CN114686438A - Ace2人源化猪的构建方法及应用 - Google Patents
Ace2人源化猪的构建方法及应用 Download PDFInfo
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Abstract
本发明提供了一种表达人ACE2的猪细胞或ACE2人源化猪及其构建方法和应用。其中包括将人ACE2基因插入猪安全港位点，获得表达SEQ ID NO：12所示人ACE2的猪细胞，所述的猪安全港位点选自猪ROSA26、AAVS1、H11或COL1A1安全港位点。进一步用所得到的表达人ACE2的猪细胞作为核移植细胞供体，得到了表达人ACE2的克隆猪。所得到的ACE2人源化猪可用于进行治疗ACE2所介导疾病的药物筛选、药效评价、疫苗效果测试以及病毒感染机制等研究。
Description
技术领域
本发明涉及基因编辑技术领域，具体涉及一种采用CRISPR/Cas9系统构建的表达人ACE2的猪细胞或ACE2人源化猪及其在生物医药领域中的应用。
背景技术
人类血管紧张素I转化酶2(hACE2)为人类血管紧张素转换酶同源物，是一种锌金属蛋白酶，属于1型跨膜蛋白，其作为SARS-2-CoV和SARS-CoV等冠状病毒在人体内的受体蛋白，广泛存在于人体各个组织中，是目前已知导致SARS-2-CoV进入宿主细胞并造成宿主发病的唯一关键蛋白。
目前，尚未有针对SARS-2-CoV的有效药物，世界各国都在争分夺秒地开展SARS-2-CoV疫苗的研发。而动物感染模型对于阐明SARS-2-CoV的感染与发病机制、传播途径以及抗病毒药物和疫苗的评价至关重要。目前，已建立了ACE2人源化的转基因小鼠模型和恒河猴模型，例如：专利文献 CN111979273A公开了一种制备人源化ACE2小鼠模型的方法，包括针对小鼠ACE2基因组序列的 exon2与exon15的序列分别设计上下游两条sgRNA引物序列；针对小鼠ACE2 exon2与人类ACE2编码区域5’端同源序列以及人类ACE2编码区域3’端与小鼠ACE2 exon15同源序列设计上下游两条 ssODN序列。专利文献CN111621523A公开了一种ACE2细胞人源化的小鼠模型，该小鼠模型以免疫缺陷小鼠为母体，且体内含有过表达ACE2受体的人源细胞。专利文献CN111549064A公开了应用腺病毒转导制备能够表达人源ACE2转基因非人动物的方法，其中该腺病毒由pShuttle-hACE2与 pAdEasy-1重组得到的重组载体在AD293细胞中拯救得到，所述pShuttle-hACE2为插入有所述人源 ACE2的DNA序列的pShuttle；所述腺病毒在转导所述动物后，所述动物的呼吸道细胞表达人源 ACE2蛋白。但是小鼠不论从体型、器官大小、生理、病理等方面都与人相差巨大，不能真实地模拟人类正常的生理、病理状态。虽然恒河猴是与人亲缘关系最近的动物，但其体型小、性成熟晚(6-7岁开始交配)，且为单胎动物，群体扩繁速度极慢，饲养成本很高。另外，灵长类动物克隆效率低、难度大、成本高。而猪作为大动物，是人类长期以来主要的肉食供应动物，其体型大小和生理功能与人类近似，易于大规模繁殖饲养，而且在伦理道德及动物保护等方面要求较低，是理想的人类疾病模型动物。
同源重组(HDR)是通过序列同源性交换DNA序列信息：即外源供体DNA中包含所需插入片段即目标片段，供体DNA的两端则是与插入位点两侧具有序列同源性的重组臂。通过细胞的同源重组，可将外源供体DNA中的目标片段插入细胞的基因组中。
基因编辑是近年来不断取得重大发展的一种生物技术，其包括从基于同源重组的基因编辑到基于核酸酶的ZFN、TALEN、CRISPR/Cas9等编辑技术，其中CRISPR/Cas9技术是当前最先进的基因编辑技术。目前，基因编辑技术被越来越多地应用到动物模型的制作上。
本申请采用基因编辑及同源重组技术构建了表达人ACE2的猪细胞，并以该细胞为核移植供体细胞生产出了ACE2人源化克隆猪。
发明内容
本发明采用CRISPR/Cas9基因编辑及同源重组技术，将人ACE2基因(hACE2)定点插入猪安全港位点，制备出人源化的hACE2转基因猪重组细胞，为进一步的通过体细胞克隆技术生产人源化 hACE2转基因克隆猪奠定坚实基础，进而为SARS-2-CoV和SARS-CoV等冠状病毒的致病机制及疾病治疗等研究提供有力的实验工具。
本发明的第一方面，提供了一种表达人ACE2的猪细胞，将人ACE2基因插入猪安全港位点，获得表达SEQ ID NO：12所示人ACE2的猪细胞，所述的猪安全港位点选自猪ROSA26、AAVS1、H11 或COL1A1安全港位点。
优选的，所述插入的人ACE2基因的核苷酸序列为编码SEQ ID NO：12的核苷酸序列。
优选的，所述插入的人ACE2基因的核苷酸序列如SEQ ID NO：13所示。
优选的，ROSA26安全港位点区域及其上下游各500bp的核苷酸序列如SEQ ID NO：14所示， AAVS1安全港位点区域及其上下游各500bp的核苷酸序列如SEQ ID NO：15所示，H11安全港位点区域及其上下游各500bp的核苷酸序列如SEQ ID NO：16所示，COL1A1安全港位点区域及其上下游各500bp的核苷酸序列如SEQ ID NO：17所示。
优选的，所述的猪细胞为猪成纤维细胞。所述的猪细胞还可以选自胚胎干细胞、成体干细胞、造血干细胞、骨髓间充质干细胞、神经干细胞、肝干细胞、肌肉卫星细胞、皮肤表皮干细胞、肠上皮干细胞、视网膜干细胞、胰腺干细胞、体细胞、成纤维细胞、肌细胞、胶质细胞、脂肪细胞或生殖细胞等等。
优选的，所述猪细胞不能发育为动物个体。
本发明的第二方面，提供了一种上述猪细胞的构建方法，使用安全港位点载体将人ACE2基因插入猪安全港位点，所述的安全港位点载体包含人ACE2基因的核苷酸序列和安全港位点载体骨架，所述的安全港位点载体骨架包含安全港插入位点的5’同源臂和3’同源臂，所述人ACE2 基因的核苷酸序列位于5’同源臂与3’同源臂之间，所述的安全港位点载体骨架选自下列任一项所示：
A)ROSA26安全港位点载体骨架，其5’同源臂如SEQ ID NO：18所示，3’同源臂如SEQID NO：19所示；
B)AAVS1安全港位点载体骨架，其5’同源臂如SEQ ID NO：5所示，3’同源臂如SEQID NO：6所示；
C)H11安全港位点载体骨架，其5’同源臂如SEQ ID NO：7所示，3’同源臂如SEQ IDNO：8所示；
或D)COL1A1安全港位点载体骨架，其5’同源臂如SEQ ID NO：9所示，3’同源臂如SEQ ID NO：10所示；
优选的，所述的安全港位点载体还包含启动子、信号分子以及编码EGFP蛋白、mCherry蛋白和puro抗性蛋白的核苷酸序列。其中，所述的启动子为EF-1α启动子、PGK启动子和/或pCAG 启动子。所述的信号分子为EF-1αpoly(A)信号、bGH poly(A)信号和/或β-globin poly(A)信号。进一步优选还包含绝缘子区域。
在本发明的一个具体实施方式中，所述的安全港位点载体骨架从5’至3’依次包括5’同源臂、绝缘子区域、EF-1αpoly(A)信号、编码EGFP的核苷酸序列、EF-1α启动子、绝缘子区域、 PGK启动子、编码mCherry的核苷酸序列、bGH poly(A)信号、loxP-puro-loxP表达框区域、绝缘子区域、β-globin poly(A)信号、pCAG启动子、绝缘子区域、3’同源臂。
在本发明的一个具体实施方式中，所述的ROSA26安全港位点载体的核苷酸序列如SEQ ID NO：4所示。
在本发明的一个具体实施方式中，所述的AAVS1安全港位点载体的核苷酸序列如SEQ ID NO：20所示。
在本发明的一个具体实施方式中，所述的H11安全港位点载体的核苷酸序列如SEQID NO： 21所示。
在本发明的一个具体实施方式中，所述的COL1A1安全港位点载体的核苷酸序列如SEQ ID NO：22所示。
优选的，使用sgRNA载体进行猪细胞的构建，所述的sgRNA载体包含靶向ROSA26、AAVS1、 H11或COL1A1安全港位点的sgRNA，其中：
靶向ROSA26的sgRNA的核苷酸序列如SEQ ID NO：28所示，靶向AAVS1的sgRNA的核苷酸序列如SEQ ID NO：29所示，靶向H11的sgRNA的核苷酸序列如SEQ ID NO：30所示，靶向COL1A1的sgRNA的核苷酸序列如SEQ ID NO：31所示。
进一步优选的，所述的sgRNA载体还包含骨架载体，所述的骨架载体的核苷酸序列为SEQ ID NO：3。
优选的，所述的构建方法包括将安全港位点载体、sgRNA载体和Cas载体共转染至猪细胞，所述的Cas载体包含编码Cas蛋白、EGFP和Puro抗性蛋白的核苷酸序列，所述的Cas蛋白选自 Casl、CaslB、Cas2、Cas3、Cas4、Cas5、Cas5d、Cas5t、Cas5h、Cas5a、Cas6、Cas7、Cas8、 Cas9、CaslO、Csyl、Csy2、Csy3、Csy4、Csel、Cse2、Cse3、Cse4、Cse5e、Cscl、Csc2、Csa5、 Csnl、Csn2、Csml、Csm2、Csm3、Csm4、Csm5、Csm6、Cmrl、Cmr3、Cmr4、Cmr5、Cmr6、 Csbl、Csb2、Csb3、Csx17、Csx14、CsxlO、Csx16、CsaX、Csx3、Csxl、CsxlS、Csfl、Csf2、 CsO、Csf4、Csdl、Csd2、Cstl、Cst2、Cshl、Csh2、Csal、Csa2、Csa3、Csa4、Csa5、C2cl、 C2c2、C2c3、Cpfl、CARF、DinG、其同源物或其修饰形式，优选为Cas9(Cas9表达载体)，优选的，所述的Cas载体的核苷酸序列如SEQ ID NO：1或2所示。
为了增加Cas9质粒的基因编辑能力，本发明在购自addgene(Plasmid#42230，fromZhang Feng lab)pX330-U6-Chimeric_BB-CBh-hSpCas9(简称PX330)载体的基础上进行改造得到pU6gRNA- eEF1a-mNLS-hSpCas9-EGFP-PURO(简称粒pKG-GE3)。PX330的图谱如图1，改造方式如下：
1)去除原载体gRNA骨架中多余无效的序列；
2)改造启动子：将原有启动子(chickenβ-actin启动子)改造为具更高表达活性的EF1a启动子，增加Cas9基因的蛋白表达能力；
3)增加核定位信号：在Cas9的N端及C端均增加核定位信号编码序列(NLS)，增加Cas9的核定位能力；
4)增加双筛选标记：原载体无任何筛选标记，不利于阳性转化细胞的筛选和富集，在Cas9的C 端，插入P2A-EGFP-T2A-PURO，赋予载体荧光和抗性筛选能力；
5)插入WPRE和3’LTR等调控基因表达的序列：在基因读码框最后插入WPRE、3’LTR等序列，可增强Cas9基因的蛋白翻译能力。
改造后载体pU6gRNA-eEF1a-mNLS-hSpCas9-EGFP-PURO(简称pKG-GE3)及改造位点如图2，质粒全序列如SEQ ID NO：2所示；pKG-GE3的主要元件有：
1)gRNA表达元件：U6 gRNA scaffold；
2)启动子：EF1a启动子和CMV增强子；
3)含多个NLS的Cas9基因：含N端和C端多核定位信号(NLS)的Cas9基因；
4)筛选标记基因：荧光和抗性双筛选标记元件P2A-EGFP-T2A-PURO；
5)增强翻译的元件：WPRE和3’LTR增强Cas9及筛选标记基因的翻译效率；
6)转录终止信号：bGHpolyA signal；
7)载体骨架：包括Amp抗性元件和ori复制子等。
质粒pKG-GE3中，具有特异融合基因；所述特异融合基因编码特异融合蛋白；
所述特异融合蛋白自N端至C端依次包括如下元件：两个核定位信号(NLS)、Cas9蛋白、两个核定位信号、自剪切多肽P2A、荧光报告蛋白、自裂解多肽T2A、抗性筛选标记蛋白；
质粒pKG-GE3中，由EF1a启动子启动所述特异融合基因的表达；
质粒pKG-GE3中，所述特异融合基因下游具有WPRE序列元件、3’LTR序列元件和bGHpoly(A) signal序列元件。
质粒pKG-GE3中，依次具有如下元件：CMV增强子、EF1a启动子、所述特异融合基因、WPRE序列元件、3’LTR序列元件、bGH poly(A)signal序列元件。
所述特异融合蛋白中，Cas9蛋白上游的两个核定位信号为SV40核定位信号，Cas9蛋白下游的两个核定位信号为nucleoplasmin核定位信号。
所述特异融合蛋白中，荧光报告蛋白具体可为EGFP蛋白。
所述特异融合蛋白中，抗性筛选标记蛋白具体可为Puromycin蛋白。
自剪切多肽P2A的氨基酸序列为“ATNFSLLKQAGDVEENPGP”(发生自剪切的断裂位置为C端开始第一个氨基酸残基和第二个氨基酸残基之间)。
自裂解多肽T2A的氨基酸序列为“EGRGSLLTCGDVEENPGP”(发生自裂解的断裂位置为C端开始第一个氨基酸残基和第二个氨基酸残基之间)。
特异融合基因具体如SEQ ID NO：2中第911-6706位核苷酸所示。
CMV增强子如SEQ ID NO：2中第395-680位核苷酸所示。
EF1a启动子如SEQ ID NO：2中第682-890位核苷酸所示。
WPRE序列元件如SEQ ID NO：2第6722-7310位核苷酸所示。
3’LTR序列元件如SEQ ID NO：2中第7382-7615位核苷酸所示。
bGH poly(A)signal序列元件如SEQ ID NO：2中第7647-7871位核苷酸所示。
优选的，所述的安全港位点载体、sgRNA载体或Cas载体均为环状质粒。
本发明的第三方面，提供了一种表达人ACE2的人源化猪的构建方法，将人ACE2基因插入猪安全港位点，获得表达SEQ ID NO：12所示人ACE2的猪，所述的猪安全港位点选自猪ROSA26、 AAVS1、H11或COL1A1安全港位点。
优选的，使用安全港位点载体将人ACE2基因插入猪安全港位点，所述的安全港位点载体包含人ACE2基因的核苷酸序列和安全港位点载体骨架，所述的安全港位点载体骨架包含安全港插入位点的5’同源臂和3’同源臂，所述人ACE2基因的核苷酸序列位于5’同源臂与3’同源臂之间，所述的安全港位点载体骨架选自下列任一项所示：
A)ROSA26安全港位点载体骨架，其5’同源臂如SEQ ID NO：18所示，3’同源臂如SEQID NO：19所示；优选的，所述的ROSA26安全港位点载体的核苷酸序列如SEQ ID NO：4所示；
B)AAVS1安全港位点载体骨架，其5’同源臂如SEQ ID NO：5所示，3’同源臂如SEQID NO：6所示；优选的，所述的AAVS1安全港位点载体的核苷酸序列如SEQ ID NO：20所示；
C)H11安全港位点载体骨架，其5’同源臂如SEQ ID NO：7所示，3’同源臂如SEQ IDNO：8所示；优选的，所述的H11安全港位点载体的核苷酸序列如SEQ ID NO：21所示；
或D)COL1A1安全港位点载体骨架，其5’同源臂如SEQ ID NO：9所示，3’同源臂如SEQ ID NO：10所示；优选的，所述的COL1A1安全港位点载体的核苷酸序列如SEQ ID NO：22所示。
优选的，使用sgRNA载体进行猪细胞的构建，所述的sgRNA载体包含靶向ROSA26、AAVS1、 H11或COL1A1安全港位点的sgRNA，其中：
靶向ROSA26的sgRNA的核苷酸序列如SEQ ID NO：28所示，靶向AAVS1的sgRNA的核苷酸序列如SEQ ID NO：29所示，靶向H11的sgRNA的核苷酸序列如SEQ ID NO：30所示，靶向COL1A1的sgRNA的核苷酸序列如SEQ ID NO：31所示。
进一步优选的，所述的sgRNA载体还包含骨架载体，所述的骨架载体的核苷酸序列为SEQ ID NO：3。
在本发明的一个具体实施方式中，所述的构建方法包括将安全港位点载体、sgRNA载体和 Cas载体共转染至猪细胞。
优选的，所述的Cas载体包含编码Cas蛋白、EGFP和Puro抗性蛋白的核苷酸序列，所述的 Cas蛋白选自Casl、CaslB、Cas2、Cas3、Cas4、Cas5、Cas5d、Cas5t、Cas5h、Cas5a、Cas6、Cas7、Cas8、Cas9、CaslO、Csyl、Csy2、Csy3、Csy4、Csel、Cse2、Cse3、Cse4、Cse5e、Cscl、Csc2、Csa5、Csnl、Csn2、Csml、Csm2、Csm3、Csm4、Csm5、Csm6、Cmrl、Cmr3、Cmr4、 Cmr5、Cmr6、Csbl、Csb2、Csb3、Csx17、Csx14、CsxlO、Csx16、CsaX、Csx3、Csxl、CsxlS、Csfl、Csf2、CsO、Csf4、Csdl、Csd2、Cstl、Cst2、Cshl、Csh2、Csal、Csa2、Csa3、Csa4、 Csa5、C2cl、C2c2、C2c3、Cpfl、CARF、DinG、其同源物或其修饰形式，进一步优选为Cas9。
优选的，所述的Cas载体的核苷酸序列如SEQ ID NO：1或2所示。
优选的，所述的安全港位点载体、sgRNA载体或Cas载体均为环状质粒。
在本发明的一个具体实施方式中，将制备的猪细胞进行体细胞核移植动物克隆，获得 hACE2基因纯合敲入的克隆猪。
本发明的第四方面，提供一种上述表达人ACE2的猪细胞在构建ACE2人源化猪中的用途。
本发明的第五方面，提供了一种ACE2人源化猪的构建方法，所述构建方法包括：
A、卵母细胞体外成熟；B、将上述获得的任一表达人ACE2的猪细胞进行体细胞核移植 (SCNT)构建重构胚；C、胚胎移植。
优选的，所述步骤A包括：(1)获得卵丘卵母细胞复合体(Cumulus-oocytecomplexes,COCs)；(2)将COCs在体外成熟培养基中进行培养。
更优选的，所述体外成熟培养基包括以TCM-199培养基为基础添加生长因子、猪卵泡液、抗生素及促卵泡成熟的激素等。
进一步优选的，所述体外成熟培养基包括以TCM-199培养基为基础添加0.1mg/mL丙酮酸、0.1mg/mL盐酸半胱氨酸、10ng/mL表皮生长因子、10％(v/v)猪卵泡液、75mg/mL青霉素， 50mg/mL链霉素，10IU/mL eCG和hCG。
优选的，所述步骤B包括：(1)去除卵母细胞周围扩张的卵丘细胞；(2)去除卵母细胞的核和极体；(3)将上述获得的任一表达人ACE2的猪细胞作为核供体，注入已去核卵母细胞的卵周隙；(4)将核供体细胞与受体卵母细胞融合获得重构胚，并培养使其进行细胞核重编程；(5) 激活重构胚。
更优选的，所述步骤(4)中重构胚在PZM-3培养基中培养2h进行细胞核重编程。
更优选的，所述步骤(5)先后采用电激活及化学激活的方法进行重构胚的激活。
优选的，所述步骤C包括将步骤B获得的重构胚移植到受体母猪输卵管中，每头母猪移植 300-350个重构胚。
本发明的第六方面，提供了上述任一构建方法获得的表达人ACE2的人源化猪。
本发明的第七方面，提供了一种安全港位点载体，所述的安全港位点载体包含人ACE2基因的核苷酸序列和安全港位点载体骨架，所述的人ACE2基因的核苷酸序列如SEQ IDNO：13所示，所述的安全港位点载体骨架包含安全港插入位点的5’同源臂和3’同源臂，所述人ACE2基因的核苷酸序列位于5’同源臂与3’同源臂之间，所述的安全港位点载体骨架选自下列任一项所示：
A)ROSA26安全港位点载体骨架，其5’同源臂如SEQ ID NO：18所示，3’同源臂如SEQID NO：19所示；优选的，所述的ROSA26安全港位点载体的核苷酸序列如SEQ ID NO：4所示；
B)AAVS1安全港位点载体骨架，其5’同源臂如SEQ ID NO：5所示，3’同源臂如SEQID NO：6所示；优选的，所述的AAVS1安全港位点载体的核苷酸序列如SEQ ID NO：20所示；
C)H11安全港位点载体骨架，其5’同源臂如SEQ ID NO：7所示，3’同源臂如SEQ IDNO：8所示；优选的，所述的H11安全港位点载体的核苷酸序列如SEQ ID NO：21所示；
或D)COL1A1安全港位点载体骨架，其5’同源臂如SEQ ID NO：9所示，3’同源臂如SEQ ID NO：10所示；优选的，所述的COL1A1安全港位点载体的核苷酸序列如SEQ ID NO：22所示。
本发明的第八方面，提供了一种上述的安全港位点载体、上述的sgRNA或者上述的sgRNA 载体在制备表达人ACE2的猪或猪细胞中的应用。
本发明的第九方面，提供了一种上述的ACE2人源化猪的猪器官、猪组织或猪细胞，即 ACE2所介导疾病的器官模型、组织模型或细胞模型。
优选的，所述猪器官、猪组织或猪细胞不能发育为动物个体。
本发明的第十方面，提供了一种上述的表达人ACE2的猪细胞、上述构建方法获得的ACE2 人源化猪的猪器官、猪组织、猪细胞，或者用上述构建方法获得的人源化猪的应用，所述应用包括
(1)筛选治疗ACE2所介导疾病的药物；
(2)进行ACE2所介导疾病药物的药效评价；
(3)进行ACE2所介导疾病的疫苗效果测试；或，
(4)进行ACE2所介导的病毒感染机制研究。
优选的，所述药物包括化学药，例如化合物、组合物，生物药，例如抗体、基因或者细胞治疗药物等等。
优选的，所述动物模型是动物疾病模型，更优选的，所述疾病是ACE2所介导疾病。
进一步优选的，所述ACE2所介导疾病包括在呼吸系统、心血管系统、泌尿系统、消化系统、生殖系统、神经系统、免疫系统等ACE2介导的疾病，例如冠状病毒感染、高血压、动脉粥样硬化、心肌梗塞、心力衰竭、肾炎、任何原因导致的肾脏损伤、肾纤维化、胰腺炎、糖尿病、肝炎、胆道纤维化、生殖系统发育、睾丸炎、脑损伤、阿尔茨海默症等等。
优选的，所述的冠状病毒选自SARS-CoV-2或SARS-CoV。
术语“载体”是细胞内能够在自身控制下复制的多核苷酸，或者通过插入到宿主细胞染色体进行复制和/或表达的遗传元件，例如质粒、染色体、病毒、转座子。合适的载体包括但不限于质粒、转座子、细菌噬菌体和粘粒。
本发明所述的“gRNA”，也称指导RNA，是由sgRNA载体在细胞中转录得到的，对细胞中的靶序列具有特异性并且可与Cas蛋白形成复合体的RNA。
与现有技术相比，本发明至少具有如下有益效果：
(1)本发明研究对象(猪)比其他动物(大小鼠、灵长类)具有更好的应用性。
大小鼠等啮齿类动物不论从体型、器官大小、生理、病理等方面都与人相差巨大，无法真实地模拟人类正常的生理、病理状态。研究表明，95％以上在大小鼠中验证有效的药物在人类临床试验中是无效的。就大动物而言，灵长类是与人亲缘关系最近的动物，但其体型小、性成熟晚(6-7岁开始交配)，且为单胎动物，群体扩繁速度极慢，饲养成本很高。另外，灵长类动物克隆效率低、难度大、成本高。
而猪作为模型动物就没有上述缺点，猪是除灵长类外与人亲缘关系最近的动物，其体型、体重、器官大小等与人相近，在解剖学、生理学、免疫学、营养代谢、疾病发病机制等方面与人类极为相似。同时，猪的性成熟早(4-6个月)，繁殖力高，一胎多仔，在2-3年内即可形成一个较大群体。另外，猪的克隆技术非常成熟，克隆及饲养成本也较灵长类低得多。
(2)本发明针对猪基因组进行了4个安全港位点基因敲入后表达情况的摸索，从中筛选出了最佳的供外源基因插入的猪基因组安全港位点，可有效改善基因敲入后目的基因的表达情况。
(3)本发明中经过实验验证改造的pU6gRNA-eEF1a-mNLS-hSpCas9-EGFP-PURO(简称pKG- GE3)载体相对改造前的pX330载体，更换了更强的启动子及添加了增强蛋白翻译的元件，提高了 Cas9的表达，并且增加了核定位信号个数，提高了Cas9蛋白的核定位能力，具有更高的基因编辑效率。本发明还在载体中加入了荧光标记及抗性标记，使其更方便运用于载体阳性转化细胞的筛选及富集。采用本发明改造的Cas9高效表达载体进行基因编辑，编辑效率比原载体提高100％以上。
(4)利用本发明所得到的hACE2基因纯合敲入单细胞克隆株进行体细胞核移植动物克隆可直接得到hACE2基因纯合敲入的克隆猪，并且该纯合插入基因可稳定遗传。
在小鼠模型制作中，通常采用受精卵显微注射基因编辑材料后再进行胚胎移植，因其直接获得纯合突变后代的概率非常低(低于5％)，需要进行后代的杂交选育，这不太适用于妊娠期较长的大动物(如猪)模型制作。因此，本发明采用技术难度大、挑战性高的原代细胞体外编辑并筛选阳性编辑单细胞克隆的方法，然后通过体细胞核移植动物克隆技术直接获得了ACE2人源化猪，大大缩短了模型猪制作周期，并节省人力、物力、财力。
(5)本发明利用体细胞核移植技术可以比较稳定，并具有较高成功率地获得hACE2纯合插入的病毒感染模型猪。其中，hACE2可以很好的在细胞膜表面表达，例如在肺泡巨噬细胞中表达量可高达管家基因β-actin表达量的0.12倍，而未改造的对照猪几乎不表达(为管家基因β-actin表达量的2×10-7倍)。所述hACE2人源化猪可进一步用于制备动物感染模型，为揭示SARS-CoV-2等冠状病毒的感染及致病机制提供有力的活体研究工具。
本发明通过基因编辑及体细胞克隆技术获得了ACE2人源化猪，将有助于研究并揭示由hACE2 介导引发的SARS-CoV-2及SARS-CoVs等冠状病毒的感染机制，并可用于进行药物筛选、药效评价、疾病病理、疫苗效果测试等研究，能够为进一步的临床应用提供有效的实验数据，进而为预防和治疗人类SARS-CoV-2及SARS-CoVs等冠状病毒感染疾病提供有力的实验手段。本发明对于人类SARS- CoV-2及SARS-CoVs治疗药物及疫苗的研发及揭示该类疾病的发病机制具有重大应用价值。
附图说明
以下，结合附图来详细说明本发明的实施例，其中：
图1为质粒pX330的结构示意图。
图2为质粒pKG-GE3的结构示意图。
图3为pU6gRNA载体的结构示意图。
图4为将20bp左右的DNA分子(用于转录形成能结合靶序列的gRNA)插入质粒pKG-U6gRNA 的示意图。
图5为含ROSA26插入位点的荧光donor质粒的结构示意图。
图6为含AAVS1插入位点的荧光donor质粒的结构示意图。
图7为含H11插入位点的荧光donor质粒的结构示意图。
图8为含COL1A1插入位点的荧光donor质粒的结构示意图。
图9为含COL1A1插入位点的hACE2基因donor质粒的结构示意图。
图10为质粒配比优化测试的测序结果。
图11为质粒pX330和质粒pKG-GE3编辑效果的测序结果。
图12为不同安全港位点调控GFP绿色荧光表达图。
图13为不同安全港位点调控GFP转录水平荧光定量PCR结果。
图14为不同安全港位点调控GFP蛋白表达的FACS检测结果。
图15为鉴定猪COL1A1安全港插入位点5’端hACE2表达框是否重组成功的电泳图。
图16为鉴定猪COL1A1安全港插入位点3’端hACE2表达框是否重组成功的电泳图。
图17为鉴定hACE2表达框是否纯合插入猪COL1A1安全港位点的电泳图。
图18为猪COL1A1安全港位点调控hACE2转录水平荧光定量PCR结果。
图19为猪COL1A1安全港位点调控hACE2蛋白表达的FACS检测结果。
图20为6头hACE2人源化小猪图片。
图21为hACE2基因在hACE2人源化猪和野生型猪肺泡巨噬细胞中的转录水平检测结果。
图22为hACE2人源化猪和野生型猪肺泡巨噬细胞的hACE2抗体结合情况检测结果。A、B分别为hACE2人源化猪、野生型猪肺泡巨噬细胞的hACE2抗体结合情况的共聚焦显微镜照片；C、D分别为hACE2人源化猪、野生型猪肺泡巨噬细胞的hACE2抗体结合情况的倒置荧光显微镜照片。
图23为质粒pMD2.G-SARS-C19的结构示意图。
图24为质粒Lenti-mCherry的结构示意图。
图25为hACE2人源化猪原代成纤维细胞与SARS-CoV-2假病毒结合情况的检测结果。其中图右侧为明场，图左侧为明场相同视野下倒置荧光显微镜的荧光观察结果。
图26为野生型猪原代成纤维细胞与SARS-CoV-2假病毒结合情况的检测结果。其中图右侧为明场，图左侧为明场相同视野下倒置荧光显微镜的荧光观察结果。
具体实施方式
下面结合具体实施方式对本发明进行进一步的详细描述，给出的实施例仅为了阐明本发明，而不是为了限制本发明的范围。以下提供的实施例可作为本技术领域普通技术人员进行进一步改进的指南，并不以任何方式构成对本发明的限制。
下述实施例中的实验方法，如无特殊说明，均为常规方法，按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。实施例中构建的重组质粒，均已进行测序验证。完全培养液(％为体积比)：15％胎牛血清 (Gibco)+83％DMEM培养基(Gibco)+1％Penicillin-Streptomycin(Gibco)+1％HEPES(Solarbio)。细胞培养条件：37℃，5％CO2、5％O2的恒温培养箱。
制备猪原代成纤维细胞的方法：①取猪耳组织0.5g，除毛，然后用75﹪酒精浸泡30-40s，然后用含5％(体积比)Penicillin-Streptomycin(Gibco)的PBS缓冲液洗涤5次，然后用PBS缓冲液洗涤一次；②用剪刀将组织剪碎，采用5mL0.1％胶原酶溶液(Sigma)，37℃消化1h，然后500g离心 5min，弃上清；③将沉淀用1mL完全培养液重悬，然后铺入含10mL完全培养基并已用0.2％明胶(VWR) 封盘的直径为10的细胞培养皿中，培养至细胞长满皿底60％左右；④完成步骤③后，采用胰蛋白酶消化并收集细胞，然后重悬于完全培养液。
实施例1、质粒的构建
出发商品质粒为：pX330-U6-Chimeric_BB-CBh-hSpCas9，简称质粒pX330，如SEQID NO：1所示。
以pX330质粒为基础，构建质粒pU6gRNAeEF1a-mNLS-hSpCas9-EGFP-PURO，简称质粒pKG-GE3，如 SEQ ID NO：2所示。
构建质粒pKG-U6gRNA，如SEQ ID NO：3所示。
质粒pX330、质粒pKG-GE3、质粒pKG-U6gRNA均为环形质粒。
质粒pX330的结构示意图见图1。SEQ ID NO：1中，第440-725位核苷酸组成CMV增强子，第727- 1208位核苷酸组成chickenβ-actin启动子，第1304-1324位核苷酸编码SV40核定位信号(NLS)，第 1325-5449位核苷酸编码Cas9蛋白，第5450-5497位核苷酸编码nucleoplasmin核定位信号(NLS)。
质粒pKG-GE3的结构示意图见图2。SEQ ID NO：2中，第395-680位核苷酸组成CMV增强子，第682- 890位核苷酸组成EF1a启动子，第986-1006位核苷酸编码核定位信号(NLS)，第1016-1036位核苷酸编码核定位信号(NLS)，第1037-5161位核苷酸编码Cas9蛋白，第5162-5209位核苷酸编码核定位信号 (NLS)，第5219-5266位核苷酸编码核定位信号(NLS)，第5276-5332位核苷酸编码自剪切多肽P2A (自剪切多肽P2A的氨基酸序列为“ATNFSLLKQAGDVEENPGP”，发生自剪切的断裂位置为C端开始第一个氨基酸残基和第二个氨基酸残基之间)，第5333-6046位核苷酸编码EGFP蛋白，第6056-6109位核苷酸编码自裂解多肽T2A(自裂解多肽T2A的氨基酸序列为“EGRGSLLTCGDVEENPGP”，发生自裂解的断裂位置为C端开始第一个氨基酸残基和第二个氨基酸残基之间)，第6110-6703位核苷酸编码Puromycin蛋白 (简称Puro蛋白)，第6722-7310位核苷酸组成WPRE序列元件，第7382-7615位核苷酸组成3’LTR序列元件，第7647-7871位核苷酸组成bGH poly(A)signal序列元件。SEQ ID NO：2中，第911-6706形成融合基因，表达融合蛋白。由于自剪切多肽P2A和自裂解多肽T2A的存在，融合蛋白自发形成如下三个蛋白：具有Cas9蛋白的蛋白、具有EGFP蛋白的蛋白和具有Puro蛋白的蛋白。
与质粒pX330相比，质粒pKG-GE3主要进行了如下改造：①去除残留的gRNA骨架序列 (GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTTT)，降低干扰；②将原有chickenβ-actin启动子改造为具更高表达活性的EF1a启动子，增加Cas9基因的蛋白表达能力；③在Cas9基因的上游和下游均增加核定位信号编码基因(NLS)，增加Cas9蛋白的核定位能力；④原质粒无任何真核细胞筛选标记，不利于阳性转化细胞的筛选和富集，依次在Cas9基因的下游插入P2A-EGFP-T2A-PURO编码基因，赋予载体荧光和真核细胞抗性筛选能力；⑤插入WPRE元件和3’LTR序列元件，增强Cas9基因的蛋白翻译能力。
质粒pKG-U6gRNA的结构示意图见图3。SEQ ID NO：3中，第2280-2539位核苷酸组成hU6启动子，第2558-2637位核苷酸用于转录形成gRNA骨架。使用时，将20bp左右的DNA分子(用于转录形成gRNA的靶序列结合区)插入质粒pKG-U6gRNA，形成重组质粒，示意图见图4,在细胞中重组质粒转录得到gRNA。
构建质粒PB-1G 2R 3-puro-ROSA26、PB-1G 2R 3-puro-AAVS1、PB-1G 2R 3-puro-H11和PB-1G 2R 3-puro-COL1A1。
质粒PB-1G 2R 3-puro-ROSA26结构示意图见图5。SEQ ID NO：4中，第1-345位核苷酸组成ROSA26 安全港插入位点5’端猪基因组区域(SH1左臂，如SEQ ID NO：18所示)，第9184-10195位核苷酸组成 ROSA26安全港插入位点3’端猪基因组区域(SH1右臂，如SEQ IDNO：19所示)，第346-546、3132- 3531、6506-6707、8975-9175位核苷酸分别组成4个不同的绝缘子区域，第1954-3131位核苷酸组成EF- 1α启动子，第1216-1935位核苷酸编码EGFP蛋白，第637-1209位核苷酸组成EF-1αpoly(A)信号,第 3543-4042位核苷酸组成PGK启动子，第4059-4769位核苷酸编码mCherry蛋白，第4791-5015位核苷酸组成bGH poly(A)信号，第5054-6504位核苷酸为loxP-puro-loxP表达框区域，第7259-8974位核苷酸组成pCAG启动子，第6969-7233位核苷酸组成β-globin poly(A)信号。
质粒PB-1G 2R 3-puro-AAVS1结构示意图见图6。仅将SEQ ID NO：4中的1-345位核苷酸替换为 AAVS1安全港插入位点5’端猪基因组区域(SH2左臂)，见SEQ ID NO：5；第9184-10195位核苷酸替换为AAVS1安全港插入位点3’端猪基因组区域(SH2右臂)，见SEQ ID NO：6。其他序列与SEQ ID NO： 4一致，具体为SEQ ID NO：20。
质粒PB-1G 2R 3-puro-H11结构示意图见图7。仅将SEQ ID NO：4中的1-345位核苷酸替换为H11安全港插入位点5’端猪基因组区域(SH3左臂)，见SEQ ID NO：7；第9184-10195位核苷酸替换为H11安全港插入位点3’端猪基因组区域(SH3右臂)，见SEQ ID NO：8。其他序列与SEQ ID NO：4一致，具体为SEQ ID NO：21。
质粒PB-1G 2R 3-puro-COL1A1结构示意图见图8。仅将SEQ ID NO：4中的1-345位核苷酸替换为 COL1A1安全港插入位点5’端猪基因组区域(SH4左臂)，见SEQ ID NO：9；第9184-10195位核苷酸替换为COL1A1安全港插入位点3’端猪基因组区域(SH4右臂)，见SEQID NO：10。其他序列与SEQ ID NO：4一致，具体为SEQ ID NO：22。
构建质粒pKG-hACE2：质粒pKG-hACE2的结构示意图见图9。SEQ ID NO：11中，第9-880位核苷酸为猪基因组COL1A1安全港插入位点5’端同源序列，第887-1286位核苷酸为绝缘子2(Insulator 2) 序列，第1287-2464位核苷酸为EF1a启动子序列，第2483-4897位核苷酸为hACE2的编码序列，来自于质粒ACE2-pENTER(Addgene),第4907-5479位核苷酸为EF1aPoly(A)序列，第5588-5917位核苷酸为 SV40启动子序列，第5966-6562位核苷酸为Puromycin抗性蛋白(简称PuroR蛋白)编码序列，第6742- 6863位核苷酸为SV40 Poly(A)序列，第5512-5545及6908-6941位核苷酸分别为同向相同的LoxP序列，第6950-7150位核苷酸为绝缘子3(Insulator 3)序列，第7153-7879位核苷酸为猪基因组COL1A1安全港插入位点3’端同源序列。
实施例2、质粒pX330和质粒pKG-GE3的效果比较
选择位于RAG1基因的高效gRNA靶点：
RAG1-gRNA4的靶点：5’-AGTTATGGCAGAACTCAGTG-3’(SEQ ID NO：23)。
用于扩增包含靶点的片段的引物如下：
RAG1-nF126：5’-CCCCATCCAAAGTTTTTAAAGGA-3’(SEQ ID NO：24)；
RAG1-nR525：5’-TGTGGCAGATGTCACAGTTTAGG-3’(SEQ ID NO：25)。
初生从江香猪(雌性，血型AO)，制备猪原代成纤维细胞。
一、制备重组质粒
取质粒pKG-U6gRNA，用限制性内切酶BbsI进行酶切，回收载体骨架(约3kb的线性大片段)。分别合成RAG1-4S和RAG1-4A，然后混合并进行退火，得到具有粘性末端的双链DNA分子。将具有粘性末端的双链DNA分子和载体骨架连接，得到质粒pKG-U6gRNA(RAG1-gRNA4)。
RAG1-4S：5’-caccgAGTTATGGCAGAACTCAGTG-3’(SEQ ID NO：26)；
RAG1-4A：5’-aaacCACTGAGTTCTGCCATAACTc-3’(SEQ ID NO：27)。
RAG1-4S和RAG1-4A均为单链DNA分子。
二、质粒配比优化
第一组：将质粒pKG-U6gRNA(RAG1-gRNA4)和质粒pKG-GE3共转染猪原代成纤维细胞。配比：约 20万个猪原代成纤维细胞：0.44μg质粒pKG-U6gRNA(RAG1-gRNA4)：1.56μg质粒pKG-GE3。即质粒 pKG-U6gRNA(RAG1-gRNA4)和质粒pKG-GE3的摩尔配比为：1：1。
第二组：将质粒pKG-U6gRNA(RAG1-gRNA4)和质粒pKG-GE3共转染猪原代成纤维细胞。配比：约 20万个猪原代成纤维细胞：0.72μg质粒pKG-U6gRNA(RAG1-gRNA4)：1.28μg质粒pKG-GE3。即质粒 pKG-U6gRNA(RAG1-gRNA4)和质粒pKG-GE3的摩尔配比为：2：1。
第三组：将质粒pKG-U6gRNA(RAG1-gRNA4)和质粒pKG-GE3共转染猪原代成纤维细胞。配比：约 20万个猪原代成纤维细胞：0.92μg质粒pKG-U6gRNA(RAG1-gRNA4)：1.08μg质粒pKG-GE3。即质粒 pKG-U6gRNA(RAG1-gRNA4)和质粒pKG-GE3的摩尔配比为：3：1。
第四组：将质粒pKG-U6gRNA(RAG1-gRNA4)转染致猪原代成纤维细胞。配比：约20万个猪原代成纤维细胞：1μg质粒pKG-U6gRNA(RAG1-gRNA4)。
共转染采用电击转染的方式，采用哺乳动物核转染试剂盒(Neon kit,Thermofisher)与Neon TM transfection system电转仪(参数设置为：1450V、10ms、3pulse)。
2、完成步骤1后，采用完全培养液培养16-18小时，然后更换新的完全培养液进行培养。培养总时间为48小时。
3、完成步骤2后，采用胰蛋白酶消化并收集细胞，提取基因组DNA，采用RAG1-nF126和RAG1- nR525组成的引物对进行PCR扩增，然后进行电泳。
电泳后回收目的条带并进行测序，测序结果见图10。
通过利用Synthego ICE工具分析测序峰图得出不同靶点的编辑效率。第一组至第三组的基因编辑效率依次为9％、53％、66％。第四组不发生基因编辑。结果表明，第三组编辑效率最高，确定单 gRNA质粒与Cas9质粒最适用量为摩尔比3：1，质粒实际用量为0.92μg：1.08μg。
三、质粒pX330和质粒pKG-GE3的效果比较
1、共转染
RAG1-B组：将质粒pKG-U6gRNA(RAG1-gRNA4)转染猪原代成纤维细胞。配比：约20万个猪原代成纤维细胞：0.92μg质粒pKG-U6gRNA(RAG1-gRNA4)。
RAG1-330组：将质粒pKG-U6gRNA(RAG1-gRNA4)和质粒pX330共转染猪原代成纤维细胞。配比：约20万个猪原代成纤维细胞：0.92μg质粒pKG-U6gRNA(RAG1-gRNA4)：1.08μg质粒pX330。
RAG1-KG组：将质粒pKG-U6gRNA(RAG1-gRNA4)和质粒pKG-GE3共转染猪原代成纤维细胞。配比：约20万个猪原代成纤维细胞：0.92μg质粒pKG-U6gRNA(RAG1-gRNA4)：1.08μg质粒pKG-GE3。
共转染采用电击转染的方式，采用哺乳动物核转染试剂盒(Neon kit,Thermofisher)与Neon TM transfection system电转仪(参数设置为：1450V、10ms、3pulse)。
2、完成步骤1后，采用完全培养液培养16-18小时，然后更换新的完全培养液进行培养。培养总时间为48小时。
3、完成步骤2后，采用胰蛋白酶消化并收集细胞，提取基因组DNA，采用RAG1-nF126和RAG1- nR525组成的引物对进行PCR扩增，将产物进行测序。
通过利用Synthego ICE工具分析测序峰图得出不同靶点的编辑效率。RAG1-B组不发生基因编辑。 RAG1-330组、RAG1-KG组的编辑效率依次为28％、68％。测序结果示例性峰图见图11。结果表明，与采用质粒pX330相比，采用质粒pKG-GE3使得基因编辑效率显著提高。
实施例3、筛选供外源基因定点插入的猪基因组最佳安全港位点
一、猪ROSA26、AAVS1、H11、COL1A1基因组安全港位点的高效切割靶点筛选
通过前期筛选，ROSA26、H11、AAVS1、COL1A1安全港位点的高效切割靶点分别为sgRNAROSA26-g3(切割效率38％)、sgRNAAAVS1-g4(切割效率30％)、sgRNAH11-g1(切割效率60％)、sgRNACOL1A1-g3(切割效率 56％),靶点序列如下：
sgRNAROSA26-g3靶点：5’-GAAGGAGCAAACTGACATGG-3’(SEQ ID NO：28)；
sgRNAAAVS1-g4靶点：5’-TGCAGTGGGTCTTTGGGGAC-3’(SEQ ID NO：29)；
sgRNAH11-g1靶点：5’-TTCCAGGAACATAAGAAAGT-3’(SEQ ID NO：30)；
sgRNACOL1A1-g3靶点：5’-GCAGTCTCAGCAACCACTGA-3’(SEQ ID NO：31)。
这4个gRNA靶点对应的gRNA质粒分别为pKG-U6gRNA(ROSA26-g3)、pKG-U6gRNA(AAVS1-g4)、pKG- U6gRNA(H11-g1)、pKG-U6gRNA(COL1A1-g3)，其中，骨架载体均为pKG-U6gRNA(SEQ ID NO：3)，质粒构建方法同实施例2。
二、含不同安全港插入位点两侧同源臂的荧光载体(包含外源基因GFP的不同安全港位点载体)、 sgRNA载体和Cas9载体混合电转猪原代成纤维细胞
分别将PB-1G 2R 3-puro-不同安全港插入位点荧光载体与对应的高效sgRNA载体以及高效Cas9 表达载体共转染猪原代成纤维细胞。使用哺乳动物核转染试剂盒(Neonkit,Thermofisher)与Neon TM transfection system电转仪进行电转实验(参数设置为：1450V、10ms、3pulse)。
共转染质粒组合及配比：
第一组：将质粒PB-1G 2R 3-puro-ROSA26、质粒pKG-U6gRNA(ROSA26-g3)和质粒pKG-GE3共转染猪原代成纤维细胞。配比：约20万个猪原代成纤维细胞：1.26μg质粒PB-1G2R 3-puro-ROSA26、 0.82μg质粒pKG-U6gRNA(ROSA26-g3)：0.92μg质粒pKG-GE3。
第二组：将质粒PB-1G 2R 3-puro-AAVS1、质粒pKG-U6gRNA(AAVS1-g4)和质粒pKG-GE3共转染猪原代成纤维细胞。配比：约20万个猪原代成纤维细胞：1.26μg质粒PB-1G 2R 3-puro-AAVS1、 0.82μg质粒pKG-U6gRNA(AAVS1-g4)：0.92μg质粒pKG-GE3。
第三组：将质粒PB-1G 2R 3-puro-H11、质粒pKG-U6gRNA(H11-g1)和质粒pKG-GE3共转染猪原代成纤维细胞。配比：约20万个猪原代成纤维细胞：1.26μg质粒PB-1G 2R 3-puro-H11、0.82μg 质粒pKG-U6gRNA(H11-g1)：0.92μg质粒pKG-GE3。
第四组：将质粒PB-1G 2R 3-puro-COL1A1、质粒pKG-U6gRNA(COL1A1-g3)和质粒pKG-GE3共转染猪原代成纤维细胞。配比：约20万个猪原代成纤维细胞：1.26μg质粒PB-1G2R 3-puro-COL1A1、 0.82μg质粒pKG-U6gRNA(COL1A1-g3)：0.92μg质粒pKG-GE3。
第五组：猪原代成纤维细胞，同等电转参数不加任何质粒进行电转染操作。
具体实施方法：
细胞：电转前猪原代成纤维细胞融合度达到60％，0.25％胰蛋白酶消化，台盼蓝染色计数，取等量细胞进行五组电转。
猪原代细胞电转：
(1)将细胞用胰酶消化，得到的细胞悬液用PBS磷酸缓冲液(Solarbio)洗一遍，600g离心6min，弃去上清，使用58μL电转基本溶液opti重悬细胞(11μL/个)，重悬过程中要避免气泡的产生；
(2)吸取10μL细胞悬液与质粒电转反应液混匀，混匀过程中注意切勿产生气泡；
(3)将试剂盒带有的电转杯放置于Neon TM transfection system电转仪杯槽内，加入3mL E Buffer；
(4)用电转枪吸取10μL步骤2)得到的混合液，插入电击杯内，选择电转程序(1450V10ms 3pulse)，电击转染后立即在超净台内将电转枪中混合液转入到6孔板中，每孔含3mL15％胎牛血清(Gibco)+83％ DMEM培养基(Gibco)+1％P/S(Gibco Penicillin-Streptomycin)+1％HEPES(Solarbio)的完全培养液；
(5)混匀后放置于37℃，5％CO2、5％O2的恒温培养箱中进行培养；
(6)电转12-24h换液，电转48h使用嘌呤霉素加压，筛选阳性细胞。
三、嘌呤霉素加压筛选
细胞经质粒电转48h，加入1.5μg/ml嘌呤霉素筛选，每两天更换含有相同浓度嘌呤霉素的培养基，同时进行GFP绿色荧光拍照，连续筛选两周，待细胞内质粒完全降解后再继续加压筛选一周。通过GFP荧光表达的强弱判断安全港位点效率的高低。
嘌呤霉素筛选一周后，ROSA26、COL1A1安全港位点实验组荧光强度明显强于AAVS1、H11实验组；嘌呤霉素筛选两周后，荧光强度由强到弱依次为：COL1A1>ROSA26>H11>AAVS1，其中H11组部分荧光弱整体荧光强,ROSA26组整体荧光强度较均一,AAVS1组细胞荧光表达最弱,COL1A1组荧光细胞数最多, 荧光最强；嘌呤霉素继续筛选三周后，荧光强度由强到弱依次为：COL1A1>ROSA26>H11>AAVS1，结果如图12。
四、GFP基因转录水平检测
为了比较GFP基因整合入四个不同安全港位点后mRNA转录水平的差异性，能否参与GFP的表达调控及对表达量的影响。在GFP基因外显子处设计一对引物，取嘌呤霉素筛选三周后的细胞，提取总
RNA,反转录成cDNA,用于检测原代细胞在四个不同安全港位点整合GFP基因后的转录水平，同时用野生型原代细胞作为对照。以GAPDH为内参基因按照2-ΔCt法进行计算。
(1)引物信息(表1)
表1：荧光定量PCR引物信息
(2)细胞总RNA提取
根据Bio Flux的Simply P总RNA提取试剂盒进行细胞总RNA提取
(3)cDNA第一链获得
根据Vazyme反转录试剂盒
II 1st Strand cDNA Synthesis Kit(R211-01/02)说明书合成 cDNA第一链，具体步骤和程序如下：
1)配制第一链cDNA合成反应液
在RNase-free离心管中配制如下表2混合液
表2
用移液枪轻轻吹打混匀。
2)按下列条件进行第一链cDNA合成反应，反应条件见表3。
表3
产物立即用于qPCR反应，或存放于-80℃保存，避免反复冻融。
(4)荧光定量PCR
利用实时荧光定量PCR法检测插入四组不同安全港位点(ROSA26、AAVS1、H11、COL1A1)猪原代成纤维细胞中GFP的表达量，GAPDH作为内参基因。操作步骤及程序如下:
1)反应体系配制见表4
表4
2)qPCR反应程序如下表5
表5
3)统计与分析
用SPSS统计学软件进行数据分析，以(平均数±标准差)表示，采用双因素方差分析进行统计学分析。2-ΔCt值结果显示嘌呤霉素筛选三周后AAVS1、H11组GFP表达量较低，ROSA26、COL1A1组GFP 表达量较高，且COL1A1组和ROSA26组相对于AAVS1和H11组GFP转录水平差异极显著(P<0.01)， 2-ΔCt值(表6)，差异显著性分析结果如图13。
表6：2-ΔCt值信息
综上，根据培养细胞三周后的荧光信号强度与GFP基因实时荧光定量PCR的结果，可以得出如下结论，在ROSA26、AAVS1、H11、COL1A1这四个基因组安全港位点中，COL1A1位点插入基因后表达效果最好。
五、GFP基因的蛋白表达水平FACS检测
为了比较GFP基因整合入四个不同安全港位点后GFP蛋白的表达情况。分别用胰蛋白酶消化细胞， 400g离心4min后，弃上清。以1mL培养基重悬细胞，并将细胞悬液分别转移至流式管内。在BD FACSMelody流式细胞仪的FITC通道内检测GFP信号，并以野生型细胞作为阴性对照，收集5×104个细胞进行分析，结果如图14所示。结果显示GFP荧光信号COL1A1>ROSA26>H11>AAVS1。
因此，综合上述结果，COL1A1位点是ROSA26、AAVS1、H11、COL1A1四个安全港位点中最高效的猪原代细胞安全港位点。
实施例4、制备hACE2定点插入猪COL1A1安全港位点的单细胞克隆
人(h)ACE2基因(如SEQ ID NO：13所示)信息：编码人angiotensin I convertingenzyme 2 蛋白；位于人X号染色体；GeneID为59272。hACE2是目前SARS-2-CoV及SARS-CoV等冠状病毒公认的受体蛋白，其编码的蛋白质片段NP_975010.1如SEQ ID NO：12所示。
一、共转染
将质粒pKG-U6gRNA(COL1A1-g3)、质粒pKG-GE3和质粒pKG-hACE2(如SEQ ID NO：11所示)共转染猪原代成纤维细胞。配比：约20万个猪原代成纤维细胞：0.89μg质粒pKG-U6gRNA(COL1A1- g3)：0.99μg质粒pKG-GE3：1.12μg质粒pKG-hACE2。
共转染采用电击转染的方式，采用哺乳动物核转染试剂盒(Neon kit,Thermofisher)与Neon TM transfection system电转仪(参数设置为：1450V、10ms、3pulse)。
二、嘌呤霉素加压筛选
1.嘌呤霉素筛选hACE2基因阳性插入细胞
细胞经质粒电转48h后，加入1.5μg/ml嘌呤霉素筛选，每天更换含有相同浓度嘌呤霉素的培养基，连续筛选一周后野生对照孔细胞全部死亡，因电转效率较低，ACE2质粒电转孔筛选一周后细胞也出现大量死亡；继续加入嘌呤霉素筛选一周，细胞只有零星死亡，部分阳性克隆开始分裂增殖，细胞数不断增多；继续加压筛选一周使细胞内质粒降解完全以排除假阳性细胞克隆。加压筛选三周后停止加压，培养两代待细胞状态恢复后分选至96孔板继续培养。
2.单克隆分选，放大培养
(1)将嘌呤霉素筛选三周后的群体细胞，进行单克隆分选，使用胰蛋白酶进行消化，完全培养基中和，500g离心5min，去上清，将沉淀用1mL完全培养基重悬，并适当稀释，用口吸管挑取单克隆转移到含100μl完全培养基的96孔板中，每组细胞挑取一板96个单克隆，放置于37℃，5％CO2、5％ O2的恒温培养箱中进行培养，每2～3天换一次细胞培养基(含1.5％嘌呤霉素)，期间用显微镜观察每孔细胞生长情况，排除无细胞及非单细胞克隆的孔；
(2)待96孔板的孔中细胞长满孔底(大约2周左右)，使用胰蛋白酶消化并收集细胞，其中2/3 细胞接种到含有完全培养基的6孔板中，剩余的1/3的细胞收集在1.5mL离心管中；
(3)待6孔板细胞长至50％丰满度时使用0.25％(Gibco)的胰蛋白酶消化并收集细胞，使用细胞冻存液(90％完全培养基+10％DMSO，体积比)将细胞冻存。
三、猪COL1A1安全港位点定点插入hACE2的基因组水平鉴定
为了检测猪COL1A1安全港位点是否成功定点插入了hACE2。取嘌呤霉素加压筛选完毕后的单细胞克隆，提取基因组DNA，进行PCR扩增(分别采用sh4-ace2-Lr-JDF1323和sh4-ace2-Lr-JDR5988组成的引物对、sh4-ace2-Rr-JDF7997和sh4-ace2-Rr-JDR12953组成的引物对、sh4-Lwt-JDF1085和sh4- Rwt-JDR1560组成的引物对)，然后进行电泳。将猪原代成纤维细胞作为野生型对照。sh4-ace2-Lr- JDF1323和sh4-ace2-Lr-JDR5988组成的引物对用来鉴定猪COL1A1安全港插入位点5’端hACE2表达框是否重组成功；sh4-ace2-Rr-JDF7997和sh4-ace2-Rr-JDR12953组成的引物对用来鉴定猪COL1A1安全港插入位点3’端hACE2表达框是否重组成功；sh4-Lwt-JDF1085和sh4-Rwt-JDR1560组成的引物对用来鉴定猪COL1A1安全港位点定点插入的为纯合型还是杂合型。
sh4-ace2-Lr-JDF1323：GCTCTCTCTGACCAGGATCTAAC(SEQ ID NO：36)
sh4-ace2-Lr-JDR5988：GACACTGGGACACTTTGTTTCAGG(SEQ ID NO：37)
sh4-ace2-Rr-JDF7997：CAGCTGAGGCCATTATATGAAGAG(SEQ ID NO：38)
sh4-ace2-Rr-JDR12953:GAGTCACCAAAGACGGTGTCAG(SEQ ID NO：39)
sh4-Lwt-JDF1085:TGCTGAGTTCTGGCTTCCTG(SEQ ID NO：40)
sh4-Rwt-JDR1560:TCTACCAAGAGAGTGACCAGCAG(SEQ ID NO：41)
电泳图分别见图15、图16和图17。通过电泳的结果，我们初步判定编号为2、7、9、23、26、 36、38、39、40、54、58、59的单细胞克隆为成功在猪COL1A1安全港位点定点插入hACE2的克隆，其中7、36、39、40号单细胞克隆初步判定为纯合定点插入，2、9、23、26、38、40、54、58、59为杂合定点插入。
四、hACE2基因的转录水平检测
为了检测猪COL1A1安全港位点定点插入hACE2的阳性单细胞克隆能否表达hACE2。我们在hACE2 基因外显子处设计一对引物，取嘌呤霉素筛选三周后的细胞，提取总RNA,反转录成cDNA,用于检测猪原代细胞hACE2基因的转录水平，同时用猪野生型原代细胞作为对照。以GAPDH为内参基因按照2-ΔCt法进行计算。
(1)引物信息(表7)
表7荧光定量PCR引物信息
(2)细胞总RNA提取
根据Bio Flux的Simply P总RNA提取试剂盒进行细胞总RNA提取
(3)cDNA第一链获得
根据Vazyme反转录试剂盒
II 1st Strand cDNA Synthesis Kit(R211-01/02)说明书合成 cDNA第一链，具体步骤和程序如下：
3)配制第一链cDNA合成反应液
在RNase-free离心管中配制如下表8混合液：
表8
用移液枪轻轻吹打混匀。
4)按下列条件进行第一链cDNA合成反应，反应条件见表9
表9
产物立即用于qPCR反应，或存放于-80℃保存，避免反复冻融。
(4)荧光定量PCR
利用实时荧光定量PCR法检测猪原代成纤维细胞中hACE2的表达量，GAPDH作为内参基因。操作步骤及程序如下:
3)反应体系配制如下表10
表10
4)qPCR反应程序如下表11
表11
3)统计与分析
用SPSS统计学软件进行数据分析，以(平均数±标准差)表示，采用单因素方差分析进行统计学分析。2-ΔCt值结果显示嘌呤霉素筛选三周后，改造后的猪原代成纤维细胞单克隆的hACE2基因表达量在统计学水平显著高于野生型猪原代成纤维细胞的hACE2基因的相对表达量(图18)。
综上，根据hACE2基因实时荧光定量PCR的结果，hACE2基因经过改造的猪原代成纤维细胞中有较好的表达。
五、hACE2基因的蛋白表达水平FACS检测
为了比较hACE2基因在编辑后的猪原代细胞中的表达情况。分别用胰蛋白酶消化人hACE2转染的人HEK293细胞(发明人之前构建)、猪野生型成纤维细胞和猪COL1A1安全位点定点插入hACE2的成纤维细胞，400g离心4min后，弃上清。加入PBS洗涤细胞，离心后，弃上清。加入90％的-20℃预冷甲醇充分重悬细胞，固定20min。固定结束后，离心，弃去固定液。加入3％BSA封闭1h。封闭结束后，离心，弃去封闭液。并用完全培养基洗涤。洗涤结束，以人特异性的ACE2抗体(Novus Biologicals， NBP2-80038)稀释液重悬细胞，室温孵育2h。抗体孵育结束后，用完全培养基充分洗涤后，加入 500μL完全培养基重悬细胞，并将细胞悬液转移至流式管内。在BD FACSMelody流式细胞仪的FITC 通道内检测hACE2抗体荧光信号，收集5×104个细胞进行分析，改造的猪成纤维细胞hACE2-2号单克隆的结果示例如图19。结果显示在hACE2转染的人HEK293细胞(hACE2-HEK293)和猪COL1A1安全港位点定点插入hACE2的成纤维细胞(hACE2-pig fibroblast)中均检测到hACE2的抗体荧光信号，并发现hACE2-pigfibroblast的hACE2抗体荧光信号强度低于hACE2-HEK293的抗体荧光信号强度；在猪野生型成纤维细胞(WTpig fibroblast)中没有检测到hACE2抗体荧光信号。
实施例5、利用体细胞核移植技术克隆生产ACE2人源化猪
1、卵母细胞体外成熟
从屠宰场采集新鲜的离体猪卵巢，卵巢在含75mg/mL青霉素和50mg/mL链霉素的0.9％(w/v)氯化钠溶液中保存，于25–30℃温度下运输至实验室。从直径3～6mm的卵泡中抽取卵丘卵母细胞复合体(Cumulus-oocyte complexes,COCs)，选择至少具有三层致密卵丘细胞的COCs，接种至4孔板中，每孔装有200μL猪卵母细胞体外成熟(IVM)培养基(即以TCM-199培养基为基础，内含 0.1mg/mL丙酮酸、0.1mg/mL盐酸半胱氨酸、10ng/mL表皮生长因子、10％(v/v)猪卵泡液、75mg/mL 青霉素，50mg/mL链霉素，10IU/mLeCG和hCG)，每孔接种50个，每次移植需培养300-400个COCs。将含COCs的培养板在38.5℃、5％CO2和饱和湿度的培养箱中培养42-44小时。
2、体细胞核移植(SCNT)与胚胎移植
(1)体细胞核移植
体外成熟42小时后，用0.1％(w/v)透明质酸酶反复吹打去除COCs的扩张卵丘细胞。将具有完整膜且含排出的第一极体的卵母细胞在含有0.1mg/mL地美可辛、0.05M蔗糖和4mg/mL牛血清白蛋白 (BSA)的NCSU23培养基中培养0.5-1h，促使卵母细胞核突起，然后使用尖部倾斜的显微注射针(直径约20μm)在含有10μm HEPES、0.3％(w/v)聚乙烯吡咯烷酮、10％FBS，0.1mg/mL地美可辛和 5mg/mL细胞松弛素B的Tyrode乳酸培养基中去除突起的细胞核和极体。以纯合插入hACE2的单细胞克隆株作为核供体，将单个供体细胞注入去核卵母细胞的卵周隙。使用胚胎细胞融合仪(ET3， Fujihira Industry)在含有0.25M D-山梨醇、0.05mM Mg(C2H3O2)2、20mg/mL BSA和0.5mM HEPES (acid-free)的融合培养基中用200V/mm的直流脉冲将供体细胞与受体卵母细胞融合20μs。将重构胚在PZM-3溶液(配方见下表12)中培养2h以允许细胞核重编程，然后在含有0.25M D-山梨醇、 0.01mM Ca(C2H3O2)2、0.05mMMg(C2H3O2)2和0.1mg/mL BSA的激活培养基中用150V/mm的单脉冲激活 100μs。然后将激活的胚胎在含5μg/mL细胞松弛素B的PZM-3中，于38.5℃、5％CO2、5％O2、90％N2和饱和湿度的培养箱中培养2小时，以进一步激活胚胎。最后将小部分重构胚移入PZM-3培养基中，在38.5℃、5％CO2、5％O2、90％N2和饱和湿度的培养箱中培养2d和7d，分别检测胚胎卵裂率和囊胚发育率。大部分重构胚在激活后培养6h即可用于后续的胚胎移植。
PZM-3溶液配方见表12：
表12
*使用前添加
(2)胚胎移植
选择5头处于发情期的杂交母猪(大白猪/长白猪)作为重构胚的代孕母猪，将激活后培养6h的重构胚移植到受体母猪的输卵管中，每头母猪移植300-350个重构胚，每次移植1-2头母猪。在胚胎移植后约28天，使用超声波扫描仪(HS-101V，日本本田电子)检查妊娠情况，确认受体母猪是否怀孕，克隆猪在胚胎移植后第116-117天左右出生。
5头代孕母猪中，3头成功怀孕的母猪共生产6头克隆猪(参见图20)，该克隆猪即为hACE2纯合插入的hACE2人源化猪。
实施例6从猪肺脏中分离肺泡巨噬细胞
(1)分离hACE2人源化猪和野生型猪完整气管和肺脏，并用含0.3％链霉素/青霉素的PBS进行清洗数次，最后一次用PBS洗涤，然后往气管中灌入50mL无菌PBS，轻轻拍打肺后倒出液体，1500g 离心4分钟。重复上述步骤2次。
(2)加入10mL红细胞裂解液重悬细胞沉淀以去除溶液中的红细胞，静置4分钟，加入2倍体积PBS，1500g离心10分钟，弃上清，沉淀即为肺泡巨噬细胞。
实施例7、hACE2人源化猪hACE2基因的转录水平检测
为了检测猪COL1A1安全港位点定点插入的hACE2的表达情况。针对hACE2基因设计了一对特异引物，对hACE2人源化克隆猪和未改造的对照克隆猪(相同细胞来源)分别分离肺泡巨噬细胞，提取总RNA,反转录成cDNA,用于检测猪肺泡巨噬细胞中hACE2基因的转录水平，同时用野生型猪肺泡巨噬细胞作为对照。以β-actin基因为内参基因按照2-ΔCt法进行计算。
引物信息见表13：
表13荧光定量PCR引物信息
用SPSS统计学软件进行数据分析，以(平均数±标准差)表示，采用单因素方差分析进行统计学分析。结果如图21显示，改造后的克隆猪(hACE2-pig)肺泡巨噬细胞的hACE2基因的表达量高达管家基因β-actin表达量的0.12倍，显著高于未改造克隆猪(WT-pig)肺泡巨噬细胞的hACE2基因的表达量(为管家基因β-actin表达量的2×10-7倍)。
综上，根据hACE2基因实时荧光定量PCR的结果，hACE2基因在经过改造后的hACE2人源化猪的肺泡巨噬细胞中有较强的表达。
实施例8、hACE2人源化猪的hACE2抗体结合情况检测
(1)使用无菌PBS重悬猪肺泡巨噬细胞沉淀，以60％-80％密度接种于细胞爬片上，于37℃培养箱中静置2小时后取出。
(2)使用预温的1×PBS清洗3次，每次10分钟。
(3)以4％的多聚甲醛室温固定20分钟后。
(4)用1×PBS清洗3次，每次10分钟。
(5)5％BSA室温封闭30分钟。
(6)加入1：200稀释的一抗(hACE2抗体，Novus Biologicals，NBP2-80038)，稀释液为含 1％BSA的PBS溶液，4℃孵育过夜。
(7)使用1×PBS清洗3次，每次10分钟。
(8)加入1:1000稀释的二抗(abcam,ab150113))，稀释液为含1％BSA的PBS溶液，4℃闭光孵育30分钟。
(9)用1×PBS清洗3次，每次10分钟。
(10)加入Hoechst 33342(用含1％BSA的PBS进行1:2000稀释)，室温闭光孵育10分钟。
(11)用1×PBS清洗3次，每次10分钟。
(12)95％甘油封片，分别于倒置荧光显微镜和共聚焦显微镜下观察，并拍照。
结果显示，hACE2人源化猪肺泡巨噬细胞(图22A，22C)相较于野生型猪肺泡巨噬细胞(图 22B,22D)，其细胞膜表面有更加显著的hACE2抗体荧光信号。
实施例9、SARS-CoV-2-Spike假病毒感染hACE2人源化猪成纤维细胞
假病毒不具备复制能力，可以最大程度降低SARS病毒研究过程中的各种风险。另外，由于假病毒的感染过程与真病毒相同，因此可以模拟病毒感染的早期过程，且假病毒内携带有报告基因，可以方便地进行各种检测分析。
9.1SARS-CoV-2-Spike假病毒的制备
9.1.1SARS-CoV-2-Spike假病毒制备所需质粒的构建
质粒pMD2.G-SARS-C19的结构示意图见图23。初始质粒为pMD2.G商品质粒，将pMD2.G质粒的 VSV-G区域删除，并将SARS-CoV-2病毒的刺突蛋白(Spike，为SARS-CoV-2的膜蛋白)进行胞内C端 19个氨基酸缺失突变，然后插入已删除VSV-G区域的pMD2.G载体中。改造后的载体序列如SEQ ID NO：50所示，其中第161-540位核苷酸为CMV增强子，第541-744位核苷酸为CMV启动子序列，第 878-1353位核苷酸为β-globin内含子序列，第1415-5188位核苷酸为SARS-CoV-2刺突蛋白(Spike) 的编码序列。第5264-5648位核苷酸为β-globinpoly(A)signal序列。
质粒Lenti-mCherry的结构示意图见图24。初始质粒为商品质粒Lenti-CRISPRV2，去除该质粒中的gRNA骨架与Cas9蛋白区域，并将报告基因mCherry插入相应区域，同时保留原质粒所携带的嘌呤霉素抗性基因。这将使利用该质粒联合配套质粒所构建的假病毒的基因组带有mCherry荧光标签及嘌呤霉素抗性标签。改造后的载体序列如SEQ ID NO：51所示，其中第2602-2813位核苷酸为EF1a 核心启动子元件序列，第2844-3551位核苷酸编码mCherry荧光蛋白，第3567-3623位核苷酸编码自剪切多肽P2A(自剪切多肽P2A的氨基酸序列为“ATNFSLLKQAGDVEENPGP”，发生自剪切的断裂位置为 C端开始第一个氨基酸残基和第二个氨基酸残基之间)，第3624-4220位核苷酸编码Puromycin抗性蛋白(简称PuroR蛋白)，第5161-5385位核苷酸为bGH poly(A)signal序列。
9.1.2SARS-CoV-2-Spike假病毒的制备
(1)将质粒pMD2.G-SARS-C19、Lenti-mCherry与psPAX2慢病毒包装质粒按6μg:4μg:5μg比例混合。
(2)加入750μL 
Medium，轻柔混合。
(3)加入24μL Lipo8000TM(碧云天,ST483)转染试剂，轻柔混合。
(4)滴入10cm的HEK293T细胞培养盘中，细胞密度在70％-80％。
(5)转染后6小时换液(含10％FBS的完全培养基)。
(6)转染后48小时收集细胞上清液，用0.45μm滤头过滤，去除细胞碎片。
(7)每10mL病毒过滤液加入3.3ml的Lenti-X Concentrator(Clontech，631231)，轻柔混合， 4℃放置过夜。
(8)4℃，1500g离心45分钟，弃上清，加入100μL DMEM溶解沉淀，即为浓缩后的SARS-CoV- 2假病毒。
9.2假病毒感染猪成纤维细胞
(1)于感染实验24小时前，取同等数量hACE2人源化猪和未改造猪的原代成纤维细胞接种于96 孔板中，使接种密度为30％-50％。
(2)向96孔板细胞中加入500μl浓缩病毒原液，同时加入0.8μL polybrene助感染试剂。
(3)感染后6小时换液(含10％FBS的完全培养基)。
(4)感染后48小时进行8μg/mL嘌呤霉素抗性筛选，连续筛选两天后，在倒置荧光显微镜下分别观察hACE2人源化猪和野生型猪原代成纤维细胞的mCherry荧光信号。
结果显示，在hACE2人源化猪原代成纤维细胞中观察到了带荧光的合胞体细胞(图25)，表明病毒能够感染细胞，而且促使被感染的细胞与邻近细胞形成合胞体，而未改造猪原代成纤维细胞中则未见有带荧光的合胞体细胞(图26)。
进一步的，本申请制备的hACE2人源化猪可用于下一步的药物筛选、药效评价、疫苗效果测试及病毒感染机制等生物医药领域研究。
以上详细描述了本发明的优选实施方式，但是，本发明并不限于上述实施方式中的具体细节，在本发明的技术构思范围内，可以对本发明的技术方案进行多种简单变型，这些简单变型均属于本发明的保护范围。
另外需要说明的是，在上述具体实施方式中所描述的各个具体技术特征，在不矛盾的情况下，可以通过任何合适的方式进行组合。
序列表
<110> 南京启真基因工程有限公司
<120> ACE2人源化猪的构建方法及应用
<130> 1
<160> 51
<170> SIPOSequenceListing 1.0
<210> 1
<211> 8484
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 60
ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 120
aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 180
atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 240
cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag 300
gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct 360
agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac 420
aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 480
ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc 540
aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 600
caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt 660
acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 720
ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac 780
ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg 840
ggggggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc 900
agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata 960
aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct 1020
ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga 1080
gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg taagggttta 1140
agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg cctgaaatca 1200
ctttttttca ggttggaccg gtgccaccat ggactataag gaccacgacg gagactacaa 1260
ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga agaagcggaa 1320
ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc tggacatcgg 1380
caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt 1440
caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct 1500
gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata 1560
caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa 1620
ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa 1680
gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta 1740
ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg 1800
gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg 1860
cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta 1920
caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct 1980
gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga 2040
gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt 2100
caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga 2160
cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc 2220
cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat 2280
caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct 2340
gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt 2400
cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt 2460
ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa 2520
gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca 2580
ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt 2640
cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt 2700
gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac 2760
catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat 2820
cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag 2880
cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga 2940
gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct 3000
gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat 3060
cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg 3120
cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa 3180
cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat 3240
cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa 3300
gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga 3360
caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa 3420
cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca 3480
ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc 3540
cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg 3600
ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa 3660
gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg 3720
cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta 3780
cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg 3840
gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat 3900
cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc 3960
cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat 4020
tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga 4080
taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc 4140
acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga 4200
agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt 4260
ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt 4320
cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga 4380
ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc 4440
taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct 4500
ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat 4560
cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt 4620
gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc 4680
caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg 4740
cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg 4800
caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag 4860
cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa 4920
ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag 4980
aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata 5040
tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa 5100
tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca 5160
gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc 5220
cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct 5280
gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga 5340
ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat 5400
caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca aaaggccggc 5460
ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa gaattcctag agctcgctga 5520
tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 5580
tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 5640
tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 5700
ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga acccctagtg 5760
atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag 5820
gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc 5880
ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc 5940
atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg 6000
tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt 6060
tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc 6120
tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg 6180
gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg 6240
agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct 6300
cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg 6360
agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat 6420
ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc 6480
caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag 6540
ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg 6600
cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg 6660
tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat 6720
ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc 6780
aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct 6840
tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag 6900
atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta 6960
agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc 7020
tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca 7080
tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg 7140
atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg 7200
ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca 7260
tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa 7320
acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa 7380
ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata 7440
aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat 7500
ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc 7560
cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata 7620
gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt 7680
actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga 7740
agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 7800
cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 7860
tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 7920
agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 7980
ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 8040
acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 8100
ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 8160
gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 8220
gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 8280
gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 8340
tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 8400
caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 8460
tttgctggcc ttttgctcac atgt 8484
<210> 2
<211> 10476
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 60
ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 120
aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 180
atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 240
cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag 300
gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttc tagcgcgtgc 360
gccaattctg cagacaaatg gctctagagg tacccgttac ataacttacg gtaaatggcc 420
cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga 480
ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 540
aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 600
ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 660
tagtcatcgc tattaccatg ggggcagagc gcacatcgcc cacagtcccc gagaagttgg 720
ggggaggggt cggcaattga tccggtgcct agagaaggtg gcgcggggta aactgggaaa 780
gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg 840
cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggttggac 900
cggtgccacc atggactata aggaccacga cggagactac aaggatcatg atattgatta 960
caaagacgat gacgataaga tggcccccaa aaagaaacga aaggtgggtg ggtccccaaa 1020
gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt acagcatcgg 1080
cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc 1140
cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat 1200
cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc 1260
cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa 1320
cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga 1380
agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta 1440
ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa 1500
ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt 1560
cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct 1620
ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc 1680
caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca 1740
gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct 1800
gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa 1860
ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga 1920
cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt 1980
gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca 2040
ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa 2100
agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag 2160
ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga 2220
actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg 2280
cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga 2340
tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat 2400
cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa 2460
gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc 2520
ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct 2580
gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa 2640
atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat 2700
cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta 2760
cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa 2820
cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga 2880
caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga 2940
cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat 3000
gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa 3060
cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt 3120
cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat 3180
ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc 3240
cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt 3300
gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca 3360
gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat 3420
caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa 3480
cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact 3540
ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa 3600
ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga 3660
caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa 3720
cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct 3780
gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac 3840
aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa 3900
gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa 3960
ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta 4020
cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt 4080
cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga 4140
aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac 4200
cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga 4260
aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag 4320
catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga 4380
gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc 4440
taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa 4500
agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat 4560
catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa 4620
agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa 4680
cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct 4740
gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc 4800
ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga 4860
gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga 4920
caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa 4980
tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga 5040
caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat 5100
ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga 5160
caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg gcggctccaa 5220
gcggcctgcc gcgacgaaga aagcgggaca ggccaagaaa aagaaaggat ccggcgcaac 5280
aaacttctct ctgctgaaac aagccggaga tgtcgaagag aatcctggac cggtgagcaa 5340
gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa 5400
cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac 5460
cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac 5520
cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt 5580
cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga 5640
cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat 5700
cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta 5760
caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt 5820
gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca 5880
gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac 5940
ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt 6000
cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggct ccggcgaggg 6060
caggggaagt cttctaacat gcggggacgt ggaggaaaat cccggcccaa ccgagtacaa 6120
gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc 6180
cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg 6240
ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg 6300
ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg 6360
ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc 6420
gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct 6480
ggccaccgtc ggagtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct 6540
ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc 6600
ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc 6660
cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaacgc gttaagtcga 6720
caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 6780
tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 6840
tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 6900
gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 6960
tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 7020
tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 7080
gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 7140
cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 7200
caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 7260
tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta 7320
agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 7380
ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct 7440
ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 7500
aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 7560
tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagggcc 7620
cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 7680
cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 7740
aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 7800
ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 7860
gggctctatg gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt 7920
atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg 7980
cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg 8040
ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 8100
taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 8160
aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 8220
ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 8280
tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt 8340
ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt 8400
ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc 8460
cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg 8520
cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat 8580
caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca 8640
tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc 8700
ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 8760
gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg 8820
cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg 8880
tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc 8940
tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca 9000
cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac 9060
tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa 9120
agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg 9180
ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt 9240
ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg 9300
aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc 9360
gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga 9420
tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta 9480
ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc 9540
cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg 9600
atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt 9660
cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa 9720
ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 9780
cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 9840
ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 9900
tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 9960
taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 10020
caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 10080
agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 10140
gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 10200
gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 10260
ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 10320
acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 10380
tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 10440
ggttcctggc cttttgctgg ccttttgctc acatgt 10476
<210> 3
<211> 3120
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 60
cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 120
tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 180
aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt 240
ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg 300
ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga 360
tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc 420
tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac 480
actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg 540
gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca 600
acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg 660
gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg 720
acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg 780
gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag 840
ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg 900
gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct 960
cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac 1020
agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact 1080
catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 1140
tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 1200
cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 1260
gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 1320
taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 1380
ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 1440
tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 1500
ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 1560
cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 1620
agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 1680
gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 1740
atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 1800
gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 1860
gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 1920
ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt 1980
cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc 2040
cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca 2100
acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc 2160
cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg 2220
accatgatta cgccaagctt gcatgcaggc ctctgcagtc gacgggcccg ggatccgatg 2280
ataaacatgt gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc 2340
tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac 2400
gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat 2460
ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt 2520
gtggaaagga cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag 2580
ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttc 2640
tagcgcgtgc gccaattctg cagacaaatg gctctagagg tacccataga tctagatgca 2700
ttcgcgaggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa 2760
accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta 2820
atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat 2880
ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt 2940
gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa 3000
cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg 3060
tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga 3120
<210> 4
<211> 14138
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
ggcgcgccct ctacctgctc tcggacccgt gggggtgggg ggtggaggaa ggagtggggg 60
gtcggtcctg ctggcttgtg ggtgggaggc gcatgttctc caaaaacccg cgcgagctgc 120
aatcctgagg gagctgcagt ggaggaggcg gagagaaggc cgcacccttc tccgcagggg 180
gaggggagtg ccgcaatacc tttatgggag ttctctgctg cctccttttc ctaaggaccg 240
ccctgggcct agaaaaatcc ctccctcccc cgcgatctcg tcatcgcctc catgtcagtt 300
tgctccttct cgattatggg cgggattctt ttgccctggc gcgccccaga cccgggcctg 360
gggggcaagt cggggggcgg ggggaggtcg ggcagggtcc cctgggagga tggggacgtg 420
ctgtgcccct agcggccacc agagggcacc aggacaccac tgcggtcggc tcagcggctc 480
ctgccctggt cagggggcgc caggtcctgc ccctcctggg gagggcgggg ggcgagaagg 540
gcgattttaa ttaacccacg tttcaacatg cacatcccag taatttggaa acattttgtt 600
tccaaagatt cacttaacat tggtttagca acatgaagct ttctatgcaa cccaaggact 660
cagtttttgg cctgttttag tgacaggcaa tcagcaacat gctgcatttc tctccagtgt 720
tgtaatcaaa gaaaccctcc catagcttta aatgatattc cttccccttc caattatgtg 780
gggggaaaac aaccctattc tccacccaga agtgttaact caagaattac attttcaaga 840
agtttccaga ttcgtaaaac cagaattaga tgtctttcac ctaaatgtct cggtgttgac 900
caaaggaaca cacaggtttc tcatttaact tttttaatgg gtctcaaaat tctgtgacaa 960
atttttggtc aagttgtttc cattaaaaag tactgatttt aaaaactaat aacttaaaac 1020
tgccacacgc aaaaaagaaa accaaagtgg tccacaaaac attctccttt ccttctgaag 1080
gttttacgat gcattgttat cattaaccag tcttttacta ctaaacttaa atggccaatt 1140
gaaacaaaca gttctgagac cgttcttcca ccactgatta agagtggggt ggcaggtatt 1200
agggataatg ctagcttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt 1260
cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga 1320
ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt 1380
ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa 1440
gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta 1500
gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag 1560
ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt 1620
gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt 1680
gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt 1740
cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt 1800
cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg 1860
gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc 1920
gcccttgctc accatggtgg cgtcgaccgt acgtcacgac acctgaaatg gaagaaaaaa 1980
actttgaacc actgtctgag gcttgagaat gaaccaagat ccaaactcaa aaagggcaaa 2040
ttccaaggag aattacatca agtgccaagc tggcctaact tcagtctcca cccactcagt 2100
gtggggaaac tccatcgcat aaaacccctc cccccaacct aaagacgacg tactccaaaa 2160
gctcgagaac taatcgaggt gcctggacgg cgcccggtac tccgtggagt cacatgaagc 2220
gacggctgag gacggaaagg cccttttcct ttgtgtgggt gactcacccg cccgctctcc 2280
cgagcgccgc gtcctccatt ttgagctccc tgcagcaggg ccgggaagcg gccatctttc 2340
cgctcacgca actggtgccg accgggccag ccttgccgcc cagggcgggg cgatacacgg 2400
cggcgcgagg ccaggcacca gagcaggccg gccagcttga gactaccccc gtccgattct 2460
cggtggccgc gctcgcaggc cccgcctcgc cgaacatgtg cgctgggacg cacgggcccc 2520
gtcgccgccc gcggccccaa aaaccgaaat accagtgtgc agatcttggc ccgcatttac 2580
aagactatct tgccagaaaa aaagcgtcgc agcaggtcat caaaaatttt aaatggctag 2640
agacttatcg aaagcagcga gacaggcgcg aaggtgccac cagattcgca cgcggcggcc 2700
ccagcgccca ggccaggcct caactcaagc acgaggcgaa ggggctcctt aagcgcaagg 2760
cctcgaactc tcccacccac ttccaacccg aagctcggga tcaagaatca cgtactgcag 2820
ccagtggaag taattcaagg cacgcaaggg ccataacccg taaagaggcc aggcccgcgg 2880
gaaccacaca cggcacttac ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc 2940
acggcgacta ctgcacttat atacggttct cccccaccct cgggaaaaag gcggagccag 3000
tacacgacat cactttccca gtttaccccg cgccaccttc tctaggcacc ggttcaattg 3060
ccgacccctc cccccaactt ctcggggact gtgggcgatg tgcgctctgc ccactgacgg 3120
gcaccggagc cctagattcg attccctttg gggcaaaact caccgcctaa tcccctataa 3180
ctctaccggg gagcccggtg gagagcagac gggctgacgc tgccacctgc cggccatccc 3240
aggataggac cgccgtattc aagtcgccct caggaaggac cctcggggca ccagaggcct 3300
tcgaagcccc aatgagtgag gcaactgagg gtcgcgggtg ccattacaag gcccagccaa 3360
ggcctagagc caaggcttga accgtggggg acccccaagc cccacctgcc caggaacagc 3420
agacactggg acactttgtt tcaggtcctg cccaggcccc tcccactgtg aggctgggat 3480
ttgtcgccca gggtgcagat gagaagagtg gggaaagcag tcctgagcca ggaaattcta 3540
ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct 3600
gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag 3660
gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt 3720
caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc 3780
acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct 3840
ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag 3900
gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc 3960
ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc 4020
ttcctcatct ccgggccttt cgacctccta gggccaccat ggtgagcaag ggcgaggacg 4080
acaacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga 4140
acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga 4200
ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc 4260
ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact 4320
tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg 4380
gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga 4440
agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct 4500
gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc 4560
agaggctgaa gctgaaggac ggcggccact acgacgccga ggtcaagacc acctacaagg 4620
ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagctg gacatcacct 4680
cccacaacga ggactacacc atcgtggaac agtacgagcg cgccgagggc cgccactcca 4740
ccggcggcat ggacgagctg tacaagtgag gatccgctga tcagcctcga ctgtgccttc 4800
tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 4860
cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 4920
tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 4980
tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaacccttct 5040
gaggcggaaa gaaccagctg ccttaatata acttcgtata atgtatgcta tacgaagtta 5100
ttaggtctga agaggagttt acgtccagcc aattctgtgg aatgtgtgtc agttagggtg 5160
tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 5220
agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 5280
tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc 5340
gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc 5400
cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 5460
aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggcggcc gcgccaccat 5520
gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg 5580
caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg 5640
ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 5700
cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 5760
cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 5820
ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 5880
cgcgtggttc ctggccaccg tcggagtctc gcccgaccac cagggcaagg gtctgggcag 5940
cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 6000
gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 6060
cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaga 6120
attcgcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 6180
gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 6240
gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac 6300
ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 6360
aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 6420
catgtctgta taccgctcga ctagagcttg cggaaccctt aatataactt cgtataatgt 6480
atgctatacg aagttattag gtccgctggc catctacgag ccaaagactt tcaaatcttt 6540
ggctgccttg gccagtagga ggcgacacga aggatttgct gctgccttgg gggatgggaa 6600
ggaacctgaa ggcatttttt ccagagtggt gcagtaccac tgaggactgt tgctgtattg 6660
attaggaaaa gagacagagt aatttgcagt ttgtttgatt tatactgggc tgcaggtcga 6720
gggatcttca taagagaaga gggacagcta tgactgggag tagtcaggag aggaggaaaa 6780
atctggctag taaaacatgt aaggaaaatt ttagggatgt taaagaaaaa aataacacaa 6840
aacaaaatat aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga 6900
cagcaggggg ctgtttcata tactgatgac ctctttatag ccacctttgt tcatggcagc 6960
cagcatatgg catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga 7020
tttgccctcc catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca 7080
atgaaaataa atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc 7140
ccataatttt tggcagaggg aaaaagatct cagtggtatt tgtgagccag ggcattggcc 7200
acaccagcca ccaccttctg ataggcagcc tgcggtacct tacatggtgg cgaattcgtt 7260
tgccaaaatg atgagacagc acaataacca gcacgttgcc caggagctgt aggaaaaaga 7320
agaaggcatg aacatggtta gcagaggctc tagagccgcc ggtcacacgc cagaagccga 7380
accccgccct gccccgtccc ccccgaaggc agccgtcccc ctgcggcagc cccgaggctg 7440
gagatggaga aggggacggc ggcgcggcga cgcacgaagg ccctccccgc ccatttcctt 7500
cctgccggcg ccgcaccgct tcgcccgcgc ccgctagagg gggtgcggcg gcgcctccca 7560
gatttcggct ccgccagatt tgggacaaag gaagtccctg cgccctctcg cacgattacc 7620
ataaaaggca atggctgcgg ctcgccgcgc ctcgacagcc gccggcgctc cggggccgcc 7680
gcgcccctcc cccgagccct ccccggcccg aggcggcccc gccccgcccg gcacccccac 7740
ctgccgccac cccccgcccg gcacggcgag ccccgcgcca cgccccgcac ggagccccgc 7800
acccgaagcc gggccgtgct cagcaactcg gggagggggg tgcagggggg ggttacagcc 7860
cgaccgccgc gcccacaccc cctgctcacc cccccacgca cacaccccgc acgcagcctt 7920
tgttcccctc gcagcccccc cgcaccgcgg ggcaccgccc ccggccgcgc tcccctcgcg 7980
cacacgcgga gcgcacaaag ccccgcgccg cgcccgcagc gctcacagcc gccgggcagc 8040
gcgggccgca cgcggcgctc cccacgcaca cacacacgca cgcacccccc gagccgctcc 8100
cccccgcaca aagggccctc ccggagccct ttaaggcttt cacgcagcca cagaaaagaa 8160
acgagccgtc attaaaccaa gcgctaatta cagcccggag gagaagggcc gtcccgcccg 8220
ctcacctgtg ggagtaacgc ggtcagtcag agccggggcg ggcggcgcga ggcggcgcgg 8280
agcggggcac ggggcgaagg caacgcagcg actcccgccc gccgcgcgct tcgcttttta 8340
tagggccgcc gccgccgccg cctcgccata aaaggaaact ttcggagcgc gccgctctga 8400
ttggctgccg ccgcacctct ccgcctcgcc ccgccccgcc cctcgccccg ccccgccccg 8460
cctggcgcgc gccccccccc cccccgcccc catcgctgca caaaataatt aaaaaataaa 8520
taaatacaaa attgggggtg gggagggggg ggagatgggg agagtgaagc agaacgtggg 8580
gctcacctcg acccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa 8640
agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt 8700
caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc 8760
gtaaatagtc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat 8820
acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta 8880
ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg 8940
attactatta ataactagtc aataatcaat gtcgtaaatg tcgtaaatgt ctcagctagt 9000
caggtagtaa aaggtgtcaa ctaggcagtg gcagagcagg attcaaattc agggctgttg 9060
tgatgcctcc gcagactctg agcgccacct ggtggtaatt tgtctgtgcc tcttctgacg 9120
tggaagaaca gcaactaaca cactaacacg gcatttacta tgggccagcc attgtacgcg 9180
ttgcttaacc tgattcttgg gcgttgtcct gcaggggatt gagcaggtgt acgaggacga 9240
gcccaatttc tctatattcc cacagtcttg agtttgtgtc acaaaataat tatagtgggg 9300
tggagatggg aaatgagtcc aggcaacacc taagcctgat tttatgcatt gagactgcgt 9360
gttattacta aagatctttg tgtcgcaatt tcctgatgaa gggagatagg ttaaaaagca 9420
cggatctact gagttttaca gtcatcccat ttgtagactt ttgctacacc accaaagtat 9480
agcatctgag attaaatatt aatctccaaa ccttaggccc cctcacttgc atccttacgg 9540
tcagataact ctcactcata ctttaagccc attttgtttg ttgtacttgc tcatccagtc 9600
ccagacatag cattggcttt ctcctcacct gttttaggta gccagcaagt catgaaatca 9660
gataagttcc accaccaatt aacactaccc atcttgagca taggcccaac agtgcattta 9720
ttcctcattt actgatgttc gtgaatattt accttgattt tcattttttt ctttttctta 9780
agctgggatt ttactcctga ccctattcac agtcagatga tcttgactac cactgcgatt 9840
ggacctgagg ttcagcaata ctccccttta tgtcttttga atacttttca ataaatctgt 9900
ttgtattttc attagttagt aactgagctc agttgccgta atgctaatag cttccaaact 9960
agtgtctctg tctccagtat ctgataaatc ttaggtgttg ctgggacagt tgtcctaaaa 10020
ttaagataaa gcatgaaaat aactgacaca actccattac tggctcctaa ctacttaaac 10080
aatgcattct atcatcacaa atgtgaaaaa ggagttccct cagtggacta accttatctt 10140
ttctcaacac ctttttcttt gcacaatttt ccacacatgc ctacaaaaag tacttatgcg 10200
gccgccataa aagttttgtt actttataga agaaattttg agtttttgtt ttttttaata 10260
aataaataaa cataaataaa ttgtttgttg aatttattat tagtatgtaa gtgtaaatat 10320
aataaaactt aatatctatt caaattaata aataaacctc gatatacaga ccgataaaac 10380
acatgcgtca attttacaca tgattatctt taacgtacgt cacaatatga ttatctttct 10440
agggttaatc tagctgcgtg ttctgcagcg tgtcgagcat cttcatctgc tccatcacgc 10500
tgtaaaacac atttgcaccg cgagtctgcc cgtcctccac gggttcaaaa acgtgaatga 10560
acgaggcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta 10620
cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg 10680
cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct 10740
gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 10800
ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 10860
gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 10920
ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 10980
gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 11040
tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 11100
tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 11160
ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac 11220
ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 11280
ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt 11340
cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct 11400
ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga 11460
tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag 11520
cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca 11580
actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga 11640
aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag 11700
tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc 11760
ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa 11820
tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt 11880
gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg 11940
gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt 12000
tattgctgat aaatctggag ccggtgagcg tggttcacgc ggtatcattg cagcactggg 12060
gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat 12120
ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact 12180
gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa 12240
aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 12300
ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 12360
ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 12420
tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 12480
gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt 12540
agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 12600
taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 12660
gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 12720
gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 12780
caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 12840
aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 12900
tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 12960
acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 13020
ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 13080
gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc 13140
tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa 13200
agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc 13260
tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca 13320
cacaggaaac agctatgacc atgattacgc caagcgcgcc cgccgggtaa ctcacggggt 13380
atccatgtcc atttctgcgg catccagcca ggatacccgt cctcgctgac gtaatatccc 13440
agcgccgcac cgctgtcatt aatctgcaca ccggcacggc agttccggct gtcgccggta 13500
ttgttcgggt tgctgatgcg cttcgggctg accatccgga actgtgtccg gaaaagccgc 13560
gacgaactgg tatcccaggt ggcctgaacg aacagttcac cgttaaaggc gtgcatggcc 13620
acaccttccc gaatcatcat ggtaaacgtg cgttttcgct caacgtcaat gcagcagcag 13680
tcatcctcgg caaactcttt ccatgccgct tcaacctcgc gggaaaaggc acgggcttct 13740
tcctccccga tgcccagata gcgccagctt gggcgatgac tgagccggaa aaaagacccg 13800
acgatatgat cctgatgcag ctagattaac cctagaaaga tagtctgcgt aaaattgacg 13860
catgcattct tgaaatattg ctctctcttt ctaaatagcg cgaatccgtc gctgtgcatt 13920
taggacatct cagtcgccgc ttggagctcc cgtgaggcgt gcttgtcaat gcggtaagtg 13980
tcactgattt tgaactataa cgaccgcgtg agtcaaaatg acgcatgatt atcttttacg 14040
tgacttttaa gatttaactc atacgataat tatattgtta tttcatgttc tacttacgtg 14100
ataacttatt atatatatat tttcttgtta tagatatc 14138
<210> 5
<211> 1073
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
gtgctgagtc cttttcccat cccacccacc tggagctccc ctcttccagt cctgagccac 60
ttgaactggc ctggtttttg ccatcctgcg ctgccctctc tccggactcg agccactgct 120
gagggcctca ggccagtcca tcctcgtctt gtctctttcg ccctgctctt tccccacctt 180
gagcgctctt aaccagcctg gcccgtgcca cctctactct gccatcgaat gctgccccac 240
tttctcgagt ccgccacttc tcccagcttc accggtaccc actgtttccc ctagtccagg 300
caggtaccac tttccctgag cgtcctcctc ctctctcctg ggcctgtgct gcttcttttc 360
ccgctctctg gcctgggccg tttcttcggc cagcccccga gccttccatg ccctttcctt 420
caggtttctg ctcttcatcc ttggtctctg ccatctgttg ccatgtaagg gtgctctttc 480
ctgagccatc gccctcaagg cgctctgctc ctcaagtgga tgcttccctc gcctggctca 540
cctcctgctc tctctcctgc ccccttcacc tgcgtgccct cctcattctc cctctgtgcc 600
acctctggcc ttgcactgta ggctctctct tggggatgtt tctccttctc cacacacttc 660
tctttcactc tgtcctcttg ctttgtgtgg gcctgcagcg ttaccctttt ttctgggcac 720
actcagagca ccctcctctt tctggttctg ggccacctgt ctgtcctcgg gtcatcttgc 780
tctctctgcc tggatgccct cctgtggctt tgggcagctt ctccctcctt cagagtgcac 840
cgccagttct cctaggcccg gtcacttccc cttcccaggg gacctagagc cctgctaggt 900
cctctctctc cacaacctgg gcccccaaac ctttccaaaa caccttgctt tctgcctcca 960
ttggtcttgt gttccagagc cagagtcact atatgtccca gaaccaggat tccctctggt 1020
tctgagggct tttatcgcat cccctgcctg gctgcagtgg gtctttgggc gcc 1073
<210> 6
<211> 260
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
gacaggccac agaagagcct ctactcctcc ctctgtcccc gaggctgtct ccctcccagt 60
cttcccagct caggccagtc cccaggcctc tcttccctgc cagagcccgt caggttcggt 120
tactttgggg cccagagagg accctgtgaa ggaagcgtgg gtaggggcac gggaatgggg 180
aggatgcctg aagaggcccc cttagccaga agaggagcag aagaggagca ggtacccaga 240
agaggagcag ttcagggaaa 260
<210> 7
<211> 546
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
aaatacccac gtttattggg acaaaagttg ttagggaaaa tggggcctca gagttatgat 60
tcaagtcata attctttcca tttataattt cactcgagac tctgttaact gattccttgt 120
gtgttgtatc ttactcctca gctcacaatt acttttagtt attcacctta actgtatgaa 180
taacagtgga gaaaaggatt ctaccagaat actctaatta tggttttgag tcccctttcc 240
agactgaaga tttttcagtc tttttgatct gaggtgattt ttcagtcttt tcgatctgag 300
gtgacagtct caagctcctc aattcaccca gtctcttgat acttgtccat ttagggccac 360
caaagctact ttgacttcat actagagagt caattaatga ggccattctc tgatggacag 420
gtgaagcagg caaggtgact atattttgac taaacggtag aaaacagcct gagtgttaac 480
agtgtagcct ataaaaccca gagctgccca ccctgatcta aacttccagg aacataagaa 540
cgcgcc 546
<210> 8
<211> 1009
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
agtaggtcac atttcagtaa aacctggctt tgtggattga gcatggtctg tctcttcctg 60
gtacttcatt agtcccctaa gtgggatttg ctgagcaaga ctcctcaatt acagaaatac 120
tccagtttag aattctcgca aaggcttttt gtttccacaa gtagaatcta gaaagcaatc 180
tcaagtaaca acagcagaga cctgaatccc aatccatctt tcctgtgtgt cctcttttac 240
ctccttccct ttcatgttga accaacagtc ctttttcagt ctagaagcta gtacgaaaga 300
aatgtacaga tgtaggtacc aagcaaagcc attagccaat aactggtgag atggagctaa 360
gaggaaataa aagtgttcct aagaatagca cagcagaagc tagatccaca gatcttaaaa 420
caattttggt tgagtaagag tagaggcaaa agaggaagct aataatgcag tttttaggag 480
ctaagagcca gataaagggt aagggcagga ggaagtgcta tctcagctaa cgagatacat 540
gaaacaacgg tggaagtcca gcaggcacaa gatgagttga gaagcaatca gggccagaag 600
gatgtgcaag gcctcaaaat aaaaaagcac agggccacag ggaaccttat ggaaattaaa 660
aggaagagga tgcagtcagg agaggaaaaa atagtgctcc ctcccccatg cccaaggaag 720
cagctgagca gccagtactt gggaagttag tagtaataag ttggtaagag ggagttctgt 780
tcgtggctca atggttaaca aatcagacta gaaaccgtga ggttgcgggt ttgatccctg 840
gccttgctca gtgggttaag gatccggcat tgccgtgacc tgtggtgtag gtcacagacg 900
tggctcagtt cccgcattcc tgtggctctg gtgtaggctg gtggctacag ctctgattag 960
acccctaggc tgggaacctc catatgccct ggaagtggcc gtagaaaag 1009
<210> 9
<211> 878
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
ggatggggac tcatgtgaat tttctaaagg tgctatttaa acggggggca cgagtgccgg 60
ctttggacag ggccgctcgc tctccaccct ttcttcttcc ccctcggccg cctctcaccc 120
cctgaggcct ctctcccccc acgacctcct ctctctcctc tgaaaccctc tcctcctcag 180
ctgcatccca ccctcgtggc ctctctctct ctctgtctgt cctgtgtcct ctctcactgg 240
gtttcagagc acagatgccc aaagcacaaa agcagttttc ccctggggtg ggaggaagca 300
agagactttg tacctatttt gtatgtgtat aataatttga gatgttttta attattttga 360
ttgctggaat aaagcatgtg gaaatgaccc aaaccaatct tgcactggcc tcctgatttc 420
cttccttgga gacggaggga gggggagacc tgggggaggg cgcttggggg ggggtgggct 480
ctcttctttc tgcgctcccc ccccccacct ccaacacctt gacgacccct cctgcttccg 540
cttgcctttc tcaggcttta acactttctc ctcgccctct cagcatgcgc atgcgcgtgc 600
ctctacctcc cccgcacatc ctggcctgcc caccctgaat ggcctggccc agcgatgcca 660
ccaactctct cgctccgtcc acggctgggg aggggggcac tctgcagggt tggggggcac 720
tgggaggctg ggttgggtga gggaggggtg cctgggcccc caccccccag caagttctct 780
ccctaggcga actggagggt cgtctggcct cttgagcctt gttgctggct ctgagctcta 840
ccaagagagt gaccagcagg accgcaccat cacgcgcc 878
<210> 10
<211> 727
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
gtggttgctg agactgcgtg ggggcccaag gagacctgga gaaaggaatg cttcctgctc 60
cttcttctgg ggccccagga gagccttccc agggccttgg agaggtgctg tccagggact 120
aaccctgtgc tctaggaagg ctgcaggccc tgaccagctg ggcaggtcct gggtccctcc 180
tggccttcta agttccccaa acatgagacc tctgggtgtg gggtggcctg gggaggtcat 240
tttgcccagg ccctacctcc tgcccattcc taaccctttt taaaaatctg tgcgtcctct 300
tcttccttct tctccctccc ttcccttttc gctcaccctc tgctgctggc ctgagagccg 360
gaggccccca gggggaaggc gactggtctc ctccccagtc tcagggaagg gagacagaga 420
atccaggaag ccagaactca gcagacgaag cacccaggga cctagagatg ggttgaaaag 480
ttgacagctg tcccacctgc ctcccaaggt ctcagggcct aaacctccaa ggcaggaaag 540
gcccctgtcc ctccctgggg tccatagaaa gagggacaag tctgcacgga ccatttgctg 600
taatattaac accttggctg tcattaggta gtcttggctg ttaattatgt cctgtgataa 660
tgtattatta gcacgccgac cacatagggt agggaactgc agctagtaaa caaaagtttg 720
ttcctat 727
<210> 11
<211> 11822
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
ggcgcgccgg atggggactc atgtgaattt tctaaaggtg ctatttaaac ggggggcacg 60
agtgccggct ttggacaggg ccgctcgctc tccacccttt cttcttcccc ctcggccgcc 120
tctcaccccc tgaggcctct ctccccccac gacctcctct ctctcctctg aaaccctctc 180
ctcctcagct gcatcccacc ctcgtggcct ctctctctct ctgtctgtcc tgtgtcctct 240
ctcactgggt ttcagagcac agatgcccaa agcacaaaag cagttttccc ctggggtggg 300
aggaagcaag agactttgta cctattttgt atgtgtataa taatttgaga tgtttttaat 360
tattttgatt gctggaataa agcatgtgga aatgacccaa accaatcttg cactggcctc 420
ctgatttcct tccttggaga cggagggagg gggagacctg ggggagggcg cttggggggg 480
ggtgggctct cttctttctg cgctcccccc ccccacctcc aacaccttga cgacccctcc 540
tgcttccgct tgcctttctc aggctttaac actttctcct cgccctctca gcatgcgcat 600
gcgcgtgcct ctacctcccc cgcacatcct ggcctgccca ccctgaatgg cctggcccag 660
cgatgccacc aactctctcg ctccgtccac ggctggggag gggggcactc tgcagggttg 720
gggggcactg ggaggctggg ttgggtgagg gaggggtgcc tgggccccca ccccccagca 780
agttctctcc ctaggcgaac tggagggtcg tctggcctct tgagccttgt tgctggctct 840
gagctctacc aagagagtga ccagcaggac cgcaccatca cgcgccctgg ctcaggactg 900
ctttccccac tcttctcatc tgcaccctgg gcgacaaatc ccagcctcac agtgggaggg 960
gcctgggcag gacctgaaac aaagtgtccc agtgtctgct gttcctgggc aggtggggct 1020
tgggggtccc ccacggttca agccttggct ctaggccttg gctgggcctt gtaatggcac 1080
ccgcgaccct cagttgcctc actcattggg gcttcgaagg cctctggtgc cccgagggtc 1140
cttcctgagg gcgacttgaa tacggcggtc ctatcctggg atggccggca ggtggcagcg 1200
tcagcccgtc tgctctccac cgggctcccc ggtagagtta taggggatta ggcggtgagt 1260
tttgccccaa agggaatcga atctagggct ccggtgcccg tcagtgggca gagcgcacat 1320
cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa 1380
ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg 1440
gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt 1500
ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg 1560
gttatggccc ttgcgtgcct tgaattactt ccactggctg cagtacgtga ttcttgatcc 1620
cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc 1680
gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg 1740
gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg 1800
acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca 1860
cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac 1920
atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca 1980
agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc 2040
ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc 2100
tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc 2160
cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta 2220
ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg 2280
ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt 2340
taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc 2400
ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc 2460
gtgacgtacg gtcgacgcca ccatgtcaag ctcttcctgg ctccttctca gccttgttgc 2520
tgtaactgct gctcagtcca ccattgagga acaggccaag acatttttgg acaagtttaa 2580
ccacgaagcc gaagacctgt tctatcaaag ttcacttgct tcttggaatt ataacaccaa 2640
tattactgaa gagaatgtcc aaaacatgaa taatgctggg gacaaatggt ctgccttttt 2700
aaaggaacag tccacacttg cccaaatgta tccactacaa gaaattcaga atctcacagt 2760
caagcttcag ctgcaggctc ttcagcaaaa tgggtcttca gtgctctcag aagacaagag 2820
caaacggttg aacacaattc taaatacaat gagcaccatc tacagtactg gaaaagtttg 2880
taacccagat aatccacaag aatgcttatt acttgaacca ggtttgaatg aaataatggc 2940
aaacagttta gactacaatg agaggctctg ggcttgggaa agctggagat ctgaggtcgg 3000
caagcagctg aggccattat atgaagagta tgtggtcttg aaaaatgaga tggcaagagc 3060
aaatcattat gaggactatg gggattattg gagaggagac tatgaagtaa atggggtaga 3120
tggctatgac tacagccgcg gccagttgat tgaagatgtg gaacatacct ttgaagagat 3180
taaaccatta tatgaacatc ttcatgccta tgtgagggca aagttgatga atgcctatcc 3240
ttcctatatc agtccaattg gatgcctccc tgctcatttg cttggtgata tgtggggtag 3300
attttggaca aatctgtact ctttgacagt tccctttgga cagaaaccaa acatagatgt 3360
tactgatgca atggtggacc aggcctggga tgcacagaga atattcaagg aggccgagaa 3420
gttctttgta tctgttggtc ttcctaatat gactcaagga ttctgggaaa attccatgct 3480
aacggaccca ggaaatgttc agaaagcagt ctgccatccc acagcttggg acctggggaa 3540
gggcgacttc aggatcctta tgtgcacaaa ggtgacaatg gacgacttcc tgacagctca 3600
tcatgagatg gggcatatcc agtatgatat ggcatatgct gcacaacctt ttctgctaag 3660
aaatggagct aatgaaggat tccatgaagc tgttggggaa atcatgtcac tttctgcagc 3720
cacacctaag catttaaaat ccattggtct tctgtcaccc gattttcaag aagacaatga 3780
aacagaaata aacttcctgc tcaaacaagc actcacgatt gttgggactc tgccatttac 3840
ttacatgtta gagaagtgga ggtggatggt ctttaaaggg gaaattccca aagaccagtg 3900
gatgaaaaag tggtgggaga tgaagcgaga gatagttggg gtggtggaac ctgtgcccca 3960
tgatgaaaca tactgtgacc ccgcatctct gttccatgtt tctaatgatt actcattcat 4020
tcgatattac acaaggaccc tttaccaatt ccagtttcaa gaagcacttt gtcaagcagc 4080
taaacatgaa ggccctctgc acaaatgtga catctcaaac tctacagaag ctggacagaa 4140
actgttcaat atgctgaggc ttggaaaatc agaaccctgg accctagcat tggaaaatgt 4200
tgtaggagca aagaacatga atgtaaggcc actgctcaac tactttgagc ccttatttac 4260
ctggctgaaa gaccagaaca agaattcttt tgtgggatgg agtaccgact ggagtccata 4320
tgcagaccaa agcatcaaag tgaggataag cctaaaatca gctcttggag ataaagcata 4380
tgaatggaac gacaatgaaa tgtacctgtt ccgatcatct gttgcatatg ctatgaggca 4440
gtacttttta aaagtaaaaa atcagatgat tctttttggg gaggaggatg tgcgagtggc 4500
taatttgaaa ccaagaatct cctttaattt ctttgtcact gcacctaaaa atgtgtctga 4560
tatcattcct agaactgaag ttgaaaaggc catcaggatg tcccggagcc gtatcaatga 4620
tgctttccgt ctgaatgaca acagcctaga gtttctgggg atacagccaa cacttggacc 4680
tcctaaccag ccccctgttt ccatatggct gattgttttt ggagttgtga tgggagtgat 4740
agtggttggc attgtcatcc tgatcttcac tgggatcaga gatcggaaga agaaaaataa 4800
agcaagaagt ggagaaaatc cttatgcctc catcgatatt agcaaaggag aaaataatcc 4860
aggattccaa aacactgatg atgttcagac ctccttttga gctagcatta tccctaatac 4920
ctgccacccc actcttaatc agtggtggaa gaacggtctc agaactgttt gtttcaattg 4980
gccatttaag tttagtagta aaagactggt taatgataac aatgcatcgt aaaaccttca 5040
gaaggaaagg agaatgtttt gtggaccact ttggttttct tttttgcgtg tggcagtttt 5100
aagttattag tttttaaaat cagtactttt taatggaaac aacttgacca aaaatttgtc 5160
acagaatttt gagacccatt aaaaaagtta aatgagaaac ctgtgtgttc ctttggtcaa 5220
caccgagaca tttaggtgaa agacatctaa ttctggtttt acgaatctgg aaacttcttg 5280
aaaatgtaat tcttgagtta acacttctgg gtggagaata gggttgtttt ccccccacat 5340
aattggaagg ggaaggaata tcatttaaag ctatgggagg gtttctttga ttacaacact 5400
ggagagaaat gcagcatgtt gctgattgcc tgtcactaaa acaggccaaa aactgagtcc 5460
ttgggttgca tagaaagctc ttctgaggcg gaaagaacca gctgccttaa tataacttcg 5520
tataatgtat gctatacgaa gttattaggt ctgaagagga gtttacgtcc agccaattct 5580
gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 5640
gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 5700
aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac 5760
tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 5820
aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta 5880
gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc 5940
cattttcggc ggccgcgcca ccatgaccga gtacaagccc acggtgcgcc tcgccacccg 6000
cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac 6060
gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt 6120
cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt 6180
ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc 6240
gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct 6300
ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga 6360
ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg 6420
cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg 6480
gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat 6540
gacccgcaag cccggtgcct gagaattcgc gggactctgg ggttcgaaat gaccgaccaa 6600
gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 6660
ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 6720
ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 6780
aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 6840
tccaaactca tcaatgtatc ttatcatgtc tgtataccgc tcgactagag cttgcggaac 6900
ccttaatata acttcgtata atgtatgcta tacgaagtta ttaggtccgc tggccatcta 6960
cgagccaaag actttcaaat ctttggctgc cttggccagt aggaggcgac acgaaggatt 7020
tgctgctgcc ttgggggatg ggaaggaacc tgaaggcatt ttttccagag tggtgcagta 7080
ccactgagga ctgttgctgt attgattagg aaaagagaca gagtaatttg cagtttgttt 7140
gatttatact tggtggttgc tgagactgcg tgggggccca aggagacctg gagaaaggaa 7200
tgcttcctgc tccttcttct ggggccccag gagagccttc ccagggcctt ggagaggtgc 7260
tgtccaggga ctaaccctgt gctctaggaa ggctgcaggc cctgaccagc tgggcaggtc 7320
ctgggtccct cctggccttc taagttcccc aaacatgaga cctctgggtg tggggtggcc 7380
tggggaggtc attttgccca ggccctacct cctgcccatt cctaaccctt tttaaaaatc 7440
tgtgcgtcct cttcttcctt cttctccctc ccttcccttt tcgctcaccc tctgctgctg 7500
gcctgagagc cggaggcccc cagggggaag gcgactggtc tcctccccag tctcagggaa 7560
gggagacaga gaatccagga agccagaact cagcagacga agcacccagg gacctagaga 7620
tgggttgaaa agttgacagc tgtcccacct gcctcccaag gtctcagggc ctaaacctcc 7680
aaggcaggaa aggcccctgt ccctccctgg ggtccataga aagagggaca agtctgcacg 7740
gaccatttgc tgtaatatta acaccttggc tgtcattagg tagtcttggc tgttaattat 7800
gtcctgtgat aatgtattat tagcacgccg accacatagg gtagggaact gcagctagta 7860
aacaaaagtt tgttcctata tgcggccgcc ataaaagttt tgttacttta tagaagaaat 7920
tttgagtttt tgtttttttt aataaataaa taaacataaa taaattgttt gttgaattta 7980
ttattagtat gtaagtgtaa atataataaa acttaatatc tattcaaatt aataaataaa 8040
cctcgatata cagaccgata aaacacatgc gtcaatttta cacatgatta tctttaacgt 8100
acgtcacaat atgattatct ttctagggtt aatctagctg cgtgttctgc agcgtgtcga 8160
gcatcttcat ctgctccatc acgctgtaaa acacatttgc accgcgagtc tgcccgtcct 8220
ccacgggttc aaaaacgtga atgaacgagg cgcgctcact ggccgtcgtt ttacaacgtc 8280
gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg 8340
ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc 8400
tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 8460
cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 8520
cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 8580
tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg 8640
gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 8700
cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct 8760
attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga 8820
tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac 8880
ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 8940
gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 9000
tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc 9060
tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc 9120
acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc 9180
cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc 9240
ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt 9300
ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt 9360
atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat 9420
cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct 9480
tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat 9540
gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc 9600
ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg 9660
ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggttc 9720
acgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta 9780
cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc 9840
ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga 9900
tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 9960
gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 10020
caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 10080
accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 10140
ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt 10200
aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 10260
accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 10320
gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 10380
ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 10440
gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 10500
gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 10560
ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 10620
aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 10680
gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc 10740
tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga 10800
agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg 10860
gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta 10920
gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg 10980
aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg 11040
cgcccgccgg gtaactcacg gggtatccat gtccatttct gcggcatcca gccaggatac 11100
ccgtcctcgc tgacgtaata tcccagcgcc gcaccgctgt cattaatctg cacaccggca 11160
cggcagttcc ggctgtcgcc ggtattgttc gggttgctga tgcgcttcgg gctgaccatc 11220
cggaactgtg tccggaaaag ccgcgacgaa ctggtatccc aggtggcctg aacgaacagt 11280
tcaccgttaa aggcgtgcat ggccacacct tcccgaatca tcatggtaaa cgtgcgtttt 11340
cgctcaacgt caatgcagca gcagtcatcc tcggcaaact ctttccatgc cgcttcaacc 11400
tcgcgggaaa aggcacgggc ttcttcctcc ccgatgccca gatagcgcca gcttgggcga 11460
tgactgagcc ggaaaaaaga cccgacgata tgatcctgat gcagctagat taaccctaga 11520
aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat attgctctct ctttctaaat 11580
agcgcgaatc cgtcgctgtg catttaggac atctcagtcg ccgcttggag ctcccgtgag 11640
gcgtgcttgt caatgcggta agtgtcactg attttgaact ataacgaccg cgtgagtcaa 11700
aatgacgcat gattatcttt tacgtgactt ttaagattta actcatacga taattatatt 11760
gttatttcat gttctactta cgtgataact tattatatat atattttctt gttatagata 11820
tc 11822
<210> 12
<211> 675
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 12
Met Asp Thr Lys Ser Ile Leu Glu Glu Leu Leu Leu Lys Arg Ser Gln
1 5 10 15
Gln Lys Lys Lys Met Ser Pro Asn Asn Tyr Lys Glu Arg Leu Phe Val
20 25 30
Leu Thr Lys Thr Asn Leu Ser Tyr Tyr Glu Tyr Asp Lys Met Lys Arg
35 40 45
Gly Ser Arg Lys Gly Ser Ile Glu Ile Lys Lys Ile Arg Cys Val Glu
50 55 60
Lys Val Asn Leu Glu Glu Gln Thr Pro Val Glu Arg Gln Tyr Pro Phe
65 70 75 80
Gln Ile Val Tyr Lys Asp Gly Leu Leu Tyr Val Tyr Ala Ser Asn Glu
85 90 95
Glu Ser Arg Ser Gln Trp Leu Lys Ala Leu Gln Lys Glu Ile Arg Gly
100 105 110
Asn Pro His Leu Leu Val Lys Tyr His Ser Gly Phe Phe Val Asp Gly
115 120 125
Lys Phe Leu Cys Cys Gln Gln Ser Cys Lys Ala Ala Pro Gly Cys Thr
130 135 140
Leu Trp Glu Ala Tyr Ala Asn Leu His Thr Ala Val Asn Glu Glu Lys
145 150 155 160
His Arg Val Pro Thr Phe Pro Asp Arg Val Leu Lys Ile Pro Arg Ala
165 170 175
Val Pro Val Leu Lys Met Asp Ala Pro Ser Ser Ser Thr Thr Leu Ala
180 185 190
Gln Tyr Asp Asn Glu Ser Lys Lys Asn Tyr Gly Ser Gln Pro Pro Ser
195 200 205
Ser Ser Thr Ser Leu Ala Gln Tyr Asp Ser Asn Ser Lys Lys Ile Tyr
210 215 220
Gly Ser Gln Pro Asn Phe Asn Met Gln Tyr Ile Pro Arg Glu Asp Phe
225 230 235 240
Pro Asp Trp Trp Gln Val Arg Lys Leu Lys Ser Ser Ser Ser Ser Glu
245 250 255
Asp Val Ala Ser Ser Asn Gln Lys Glu Arg Asn Val Asn His Thr Thr
260 265 270
Ser Lys Ile Ser Trp Glu Phe Pro Glu Ser Ser Ser Ser Glu Glu Glu
275 280 285
Glu Asn Leu Asp Asp Tyr Asp Trp Phe Ala Gly Asn Ile Ser Arg Ser
290 295 300
Gln Ser Glu Gln Leu Leu Arg Gln Lys Gly Lys Glu Gly Ala Phe Met
305 310 315 320
Val Arg Asn Ser Ser Gln Val Gly Met Tyr Thr Val Ser Leu Phe Ser
325 330 335
Lys Ala Val Asn Asp Lys Lys Gly Thr Val Lys His Tyr His Val His
340 345 350
Thr Asn Ala Glu Asn Lys Leu Tyr Leu Ala Glu Asn Tyr Cys Phe Asp
355 360 365
Ser Ile Pro Lys Leu Ile His Tyr His Gln His Asn Ser Ala Gly Met
370 375 380
Ile Thr Arg Leu Arg His Pro Val Ser Thr Lys Ala Asn Lys Val Pro
385 390 395 400
Asp Ser Val Ser Leu Gly Asn Gly Ile Trp Glu Leu Lys Arg Glu Glu
405 410 415
Ile Thr Leu Leu Lys Glu Leu Gly Ser Gly Gln Phe Gly Val Val Gln
420 425 430
Leu Gly Lys Trp Lys Gly Gln Tyr Asp Val Ala Val Lys Met Ile Lys
435 440 445
Glu Gly Ser Met Ser Glu Asp Glu Phe Phe Gln Glu Ala Gln Thr Met
450 455 460
Met Lys Leu Ser His Pro Lys Leu Val Lys Phe Tyr Gly Val Cys Ser
465 470 475 480
Lys Glu Tyr Pro Ile Tyr Ile Val Thr Glu Tyr Ile Ser Asn Gly Cys
485 490 495
Leu Leu Asn Tyr Leu Arg Ser His Gly Lys Gly Leu Glu Pro Ser Gln
500 505 510
Leu Leu Glu Met Cys Tyr Asp Val Cys Glu Gly Met Ala Phe Leu Glu
515 520 525
Ser His Gln Phe Ile His Arg Asp Leu Ala Ala Arg Asn Cys Leu Val
530 535 540
Asp Arg Asp Leu Cys Val Lys Val Ser Asp Phe Gly Met Thr Arg Tyr
545 550 555 560
Val Leu Asp Asp Gln Tyr Val Ser Ser Val Gly Thr Lys Phe Pro Val
565 570 575
Lys Trp Ser Ala Pro Glu Val Phe His Tyr Phe Lys Tyr Ser Ser Lys
580 585 590
Ser Asp Val Trp Ala Phe Gly Ile Leu Met Trp Glu Val Phe Ser Leu
595 600 605
Gly Lys Gln Pro Tyr Asp Leu Tyr Asp Asn Ser Gln Val Val Leu Lys
610 615 620
Val Ser Gln Gly His Arg Leu Tyr Arg Pro His Leu Ala Ser Asp Thr
625 630 635 640
Ile Tyr Gln Ile Met Tyr Ser Cys Trp His Glu Leu Pro Glu Lys Arg
645 650 655
Pro Thr Phe Gln Gln Leu Leu Ser Ser Ile Glu Pro Leu Arg Glu Lys
660 665 670
Asp Lys His
675
<210> 13
<211> 2415
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
atgtcaagct cttcctggct ccttctcagc cttgttgctg taactgctgc tcagtccacc 60
attgaggaac aggccaagac atttttggac aagtttaacc acgaagccga agacctgttc 120
tatcaaagtt cacttgcttc ttggaattat aacaccaata ttactgaaga gaatgtccaa 180
aacatgaata atgctgggga caaatggtct gcctttttaa aggaacagtc cacacttgcc 240
caaatgtatc cactacaaga aattcagaat ctcacagtca agcttcagct gcaggctctt 300
cagcaaaatg ggtcttcagt gctctcagaa gacaagagca aacggttgaa cacaattcta 360
aatacaatga gcaccatcta cagtactgga aaagtttgta acccagataa tccacaagaa 420
tgcttattac ttgaaccagg tttgaatgaa ataatggcaa acagtttaga ctacaatgag 480
aggctctggg cttgggaaag ctggagatct gaggtcggca agcagctgag gccattatat 540
gaagagtatg tggtcttgaa aaatgagatg gcaagagcaa atcattatga ggactatggg 600
gattattgga gaggagacta tgaagtaaat ggggtagatg gctatgacta cagccgcggc 660
cagttgattg aagatgtgga acataccttt gaagagatta aaccattata tgaacatctt 720
catgcctatg tgagggcaaa gttgatgaat gcctatcctt cctatatcag tccaattgga 780
tgcctccctg ctcatttgct tggtgatatg tggggtagat tttggacaaa tctgtactct 840
ttgacagttc cctttggaca gaaaccaaac atagatgtta ctgatgcaat ggtggaccag 900
gcctgggatg cacagagaat attcaaggag gccgagaagt tctttgtatc tgttggtctt 960
cctaatatga ctcaaggatt ctgggaaaat tccatgctaa cggacccagg aaatgttcag 1020
aaagcagtct gccatcccac agcttgggac ctggggaagg gcgacttcag gatccttatg 1080
tgcacaaagg tgacaatgga cgacttcctg acagctcatc atgagatggg gcatatccag 1140
tatgatatgg catatgctgc acaacctttt ctgctaagaa atggagctaa tgaaggattc 1200
catgaagctg ttggggaaat catgtcactt tctgcagcca cacctaagca tttaaaatcc 1260
attggtcttc tgtcacccga ttttcaagaa gacaatgaaa cagaaataaa cttcctgctc 1320
aaacaagcac tcacgattgt tgggactctg ccatttactt acatgttaga gaagtggagg 1380
tggatggtct ttaaagggga aattcccaaa gaccagtgga tgaaaaagtg gtgggagatg 1440
aagcgagaga tagttggggt ggtggaacct gtgccccatg atgaaacata ctgtgacccc 1500
gcatctctgt tccatgtttc taatgattac tcattcattc gatattacac aaggaccctt 1560
taccaattcc agtttcaaga agcactttgt caagcagcta aacatgaagg ccctctgcac 1620
aaatgtgaca tctcaaactc tacagaagct ggacagaaac tgttcaatat gctgaggctt 1680
ggaaaatcag aaccctggac cctagcattg gaaaatgttg taggagcaaa gaacatgaat 1740
gtaaggccac tgctcaacta ctttgagccc ttatttacct ggctgaaaga ccagaacaag 1800
aattcttttg tgggatggag taccgactgg agtccatatg cagaccaaag catcaaagtg 1860
aggataagcc taaaatcagc tcttggagat aaagcatatg aatggaacga caatgaaatg 1920
tacctgttcc gatcatctgt tgcatatgct atgaggcagt actttttaaa agtaaaaaat 1980
cagatgattc tttttgggga ggaggatgtg cgagtggcta atttgaaacc aagaatctcc 2040
tttaatttct ttgtcactgc acctaaaaat gtgtctgata tcattcctag aactgaagtt 2100
gaaaaggcca tcaggatgtc ccggagccgt atcaatgatg ctttccgtct gaatgacaac 2160
agcctagagt ttctggggat acagccaaca cttggacctc ctaaccagcc ccctgtttcc 2220
atatggctga ttgtttttgg agttgtgatg ggagtgatag tggttggcat tgtcatcctg 2280
atcttcactg ggatcagaga tcggaagaag aaaaataaag caagaagtgg agaaaatcct 2340
tatgcctcca tcgatattag caaaggagaa aataatccag gattccaaaa cactgatgat 2400
gttcagacct ccttt 2415
<210> 14
<211> 1101
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
aataaatgca ctgttgggcc tatgctcaag atgggtagtg ttaattggtg gtggaactta 60
tctgatttca tgacttgctg gctacctaaa acaggtgagg agaaagccaa tgggactggg 120
actggatgag caagtacaac aaacaaaatg ggcttaaagt atgagtgaga gttatctgac 180
cgtaaggatg caagtgaggg ggcctaaggt ttggagatta atatttaatc tcagatgcta 240
tactttggtg gtgtagcaaa agtctacaaa tgggatgact gtaaaactca gtagatccgt 300
gctttttaac ctatctccct tcatcaggaa attgcgacac aaagatcttt agtaataaca 360
cgcagtctca atgcataaaa tcaggcttag gtgttgcctg gactcatttc ccatctccac 420
cccactataa ttattttgtg acacaaactc aagactgtgg gaatatagag aaattgggct 480
cgtcctcgta cacctgctca atcccctgca ggacaacgcc caagaatcag gttaagccag 540
ggcaaaagaa tcccgcccat aatcgagaag gagcaaactg acatggaggc gatgacgaga 600
tcgcggggga gggagggatt tttctaggcc cagggcggtc cttaggaaaa ggaggcagca 660
gagaactccc ataaaggtat tgcggcactc ccctccccct gcggagaagg gtgcggcctt 720
ctctccgcct cctccactgc agctccctca ggattgcagc tcgcgcgggt ttttggagaa 780
catgcgcctc ccacccacaa gccagcagga ccgacccccc actccttcct ccacccccca 840
cccccacggg tccgagagca ggtagagagc tagtctcgtc cttcaggcgg cggacgccca 900
gggcggagcc gcagtcacca ccacccagaa gcctcggccc ggcagcccgc ccccgcctcc 960
tgcgcgcgct tcctgccacg ttgcgcaggg gcgaggggcc agacactgcg gcgctggcct 1020
cggggagggc cgtaccaaag accgcctccc tgccgactcg cgtagtggtt tcgctcattt 1080
gggacccaag ccaataacaa g 1101
<210> 15
<211> 1056
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
tgctctctct cctgccccct tcacctgcgt gccctcctca ttctccctct gtgccacctc 60
tggccttgca ctgtaggctc tctcttgggg atgtttctct ttctccacac acttctcttt 120
cactctgtcc tcttgctttg tgtgggcctg cagcgttacc cttttttctg ggcacactca 180
gagcaccctc ctctttctgg ttctgggcca cctgtctgtc ctcgggtcat cttgctctct 240
ctgcctggat gccctcctgt ggctttgggc agcttctccc tccttcagag tgcaccgcca 300
gttctcctag gcccggtcac ttccccttcc caggggacct agagccctgc taggtcctct 360
ctctccacaa cctgggcccc caaacctttc caaaacacct tgctttctgc ctccattggt 420
cttgtgttcc agagccagag tcactatatg tcccagaacc aggattccct ctggttctga 480
gggcttttat cgcatcccct gcctggctgc agtgggtctt tggggacagg ccacagaaga 540
gcctctactc ctccctctgt ccccgaggct gtctccctcc cagtcttccc agctcaggcc 600
agtccccagg cctctcttcc ctgccagagc ccgtcaggtt cggttacttt ggggcccaga 660
gaggaccctg tgaaggaagc gtgggtaggg gcacgggaat ggggaggatg cctgaagagg 720
cccccttagc cagaagagga gcagaagagg agcaggtacc cagaagagga gcagttcagg 780
gaaatagaag agtcccgagc tctttttttt tttttttttt atttcttttc ttttcttttc 840
tttttatggc agcatccgtg gtatatggag gttcccagcc taggggtcag atcatacctg 900
caactgccag cctacaccac agccacagca ctcaggatcc gagctgcatc tgcggcttac 960
gccacaggtc acagcaacgc tggatcctta acccactgaa tgaggccagg gattgaacct 1020
gcaacctcat gcacactatg ctggggtctt aatcgg 1056
<210> 16
<211> 1108
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
acttcctcct gcccttaccc tttatctggc tcttagctcc taaaaactgc attattagct 60
tcctcttttg cctctactct tactcaacca aaattgtttt aagatctgtg gatctagctt 120
ctgctgtgct attcttagga acacttttat ttcctcttag ctccatctca ccagttattg 180
gctaatggct ttgcttggta cctacatctg tacatttctt tcgtactagc ttctagactg 240
aaaaaggact gttggttcaa catgaaaggg aaggaggtaa aagaggacac acaggaaaga 300
tggattggga ttcaggtctc tgctgttgtt acttgagatt gctttctaga ttctacttgt 360
ggaaacaaaa agcctttgcg agaattctaa actggagtat ttctgtaatt gaggagtctt 420
gctcagcaaa tcccacttag gggactaatg aagtaccagg aagagacaga ccatgctcaa 480
tccacaaagc caggttttac tgaaatgtga cctactttct tatgttcctg gaagtttaga 540
tcagggtggg cagctctggg ttttataggc tacactgtta acactcaggc tgttttctac 600
cgtttagtca aaatatagtc accttgcctg cttcacctgt ccatcagaga atggcctcat 660
taattgactc tctagtatga agtcaaagta gctttggtgg ccctaaatgg acaagtatca 720
agagactggg tgaattgagg agcttgagac tgtcacctca gatcgaaaag actgaaaaat 780
cacctcagat caaaaagact gaaaaatctt cagtctggaa aggggactca aaaccataat 840
tagagtattc tggtagaatc cttttctcca ctgttattca tacagttaag gtgaataact 900
aaaagtaatt gtgagctgag gagtaagata caacacacaa ggaatcagtt aacagagtct 960
cgagtgaaat tataaatgga aagaattatg acttgaatca taactctgag gccccatttt 1020
ccctaacaac ttttgtccca ataaacgtgg gtatttgttt gggagaaact atcatataca 1080
tgattaccca gtaaacagac tgtttact 1108
<210> 17
<211> 1288
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 17
actttgtacc tattttgtat gtgtataata atttgagatg tttttaatta ttttgattgc 60
tggaataaag catgtggaaa tgacccaaac caatcttgca ctggcctcct gatttccttc 120
cttggagacg gagggagggg gagacctggg ggagggcgct tggggggggg tgggctctct 180
tctttctgcg ctcccccccc ccacctccaa caccttgacg acccctcctg cttccgcttg 240
cctttctcag gctttaacac tttctcctcg ccctctcagc atgcgcatgc gcgtgcctct 300
acctcccccg cacatcctgg cctgcccacc ctgaatgtcc tggcccagcg atgccaccaa 360
ctctctcgct ccgtccacgg ctggggaggg gggcactctg cagggttggg gggcactggg 420
aggctgggtt gggtgaggga ggggtgcctg ggcccccacc ccccagcaag ttctctccct 480
aggcgaactg gagggtcgtc tggcctcttg agccttgttg ctggctctga gctctaccaa 540
gagagtgacc agcaggaccg caccatcagt ggttgctgag actgcgtggg ggcccaagga 600
gacctggaga aaggaatgct tcctgctcct tcttctgggg ccccaggaga gccttcccag 660
ggccttggag agttgctgtc cagggactaa ccctgtgctc taggaaggct gcaggccctg 720
accagctggg caggtcctgg gtccctcctg gccttctaag ttccccaaac atgagacctc 780
tgggtgtggg gtggcctggg gaggtcattt tgcccaggcc ctacctcctg cccattccta 840
acccttttta aaaatctgtg cgtcctcttc ttccttcttc tccctccctt cccttttcgc 900
tcaccctctg ctgctggcct gagagccgga ggcccccagg gggaaggcga ctggtctcct 960
ccccagtctc agggaaggga gacagagaat ccaggaagcc agaactcagc agacgaagca 1020
cccagggacc tagagatggg ttgaaaagtt gacagctgtc ccacctgcct cccaaggtct 1080
cagggcctac acccttctcc gcagggggag gggagtgccg caataccttt atgggagttc 1140
tctgctgcct ccttttccta aggaccgccc tgggcctaga aaaatccctc cctcccccgc 1200
gatctcgtca tcgcctccat gtcagtttgc tccttctcga ttatgggcgg gattcttttg 1260
ccctggcgcg ccccagaccc gggcctgg 1288
<210> 18
<211> 345
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 18
ggcgcgccct ctacctgctc tcggacccgt gggggtgggg ggtggaggaa ggagtggggg 60
gtcggtcctg ctggcttgtg ggtgggaggc gcatgttctc caaaaacccg cgcgagctgc 120
aatcctgagg gagctgcagt ggaggaggcg gagagaaggc cgcacccttc tccgcagggg 180
gaggggagtg ccgcaatacc tttatgggag ttctctgctg cctccttttc ctaaggaccg 240
ccctgggcct agaaaaatcc ctccctcccc cgcgatctcg tcatcgcctc catgtcagtt 300
tgctccttct cgattatggg cgggattctt ttgccctggc gcgcc 345
<210> 19
<211> 1012
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 19
cttaacctga ttcttgggcg ttgtcctgca ggggattgag caggtgtacg aggacgagcc 60
caatttctct atattcccac agtcttgagt ttgtgtcaca aaataattat agtggggtgg 120
agatgggaaa tgagtccagg caacacctaa gcctgatttt atgcattgag actgcgtgtt 180
attactaaag atctttgtgt cgcaatttcc tgatgaaggg agataggtta aaaagcacgg 240
atctactgag ttttacagtc atcccatttg tagacttttg ctacaccacc aaagtatagc 300
atctgagatt aaatattaat ctccaaacct taggccccct cacttgcatc cttacggtca 360
gataactctc actcatactt taagcccatt ttgtttgttg tacttgctca tccagtccca 420
gacatagcat tggctttctc ctcacctgtt ttaggtagcc agcaagtcat gaaatcagat 480
aagttccacc accaattaac actacccatc ttgagcatag gcccaacagt gcatttattc 540
ctcatttact gatgttcgtg aatatttacc ttgattttca tttttttctt tttcttaagc 600
tgggatttta ctcctgaccc tattcacagt cagatgatct tgactaccac tgcgattgga 660
cctgaggttc agcaatactc ccctttatgt cttttgaata cttttcaata aatctgtttg 720
tattttcatt agttagtaac tgagctcagt tgccgtaatg ctaatagctt ccaaactagt 780
gtctctgtct ccagtatctg ataaatctta ggtgttgctg ggacagttgt cctaaaatta 840
agataaagca tgaaaataac tgacacaact ccattactgg ctcctaacta cttaaacaat 900
gcattctatc atcacaaatg tgaaaaagga gttccctcag tggactaacc ttatcttttc 960
tcaacacctt tttctttgca caattttcca cacatgccta caaaaagtac tt 1012
<210> 20
<211> 14114
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 20
gtgctgagtc cttttcccat cccacccacc tggagctccc ctcttccagt cctgagccac 60
ttgaactggc ctggtttttg ccatcctgcg ctgccctctc tccggactcg agccactgct 120
gagggcctca ggccagtcca tcctcgtctt gtctctttcg ccctgctctt tccccacctt 180
gagcgctctt aaccagcctg gcccgtgcca cctctactct gccatcgaat gctgccccac 240
tttctcgagt ccgccacttc tcccagcttc accggtaccc actgtttccc ctagtccagg 300
caggtaccac tttccctgag cgtcctcctc ctctctcctg ggcctgtgct gcttcttttc 360
ccgctctctg gcctgggccg tttcttcggc cagcccccga gccttccatg ccctttcctt 420
caggtttctg ctcttcatcc ttggtctctg ccatctgttg ccatgtaagg gtgctctttc 480
ctgagccatc gccctcaagg cgctctgctc ctcaagtgga tgcttccctc gcctggctca 540
cctcctgctc tctctcctgc ccccttcacc tgcgtgccct cctcattctc cctctgtgcc 600
acctctggcc ttgcactgta ggctctctct tggggatgtt tctccttctc cacacacttc 660
tctttcactc tgtcctcttg ctttgtgtgg gcctgcagcg ttaccctttt ttctgggcac 720
actcagagca ccctcctctt tctggttctg ggccacctgt ctgtcctcgg gtcatcttgc 780
tctctctgcc tggatgccct cctgtggctt tgggcagctt ctccctcctt cagagtgcac 840
cgccagttct cctaggcccg gtcacttccc cttcccaggg gacctagagc cctgctaggt 900
cctctctctc cacaacctgg gcccccaaac ctttccaaaa caccttgctt tctgcctcca 960
ttggtcttgt gttccagagc cagagtcact atatgtccca gaaccaggat tccctctggt 1020
tctgagggct tttatcgcat cccctgcctg gctgcagtgg gtctttgggc gccccagacc 1080
cgggcctggg gggcaagtcg gggggcgggg ggaggtcggg cagggtcccc tgggaggatg 1140
gggacgtgct gtgcccctag cggccaccag agggcaccag gacaccactg cggtcggctc 1200
agcggctcct gccctggtca gggggcgcca ggtcctgccc ctcctgggga gggcgggggg 1260
cgagaagggc gattttaatt aacccacgtt tcaacatgca catcccagta atttggaaac 1320
attttgtttc caaagattca cttaacattg gtttagcaac atgaagcttt ctatgcaacc 1380
caaggactca gtttttggcc tgttttagtg acaggcaatc agcaacatgc tgcatttctc 1440
tccagtgttg taatcaaaga aaccctccca tagctttaaa tgatattcct tccccttcca 1500
attatgtggg gggaaaacaa ccctattctc cacccagaag tgttaactca agaattacat 1560
tttcaagaag tttccagatt cgtaaaacca gaattagatg tctttcacct aaatgtctcg 1620
gtgttgacca aaggaacaca caggtttctc atttaacttt tttaatgggt ctcaaaattc 1680
tgtgacaaat ttttggtcaa gttgtttcca ttaaaaagta ctgattttaa aaactaataa 1740
cttaaaactg ccacacgcaa aaaagaaaac caaagtggtc cacaaaacat tctcctttcc 1800
ttctgaaggt tttacgatgc attgttatca ttaaccagtc ttttactact aaacttaaat 1860
ggccaattga aacaaacagt tctgagaccg ttcttccacc actgattaag agtggggtgg 1920
caggtattag ggataatgct agcttacttg tacagctcgt ccatgccgag agtgatcccg 1980
gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc 2040
agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg 2100
ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg 2160
atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg 2220
ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg 2280
cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc 2340
ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg 2400
gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg 2460
ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag 2520
atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg 2580
aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac 2640
agctcctcgc ccttgctcac catggtggcg tcgaccgtac gtcacgacac ctgaaatgga 2700
agaaaaaaac tttgaaccac tgtctgaggc ttgagaatga accaagatcc aaactcaaaa 2760
agggcaaatt ccaaggagaa ttacatcaag tgccaagctg gcctaacttc agtctccacc 2820
cactcagtgt ggggaaactc catcgcataa aacccctccc cccaacctaa agacgacgta 2880
ctccaaaagc tcgagaacta atcgaggtgc ctggacggcg cccggtactc cgtggagtca 2940
catgaagcga cggctgagga cggaaaggcc cttttccttt gtgtgggtga ctcacccgcc 3000
cgctctcccg agcgccgcgt cctccatttt gagctccctg cagcagggcc gggaagcggc 3060
catctttccg ctcacgcaac tggtgccgac cgggccagcc ttgccgccca gggcggggcg 3120
atacacggcg gcgcgaggcc aggcaccaga gcaggccggc cagcttgaga ctacccccgt 3180
ccgattctcg gtggccgcgc tcgcaggccc cgcctcgccg aacatgtgcg ctgggacgca 3240
cgggccccgt cgccgcccgc ggccccaaaa accgaaatac cagtgtgcag atcttggccc 3300
gcatttacaa gactatcttg ccagaaaaaa agcgtcgcag caggtcatca aaaattttaa 3360
atggctagag acttatcgaa agcagcgaga caggcgcgaa ggtgccacca gattcgcacg 3420
cggcggcccc agcgcccagg ccaggcctca actcaagcac gaggcgaagg ggctccttaa 3480
gcgcaaggcc tcgaactctc ccacccactt ccaacccgaa gctcgggatc aagaatcacg 3540
tactgcagcc agtggaagta attcaaggca cgcaagggcc ataacccgta aagaggccag 3600
gcccgcggga accacacacg gcacttacct gtgttctggc ggcaaacccg ttgcgaaaaa 3660
gaacgttcac ggcgactact gcacttatat acggttctcc cccaccctcg ggaaaaaggc 3720
ggagccagta cacgacatca ctttcccagt ttaccccgcg ccaccttctc taggcaccgg 3780
ttcaattgcc gacccctccc cccaacttct cggggactgt gggcgatgtg cgctctgccc 3840
actgacgggc accggagccc tagattcgat tccctttggg gcaaaactca ccgcctaatc 3900
ccctataact ctaccgggga gcccggtgga gagcagacgg gctgacgctg ccacctgccg 3960
gccatcccag gataggaccg ccgtattcaa gtcgccctca ggaaggaccc tcggggcacc 4020
agaggccttc gaagccccaa tgagtgaggc aactgagggt cgcgggtgcc attacaaggc 4080
ccagccaagg cctagagcca aggcttgaac cgtgggggac ccccaagccc cacctgccca 4140
ggaacagcag acactgggac actttgtttc aggtcctgcc caggcccctc ccactgtgag 4200
gctgggattt gtcgcccagg gtgcagatga gaagagtggg gaaagcagtc ctgagccagg 4260
aaattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca 4320
gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc 4380
accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct 4440
cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg 4500
gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg 4560
gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg 4620
ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc 4680
cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg 4740
ttctcctctt cctcatctcc gggcctttcg acctcctagg gccaccatgg tgagcaaggg 4800
cgaggacgac aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg 4860
ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg 4920
cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat 4980
cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc 5040
cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga 5100
ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta 5160
caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac 5220
catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga 5280
gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgccgagg tcaagaccac 5340
ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagctgga 5400
catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgcg ccgagggccg 5460
ccactccacc ggcggcatgg acgagctgta caagtgagga tccgctgatc agcctcgact 5520
gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 5580
gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 5640
agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 5700
gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga 5760
acccttctga ggcggaaaga accagctgcc ttaatataac ttcgtataat gtatgctata 5820
cgaagttatt aggtctgaag aggagtttac gtccagccaa ttctgtggaa tgtgtgtcag 5880
ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 5940
aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 6000
agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 6060
ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 6120
gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt 6180
ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggcggccgc 6240
gccaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg 6300
gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat 6360
ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg 6420
ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg 6480
ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg 6540
agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc 6600
aaggagcccg cgtggttcct ggccaccgtc ggagtctcgc ccgaccacca gggcaagggt 6660
ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc 6720
ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc 6780
accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt 6840
gcctgagaat tcgcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc 6900
catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt 6960
tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc 7020
accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 7080
tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 7140
tatcttatca tgtctgtata ccgctcgact agagcttgcg gaacccttaa tataacttcg 7200
tataatgtat gctatacgaa gttattaggt ccgctggcca tctacgagcc aaagactttc 7260
aaatctttgg ctgccttggc cagtaggagg cgacacgaag gatttgctgc tgccttgggg 7320
gatgggaagg aacctgaagg cattttttcc agagtggtgc agtaccactg aggactgttg 7380
ctgtattgat taggaaaaga gacagagtaa tttgcagttt gtttgattta tactgggctg 7440
caggtcgagg gatcttcata agagaagagg gacagctatg actgggagta gtcaggagag 7500
gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa 7560
taacacaaaa caaaatataa aaaaaatcta acctcaagtc aaggcttttc tatggaataa 7620
ggaatggaca gcagggggct gtttcatata ctgatgacct ctttatagcc acctttgttc 7680
atggcagcca gcatatggca tatgttgcca aactctaaac caaatactca ttctgatgtt 7740
ttaaatgatt tgccctccca tatgtccttc cgagtgagag acacaaaaaa ttccaacaca 7800
ctattgcaat gaaaataaat ttcctttatt agccagaagt cagatgctca aggggcttca 7860
tgatgtcccc ataatttttg gcagagggaa aaagatctca gtggtatttg tgagccaggg 7920
cattggccac accagccacc accttctgat aggcagcctg cggtacctta catggtggcg 7980
aattcgtttg ccaaaatgat gagacagcac aataaccagc acgttgccca ggagctgtag 8040
gaaaaagaag aaggcatgaa catggttagc agaggctcta gagccgccgg tcacacgcca 8100
gaagccgaac cccgccctgc cccgtccccc ccgaaggcag ccgtccccct gcggcagccc 8160
cgaggctgga gatggagaag gggacggcgg cgcggcgacg cacgaaggcc ctccccgccc 8220
atttccttcc tgccggcgcc gcaccgcttc gcccgcgccc gctagagggg gtgcggcggc 8280
gcctcccaga tttcggctcc gccagatttg ggacaaagga agtccctgcg ccctctcgca 8340
cgattaccat aaaaggcaat ggctgcggct cgccgcgcct cgacagccgc cggcgctccg 8400
gggccgccgc gcccctcccc cgagccctcc ccggcccgag gcggccccgc cccgcccggc 8460
acccccacct gccgccaccc cccgcccggc acggcgagcc ccgcgccacg ccccgcacgg 8520
agccccgcac ccgaagccgg gccgtgctca gcaactcggg gaggggggtg cagggggggg 8580
ttacagcccg accgccgcgc ccacaccccc tgctcacccc cccacgcaca caccccgcac 8640
gcagcctttg ttcccctcgc agcccccccg caccgcgggg caccgccccc ggccgcgctc 8700
ccctcgcgca cacgcggagc gcacaaagcc ccgcgccgcg cccgcagcgc tcacagccgc 8760
cgggcagcgc gggccgcacg cggcgctccc cacgcacaca cacacgcacg caccccccga 8820
gccgctcccc cccgcacaaa gggccctccc ggagcccttt aaggctttca cgcagccaca 8880
gaaaagaaac gagccgtcat taaaccaagc gctaattaca gcccggagga gaagggccgt 8940
cccgcccgct cacctgtggg agtaacgcgg tcagtcagag ccggggcggg cggcgcgagg 9000
cggcgcggag cggggcacgg ggcgaaggca acgcagcgac tcccgcccgc cgcgcgcttc 9060
gctttttata gggccgccgc cgccgccgcc tcgccataaa aggaaacttt cggagcgcgc 9120
cgctctgatt ggctgccgcc gcacctctcc gcctcgcccc gccccgcccc tcgccccgcc 9180
ccgccccgcc tggcgcgcgc cccccccccc cccgccccca tcgctgcaca aaataattaa 9240
aaaataaata aatacaaaat tgggggtggg gagggggggg agatggggag agtgaagcag 9300
aacgtggggc tcacctcgac ccatggtaat agcgatgact aatacgtaga tgtactgcca 9360
agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc 9420
attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc 9480
agtttaccgt aaatagtcca cccattgacg tcaatggaaa gtccctattg gcgttactat 9540
gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg 9600
gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc 9660
cgtaattgat tactattaat aactagtcaa taatcaatgt cgtaaatgtc gtaaatgtct 9720
cagctagtca ggtagtaaaa ggtgtcaact aggcagtggc agagcaggat tcaaattcag 9780
ggctgttgtg atgcctccgc agactctgag cgccacctgg tggtaatttg tctgtgcctc 9840
ttctgacgtg gaagaacagc aactaacaca ctaacacggc atttactatg ggccagccat 9900
tgtacgcgtt ggacaggcca cagaagagcc tctactcctc cctctgtccc cgaggctgtc 9960
tccctcccag tcttcccagc tcaggccagt ccccaggcct ctcttccctg ccagagcccg 10020
tcaggttcgg ttactttggg gcccagagag gaccctgtga aggaagcgtg ggtaggggca 10080
cgggaatggg gaggatgcct gaagaggccc ccttagccag aagaggagca gaagaggagc 10140
aggtacccag aagaggagca gttcagggaa aatgcggccg ccataaaagt tttgttactt 10200
tatagaagaa attttgagtt tttgtttttt ttaataaata aataaacata aataaattgt 10260
ttgttgaatt tattattagt atgtaagtgt aaatataata aaacttaata tctattcaaa 10320
ttaataaata aacctcgata tacagaccga taaaacacat gcgtcaattt tacacatgat 10380
tatctttaac gtacgtcaca atatgattat ctttctaggg ttaatctagc tgcgtgttct 10440
gcagcgtgtc gagcatcttc atctgctcca tcacgctgta aaacacattt gcaccgcgag 10500
tctgcccgtc ctccacgggt tcaaaaacgt gaatgaacga ggcgcgctca ctggccgtcg 10560
ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac 10620
atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac 10680
agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg 10740
gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 10800
tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 10860
gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 10920
attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 10980
cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 11040
ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 11100
aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa 11160
tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 11220
acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 11280
aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc 11340
attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga 11400
tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga 11460
gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg 11520
cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc 11580
tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac 11640
agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact 11700
tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca 11760
tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg 11820
tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact 11880
acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg 11940
accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 12000
tgagcgtggt tcacgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 12060
cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc 12120
tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 12180
actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 12240
tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 12300
cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 12360
gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 12420
tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt 12480
gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 12540
gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 12600
ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 12660
acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 12720
agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 12780
cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 12840
tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 12900
gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 12960
ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 13020
ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag 13080
cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca 13140
ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat 13200
taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg 13260
tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga 13320
ttacgccaag cgcgcccgcc gggtaactca cggggtatcc atgtccattt ctgcggcatc 13380
cagccaggat acccgtcctc gctgacgtaa tatcccagcg ccgcaccgct gtcattaatc 13440
tgcacaccgg cacggcagtt ccggctgtcg ccggtattgt tcgggttgct gatgcgcttc 13500
gggctgacca tccggaactg tgtccggaaa agccgcgacg aactggtatc ccaggtggcc 13560
tgaacgaaca gttcaccgtt aaaggcgtgc atggccacac cttcccgaat catcatggta 13620
aacgtgcgtt ttcgctcaac gtcaatgcag cagcagtcat cctcggcaaa ctctttccat 13680
gccgcttcaa cctcgcggga aaaggcacgg gcttcttcct ccccgatgcc cagatagcgc 13740
cagcttgggc gatgactgag ccggaaaaaa gacccgacga tatgatcctg atgcagctag 13800
attaacccta gaaagatagt ctgcgtaaaa ttgacgcatg cattcttgaa atattgctct 13860
ctctttctaa atagcgcgaa tccgtcgctg tgcatttagg acatctcagt cgccgcttgg 13920
agctcccgtg aggcgtgctt gtcaatgcgg taagtgtcac tgattttgaa ctataacgac 13980
cgcgtgagtc aaaatgacgc atgattatct tttacgtgac ttttaagatt taactcatac 14040
gataattata ttgttatttc atgttctact tacgtgataa cttattatat atatattttc 14100
ttgttataga tatc 14114
<210> 21
<211> 14336
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 21
aaatacccac gtttattggg acaaaagttg ttagggaaaa tggggcctca gagttatgat 60
tcaagtcata attctttcca tttataattt cactcgagac tctgttaact gattccttgt 120
gtgttgtatc ttactcctca gctcacaatt acttttagtt attcacctta actgtatgaa 180
taacagtgga gaaaaggatt ctaccagaat actctaatta tggttttgag tcccctttcc 240
agactgaaga tttttcagtc tttttgatct gaggtgattt ttcagtcttt tcgatctgag 300
gtgacagtct caagctcctc aattcaccca gtctcttgat acttgtccat ttagggccac 360
caaagctact ttgacttcat actagagagt caattaatga ggccattctc tgatggacag 420
gtgaagcagg caaggtgact atattttgac taaacggtag aaaacagcct gagtgttaac 480
agtgtagcct ataaaaccca gagctgccca ccctgatcta aacttccagg aacataagaa 540
cgcgccccag acccgggcct ggggggcaag tcggggggcg gggggaggtc gggcagggtc 600
ccctgggagg atggggacgt gctgtgcccc tagcggccac cagagggcac caggacacca 660
ctgcggtcgg ctcagcggct cctgccctgg tcagggggcg ccaggtcctg cccctcctgg 720
ggagggcggg gggcgagaag ggcgatttta attaacccac gtttcaacat gcacatccca 780
gtaatttgga aacattttgt ttccaaagat tcacttaaca ttggtttagc aacatgaagc 840
tttctatgca acccaaggac tcagtttttg gcctgtttta gtgacaggca atcagcaaca 900
tgctgcattt ctctccagtg ttgtaatcaa agaaaccctc ccatagcttt aaatgatatt 960
ccttcccctt ccaattatgt ggggggaaaa caaccctatt ctccacccag aagtgttaac 1020
tcaagaatta cattttcaag aagtttccag attcgtaaaa ccagaattag atgtctttca 1080
cctaaatgtc tcggtgttga ccaaaggaac acacaggttt ctcatttaac ttttttaatg 1140
ggtctcaaaa ttctgtgaca aatttttggt caagttgttt ccattaaaaa gtactgattt 1200
taaaaactaa taacttaaaa ctgccacacg caaaaaagaa aaccaaagtg gtccacaaaa 1260
cattctcctt tccttctgaa ggttttacga tgcattgtta tcattaacca gtcttttact 1320
actaaactta aatggccaat tgaaacaaac agttctgaga ccgttcttcc accactgatt 1380
aagagtgggg tggcaggtat tagggataat gctagcttac ttgtacagct cgtccatgcc 1440
gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt 1500
ggggtctttg ctcagggcgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg 1560
gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc 1620
gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat 1680
atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc 1740
cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac 1800
ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta 1860
gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct 1920
gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt 1980
gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc 2040
gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac 2100
caccccggtg aacagctcct cgcccttgct caccatggtg gcgtcgaccg tacgtcacga 2160
cacctgaaat ggaagaaaaa aactttgaac cactgtctga ggcttgagaa tgaaccaaga 2220
tccaaactca aaaagggcaa attccaagga gaattacatc aagtgccaag ctggcctaac 2280
ttcagtctcc acccactcag tgtggggaaa ctccatcgca taaaacccct ccccccaacc 2340
taaagacgac gtactccaaa agctcgagaa ctaatcgagg tgcctggacg gcgcccggta 2400
ctccgtggag tcacatgaag cgacggctga ggacggaaag gcccttttcc tttgtgtggg 2460
tgactcaccc gcccgctctc ccgagcgccg cgtcctccat tttgagctcc ctgcagcagg 2520
gccgggaagc ggccatcttt ccgctcacgc aactggtgcc gaccgggcca gccttgccgc 2580
ccagggcggg gcgatacacg gcggcgcgag gccaggcacc agagcaggcc ggccagcttg 2640
agactacccc cgtccgattc tcggtggccg cgctcgcagg ccccgcctcg ccgaacatgt 2700
gcgctgggac gcacgggccc cgtcgccgcc cgcggcccca aaaaccgaaa taccagtgtg 2760
cagatcttgg cccgcattta caagactatc ttgccagaaa aaaagcgtcg cagcaggtca 2820
tcaaaaattt taaatggcta gagacttatc gaaagcagcg agacaggcgc gaaggtgcca 2880
ccagattcgc acgcggcggc cccagcgccc aggccaggcc tcaactcaag cacgaggcga 2940
aggggctcct taagcgcaag gcctcgaact ctcccaccca cttccaaccc gaagctcggg 3000
atcaagaatc acgtactgca gccagtggaa gtaattcaag gcacgcaagg gccataaccc 3060
gtaaagaggc caggcccgcg ggaaccacac acggcactta cctgtgttct ggcggcaaac 3120
ccgttgcgaa aaagaacgtt cacggcgact actgcactta tatacggttc tcccccaccc 3180
tcgggaaaaa ggcggagcca gtacacgaca tcactttccc agtttacccc gcgccacctt 3240
ctctaggcac cggttcaatt gccgacccct ccccccaact tctcggggac tgtgggcgat 3300
gtgcgctctg cccactgacg ggcaccggag ccctagattc gattcccttt ggggcaaaac 3360
tcaccgccta atcccctata actctaccgg ggagcccggt ggagagcaga cgggctgacg 3420
ctgccacctg ccggccatcc caggatagga ccgccgtatt caagtcgccc tcaggaagga 3480
ccctcggggc accagaggcc ttcgaagccc caatgagtga ggcaactgag ggtcgcgggt 3540
gccattacaa ggcccagcca aggcctagag ccaaggcttg aaccgtgggg gacccccaag 3600
ccccacctgc ccaggaacag cagacactgg gacactttgt ttcaggtcct gcccaggccc 3660
ctcccactgt gaggctggga tttgtcgccc agggtgcaga tgagaagagt ggggaaagca 3720
gtcctgagcc aggaaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc 3780
atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac 3840
acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc 3900
accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag 3960
gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga 4020
gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt 4080
ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg 4140
cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac 4200
gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctcct agggccacca 4260
tggtgagcaa gggcgaggac gacaacatgg ccatcatcaa ggagttcatg cgcttcaagg 4320
tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc 4380
gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct 4440
tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc 4500
ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg 4560
tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg 4620
gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa 4680
tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg 4740
ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgccg 4800
aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca 4860
acatcaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc 4920
gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtga ggatccgctg 4980
atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 5040
ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 5100
atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 5160
gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc 5220
tgaggcggaa agaacccttc tgaggcggaa agaaccagct gccttaatat aacttcgtat 5280
aatgtatgct atacgaagtt attaggtctg aagaggagtt tacgtccagc caattctgtg 5340
gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 5400
aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 5460
cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 5520
gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 5580
tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg 5640
aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat 5700
tttcggcggc cgcgccacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 5760
cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 5820
ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 5880
cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 5940
ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 6000
catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 6060
gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca 6120
ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 6180
cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 6240
cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 6300
ccgcaagccc ggtgcctgag aattcgcggg actctggggt tcgaaatgac cgaccaagcg 6360
acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc 6420
ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg 6480
gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat 6540
agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 6600
aaactcatca atgtatctta tcatgtctgt ataccgctcg actagagctt gcggaaccct 6660
taatataact tcgtataatg tatgctatac gaagttatta ggtccgctgg ccatctacga 6720
gccaaagact ttcaaatctt tggctgcctt ggccagtagg aggcgacacg aaggatttgc 6780
tgctgccttg ggggatggga aggaacctga aggcattttt tccagagtgg tgcagtacca 6840
ctgaggactg ttgctgtatt gattaggaaa agagacagag taatttgcag tttgtttgat 6900
ttatactggg ctgcaggtcg agggatcttc ataagagaag agggacagct atgactggga 6960
gtagtcagga gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg 7020
ttaaagaaaa aaataacaca aaacaaaata taaaaaaaat ctaacctcaa gtcaaggctt 7080
ttctatggaa taaggaatgg acagcagggg gctgtttcat atactgatga cctctttata 7140
gccacctttg ttcatggcag ccagcatatg gcatatgttg ccaaactcta aaccaaatac 7200
tcattctgat gttttaaatg atttgccctc ccatatgtcc ttccgagtga gagacacaaa 7260
aaattccaac acactattgc aatgaaaata aatttccttt attagccaga agtcagatgc 7320
tcaaggggct tcatgatgtc cccataattt ttggcagagg gaaaaagatc tcagtggtat 7380
ttgtgagcca gggcattggc cacaccagcc accaccttct gataggcagc ctgcggtacc 7440
ttacatggtg gcgaattcgt ttgccaaaat gatgagacag cacaataacc agcacgttgc 7500
ccaggagctg taggaaaaag aagaaggcat gaacatggtt agcagaggct ctagagccgc 7560
cggtcacacg ccagaagccg aaccccgccc tgccccgtcc cccccgaagg cagccgtccc 7620
cctgcggcag ccccgaggct ggagatggag aaggggacgg cggcgcggcg acgcacgaag 7680
gccctccccg cccatttcct tcctgccggc gccgcaccgc ttcgcccgcg cccgctagag 7740
ggggtgcggc ggcgcctccc agatttcggc tccgccagat ttgggacaaa ggaagtccct 7800
gcgccctctc gcacgattac cataaaaggc aatggctgcg gctcgccgcg cctcgacagc 7860
cgccggcgct ccggggccgc cgcgcccctc ccccgagccc tccccggccc gaggcggccc 7920
cgccccgccc ggcaccccca cctgccgcca ccccccgccc ggcacggcga gccccgcgcc 7980
acgccccgca cggagccccg cacccgaagc cgggccgtgc tcagcaactc ggggaggggg 8040
gtgcaggggg gggttacagc ccgaccgccg cgcccacacc ccctgctcac ccccccacgc 8100
acacaccccg cacgcagcct ttgttcccct cgcagccccc ccgcaccgcg gggcaccgcc 8160
cccggccgcg ctcccctcgc gcacacgcgg agcgcacaaa gccccgcgcc gcgcccgcag 8220
cgctcacagc cgccgggcag cgcgggccgc acgcggcgct ccccacgcac acacacacgc 8280
acgcaccccc cgagccgctc ccccccgcac aaagggccct cccggagccc tttaaggctt 8340
tcacgcagcc acagaaaaga aacgagccgt cattaaacca agcgctaatt acagcccgga 8400
ggagaagggc cgtcccgccc gctcacctgt gggagtaacg cggtcagtca gagccggggc 8460
gggcggcgcg aggcggcgcg gagcggggca cggggcgaag gcaacgcagc gactcccgcc 8520
cgccgcgcgc ttcgcttttt atagggccgc cgccgccgcc gcctcgccat aaaaggaaac 8580
tttcggagcg cgccgctctg attggctgcc gccgcacctc tccgcctcgc cccgccccgc 8640
ccctcgcccc gccccgcccc gcctggcgcg cgcccccccc ccccccgccc ccatcgctgc 8700
acaaaataat taaaaaataa ataaatacaa aattgggggt ggggaggggg gggagatggg 8760
gagagtgaag cagaacgtgg ggctcacctc gacccatggt aatagcgatg actaatacgt 8820
agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg 8880
gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta 8940
ctgccaagtg ggcagtttac cgtaaatagt ccacccattg acgtcaatgg aaagtcccta 9000
ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg 9060
gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat 9120
gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa tgtcgtaaat 9180
gtcgtaaatg tctcagctag tcaggtagta aaaggtgtca actaggcagt ggcagagcag 9240
gattcaaatt cagggctgtt gtgatgcctc cgcagactct gagcgccacc tggtggtaat 9300
ttgtctgtgc ctcttctgac gtggaagaac agcaactaac acactaacac ggcatttact 9360
atgggccagc cattgtacgc gttgagtagg tcacatttca gtaaaacctg gctttgtgga 9420
ttgagcatgg tctgtctctt cctggtactt cattagtccc ctaagtggga tttgctgagc 9480
aagactcctc aattacagaa atactccagt ttagaattct cgcaaaggct ttttgtttcc 9540
acaagtagaa tctagaaagc aatctcaagt aacaacagca gagacctgaa tcccaatcca 9600
tctttcctgt gtgtcctctt ttacctcctt ccctttcatg ttgaaccaac agtccttttt 9660
cagtctagaa gctagtacga aagaaatgta cagatgtagg taccaagcaa agccattagc 9720
caataactgg tgagatggag ctaagaggaa ataaaagtgt tcctaagaat agcacagcag 9780
aagctagatc cacagatctt aaaacaattt tggttgagta agagtagagg caaaagagga 9840
agctaataat gcagttttta ggagctaaga gccagataaa gggtaagggc aggaggaagt 9900
gctatctcag ctaacgagat acatgaaaca acggtggaag tccagcaggc acaagatgag 9960
ttgagaagca atcagggcca gaaggatgtg caaggcctca aaataaaaaa gcacagggcc 10020
acagggaacc ttatggaaat taaaaggaag aggatgcagt caggagagga aaaaatagtg 10080
ctccctcccc catgcccaag gaagcagctg agcagccagt acttgggaag ttagtagtaa 10140
taagttggta agagggagtt ctgttcgtgg ctcaatggtt aacaaatcag actagaaacc 10200
gtgaggttgc gggtttgatc cctggccttg ctcagtgggt taaggatccg gcattgccgt 10260
gacctgtggt gtaggtcaca gacgtggctc agttcccgca ttcctgtggc tctggtgtag 10320
gctggtggct acagctctga ttagacccct aggctgggaa cctccatatg ccctggaagt 10380
ggccgtagaa aagatgcggc cgccataaaa gttttgttac tttatagaag aaattttgag 10440
tttttgtttt ttttaataaa taaataaaca taaataaatt gtttgttgaa tttattatta 10500
gtatgtaagt gtaaatataa taaaacttaa tatctattca aattaataaa taaacctcga 10560
tatacagacc gataaaacac atgcgtcaat tttacacatg attatcttta acgtacgtca 10620
caatatgatt atctttctag ggttaatcta gctgcgtgtt ctgcagcgtg tcgagcatct 10680
tcatctgctc catcacgctg taaaacacat ttgcaccgcg agtctgcccg tcctccacgg 10740
gttcaaaaac gtgaatgaac gaggcgcgct cactggccgt cgttttacaa cgtcgtgact 10800
gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct 10860
ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg 10920
gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 10980
gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 11040
ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt 11100
tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac 11160
gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 11220
ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt 11280
ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac 11340
aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg 11400
gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 11460
gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag 11520
tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt 11580
tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt 11640
gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga 11700
acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat 11760
tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga 11820
gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag 11880
tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg 11940
accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg 12000
ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt 12060
agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg 12120
gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc 12180
ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg gttcacgcgg 12240
tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac 12300
ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact 12360
gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa 12420
acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa 12480
aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg 12540
atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc 12600
gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac 12660
tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca 12720
ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt 12780
ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc 12840
ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg 12900
aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc 12960
cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac 13020
gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct 13080
ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc 13140
cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt 13200
tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac 13260
cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg 13320
cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga 13380
caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac 13440
tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt 13500
gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcccg 13560
ccgggtaact cacggggtat ccatgtccat ttctgcggca tccagccagg atacccgtcc 13620
tcgctgacgt aatatcccag cgccgcaccg ctgtcattaa tctgcacacc ggcacggcag 13680
ttccggctgt cgccggtatt gttcgggttg ctgatgcgct tcgggctgac catccggaac 13740
tgtgtccgga aaagccgcga cgaactggta tcccaggtgg cctgaacgaa cagttcaccg 13800
ttaaaggcgt gcatggccac accttcccga atcatcatgg taaacgtgcg ttttcgctca 13860
acgtcaatgc agcagcagtc atcctcggca aactctttcc atgccgcttc aacctcgcgg 13920
gaaaaggcac gggcttcttc ctccccgatg cccagatagc gccagcttgg gcgatgactg 13980
agccggaaaa aagacccgac gatatgatcc tgatgcagct agattaaccc tagaaagata 14040
gtctgcgtaa aattgacgca tgcattcttg aaatattgct ctctctttct aaatagcgcg 14100
aatccgtcgc tgtgcattta ggacatctca gtcgccgctt ggagctcccg tgaggcgtgc 14160
ttgtcaatgc ggtaagtgtc actgattttg aactataacg accgcgtgag tcaaaatgac 14220
gcatgattat cttttacgtg acttttaaga tttaactcat acgataatta tattgttatt 14280
tcatgttcta cttacgtgat aacttattat atatatattt tcttgttata gatatc 14336
<210> 22
<211> 14386
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 22
ggatggggac tcatgtgaat tttctaaagg tgctatttaa acggggggca cgagtgccgg 60
ctttggacag ggccgctcgc tctccaccct ttcttcttcc ccctcggccg cctctcaccc 120
cctgaggcct ctctcccccc acgacctcct ctctctcctc tgaaaccctc tcctcctcag 180
ctgcatccca ccctcgtggc ctctctctct ctctgtctgt cctgtgtcct ctctcactgg 240
gtttcagagc acagatgccc aaagcacaaa agcagttttc ccctggggtg ggaggaagca 300
agagactttg tacctatttt gtatgtgtat aataatttga gatgttttta attattttga 360
ttgctggaat aaagcatgtg gaaatgaccc aaaccaatct tgcactggcc tcctgatttc 420
cttccttgga gacggaggga gggggagacc tgggggaggg cgcttggggg ggggtgggct 480
ctcttctttc tgcgctcccc ccccccacct ccaacacctt gacgacccct cctgcttccg 540
cttgcctttc tcaggcttta acactttctc ctcgccctct cagcatgcgc atgcgcgtgc 600
ctctacctcc cccgcacatc ctggcctgcc caccctgaat ggcctggccc agcgatgcca 660
ccaactctct cgctccgtcc acggctgggg aggggggcac tctgcagggt tggggggcac 720
tgggaggctg ggttgggtga gggaggggtg cctgggcccc caccccccag caagttctct 780
ccctaggcga actggagggt cgtctggcct cttgagcctt gttgctggct ctgagctcta 840
ccaagagagt gaccagcagg accgcaccat cacgcgcccc agacccgggc ctggggggca 900
agtcgggggg cggggggagg tcgggcaggg tcccctggga ggatggggac gtgctgtgcc 960
cctagcggcc accagagggc accaggacac cactgcggtc ggctcagcgg ctcctgccct 1020
ggtcaggggg cgccaggtcc tgcccctcct ggggagggcg gggggcgaga agggcgattt 1080
taattaaccc acgtttcaac atgcacatcc cagtaatttg gaaacatttt gtttccaaag 1140
attcacttaa cattggttta gcaacatgaa gctttctatg caacccaagg actcagtttt 1200
tggcctgttt tagtgacagg caatcagcaa catgctgcat ttctctccag tgttgtaatc 1260
aaagaaaccc tcccatagct ttaaatgata ttccttcccc ttccaattat gtggggggaa 1320
aacaacccta ttctccaccc agaagtgtta actcaagaat tacattttca agaagtttcc 1380
agattcgtaa aaccagaatt agatgtcttt cacctaaatg tctcggtgtt gaccaaagga 1440
acacacaggt ttctcattta acttttttaa tgggtctcaa aattctgtga caaatttttg 1500
gtcaagttgt ttccattaaa aagtactgat tttaaaaact aataacttaa aactgccaca 1560
cgcaaaaaag aaaaccaaag tggtccacaa aacattctcc tttccttctg aaggttttac 1620
gatgcattgt tatcattaac cagtctttta ctactaaact taaatggcca attgaaacaa 1680
acagttctga gaccgttctt ccaccactga ttaagagtgg ggtggcaggt attagggata 1740
atgctagctt acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac 1800
tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactgggtg 1860
ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg 1920
tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc 1980
ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac 2040
tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg 2100
cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg 2160
tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag 2220
tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg 2280
gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc 2340
agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt 2400
acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg 2460
ctcaccatgg tggcgtcgac cgtacgtcac gacacctgaa atggaagaaa aaaactttga 2520
accactgtct gaggcttgag aatgaaccaa gatccaaact caaaaagggc aaattccaag 2580
gagaattaca tcaagtgcca agctggccta acttcagtct ccacccactc agtgtgggga 2640
aactccatcg cataaaaccc ctccccccaa cctaaagacg acgtactcca aaagctcgag 2700
aactaatcga ggtgcctgga cggcgcccgg tactccgtgg agtcacatga agcgacggct 2760
gaggacggaa aggccctttt cctttgtgtg ggtgactcac ccgcccgctc tcccgagcgc 2820
cgcgtcctcc attttgagct ccctgcagca gggccgggaa gcggccatct ttccgctcac 2880
gcaactggtg ccgaccgggc cagccttgcc gcccagggcg gggcgataca cggcggcgcg 2940
aggccaggca ccagagcagg ccggccagct tgagactacc cccgtccgat tctcggtggc 3000
cgcgctcgca ggccccgcct cgccgaacat gtgcgctggg acgcacgggc cccgtcgccg 3060
cccgcggccc caaaaaccga aataccagtg tgcagatctt ggcccgcatt tacaagacta 3120
tcttgccaga aaaaaagcgt cgcagcaggt catcaaaaat tttaaatggc tagagactta 3180
tcgaaagcag cgagacaggc gcgaaggtgc caccagattc gcacgcggcg gccccagcgc 3240
ccaggccagg cctcaactca agcacgaggc gaaggggctc cttaagcgca aggcctcgaa 3300
ctctcccacc cacttccaac ccgaagctcg ggatcaagaa tcacgtactg cagccagtgg 3360
aagtaattca aggcacgcaa gggccataac ccgtaaagag gccaggcccg cgggaaccac 3420
acacggcact tacctgtgtt ctggcggcaa acccgttgcg aaaaagaacg ttcacggcga 3480
ctactgcact tatatacggt tctcccccac cctcgggaaa aaggcggagc cagtacacga 3540
catcactttc ccagtttacc ccgcgccacc ttctctaggc accggttcaa ttgccgaccc 3600
ctccccccaa cttctcgggg actgtgggcg atgtgcgctc tgcccactga cgggcaccgg 3660
agccctagat tcgattccct ttggggcaaa actcaccgcc taatccccta taactctacc 3720
ggggagcccg gtggagagca gacgggctga cgctgccacc tgccggccat cccaggatag 3780
gaccgccgta ttcaagtcgc cctcaggaag gaccctcggg gcaccagagg ccttcgaagc 3840
cccaatgagt gaggcaactg agggtcgcgg gtgccattac aaggcccagc caaggcctag 3900
agccaaggct tgaaccgtgg gggaccccca agccccacct gcccaggaac agcagacact 3960
gggacacttt gtttcaggtc ctgcccaggc ccctcccact gtgaggctgg gatttgtcgc 4020
ccagggtgca gatgagaaga gtggggaaag cagtcctgag ccaggaaatt ctaccgggta 4080
ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact 4140
tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa 4200
ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag 4260
ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc 4320
actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc 4380
agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg 4440
tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga 4500
ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca 4560
tctccgggcc tttcgacctc ctagggccac catggtgagc aagggcgagg acgacaacat 4620
ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca 4680
cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa 4740
gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt 4800
catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct 4860
gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt 4920
gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg 4980
cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc 5040
ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct 5100
gaagctgaag gacggcggcc actacgacgc cgaggtcaag accacctaca aggccaagaa 5160
gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ctggacatca cctcccacaa 5220
cgaggactac accatcgtgg aacagtacga gcgcgccgag ggccgccact ccaccggcgg 5280
catggacgag ctgtacaagt gaggatccgc tgatcagcct cgactgtgcc ttctagttgc 5340
cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 5400
actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 5460
attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 5520
catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccct tctgaggcgg 5580
aaagaaccag ctgccttaat ataacttcgt ataatgtatg ctatacgaag ttattaggtc 5640
tgaagaggag tttacgtcca gccaattctg tggaatgtgt gtcagttagg gtgtggaaag 5700
tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 5760
aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 5820
tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 5880
tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 5940
gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 6000
tgcaaaaagc tcccgggagc ttgtatatcc attttcggcg gccgcgccac catgaccgag 6060
tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 6120
gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 6180
gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 6240
gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 6300
gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 6360
gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 6420
ttcctggcca ccgtcggagt ctcgcccgac caccagggca agggtctggg cagcgccgtc 6480
gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 6540
gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 6600
gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agaattcgcg 6660
ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 6720
attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 6780
ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 6840
ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 6900
ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 6960
gtataccgct cgactagagc ttgcggaacc cttaatataa cttcgtataa tgtatgctat 7020
acgaagttat taggtccgct ggccatctac gagccaaaga ctttcaaatc tttggctgcc 7080
ttggccagta ggaggcgaca cgaaggattt gctgctgcct tgggggatgg gaaggaacct 7140
gaaggcattt tttccagagt ggtgcagtac cactgaggac tgttgctgta ttgattagga 7200
aaagagacag agtaatttgc agtttgtttg atttatactg ggctgcaggt cgagggatct 7260
tcataagaga agagggacag ctatgactgg gagtagtcag gagaggagga aaaatctggc 7320
tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca caaaacaaaa 7380
tataaaaaaa atctaacctc aagtcaaggc ttttctatgg aataaggaat ggacagcagg 7440
gggctgtttc atatactgat gacctcttta tagccacctt tgttcatggc agccagcata 7500
tggcatatgt tgccaaactc taaaccaaat actcattctg atgttttaaa tgatttgccc 7560
tcccatatgt ccttccgagt gagagacaca aaaaattcca acacactatt gcaatgaaaa 7620
taaatttcct ttattagcca gaagtcagat gctcaagggg cttcatgatg tccccataat 7680
ttttggcaga gggaaaaaga tctcagtggt atttgtgagc cagggcattg gccacaccag 7740
ccaccacctt ctgataggca gcctgcggta ccttacatgg tggcgaattc gtttgccaaa 7800
atgatgagac agcacaataa ccagcacgtt gcccaggagc tgtaggaaaa agaagaaggc 7860
atgaacatgg ttagcagagg ctctagagcc gccggtcaca cgccagaagc cgaaccccgc 7920
cctgccccgt cccccccgaa ggcagccgtc cccctgcggc agccccgagg ctggagatgg 7980
agaaggggac ggcggcgcgg cgacgcacga aggccctccc cgcccatttc cttcctgccg 8040
gcgccgcacc gcttcgcccg cgcccgctag agggggtgcg gcggcgcctc ccagatttcg 8100
gctccgccag atttgggaca aaggaagtcc ctgcgccctc tcgcacgatt accataaaag 8160
gcaatggctg cggctcgccg cgcctcgaca gccgccggcg ctccggggcc gccgcgcccc 8220
tcccccgagc cctccccggc ccgaggcggc cccgccccgc ccggcacccc cacctgccgc 8280
caccccccgc ccggcacggc gagccccgcg ccacgccccg cacggagccc cgcacccgaa 8340
gccgggccgt gctcagcaac tcggggaggg gggtgcaggg gggggttaca gcccgaccgc 8400
cgcgcccaca ccccctgctc acccccccac gcacacaccc cgcacgcagc ctttgttccc 8460
ctcgcagccc ccccgcaccg cggggcaccg cccccggccg cgctcccctc gcgcacacgc 8520
ggagcgcaca aagccccgcg ccgcgcccgc agcgctcaca gccgccgggc agcgcgggcc 8580
gcacgcggcg ctccccacgc acacacacac gcacgcaccc cccgagccgc tcccccccgc 8640
acaaagggcc ctcccggagc cctttaaggc tttcacgcag ccacagaaaa gaaacgagcc 8700
gtcattaaac caagcgctaa ttacagcccg gaggagaagg gccgtcccgc ccgctcacct 8760
gtgggagtaa cgcggtcagt cagagccggg gcgggcggcg cgaggcggcg cggagcgggg 8820
cacggggcga aggcaacgca gcgactcccg cccgccgcgc gcttcgcttt ttatagggcc 8880
gccgccgccg ccgcctcgcc ataaaaggaa actttcggag cgcgccgctc tgattggctg 8940
ccgccgcacc tctccgcctc gccccgcccc gcccctcgcc ccgccccgcc ccgcctggcg 9000
cgcgcccccc ccccccccgc ccccatcgct gcacaaaata attaaaaaat aaataaatac 9060
aaaattgggg gtggggaggg gggggagatg gggagagtga agcagaacgt ggggctcacc 9120
tcgacccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca 9180
taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg 9240
gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata 9300
gtccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat 9360
tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag 9420
ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta 9480
ttaataacta gtcaataatc aatgtcgtaa atgtcgtaaa tgtctcagct agtcaggtag 9540
taaaaggtgt caactaggca gtggcagagc aggattcaaa ttcagggctg ttgtgatgcc 9600
tccgcagact ctgagcgcca cctggtggta atttgtctgt gcctcttctg acgtggaaga 9660
acagcaacta acacactaac acggcattta ctatgggcca gccattgtac gcgttggtgg 9720
ttgctgagac tgcgtggggg cccaaggaga cctggagaaa ggaatgcttc ctgctccttc 9780
ttctggggcc ccaggagagc cttcccaggg ccttggagag gtgctgtcca gggactaacc 9840
ctgtgctcta ggaaggctgc aggccctgac cagctgggca ggtcctgggt ccctcctggc 9900
cttctaagtt ccccaaacat gagacctctg ggtgtggggt ggcctgggga ggtcattttg 9960
cccaggccct acctcctgcc cattcctaac cctttttaaa aatctgtgcg tcctcttctt 10020
ccttcttctc cctcccttcc cttttcgctc accctctgct gctggcctga gagccggagg 10080
cccccagggg gaaggcgact ggtctcctcc ccagtctcag ggaagggaga cagagaatcc 10140
aggaagccag aactcagcag acgaagcacc cagggaccta gagatgggtt gaaaagttga 10200
cagctgtccc acctgcctcc caaggtctca gggcctaaac ctccaaggca ggaaaggccc 10260
ctgtccctcc ctggggtcca tagaaagagg gacaagtctg cacggaccat ttgctgtaat 10320
attaacacct tggctgtcat taggtagtct tggctgttaa ttatgtcctg tgataatgta 10380
ttattagcac gccgaccaca tagggtaggg aactgcagct agtaaacaaa agtttgttcc 10440
tatatgcggc cgccataaaa gttttgttac tttatagaag aaattttgag tttttgtttt 10500
ttttaataaa taaataaaca taaataaatt gtttgttgaa tttattatta gtatgtaagt 10560
gtaaatataa taaaacttaa tatctattca aattaataaa taaacctcga tatacagacc 10620
gataaaacac atgcgtcaat tttacacatg attatcttta acgtacgtca caatatgatt 10680
atctttctag ggttaatcta gctgcgtgtt ctgcagcgtg tcgagcatct tcatctgctc 10740
catcacgctg taaaacacat ttgcaccgcg agtctgcccg tcctccacgg gttcaaaaac 10800
gtgaatgaac gaggcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc 10860
tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag 10920
cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga 10980
cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 11040
tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 11100
gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 11160
tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 11220
atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 11280
actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 11340
agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 11400
cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg 11460
cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 11520
caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 11580
ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 11640
gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 11700
gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 11760
atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg 11820
caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 11880
gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 11940
accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 12000
ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 12060
gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca 12120
acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta 12180
atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct 12240
ggctggttta ttgctgataa atctggagcc ggtgagcgtg gttcacgcgg tatcattgca 12300
gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag 12360
gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat 12420
tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt 12480
taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 12540
cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 12600
gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 12660
gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 12720
agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 12780
aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 12840
agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 12900
cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 12960
accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 13020
aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 13080
ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 13140
cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 13200
gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 13260
tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 13320
agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc 13380
aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc 13440
gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca 13500
ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa 13560
caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcccg ccgggtaact 13620
cacggggtat ccatgtccat ttctgcggca tccagccagg atacccgtcc tcgctgacgt 13680
aatatcccag cgccgcaccg ctgtcattaa tctgcacacc ggcacggcag ttccggctgt 13740
cgccggtatt gttcgggttg ctgatgcgct tcgggctgac catccggaac tgtgtccgga 13800
aaagccgcga cgaactggta tcccaggtgg cctgaacgaa cagttcaccg ttaaaggcgt 13860
gcatggccac accttcccga atcatcatgg taaacgtgcg ttttcgctca acgtcaatgc 13920
agcagcagtc atcctcggca aactctttcc atgccgcttc aacctcgcgg gaaaaggcac 13980
gggcttcttc ctccccgatg cccagatagc gccagcttgg gcgatgactg agccggaaaa 14040
aagacccgac gatatgatcc tgatgcagct agattaaccc tagaaagata gtctgcgtaa 14100
aattgacgca tgcattcttg aaatattgct ctctctttct aaatagcgcg aatccgtcgc 14160
tgtgcattta ggacatctca gtcgccgctt ggagctcccg tgaggcgtgc ttgtcaatgc 14220
ggtaagtgtc actgattttg aactataacg accgcgtgag tcaaaatgac gcatgattat 14280
cttttacgtg acttttaaga tttaactcat acgataatta tattgttatt tcatgttcta 14340
cttacgtgat aacttattat atatatattt tcttgttata gatatc 14386
<210> 23
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 23
agttatggca gaactcagtg 20
<210> 24
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 24
ccccatccaa agtttttaaa gga 23
<210> 25
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 25
tgtggcagat gtcacagttt agg 23
<210> 26
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 26
caccgagtta tggcagaact cagtg 25
<210> 27
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 27
aaaccactga gttctgccat aactc 25
<210> 28
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 28
gaaggagcaa actgacatgg 20
<210> 29
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 29
tgcagtgggt ctttggggac 20
<210> 30
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 30
ttccaggaac ataagaaagt 20
<210> 31
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 31
gcagtctcag caaccactga 20
<210> 32
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 32
ggtcggagtg aacggatttg 20
<210> 33
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 33
ccatttgatg ttggcgggat 20
<210> 34
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 34
agatccgcca caacatcgag 20
<210> 35
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 35
gtccatgccg agagtgatcc 20
<210> 36
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 36
gctctctctg accaggatct aac 23
<210> 37
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 37
gacactggga cactttgttt cagg 24
<210> 38
<211> 24
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 38
cagctgaggc cattatatga agag 24
<210> 39
<211> 22
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 39
gagtcaccaa agacggtgtc ag 22
<210> 40
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 40
tgctgagttc tggcttcctg 20
<210> 41
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 41
tctaccaaga gagtgaccag cag 23
<210> 42
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 42
cacgccatcc tgcgtctgga 20
<210> 43
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 43
agcaccgtgt tggcgtagag 20
<210> 44
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 44
ctaaccagcc ccctgtttcc 20
<210> 45
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 45
ggaggcataa ggattttctc cac 23
<210> 46
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 46
cacgccatcc tgcgtctgga 20
<210> 47
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 47
agcaccgtgt tggcgtagag 20
<210> 48
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 48
ctaaccagcc ccctgtttcc 20
<210> 49
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 49
ggaggcataa ggattttctc cac 23
<210> 50
<211> 7922
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 50
ggatcccctg agggggcccc catgggctag aggatccggc ctcggcctct gcataaataa 60
aaaaaattag tcagccatga gcttggccca ttgcatacgt tgtatccata tcataatatg 120
tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt 180
tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt 240
acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg 300
tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg 360
gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt 420
acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg 480
accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg 540
gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt 600
ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac 660
tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg 720
tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat 780
ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccc ctcgaagctt 840
acatgtggta ccgagctcgg atcctgagaa cttcagggtg agtctatggg acccttgatg 900
ttttctttcc ccttcttttc tatggttaag ttcatgtcat aggaagggga gaagtaacag 960
ggtacacata ttgaccaaat cagggtaatt ttgcatttgt aattttaaaa aatgctttct 1020
tcttttaata tacttttttg tttatcttat ttctaatact ttccctaatc tctttctttc 1080
agggcaataa tgatacaatg tatcatgcct ctttgcacca ttctaaagaa taacagtgat 1140
aatttctggg ttaaggcaat agcaatattt ctgcatataa atatttctgc atataaattg 1200
taactgatgt aagaggtttc atattgctaa tagcagctac aatccagcta ccattctgct 1260
tttattttat ggttgggata aggctggatt attctgagtc caagctaggc ccttttgcta 1320
atcatgttca tacctcttat cttcctccca cagctcctgg gcaacgtgct ggtctgtgtg 1380
ctggcccatc actttggcaa agcacgtgag atctatgttt gtttttcttg ttttattgcc 1440
actagtctct agtcagtgtg ttaatcttac aaccagaact caattacccc ctgcatacac 1500
taattctttc acacgtggtg tttattaccc tgacaaagtt ttcagatcct cagttttaca 1560
ttcaactcag gacttgttct tacctttctt ttccaatgtt acttggttcc atgctataca 1620
tgtctctggg accaatggta ctaagaggtt tgataaccct gtcctaccat ttaatgatgg 1680
tgtttatttt gcttccactg agaagtctaa cataataaga ggctggattt ttggtactac 1740
tttagattcg aagacccagt ccctacttat tgttaataac gctactaatg ttgttattaa 1800
agtctgtgaa tttcaatttt gtaatgatcc atttttgggt gtttattacc acaaaaacaa 1860
caaaagttgg atggaaagtg agttcagagt ttattctagt gcgaataatt gcacttttga 1920
atatgtctct cagccttttc ttatggacct tgaaggaaaa cagggtaatt tcaaaaatct 1980
tagggaattt gtgtttaaga atattgatgg ttattttaaa atatattcta agcacacgcc 2040
tattaattta gtgcgtgatc tccctcaggg tttttcggct ttagaaccat tggtagattt 2100
gccaataggt attaacatca ctaggtttca aactttactt gctttacata gaagttattt 2160
gactcctggt gattcttctt caggttggac agctggtgct gcagcttatt atgtgggtta 2220
tcttcaacct aggacttttc tattaaaata taatgaaaat ggaaccatta cagatgctgt 2280
agactgtgca cttgaccctc tctcagaaac aaagtgtacg ttgaaatcct tcactgtaga 2340
aaaaggaatc tatcaaactt ctaactttag agtccaacca acagaatcta ttgttagatt 2400
tcctaatatt acaaacttgt gcccttttgg tgaagttttt aacgccacca gatttgcatc 2460
tgtttatgct tggaacagga agagaatcag caactgtgtt gctgattatt ctgtcctata 2520
taattccgca tcattttcca cttttaagtg ttatggagtg tctcctacta aattaaatga 2580
tctctgcttt actaatgtct atgcagattc atttgtaatt agaggtgatg aagtcagaca 2640
aatcgctcca gggcaaactg gaaagattgc tgattataat tataaattac cagatgattt 2700
tacaggctgc gttatagctt ggaattctaa caatcttgat tctaaggttg gtggtaatta 2760
taattacctg tatagattgt ttaggaagtc taatctcaaa ccttttgaga gagatatttc 2820
aactgaaatc tatcaggccg gtagcacacc ttgtaatggt gttgaaggtt ttaattgtta 2880
ctttccttta caatcatatg gtttccaacc cactaatggt gttggttacc aaccatacag 2940
agtagtagta ctttcttttg aacttctaca tgcaccagca actgtttgtg gacctaaaaa 3000
gtctactaat ttggttaaaa acaaatgtgt caatttcaac ttcaatggtt taacaggcac 3060
aggtgttctt actgagtcta acaaaaagtt tctgcctttc caacaatttg gcagagacat 3120
tgctgacact actgatgctg tccgtgatcc acagacactt gagattcttg acattacacc 3180
atgttctttt ggtggtgtca gtgttataac accaggaaca aatacttcta accaggttgc 3240
tgttctttat caggatgtta actgcacaga agtccctgtt gctattcatg cagatcaact 3300
tactcctact tggcgtgttt attctacagg ttctaatgtt tttcaaacac gtgcaggctg 3360
tttaataggg gctgaacatg tcaacaactc atatgagtgt gacataccca ttggtgcagg 3420
tatatgcgct agttatcaga ctcagactaa ttctcctcgg cgggcacgta gtgtagctag 3480
tcaatccatc attgcctaca ctatgtcact tggtgcagaa aattcagttg cttactctaa 3540
taactctatt gccataccca caaattttac tattagtgtt accacagaaa ttctaccagt 3600
gtctatgacc aagacatcag tagattgtac aatgtacatt tgtggtgatt caactgaatg 3660
cagcaatctt ttgttgcaat atggcagttt ttgtacacaa ttaaaccgtg ctttaactgg 3720
aatagctgtt gaacaagaca aaaacaccca agaagttttt gcacaagtca aacaaattta 3780
caaaacacca ccaattaaag attttggtgg ttttaatttt tcacaaatat taccagatcc 3840
atcaaaacca agcaagaggt catttattga agatctactt ttcaacaaag tgacacttgc 3900
agatgctggc ttcatcaaac aatatggtga ttgccttggt gatattgctg ctagagacct 3960
catttgtgca caaaagttta acggccttac tgttttgcca cctttgctca cagatgaaat 4020
gattgctcaa tacacttctg cactgttagc gggtacaatc acttctggtt ggacctttgg 4080
tgcaggtgct gcattacaaa taccatttgc tatgcaaatg gcttataggt ttaatggtat 4140
tggagttaca cagaatgttc tctatgagaa ccaaaaattg attgccaacc aatttaatag 4200
tgctattggc aaaattcaag actcactttc ttccacagca agtgcacttg gaaaacttca 4260
agatgtggtc aaccaaaatg cacaagcttt aaacacgctt gttaaacaac ttagctccaa 4320
ttttggtgca atttcaagtg ttttaaatga tatcctttca cgtcttgaca aagttgaggc 4380
tgaagtgcaa attgataggt tgatcacagg cagacttcaa agtttgcaga catatgtgac 4440
tcaacaatta attagagctg cagaaatcag agcttctgct aatcttgctg ctactaaaat 4500
gtcagagtgt gtacttggac aatcaaaaag agttgatttt tgtggaaagg gctatcatct 4560
tatgtccttc cctcagtcag cacctcatgg tgtagtcttc ttgcatgtga cttatgtccc 4620
tgcacaagaa aagaacttca caactgctcc tgccatttgt catgatggaa aagcacactt 4680
tcctcgtgaa ggtgtctttg tttcaaatgg cacacactgg tttgtaacac aaaggaattt 4740
ttatgaacca caaatcatta ctacagacaa cacatttgtg tctggtaact gtgatgttgt 4800
aataggaatt gtcaacaaca cagtttatga tcctttgcaa cctgaattag actcattcaa 4860
ggaggagtta gataaatatt ttaagaatca tacatcacca gatgttgatt taggtgacat 4920
ctctggcatt aatgcttcag ttgtaaacat tcaaaaagaa attgaccgcc tcaatgaggt 4980
tgccaagaat ttaaatgaat ctctcatcga tctccaagaa cttggaaagt atgagcagta 5040
tataaaatgg ccatggtaca tttggctagg ttttatagct ggcttgattg ccatagtaat 5100
ggtgacaatt atgctttgct gtatgaccag ttgctgtagt tgtctcaagg gctgttgttc 5160
ttgtggatcc tgctgcaaat ttgattaaac cccaccagtg caggctgcct atcagaaagt 5220
ggtggctggt gtggctaatg ccctggccca caagtatcac taagctcgct ttcttgctgt 5280
ccaatttcta ttaaaggttc ctttgttccc taagtccaac tactaaactg ggggatatta 5340
tgaagggcct tgagcatctg gattctgcct aataaaaaac atttattttc attgcaatga 5400
tgtatttaaa ttatttctga atattttact aaaaagggaa tgtgggaggt cagtgcattt 5460
aaaacataaa gaaatgaaga gctagttcaa accttgggaa aatacactat atcttaaact 5520
ccatgaaaga aggtgaggct gcaaacagct aatgcacatt ggcaacagcc cctgatgcct 5580
atgccttatt catccctcag aaaaggattc aagtagaggc ttgatttgga ggttaaagtt 5640
ttgctatgct gtattttaca ttacttattg ttttagctgt cctcatgaat gtcttttcac 5700
tacccatttg cttatcctgc atctctcagc cttgactcca ctcagttctc ttgcttagag 5760
ataccacctt tcccctgaag tgttccttcc atgttttacg gcgagatggt ttctcctcgc 5820
ctggccactc agccttagtt gtctctgttg tcttatagag gtctacttga agaaggaaaa 5880
acagggggca tggtttgact gtcctgtgag cccttcttcc ctgcctcccc cactcacagt 5940
gacccggaat ccctcgacat ggcagtctag cactagtgcg gccgcagatc tgcttcctcg 6000
ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 6060
gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 6120
ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 6180
cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 6240
ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 6300
accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 6360
catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 6420
gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 6480
tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 6540
agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 6600
actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 6660
gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 6720
aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 6780
gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 6840
aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 6900
atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 6960
gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 7020
atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 7080
ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 7140
cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 7200
agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 7260
cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 7320
tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 7380
agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 7440
gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 7500
gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 7560
ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 7620
tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 7680
tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 7740
gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 7800
caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 7860
atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 7920
gt 7922
<210> 51
<211> 9144
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 51
gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 60
atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 120
gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 180
tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 240
attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 300
atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 360
acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 420
tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 480
tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 540
attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 600
tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 660
ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 720
accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 780
gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 840
ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 900
aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 960
tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1020
gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1080
ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1140
ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1200
ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1260
aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1320
tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1380
caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1440
aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1500
aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg 1560
agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga 1620
gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata 1680
ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg 1740
acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg 1800
ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag 1860
ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt 1920
tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt 1980
aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt 2040
aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag 2100
aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata 2160
acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta 2220
agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta 2280
tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa 2340
gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt 2400
gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat 2460
tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa 2520
agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag 2580
agatccagtt tggttaatta agggcagagc gcacatcgcc cacagtcccc gagaagttgg 2640
ggggaggggt cggcaattga tccggtgcct agagaaggtg gcgcggggta aactgggaaa 2700
gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg 2760
cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggaccggt 2820
tctagagcgc tctcgaggcc accatggtga gcaagggcga ggaggataac atggccatca 2880
tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg 2940
agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg 3000
tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg 3060
gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc 3120
ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga 3180
cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca 3240
acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg 3300
agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga 3360
aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc 3420
agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact 3480
acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg 3540
agctgtacaa gggtaccgga tccggcgcaa caaacttctc tctgctgaaa caagccggag 3600
atgtcgaaga gaatcctgga ccgaccgagt acaagcccac ggtgcgcctc gccacccgcg 3660
acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc 3720
gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc 3780
tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg 3840
cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc 3900
gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg 3960
cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc 4020
accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg 4080
ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc 4140
tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga 4200
cccgcaagcc cggtgcctga acgcgttaag tcgacaatca acctctggat tacaaaattt 4260
gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg 4320
ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt 4380
ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg 4440
tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc 4500
agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg 4560
cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt 4620
tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc 4680
gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg 4740
gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga 4800
tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt 4860
agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg 4920
aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg 4980
ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 5040
gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 5100
cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga 5160
ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 5220
tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 5280
tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 5340
gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa 5400
gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg 5460
cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 5520
ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 5580
atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 5640
ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 5700
tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 5760
accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt 5820
taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca 5880
gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 5940
caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 6000
aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 6060
cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 6120
tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt 6180
tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga 6240
tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag 6300
gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac 6360
gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag 6420
gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac 6480
caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac 6540
gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc 6600
gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc 6660
gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc 6720
gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 6780
cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 6840
aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 6900
cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 6960
acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 7020
ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 7080
taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 7140
aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 7200
attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 7260
cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 7320
gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 7380
ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 7440
agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 7500
tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 7560
ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 7620
gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 7680
ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 7740
gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 7800
aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 7860
aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 7920
ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 7980
gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 8040
gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 8100
tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 8160
ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 8220
ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 8280
atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 8340
ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 8400
tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 8460
attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 8520
tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 8580
ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 8640
gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 8700
gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 8760
gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 8820
aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 8880
taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 8940
tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 9000
tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 9060
atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 9120
tttccccgaa aagtgccacc tgac 9144
Claims (13)
1.一种表达人ACE2的猪细胞，其特征在于，将人ACE2基因插入猪安全港位点，获得表达SEQ ID NO：12所示人ACE2的猪细胞，所述的猪安全港位点选自猪ROSA26、AAVS1、H11或COL1A1安全港位点。
2.根据权利要求1所述的猪细胞，其特征在于，所述插入的人ACE2基因的核苷酸序列如SEQ ID NO：13所示。
3.根据权利要求1或2所述的猪细胞，其特征在于，ROSA26安全港位点区域及其上下游各500bp的核苷酸序列如SEQ ID NO：14所示，AAVS1安全港位点区域及其上下游各500bp的核苷酸序列如SEQ ID NO：15所示，H11安全港位点区域及其上下游各500bp的核苷酸序列如SEQ ID NO：16所示，COL1A1安全港位点区域及其上下游各500bp的核苷酸序列如SEQ IDNO：17所示。
4.根据权利要求1-3任一所述的猪细胞，其特征在于，所述的猪细胞为猪成纤维细胞。
5.一种权利要求1-4任一所述猪细胞的构建方法，其特征在于，使用安全港位点载体将人ACE2基因插入猪安全港位点，所述的安全港位点载体包含人ACE2基因的核苷酸序列和安全港位点载体骨架，所述的安全港位点载体骨架包含安全港插入位点的5’同源臂和3’同源臂，所述人ACE2基因的核苷酸序列位于5’同源臂与3’同源臂之间，所述的安全港位点载体骨架选自下列任一项所示：
A)ROSA26安全港位点载体骨架，其5’同源臂如SEQ ID NO：18所示，3’同源臂如SEQ IDNO：19所示；优选的，所述的ROSA26安全港位点载体的核苷酸序列如SEQ ID NO：4所示；
B)AAVS1安全港位点载体骨架，其5’同源臂如SEQ ID NO：5所示，3’同源臂如SEQ IDNO：6所示；优选的，所述的AAVS1安全港位点载体的核苷酸序列如SEQ ID NO：20所示；
C)H11安全港位点载体骨架，其5’同源臂如SEQ ID NO：7所示，3’同源臂如SEQ ID NO：8所示；优选的，所述的H11安全港位点载体的核苷酸序列如SEQ ID NO：21所示；
或D)COL1A1安全港位点载体骨架，其5’同源臂如SEQ ID NO：9所示，3’同源臂如SEQ IDNO：10所示；优选的，所述的COL1A1安全港位点载体的核苷酸序列如SEQ ID NO：22所示。
6.根据权利要求5所述的构建方法，其特征在于，使用sgRNA载体进行猪细胞的构建，所述的sgRNA载体包含靶向ROSA26、AAVS1、H11或COL1A1安全港位点的sgRNA，其中：
靶向ROSA26的sgRNA的核苷酸序列如SEQ ID NO：28所示，靶向AAVS1的sgRNA的核苷酸序列如SEQ ID NO：29所示，靶向H11的sgRNA的核苷酸序列如SEQ ID NO：30所示，靶向COL1A1的sgRNA的核苷酸序列如SEQ ID NO：31所示。
7.根据权利要求5或6所述的构建方法，其特征在于，所述的构建方法包括将安全港位点载体、sgRNA载体和Cas载体共转染至猪细胞，所述的Cas载体包含编码Cas蛋白、EGFP和Puro抗性蛋白的核苷酸序列，所述的Cas蛋白选自Casl、CaslB、Cas2、Cas3、Cas4、Cas5、Cas5d、Cas5t、Cas5h、Cas5a、Cas6、Cas7、Cas8、Cas9、CaslO、Csyl、Csy2、Csy3、Csy4、Csel、Cse2、Cse3、Cse4、Cse5e、Cscl、Csc2、Csa5、Csnl、Csn2、Csml、Csm2、Csm3、Csm4、Csm5、Csm6、Cmrl、Cmr3、Cmr4、Cmr5、Cmr6、Csbl、Csb2、Csb3、Csx17、Csx14、CsxlO、Csx16、CsaX、Csx3、Csxl、CsxlS、Csfl、Csf2、CsO、Csf4、Csdl、Csd2、Cstl、Cst2、Cshl、Csh2、Csal、Csa2、Csa3、Csa4、Csa5、C2cl、C2c2、C2c3、Cpfl、CARF、DinG、其同源物或其修饰形式，优选为Cas9，优选的，所述的Cas载体的核苷酸序列如SEQ ID NO：1或2所示。
8.一种表达人ACE2的人源化猪的构建方法，其特征在于，将人ACE2基因插入猪安全港位点，获得表达SEQ ID NO：12所示人ACE2的猪，所述的猪安全港位点选自猪ROSA26、AAVS1、H11或COL1A1安全港位点。
9.一种安全港位点载体，其特征在于，所述的安全港位点载体包含人ACE2基因的核苷酸序列和安全港位点载体骨架，所述的人ACE2基因的核苷酸序列如SEQ ID NO：13所示，所述的安全港位点载体骨架包含安全港插入位点的5’同源臂和3’同源臂，所述人ACE2基因的核苷酸序列位于5’同源臂与3’同源臂之间，所述的安全港位点载体骨架选自下列任一项所示：
A)ROSA26安全港位点载体骨架，其5’同源臂如SEQ ID NO：18所示，3’同源臂如SEQ IDNO：19所示；优选的，所述的ROSA26安全港位点载体的核苷酸序列如SEQ ID NO：4所示；
B)AAVS1安全港位点载体骨架，其5’同源臂如SEQ ID NO：5所示，3’同源臂如SEQ IDNO：6所示；优选的，所述的AAVS1安全港位点载体的核苷酸序列如SEQ ID NO：20所示；
C)H11安全港位点载体骨架，其5’同源臂如SEQ ID NO：7所示，3’同源臂如SEQ ID NO：8所示；优选的，所述的H11安全港位点载体的核苷酸序列如SEQ ID NO：21所示；
或D)COL1A1安全港位点载体骨架，其5’同源臂如SEQ ID NO：9所示，3’同源臂如SEQ IDNO：10所示；优选的，所述的COL1A1安全港位点载体的核苷酸序列如SEQ ID NO：22所示。
10.一种权利要求9所述的安全港位点载体、权利要求6所述的sgRNA或者权利要求6所述的sgRNA载体在制备表达人ACE2的猪或猪细胞中的应用。
11.一种ACE2人源化猪的构建方法，其特征在于，所述构建方法包括：
A、卵母细胞体外成熟；B、将权利要求1-4任一所述的猪细胞进行体细胞核移植(SCNT)构建重构胚；C、胚胎移植。
12.权利要求8或11所述构建方法获得的ACE2人源化猪的猪器官、猪组织或猪细胞。
13.一种权利要求1-4任一所述的猪细胞、权利要求5-7任一所述构建方法获得的猪细胞、权利要求12所述的ACE2人源化猪的猪器官、猪组织或猪细胞，或者权利要求8或11所述构建方法获得的ACE2人源化猪的应用，其特征在于，所述应用包括：
(1)筛选治疗ACE2所介导疾病的药物；
(2)进行ACE2所介导疾病药物的药效评价；
(3)进行ACE2所介导疾病的疫苗效果测试；或，
(4)进行ACE2所介导的病毒感染机制研究。
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