 4920
ctctcctggt gctcctgccg ctcaccctga aggacaacct ccacctgagc ctgagtgtcc 4980
tggacctcag ctgaataccg gcacacagct ggttctgcag cacgtgcaag ccctgctggt 5040
caagagattc cagcacacca tcagaagcca caaggacttt ctggcccaga ttgtgctgcc 5100
cgccaccttc gtttttctgg ctctgatgct gagcatcgtg atccctccat tcggcgagta 5160
ccccgctctg acactgcacc cttggatcta cggccagcag tacaccttct tcagcatgga 5220
cgagcctggc agcgagcagt ttacagtgct ggctgatgtg ctgctgaaca agcccggctt 5280
cggcaaccgc tgtctgaaag aaggatggct gcctgagtac ccctgcggca atagcacccc 5340
ttggaaaacc cctagcgtgt cccctaacat cacccagctg ttccagaaac agaaatggac 5400
ccaagtgaac ccctctccat cctgccggtg ctccaccaga gaaaagctga ccatgctgcc 5460
cgagtgtcca gaaggtgcag gcggacttcc tccacctcag agaacccaga gaagcaccga 5520
gattctgcag gacctgaccg accggaatat cagcgacttc ctggttaaga cataccccgc 5580
actgatccgg tccagcctga agtccaagtt ctgggtcaac gagcagagat acggcggcat 5640
ctctatcggc ggaaagctgc ctgtggtgcc tatcacaggc gaagccctcg tgggctttct 5700
gtctgacctg ggcagaatca tgaacgtgtc cggcggaccc atcaccagag aggccagcaa 5760
agagatcccc gatttcctga agcacctgga aaccgaggat aacatcaaag tgtggttcaa 5820
caacaaaggc tggcacgccc tggtgtcctt tctgaacgtg gcccacaacg ccatcctgag 5880
agccagcctg cctaaggaca gaagccctga ggaatacggc atcaccgtga tctcccagcc 5940
actgaacctg accaaagagc agctgagcga gatcaccgtg ctgaccacat ctgtggatgc 6000
cgtggtggcc atctgcgtga tcttctccat gagcttcgtg cctgcctcct tcgtgctgta 6060
tctgatccaa gagagagtga acaagagcaa gcacctccag tttatcagcg gagtgtcccc 6120
aaccacctac tgggtcacca actttctgtg ggacatcatg aactactccg tgtctgccgg 6180
cctggtcgtg ggcatcttca tcggctttca gaagaaggcc tacacaagcc ccgagaacct 6240
gcctgctctg gttgctctgc tgctgctgta tggctgggcc gtgattccca tgatgtaccc 6300
cgccagcttc ctgttcgatg tgcctagcac agcctacgtg gccctgagct gcgccaatct 6360
gttcatcggc atcaacagca gcgccatcac cttcatcctg gaactgttcg agaacaacag 6420
aaccctgctg cggttcaacg ccgtgctgag aaagctgctg atcgtgttcc ctcacttctg 6480
cctcggcaga ggcctgattg atctggccct gtctcaggcc gtgaccgacg tgtacgctag 6540
atttggcgag gaacacagcg ccaatccttt ccactgggac ctgatcggca agaacctgtt 6600
cgccatggtg gtggaaggcg tggtgtattt cctgctgact ctgctggtgc agcggcactt 6660
ctttctgtcc caatggatcg ccgagcctac caaagaaccc atcgtggacg aggacgacga 6720
cgtggccgaa gagagacaga gaatcatcac cggcggcaac aagaccgaca tcctgaggct 6780
gcacgagctg accaagatct accctggcac atctagccct gccgtggaca gactgtgtgt 6840
gggagttaga cctggcgagt gctttggcct gctgggagtt aatggcgccg gaaagaccac 6900
caccttcaag atgctgaccg gcgacaccac agtgaccagc ggagatgcta ctgtggccgg 6960
caagagcatc ctgaccaaca tcagcgaggt gcaccagaac atgggctact gccctcagtt 7020
cgacgccatc gacgaactgc tgacaggcag agagcacctg tatctgtacg ccagactgag 7080
aggcgtgccc gccgaagaaa ttgagaaggt ggccaactgg tccatcaaga gcctgggact 7140
gaccgtgtac gccgattgtc tggccggcac atatagcggc ggaaacaagc ggaagctgag 7200
caccgccatt gctctgatcg gatgccctcc tctggtcctg ctggatgagc ctacaaccgg 7260
catggatccc caggctagac ggatgctgtg gaacgtgatc gtgtccatca tccgcgaagg 7320
cagagccgtg gtcctgacaa gccacagcat ggaagagtgc gaggccctgt gtaccagact 7380
ggccatcatg gtcaagggcg ccttcagatg catgggcacc attcagcacc tgaaaagcaa 7440
gttcggcgac ggctacatcg tgaccatgaa gatcaagtcc ccaaaggacg acctgctgcc 7500
agatctgaac cccgtggaac agttcttcca gggcaacttc cctggctccg tgcagcggga 7560
aagacactac aacatgctgc agtttcaggt gtccagcagc tccctggcca gaatctttca 7620
gctgctgctc tcccacaagg atagcctgct gattgaagag tacagcgtga cccagaccac 7680
actggaccag gtgttcgtga acttcgccaa gcagcagacc gagagccacg acttgcctct 7740
gcaccctaga gctgccggcg cttctagaca ggcccaggat taagcggccg cgataatcaa 7800
cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 7860
acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 7920
ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga actcatcgcc 7980
gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 8040
cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga 8100
ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt 8160
gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg 8220
attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct cgagtagata 8280
agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc 8340
cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg 8400
gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ccttaattaa cctaattcac 8460
tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc 8520
ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc 8580
cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa 8640
gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 8700
ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 8760
ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 8820
aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 8880
gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 8940
cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 9000
attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 9060
cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 9120
tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 9180
ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt 9240
ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga 9300
tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa 9360
gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct 9420
gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat 9480
acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga 9540
tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc 9600
caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat 9660
gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa 9720
cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac 9780
tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa 9840
agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc 9900
tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc 9960
ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag 10020
acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta 10080
ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 10140
gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 10200
gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 10260
ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 10320
gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 10380
tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 10440
cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 10500
cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 10560
ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 10620
tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 10680
cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 10740
ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 10800
aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 10860
ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 10920
tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga 10980
gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg 11040
gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg 11100
caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct 11160
tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta 11220
tgaccatgat tacgccagat ttaattaagg 11250
<210> 17
<211> 7534
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 17
ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 60
ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120
aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct 180
aggaagatcg gaattcgccc ttaagggccc cagaagcctg gtggttgttt gtccttctca 240
ggggaaaagt gaggcggccc cttggaggaa ggggccgggc agaatgatct aatcggattc 300
caagcagctc aggggattgt ctttttctag caccttcttg ccactcctaa gcgtcctccg 360
tgaccccggc tgggatttag cctggtgctg tgtcagcccc ggtctcccag gggcttccca 420
gtggtcccca ggaaccctcg acagggcccg gtctctctcg tccagcaagg gcagggacgg 480
gccacaggcc aagggcggta ctctagagga tccggtactc gaggaactga aaaaccagaa 540
agttaactgg taagtttagt ctttttgtct tttatttcag gtcccggatc cggtggtggt 600
gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct gtacggaagt 660
gttacttctg ctctaaaagc tgcggaattg tacccgcggc cgccaccatg ggctttgtgc 720
gacagattca gctgctgctg tggaagaact ggaccctgcg gaagcggcag aaaatcagat 780
tcgtggtgga actcgtgtgg cccctgagcc tgtttctggt gctgatctgg ctgcggaacg 840
ccaatcctct gtacagccac cacgagtgtc acttccccaa caaggccatg ccttctgccg 900
gaatgctgcc ttggctgcag ggcatcttct gcaacgtgaa caacccctgc tttcagagcc 960
ccacacctgg cgaaagccct ggcatcgtgt ccaactacaa caacagcatc ctggccagag 1020
tgtaccggga cttccaagag ctgctgatga acgcccctga gtctcagcac ctgggcagaa 1080
tctggaccga gctgcacatc ctgagccagt tcatggacac cctgagaaca caccccgaga 1140
gaatcgccgg caggggcatc agaatccggg acatcctgaa ggacgaggaa accctgacac 1200
tgttcctcat caagaacatc ggcctgagcg acagcgtggt gtacctgctg atcaacagcc 1260
aagtgcggcc cgagcagttt gctcatggcg tgccagatct cgccctgaag gatatcgcct 1320
gttctgaggc cctgctggaa cggttcatca tcttcagcca gcggagaggc gccaagaccg 1380
tcagatatgc cctgtgcagt ctgagccagg gaaccctgca gtggatcgag gataccctgt 1440
acgccaacgt ggacttcttc aagctgttcc gggtgctgcc cacactgctg gattctagat 1500
cccagggcat caacctgaga agctggggcg gcatcctgtc cgacatgagc ccaagaatcc 1560
aagagttcat ccaccggcct agcatgcagg acctgctgtg ggttaccaga cctctgatgc 1620
agaacggcgg acccgagaca ttcaccaagc tgatgggaat tctgagcgat ctgctgtgcg 1680
gctaccctga aggcggagga tctagagtgc tgagcttcaa ttggtacgag gacaacaact 1740
acaaggcctt cctgggcatc gactccacca gaaaggaccc catctacagc tacgaccggc 1800
ggacaaccag cttctgcaat gccctgatcc agagcctgga aagcaaccct ctgaccaaga 1860
tcgcttggag ggccgccaaa cctctgctga tgggaaagat cctgtacacc cctgacagcc 1920
ctgccgccag aagaatcctg aagaacgcca acagcacctt cgaggaactg gaacacgtgc 1980
gcaagctggt caaggcctgg gaagaagtgg gacctcagat ctggtacttc ttcgacaata 2040
gcacccagat gaacatgatc agagacaccc tgggcaaccc taccgtgaag gacttcctga 2100
acagacagct gggcgaagag ggcattaccg ccgaggccat cctgaacttt ctgtacaagg 2160
gccccagaga gtcccaggcc gacgacatgg ccaacttcga ttggcgggac atcttcaaca 2220
tcaccgacag aaccctgcgg ctggtcaacc agtacctgga atgcctggtg ctggacaagt 2280
tcgagagcta caacgacgag acacagctga cccagagagc cctgtctctg ctggaagaga 2340
atatgttctg ggctggcgtg gtgttccccg acatgtaccc ttggacaagc agcctgcctc 2400
ctcacgtgaa gtacaagatc cggatggaca tcgacgtggt cgaaaagacc aacaagatca 2460
aggatcggta ctgggacagc ggccctagag ctgatcccgt ggaagatttt cggtacatct 2520
ggggcggatt cgcatacctg caggacatgg tggaacaggg aatcacacgg tcccaggtgc 2580
aggctgaagc tcctgtggga atctacctgc agcagatgcc ttatccttgc ttcgtggacg 2640
acagcttcat gatcatcctg aatcggtgct tccccatctt catggtgctg gcctggatct 2700
actccgtgtc tatgaccgtg aagtccatcg tgctggaaaa agagctgcgg ctgaaagaga 2760
cactgaagaa ccagggcgtg tccaatgccg tgatctggtg cacctggttt ctggacagct 2820
tctccattat gagcatgagc atctttctgc tgacgatctt catcatgcac ggccggatcc 2880
tgcactacag cgaccccttt atcctcttcc tgttcctgct ggccttcagc accgctacaa 2940
tcatgctgtg ttttctgctg tccaccttct tcagcaaggc ctctctggcc gctgcttgta 3000
gcggcgtgat ctacttcacc ctgtacctgc ctcacatcct gtgcttcgca tggcaggaca 3060
gaatgaccgc cgagctgaag aaagctgtgt ccctgctgag ccctgtggcc tttggctttg 3120
gcaccgagta cctcgtcaga tttgaggaac aaggactggg actgcagtgg tccaacatcg 3180
gcaatagccc tacagagggc gacgagttca gcttcctgct gtctatgcag atgatgctgc 3240
tggacgccgc cgtgtatgga ctgctggctt ggtatctgga ccaggtgttc ccaggcgatt 3300
acggcactcc tctgccttgg tatttcctgc tgcaagagag ctactggctc ggcggcgagg 3360
gatgtagcac cagagaagaa agagccctgg aaaagaccga gcctctgacc gaggaaacag 3420
aggaccctga acacccagag ggcatccacg atagcttttt cgagagagaa caccccggct 3480
gggtgccagg cgtgtgtgtg aagaatctgg tcaagatctt cgagccctgc ggcagacctg 3540
ccgtggacag actgaacatc accttctacg agaaccagat taccgccttt ctgggccaca 3600
acggcgctgg caagacaacc acactgagca tcctcacagg cctgctgcct ccaacaagcg 3660
gcacagttct cgttggcggc agagacatcg agacaagcct ggatgccgtc agacagtccc 3720
tgggcatgtg ccctcagcac aacatcctgt ttcaccacct gaccgtggcc gagcacatgc 3780
tgttttatgc ccagctgaag ggcaagagcc aagaagaggc tcagctggaa atggaagcca 3840
tgctcgagga caccggcctg caccacaaga gaaatgagga agcccaggat ctgagcggcg 3900
gcatgcagag aaaactgagc gtggccattg ccttcgtggg cgacgccaag gttgtgatcc 3960
tggatgagcc tacaagcggc gtggaccctt acagcagaag atccatctgg gatctgctgc 4020
tgaagtacag aagcggccgg accatcatca tgagcaccca ccacatggac gaggccgatc 4080
tgctcggaga cagaatcgcc atcattgctc agggcagact gtactgcagc ggcaccccac 4140
tgtttctgaa gaactgtctg gctggcgata ctctcattac cctggccgat ggacgacgag 4200
tgcctattag agaactggtg tcacagcaga atttttccgt gtgggctctg aatcctcaga 4260
cttaccgcct ggagagggct agagtgagta gagctttctg taccggcatc aaacctgtgt 4320
accgcctcac cactagactg gggagatcca ttagggccac tgccaaccac cgatttctca 4380
cacctcaggg ctggaaacga gtcgatgaac tccagcctgg agattacctg gctctgccta 4440
ggagaatccc tactgcctcc taaacgcgtc ctcgactgtg ccttctagtt gccagccatc 4500
tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 4560
ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 4620
gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 4680
ggatgcggtg ggctctatgg cctcgagtta agggcgaatt cccgataagg atcttcctag 4740
agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag 4800
tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa 4860
aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc 4920
ttaattaacc taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg 4980
ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctgcatt aatgaatcgg 5040
ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 5100
ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 5160
acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 5220
aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 5280
tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 5340
aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 5400
gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 5460
acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 5520
accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 5580
ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 5640
gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 5700
aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 5760
ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 5820
gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 5880
cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 5940
cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 6000
gtaaacttgg tctgacagtt agaaaaactc atcgagcatc aaatgaaact gcaatttatt 6060
catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa 6120
ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg 6180
tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa 6240
atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca 6300
gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc 6360
gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca 6420
attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt 6480
ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccag ggatcgcagt 6540
ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat 6600
aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc 6660
tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt 6720
cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat 6780
gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt 6840
cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 6900
tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 6960
acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac 7020
gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 7080
cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 7140
cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 7200
tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 7260
ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 7320
ggcctcttcg ctattacgcc agctggcacg acaggtttcc cgactggaaa gcgggcagtg 7380
agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta 7440
tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca 7500
gctatgacca tgattacgcc agatttaatt aagg 7534
<210> 18
<211> 7312
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 18
ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 60
ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120
aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct 180
aggaagatcg gaattcgccc ttaagggccc cagaagcctg gtggttgttt gtccttctca 240
ggggaaaagt gaggcggccc cttggaggaa ggggccgggc agaatgatct aatcggattc 300
caagcagctc aggggattgt ctttttctag caccttcttg ccactcctaa gcgtcctccg 360
tgaccccggc tgggatttag cctggtgctg tgtcagcccc ggtctcccag gggcttccca 420
gtggtcccca ggaaccctcg acagggcccg gtctctctcg tccagcaagg gcagggacgg 480
gccacaggcc aagggcggta ctctagagga tccggtactc gaggaactga aaaaccagaa 540
agttaactgg taagtttagt ctttttgtct tttatttcag gtcccggatc cggtggtggt 600
gcaaatcaaa gaactgctcc tcagtggatg ttgcctttac ttctaggcct gtacggaagt 660
gttacttctg ctctaaaagc tgcggaattg tacccgcggc cgccaccatg gcggcggcgt 720
gcccggaact gcgtcagctg gcgcagagcg atgtgtattg ggatccgatt gtgagcattg 780
aaccggatgg cgtggaagaa gtgtttgatc tgaccgtgcc gggcccgcat aactttgtgg 840
cgaacgatat tattgcgcat aactgttttg gcaccggcct gtacctgaca ctcgtgcgga 900
agatgaagaa catccagagc cagcggaagg gcagcgaggg cacatgtagc tgtagcagca 960
agggcttcag caccacctgt cctgctcacg tggacgatct gacccctgaa caggtgctgg 1020
atggcgacgt gaacgagctg atggacgtgg tgctgcacca cgtgcctgag gccaagctgg 1080
tggaatgcat cggccaagag ctgatcttcc tgctgcctaa caagaacttc aagcaccggg 1140
cctacgccag cctgttcaga gagctggaag agacactggc cgatctgggc ctgagcagct 1200
ttggcatcag cgacacccct ctggaagaaa tcttcctgaa agtgaccgag gacagcgaca 1260
gcggcccact ttttgctggc ggagcacagc agaaacgcga gaacgtgaac cctagacacc 1320
cctgtctggg ccctagagag aaagccggac agacccctca ggacagcaat gtgtgctctc 1380
ctggtgctcc tgccgctcac cctgaaggac aacctccacc tgagcctgag tgtcctggac 1440
ctcagctgaa taccggcaca cagctggttc tgcagcacgt gcaagccctg ctggtcaaga 1500
gattccagca caccatcaga agccacaagg actttctggc ccagattgtg ctgcccgcca 1560
ccttcgtttt tctggctctg atgctgagca tcgtgatccc tccattcggc gagtaccccg 1620
ctctgacact gcacccttgg atctacggcc agcagtacac cttcttcagc atggacgagc 1680
ctggcagcga gcagtttaca gtgctggctg atgtgctgct gaacaagccc ggcttcggca 1740
accgctgtct gaaagaagga tggctgcctg agtacccctg cggcaatagc accccttgga 1800
aaacccctag cgtgtcccct aacatcaccc agctgttcca gaaacagaaa tggacccaag 1860
tgaacccctc tccatcctgc cggtgctcca ccagagaaaa gctgaccatg ctgcccgagt 1920
gtccagaagg tgcaggcgga cttcctccac ctcagagaac ccagagaagc accgagattc 1980
tgcaggacct gaccgaccgg aatatcagcg acttcctggt taagacatac cccgcactga 2040
tccggtccag cctgaagtcc aagttctggg tcaacgagca gagatacggc ggcatctcta 2100
tcggcggaaa gctgcctgtg gtgcctatca caggcgaagc cctcgtgggc tttctgtctg 2160
acctgggcag aatcatgaac gtgtccggcg gacccatcac cagagaggcc agcaaagaga 2220
tccccgattt cctgaagcac ctggaaaccg aggataacat caaagtgtgg ttcaacaaca 2280
aaggctggca cgccctggtg tcctttctga acgtggccca caacgccatc ctgagagcca 2340
gcctgcctaa ggacagaagc cctgaggaat acggcatcac cgtgatctcc cagccactga 2400
acctgaccaa agagcagctg agcgagatca ccgtgctgac cacatctgtg gatgccgtgg 2460
tggccatctg cgtgatcttc tccatgagct tcgtgcctgc ctccttcgtg ctgtatctga 2520
tccaagagag agtgaacaag agcaagcacc tccagtttat cagcggagtg tccccaacca 2580
cctactgggt caccaacttt ctgtgggaca tcatgaacta ctccgtgtct gccggcctgg 2640
tcgtgggcat cttcatcggc tttcagaaga aggcctacac aagccccgag aacctgcctg 2700
ctctggttgc tctgctgctg ctgtatggct gggccgtgat tcccatgatg taccccgcca 2760
gcttcctgtt cgatgtgcct agcacagcct acgtggccct gagctgcgcc aatctgttca 2820
tcggcatcaa cagcagcgcc atcaccttca tcctggaact gttcgagaac aacagaaccc 2880
tgctgcggtt caacgccgtg ctgagaaagc tgctgatcgt gttccctcac ttctgcctcg 2940
gcagaggcct gattgatctg gccctgtctc aggccgtgac cgacgtgtac gctagatttg 3000
gcgaggaaca cagcgccaat cctttccact gggacctgat cggcaagaac ctgttcgcca 3060
tggtggtgga aggcgtggtg tatttcctgc tgactctgct ggtgcagcgg cacttctttc 3120
tgtcccaatg gatcgccgag cctaccaaag aacccatcgt ggacgaggac gacgacgtgg 3180
ccgaagagag acagagaatc atcaccggcg gcaacaagac cgacatcctg aggctgcacg 3240
agctgaccaa gatctaccct ggcacatcta gccctgccgt ggacagactg tgtgtgggag 3300
ttagacctgg cgagtgcttt ggcctgctgg gagttaatgg cgccggaaag accaccacct 3360
tcaagatgct gaccggcgac accacagtga ccagcggaga tgctactgtg gccggcaaga 3420
gcatcctgac caacatcagc gaggtgcacc agaacatggg ctactgccct cagttcgacg 3480
ccatcgacga actgctgaca ggcagagagc acctgtatct gtacgccaga ctgagaggcg 3540
tgcccgccga agaaattgag aaggtggcca actggtccat caagagcctg ggactgaccg 3600
tgtacgccga ttgtctggcc ggcacatata gcggcggaaa caagcggaag ctgagcaccg 3660
ccattgctct gatcggatgc cctcctctgg tcctgctgga tgagcctaca accggcatgg 3720
atccccaggc tagacggatg ctgtggaacg tgatcgtgtc catcatccgc gaaggcagag 3780
ccgtggtcct gacaagccac agcatggaag agtgcgaggc cctgtgtacc agactggcca 3840
tcatggtcaa gggcgccttc agatgcatgg gcaccattca gcacctgaaa agcaagttcg 3900
gcgacggcta catcgtgacc atgaagatca agtccccaaa ggacgacctg ctgccagatc 3960
tgaaccccgt ggaacagttc ttccagggca acttccctgg ctccgtgcag cgggaaagac 4020
actacaacat gctgcagttt caggtgtcca gcagctccct ggccagaatc tttcagctgc 4080
tgctctccca caaggatagc ctgctgattg aagagtacag cgtgacccag accacactgg 4140
accaggtgtt cgtgaacttc gccaagcagc agaccgagag ccacgacttg cctctgcacc 4200
ctagagctgc cggcgcttct agacaggccc aggattaata aacgcgtcct cgactgtgcc 4260
ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 4320
tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 4380
gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 4440
caatagcagg catgctgggg atgcggtggg ctctatggcc tcgagttaag ggcgaattcc 4500
cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta 4560
actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca 4620
ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga 4680
gcgagcgagc gcgcagcctt aattaaccta attcactggc cgtcgtttta caacgtcgtg 4740
actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca 4800
gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 4860
cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 4920
tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 4980
gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 5040
ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 5100
aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 5160
tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 5220
ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 5280
gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 5340
tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 5400
caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 5460
ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt 5520
cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 5580
ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 5640
cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 5700
gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 5760
aatctaaagt atatatgagt aaacttggtc tgacagttag aaaaactcat cgagcatcaa 5820
atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt 5880
ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg 5940
gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat 6000
aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag 6060
tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc 6120
actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg 6180
atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc 6240
cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt 6300
tttcccaggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt 6360
gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac 6420
atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc 6480
atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc 6540
atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg 6600
aatatggctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 6660
catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 6720
atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 6780
taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 6840
cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 6900
cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 6960
tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 7020
atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 7080
ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcacgac aggtttcccg 7140
actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac 7200
cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac 7260
aatttcacac aggaaacagc tatgaccatg attacgccag atttaattaa gg 7312

Claims (17)

1.ABCA4基因片段，其特征在于，所述的基因片段编码ABCA4全长蛋白以p1150Cys为***位点断裂得到的任意一个片段。
2.根据权利要求1所述的ABCA4基因片段，其特征在于所述的ABCA4基因片段的一端含有内含肽片段；所述的内含肽选择SEQ ID NO.3所示的Rma intein或SEQ ID NO.4SEQ IDNO.4所示的Npu intein。
3.根据权利要求2所述的ABCA4基因片段，其特征在于所述的ABCA4基因片段选自以下任意一个：
(1)如SEQ ID NO.8所示的含内含肽片段的人ABCA4-N蛋白片段的编码基因，
(2)如SEQ ID NO.9所示的含内含肽片段的人ABCA4-C蛋白片段的编码基因。
4.一种ABCA4-N或ABCA4-C端蛋白表达框，由增强子-启动子-内含子-目的基因序列-polyA信号组成，其特征在于，所述的增强子选自人214bp interphotoreceptor retinoid-binding protein enhancer增强子序列，启动子选自视网膜感光细胞特异的人RS1启动子，人Rho启动子、人RK启动子或小鼠CAR启动子；内含子选自SV40内含子；目的基因选自含内含肽片段的人ABCA4-N或ABCA4-C蛋白片段的编码基因；polyA信号序列选自SV40 PolyA，bGHpolyA，hGH polyA或rBG polyA序列；所述的增强子、启动子、内含子、目的基因序列和polyA之间通过键或核苷酸连接序列连接；所述的人ABCA4-N和ABCA4-C蛋白片段为ABCA4全长蛋白以p1150Cys为***位点断裂所得。
5.根据权利要求4所述的ABCA4-N或ABCA4-C端蛋白表达框，其特征在于，所述的内含肽选择SEQ ID NO.3所示的Rma intein或SEQ ID NO.4所示的Npu intein；优选SEQ ID NO.3所示的Rma intein。
6.根据权利要求5所述的ABCA4-N或ABCA4-C端蛋白表达框，其特征在于，所述的含内含肽片段的人ABCA4-N端氨基酸序列如SEQ ID NO.6所示，所述的含内含肽片段的人ABCA4-C端氨基酸序列如SEQ ID NO.7所示。
7.根据权利要求6所述的ABCA4-N或ABCA4-C端蛋白表达框，其特征在于，所述的含内含肽片段的人ABCA4-N蛋白片段的编码基因,其核苷酸序列如SEQ ID NO.8所示；含内含肽片段的人ABCA4-C蛋白片段的编码基因,其核苷酸序列如SEQ ID NO.9所示。
8.根据权利要求1所述的ABCA4-N或ABCA4-C端蛋白表达框，其特征在于，所述的启动子选自RK启动子，序列如SEQ ID NO.10所示；内含子序列选自SV40内含子，序列如GenBank登录号MK225672.1公开的核苷酸序列的第4863bp-4959bp所示；polyA选自bGH ployA，序列如GenBank登录号MT267334.1公开的核苷酸序列的第957bp-1181bp所示。
9.根据权利要求4-8中任一项所述的ABCA4-N或ABCA4-C端蛋白表达框，其特征在于，所述的ABCA4-N端蛋白表达框的核苷酸序列如SEQ ID NO.11；所述的ABCA4-C端蛋白表达框的核苷酸序列如SEQ ID NO.12。
10.一种载体，其特征在于含有权利要求1-3中任一项所述的ABCA4基因片段、或权利要求4-8中任一项所述的ABCA4-N或ABCA4-C端蛋白表达框。
11.根据权利要求10所述的载体，其特征在于所述载体选自以下任意一种重组腺相关病毒载体血清型选自：AAV1、AAV2、AAV3B、AAV4、AAV5、AAV6、AAV7、AAV8、AAV9、AAVrh10、AAV-LK03或AAVAnc80d，优选AAV2、AAV5、AAV8或AAV9。
12.一种质粒组合物，其特征在于包含权利要求10所述的含有所述的ABCA4-N端蛋白表达框的载体和含有所述的ABCA4-C端蛋白表达框的载体；优选包含权利要求10或11所述的载体、AAV的Rep及Cap蛋白表达质粒、辅助质粒；进一步优选包含权利要求10或11所述的载体、AAV的Rep及Cap蛋白表达质粒pAAV2/8以及辅助质粒pAdΔF6。
13.一种重组腺相关病毒，其特征在于由权利要求10或11所述的载体、AAV的Rep及Cap蛋白表达质粒以及辅助质粒共转染HEK293细胞包装制备得到。
14.根据权利要求13所述的重组腺相关病毒，其特征在于由权利要求10或11所述的含有ABCA4-N蛋白表达框的载体、AAV的Rep及Cap蛋白表达质粒pAAV2/8以及辅助质粒pAdΔF6共转染HEK293细胞包装制备得到。
15.根据权利要求13所述的重组腺相关病毒，其特征在于由权利要求10或11所述的含有ABCA4-C蛋白表达框的载体、AAV的Rep及Cap蛋白表达质粒、辅助质粒共转染HEK293细胞包装制备得到。
16.一种重组腺相关病毒组合物，其特征在于包含权利要求14和权利要求15所述的重组腺相关病毒。
17.权利要求1-3中任一项所述的ABCA4基因片段、权利要求4-8中任一项所述的ABCA4-N或ABCA4-C端蛋白表达框、权利要求10或11所述的载体、权利要求12所述的质粒组合物、权利要求13-15中任一项所述的重组腺相关病毒、权利要求16所述的重组腺相关病毒组合物在制备治疗STGDI疾病的药物中的应用。
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