CN112168958A - 基于慢病毒外壳修饰和mRNA递送的SARS-CoV-2疫苗及其制备方法 - Google Patents

基于慢病毒外壳修饰和mRNA递送的SARS-CoV-2疫苗及其制备方法 Download PDF

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CN112168958A
CN112168958A CN202010995678.3A CN202010995678A CN112168958A CN 112168958 A CN112168958 A CN 112168958A CN 202010995678 A CN202010995678 A CN 202010995678A CN 112168958 A CN112168958 A CN 112168958A
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蔡宇伽
尹笛
凌思凯
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Abstract

本发明提供了一种基于慢病毒外壳修饰和mRNA递送的SARS‑CoV‑2疫苗及其制备方法，所述疫苗是慢病毒颗粒作为外壳包裹SARS‑CoV‑2的棘突蛋白Spike的mRNA得到。通过慢病毒颗粒的递送功能将SARS‑CoV‑2的抗原，也即棘突蛋白Spike及其mRNA递送到体内。利用本发明，可以诱导机体产生针对Spike蛋白的中和抗体，从而起到预防SARS‑CoV‑2感染的作用。通过将SARS‑CoV‑2的棘突蛋白Spike替换成其他病毒的膜蛋白，本发明也可以作为其他病毒的疫苗。

Description

基于慢病毒外壳修饰和mRNA递送的SARS-CoV-2疫苗及其制备 方法
技术领域
本发明属于生物技术领域，具体涉及一种SARS-CoV-2mRNA疫苗的制备方法，尤其涉及一种慢病毒载体制备冠状病毒SARS-CoV-2mRNA疫苗的方法。
背景技术
疫苗是预防病毒、细菌等引起感染性疾病的有效方法。常见的疫苗平台有减毒疫苗、灭活病毒、重组蛋白、多肽、病毒载体递送的基因、DNA和mRNA等。传统的减毒和灭活疫苗等疫苗的制作工艺复杂，生产速度慢，安全级别要求高，无法快速应对紧急疫情。病毒载体和DNA疫苗则有整合到基因组的风险。所以亟待开发安全有效，可快速生产的疫苗制备方法。
mRNA疫苗作为新一代疫苗的方式逐渐成为研究热点。mRNA疫苗是将抗原的信使RNA输送到体内，在体内表达抗原。抗原与体内的受体结合引发特异性免疫反应并产生抗体。mRNA疫苗既可以应用于感染性疾病，也可以应用于非感染性疾病。已经有公司开展了针对肿瘤的mRNA疫苗I/II期临床。mRNA疫苗有以下优势：(1)mRNA疫苗设计简单，可以快速生产新出现病毒的抗原mRNA。(2)mRNA疫苗较为安全。与DNA疫苗相比，mRNA疫苗没有整合到基因组的风险。
mRNA需要借助载体才能进入细胞，发挥作用。现有最常用的mRNA疫苗的递送方式是脂质体递送。2019年新型冠状病毒SARS-CoV-2疫情爆发之后，Moderna公司研制的疫苗mRNA-1273是全球首个进入临床试验的mRNA疫苗。mRNA-1273是由脂质体包裹Spike蛋白的mRNA制成。然而脂质体递送方法感染细胞的效率较低，而感染效率对疫苗的成功有着关键的作用。
在先专利文献申请号为201810533437.X的中国发明专利申请公开了一种慢病毒载体及其递送外源RNA的方法；利用RNA结合蛋白和其所识别的RNA序列相互结合的原理，将RNA结合蛋白整合至慢病毒GagPol长链蛋白的骨架，并将其所识别的RNA序列连接到外源目的RNA,从而使目的RNA在慢病毒颗粒的组装过程中被包装进慢病毒颗粒。但该方法仅仅递送RNA，较难诱导产生针对构象表位的中和抗体。
发明内容
针对现有技术的不足，本发明的目的是提供一种慢病毒载体制备冠状病毒SARS-CoV-2mRNA疫苗的方法。生产一种兼具有效性和安全性的SARS-CoV-2疫苗。本发明既是一种mRNA疫苗，也是一种蛋白疫苗。本发明通过慢病毒颗粒包裹SARS-CoV-2的棘突蛋白Spike的mRNA。在此基础上，通过用Spike蛋白对慢病毒颗粒表面的装饰，从而模拟新冠病毒的外表，以三维结构的形式呈递Spike的蛋白，诱导机体产生针对构象表位的中和抗体。
本发明利用慢病毒颗粒作为递送载体，(1)慢病毒颗粒有囊膜蛋白，可将mRNA包裹在囊膜颗粒中，避免mRNA被核酸酶降解；(2)慢病毒颗粒具有高效的感染细胞的能力，大大提高了疫苗进入细胞的效率；(3)慢病毒载体进入细胞后将mRNA释放至细胞质中，高效地表达病毒抗原；(4)本发明的慢病毒颗粒外表装饰有Spike蛋白，可以将Spike蛋白以稳定的三维结构形式呈递给抗原呈递细胞。
本发明利用RNA结合蛋白与其所识别的RNA茎环结构的相互作用，将含该种可识别的RNA序列的Spike蛋白mRNA包装进慢病毒颗粒。与此同时，Spike蛋白作为一种棘突蛋白可以组装进慢病毒的包膜结构上。
本发明的目的是通过以下技术方案实现的：
本发明提供了一种基于慢病毒外壳修饰和mRNA递送的SARS-CoV-2疫苗，所述SARS-CoV-2疫苗是慢病毒颗粒作为外壳包裹SARS-CoV-2的棘突蛋白Spike的mRNA得到。
优选地，所述慢病毒颗粒的包膜上装饰有SARS-CoV-2的棘突蛋白Spike；所述棘突蛋白Spike的基因是密码子优化后和氨基酸突变后的基因，序列如SEQ ID NO.9所示。
优选地，所述慢病毒颗粒的包膜上还装饰有包膜蛋白VSV-G，表达所述包膜蛋白VSV-G的质粒的序列如SEQ ID NO.1所示。
优选地，所述棘突蛋白Spike的mRNA由SARS-CoV-2的棘突蛋白Spike基因转录得到；所述棘突蛋白Spike的基因是经过密码子优化以及氨基酸突变后的基因，序列如SEQ IDNO.9所示。
本发明还提供了一种SARS-CoV-2疫苗的制备方法，通过将疫苗生产质粒转染入病毒生产细胞；收集上清，浓缩，制备而得。
优选地，所述制备方法的具体步骤如下：
S1、将表达VSV-G膜蛋白的质粒、表达含RNA结合蛋白的慢病毒GagPol长链蛋白的质粒、表达含慢病毒GagPol长链蛋白的质粒、表达含RNA结合蛋白所识别的茎环结构和棘突蛋白Spike mRNA的质粒共转染病毒生产细胞；所述茎环结构置于棘突蛋白Spike mRNA的表达框内；所述棘突蛋白Spike mRNA由SARS-CoV-2的棘突蛋白Spike基因转录得到；
S2、收集含病毒颗粒的上清，浓缩，得到含棘突蛋白Spike的mRNA、并在外壳装饰有棘突蛋白Spike的慢病毒颗粒。
优选地，所述表达含RNA结合蛋白的慢病毒GagPol长链蛋白的质粒是通过将RNA结合蛋白整合至第三代慢病毒GagPol长链蛋白的N-端制成，所述RNA结合蛋白为MS2衣壳蛋白；
所述MS2衣壳蛋白的基因序列如SEQ ID NO.5所示，表达含RNA结合蛋白的慢病毒GagPol长链蛋白的质粒序列如SEQ ID NO.4所示。
优选地，所述表达含RNA结合蛋白所识别的茎环结构的棘突蛋白Spike mRNA的质粒是通过将RNA结合蛋白所识别的茎环结构的RNA序列与棘突蛋白Spike的mRNA序列融合而得到，所得质粒的序列如SEQ ID NO.11所示。
优选地，所述含RNA结合蛋白所识别的茎环结构在Spike mRNA的表达框中，茎环结构的RNA序列如SEQ ID NO.8所示。
优选地，所述病毒生产细胞选自293T细胞、293FTX细胞、HEK293细胞；
所述表达含慢病毒GagPol长链蛋白的质粒序列如SEQ ID NO.2所示。
本发明所述的SARS-CoV-2疫苗是由慢病毒载体递送棘突蛋白Spike的mRNA和蛋白本身。以此方法制备的慢病毒载体还可以通过递送SARS、MERS、HIV等其它病毒的抗原蛋白的mRNA来制备相应的疫苗。
本发明以慢病毒颗粒作为递送载体的SARS-CoV-2疫苗可用于实现预防新型冠状病毒SARS-CoV-2感染的用途。
与已有的脂质体作为递送介质的SARS-CoV-2mRNA疫苗相比，本发明具有如下的有益效果：
(1)本发明的SARS-CoV-2mRNA疫苗是由慢病毒颗粒携带Spike蛋白的mRNA,其感染细胞的效率增加。
(2)本发明SARS-CoV-2mRNA疫苗的慢病毒颗粒的囊膜结构上含有Spike蛋白，从而模拟新冠病毒的外表，以三维结构的形式呈递Spike的蛋白，诱导机体产生更有效的中和抗体。
(3)本发明利用慢病毒颗粒作为递送载体，慢病毒颗粒有囊膜蛋白，还可将mRNA包裹在囊膜颗粒中，避免mRNA被核酸酶降解；慢病毒载体进入细胞后将mRNA释放至细胞质中，可高效地表达病毒抗原。
附图说明
通过阅读参照以下附图对非限制性实施例所作的详细描述，本发明的其它特征、目的和优点将会变得更明显：
图1为：SARS-CoV-2疫苗的Spike mRNA表达质粒的构造；
图2为：SARS-CoV 2mRNA疫苗的包装策略；
图3为：SARS-CoV-2疫苗包装的Spike蛋白情况；
图4为：SARS-CoV-2疫苗感染293T细胞，所携带的mRNA表达的Spike蛋白情况；
图5为：SARS-CoV-2疫苗接种小鼠，产生中和抗体。
具体实施方式
下面结合具体实施例对本发明进行详细说明。以下实施例将有助于本领域的技术人员进一步理解本发明，但不以任何形式限制本发明。应当指出的是，对本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进。这些都属于本发明的保护范围。
以下对本发明中涉及的一些术语进行解释：慢病毒载体是由HIV-1改造而来的一种不能自我复制的病毒载体。慢病毒载体可以高效地感染分裂细胞和非分裂细胞，是生物学研究和基因治疗常用的载体。目前，慢病毒载体已经发展到第三代，安全性越来越高。Spike蛋白是冠状病毒的棘突蛋白，能与细胞表面受体结合而感染细胞。Spike蛋白可引起机体免疫产生对应的抗体，以此原理设计疫苗是常用的方法。
本发明的目的是：将SARS-CoV-2的Spike蛋白mRNA包装进慢病毒载体。
本发明的原理是：利用RNA结合蛋白与其所识别的RNA茎环结构的相互作用，将融合了该种可识别的RNA序列的Spike蛋白mRNA包装进慢病毒颗粒。
本发明的特点是：慢病毒载体将SARS-CoV-2的Spike蛋白mRNA递送进细胞表达Spike蛋白；Spike蛋白作为棘突蛋白可以组装进慢病毒的包膜结构上，可作为抗原与细胞受体结合。
实施例1、SARS-CoV-2mRNA疫苗的质粒构造方法及包装策略
本实施例涉及SARS-CoV-2mRNA疫苗制备方法；Spike mRNA表达质粒的构造如图1所示，SARS-CoV-2mRNA疫苗的包装策略如图2所示。具体步骤包括：
步骤1：将表达膜蛋白的质粒(如VSV-G)、表达慢病毒GagPol长链蛋白的质粒、表达含RNA结合蛋白的慢病毒GagPol长链蛋白的质粒、表达带RNA结合蛋白所识别的茎环结构和棘突蛋白Spike mRNA的质粒共转染病毒生产细胞(如293T)。其中，
(1)表达膜蛋白VSV-G的质粒是公知质粒pMD2G，其质粒序列如SEQ ID NO.1所示。
(2)表达含慢病毒GagPol长链蛋白的质粒是公知质粒pMDlg/PRRE的整合酶缺陷型，其质粒序列如SEQ ID NO.2所示。辅助质粒是公知质粒pRSV-REV的质粒序列如SEQ IDNO.3所示。辅助质粒pRSV-REV与pMDlg/PRRE的整合酶缺陷型质粒一起转染进病毒生产细胞用于疫苗的制备。
(3)表达含RNA结合蛋白的慢病毒GagPol长链蛋白的质粒是通过将MS2衣壳蛋白与慢病毒GagPol长链蛋白融合而得到。所述表达含RNA结合蛋白的慢病毒GagPol长链蛋白的质粒序列如SEQ ID NO.4所示。
所述MS2衣壳蛋白的基因序列如SEQ ID NO.5所示，其编码的蛋白可与GagPol长链蛋白相连，置于该GagPol长链蛋白的N-端。
该质粒的制备方案如下：
①根据基因序列SEQ ID NO.5所示合成MS2蛋白的基因；
②PCR扩增MS2蛋白的基因，引物序列为F:5'-tagcaccggtcgccaccatggcctctaattttactca-3'(SEQ ID NO.13),R:5'-tagctccggactgcacaa tcggatagttctggctgtatatcccgctgttggcgg-3'(SEQ ID NO.14)；
③采用限制性内切酶AgeI和AccIII分别酶切步骤②扩增的PCR产物(即插入片段)和载体质粒pGFP-PH-gagpol-D64V(序列如SEQ ID NO.6所示)；
④用T4连接酶连接步骤③酶切后的载体质粒pGFP-PH-gagpol-D64V和插入片段后转化，抽提质粒，具体转化，抽提质粒的步骤为：连接体系经过感受态XL1-Blue细胞转化、挑取单克隆和摇菌，最后用质粒抽提试剂盒抽提质粒。
⑤用引物5'-attgacgcaaatgggcggtag-3'(SEQ ID NO.15)进行一代测序验证，结果与设计的质粒序列一致时，即得表达含RNA结合蛋白的慢病毒GagPol长链蛋白的质粒，并进行后续步骤。
(4)表达含RNA结合蛋白所识别的茎环结构和棘突蛋白Spike mRNA的质粒(pCMV-nCoV-S-6XMS2-mut-flag)是通过将MS2衣壳蛋白所识别的茎环结构的RNA序列与SARS-CoV-2Spike蛋白的mRNA序列融合而得到，质粒结构参见图1，pCMV-nCoV-S-6XMS2-mut-flag序列如SEQ ID NO.11所示。
其中，MS2衣壳蛋白所识别的RNA序列如SEQ ID NO.7所示，该序列可以以单次或者多次重复的方式与外源目的RNA连接从而形成含有单个或多个茎环结构的外源目的RNA序列。本实施例用的6次重复(每个RNA序列的两端都有连接序列)的序列是由基因合成所得，序列如SEQ ID NO.8所示。
所述Spike蛋白的mRNA序列由SARS-CoV-2的棘突蛋白Spike基因转录得到；所述SARS-CoV-2Spike蛋白基因是经过了密码子优化后由基因合成所得，并引入氨基酸突变(K1003P和V1004P)。所得基因序列如SEQ ID NO.9所示。
表达含RNA结合蛋白所识别的茎环结构和棘突蛋白Spike mRNA的质粒制备方案如下：
①根据基因序列SEQ ID NO.8合成RNA结合蛋白所识别的RNA茎环结构序列；根据基因序列SEQ ID NO.9所示合成SARS-CoV-2Spike蛋白的基因，所述基因可转录棘突蛋白Spike mRNA；
②PCR扩增SARS-CoV-2Spike蛋白的基因，引物序列为F:5'-tagcccgcggacgcgtgccaccatggactggacctggatcct-3'(SEQ ID NO.16)，R：5'-ttacttatcgtcatcgtctt-3'(SEQ IDNO.17)；
PCR扩增茎环结构RNA的基因序列，引物为F：5'-aagacgatgacgataagtaaaccggtcttgcagcacatacatgag-3'(SEQ ID NO.18)和R：5'-tagcccgcgggccaagcttaaggtgcacac-3'(SEQ ID NO.19)。
③融合PCR：将步骤②扩增得到的两段基因融合成一条，引物为：F:5'-tagcacgcgtgccaccatggactggacctggatcct-3'(SEQ ID NO.20)，R：5'-tagcccgcgggccaagcttaaggtgcacac-3'(SEQ ID NO.21)。
④采用限制性内切酶SacII分别酶切步骤③融合后的PCR产物(即插入片段)和载体质粒pCMV-GFP(序列如SEQ ID NO.10所示)；
⑤用T4连接酶连接步骤④的载体质粒和插入片段后转化，抽提质粒，具体转化，抽提质粒的步骤为：连接体系经过感受态XL1-Blue细胞转化、挑取单克隆和摇菌，最后用质粒抽提试剂盒抽提质粒。
⑥用引物F:5'-attgacgcaaatgggcggtag-3'(SEQ ID NO.15)和R:5'-gtttattgcagcttataatg-3'(SEQ ID NO.22)进行一代测序验证，结果与设计的质粒序列一致时，即得表达含RNA结合蛋白所识别的茎环结构和棘突蛋白Spike mRNA的质粒(pCMV-nCoV-S-6XMS2-mut-flag)，并进行后续步骤。
步骤2：收集含病毒颗粒的上清，通过高速离心或者HPLC等方法进行浓缩，得到高滴度的含mRNA的慢病毒颗粒，即SARS-CoV-2mRNA疫苗。
本发明中介绍的mRNA疫苗制备方法可以用于制备SARS-CoV-2mRNA疫苗，也可以制备其它病毒的疫苗。
实施例2、SARS-CoV-2mRNA疫苗包装的Spike蛋白情况
具体步骤同实施例1，所用的SARS-CoV-2Spike蛋白基因C-末端含有1个拷贝的Flag蛋白的基因。得到高滴度SARS-CoV-2mRNA疫苗的慢病毒颗粒后，用慢病毒滴度试剂盒检测p24含量。取p24 300ng进行Western Blot(免疫印迹试验)。显示Spike蛋白条带位置的一抗用鼠源的anti-Flag；显示p24蛋白条带位置的一抗用鼠源的anti-p24,二抗用anti-Mouse IgG。去糖基化酶用PNGase F。结果见图3，显示本实施例方法制备的SARS-CoV-2mRNA疫苗(VSP-S-mut表示)含有Spike蛋白的S2亚基(Spike蛋白在病毒成熟时被剪切使得S1亚基和S2亚基分开，本实施例中，Flag在Spike蛋白的C-端，所以检测到S2亚基)，且Spike蛋白的含量比含有未突变Spike基因的SARS-CoV-2mRNA疫苗颗粒(VSP-S表示，其制备方法与实施例1基本相同，不同之处仅在于：表达含RNA结合蛋白所识别的茎环结构和棘突蛋白SpikemRNA的质粒制备中，采用的SARS-CoV-2Spike蛋白基因没有进行氨基酸突变)的多。
图3中还对比了核酸载体分别含有Spike蛋白基因(未突变/突变)的普通慢病毒(IDLV-S/IDLV-S-mut，其制备方法与实施例1基本相同，不同的是将表达膜蛋白的质粒(如VSV-G)、表达慢病毒GagPol长链蛋白的质粒、表达Spike蛋白基因(未突变/突变)的慢病毒载体质粒共转染进细胞生产而得。其中，表达Spike蛋白基因(氨基酸突变)的慢病毒载体质粒(是由Spike蛋白基因(氨基酸突变)替换了慢病毒载体质粒pCCL-PGK-eGFP中的GFP基因所得，基因序列如SEQ ID NO.12所示)不能或很少携带Spike蛋白。去糖基化处理后得到真实S2亚基(约80kDa)。普通慢病毒颗粒IDLV作为阴性对照。
实施例3、SARS-CoV-2疫苗感染293T细胞，所携带的mRNA表达的Spike蛋白情况
在48孔板里种293T细胞，4×104个/孔。24小时后分别感染p24 300ng SARS-CoV-2mRNA疫苗(实施例2制备的VSP-S-mut)，含有未突变Spike基因的SARS-CoV-2mRNA疫苗(实施例2制备的VSP-S)以及核酸载体含有Spike蛋白基因(未突变/突变)的普通慢病毒(IDLV-S,IDLV-S-mut)。同时293T细胞各转染2ug质粒pCMV-nCoV-S-6XMS2-flag和pCMV-nCoV-S-6XMS2-mut-flag。转染和感染各种制备好的慢病毒载体和质粒36小时后收细胞进行Western Blot(免疫印迹试验)。所用抗体同实施例2。图4显示，本实施例方法制备的SARS-CoV-2mRNA疫苗(VSP-S-mut)可以感染细胞，并表达大量的Spike蛋白。未被转染和感染的293T细胞作为空白对照。
实施例4、SARS-CoV-2疫苗接种小鼠，产生中和抗体
足垫注射1.5ug p24的SARS-CoV-2疫苗到小鼠体内(VSP-S-mut)，14天后取血清检测中和抗体。注射PBS的小鼠(Placebo)作为对照。用包装了荧光素酶(luciferase)的SARS-CoV-2假病毒(自行制备，制备方法参考文献：Lu L,Liu Q,Zhu Y,Chan K-H,Qin L,Li Y,etal.Structure-based discovery of Middle East respiratory syndrome coronavirusfusion inhibitor.2014；5(1):1-12.)的荧光信号值评价中和效果。假病毒被抗体中和后无法进入细胞表达荧光。EC50(能中和50％假病毒的血清稀释倍数)作为衡量指标。图5显示，在注射SARS-CoV-2疫苗的小鼠的高稀释浓度的血清中检测到了中和抗体。
本发明具体应用途径很多，以上所述仅是本发明的优选实施方式。应当指出，以上实施例仅用于说明本发明，而并不用于限制本发明的保护范围。对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进，这些改进也应视为本发明的保护范围。
序列表
<110> 上海交通大学
<120> 基于慢病毒外壳修饰和mRNA递送的SARS-CoV-2疫苗及其制备方法
<130> KAG43762
<160> 22
<170> SIPOSequenceListing 1.0
<210> 1
<211> 5824
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
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 atctgaattc aacagagatc gatctgtttc 1440
cttgacacta tgaagtgcct tttgtactta gcctttttat tcattggggt gaattgcaag 1500
ttcaccatag tttttccaca caaccaaaaa ggaaactgga aaaatgttcc ttctaattac 1560
cattattgcc cgtcaagctc agatttaaat tggcataatg acttaatagg cacagccata 1620
caagtcaaaa tgcccaagag tcacaaggct attcaagcag acggttggat gtgtcatgct 1680
tccaaatggg tcactacttg tgatttccgc tggtatggac cgaagtatat aacacagtcc 1740
atccgatcct tcactccatc tgtagaacaa tgcaaggaaa gcattgaaca aacgaaacaa 1800
ggaacttggc tgaatccagg cttccctcct caaagttgtg gatatgcaac tgtgacggat 1860
gccgaagcag tgattgtcca ggtgactcct caccatgtgc tggttgatga atacacagga 1920
gaatgggttg attcacagtt catcaacgga aaatgcagca attacatatg ccccactgtc 1980
cataactcta caacctggca ttctgactat aaggtcaaag ggctatgtga ttctaacctc 2040
atttccatgg acatcacctt cttctcagag gacggagagc tatcatccct gggaaaggag 2100
ggcacagggt tcagaagtaa ctactttgct tatgaaactg gaggcaaggc ctgcaaaatg 2160
caatactgca agcattgggg agtcagactc ccatcaggtg tctggttcga gatggctgat 2220
aaggatctct ttgctgcagc cagattccct gaatgcccag aagggtcaag tatctctgct 2280
ccatctcaga cctcagtgga tgtaagtcta attcaggacg ttgagaggat cttggattat 2340
tccctctgcc aagaaacctg gagcaaaatc agagcgggtc ttccaatctc tccagtggat 2400
ctcagctatc ttgctcctaa aaacccagga accggtcctg ctttcaccat aatcaatggt 2460
accctaaaat actttgagac cagatacatc agagtcgata ttgctgctcc aatcctctca 2520
agaatggtcg gaatgatcag tggaactacc acagaaaggg aactgtggga tgactgggca 2580
ccatatgaag acgtggaaat tggacccaat ggagttctga ggaccagttc aggatataag 2640
tttcctttat acatgattgg acatggtatg ttggactccg atcttcatct tagctcaaag 2700
gctcaggtgt tcgaacatcc tcacattcaa gacgctgctt cgcaacttcc tgatgatgag 2760
agtttatttt ttggtgatac tgggctatcc aaaaatccaa tcgagcttgt agaaggttgg 2820
ttcagtagtt ggaaaagctc tattgcctct tttttcttta tcatagggtt aatcattgga 2880
ctattcttgg ttctccgagt tggtatccat ctttgcatta aattaaagca caccaagaaa 2940
agacagattt atacagacat agagatgaac cgacttggaa agtaactcaa atcctgcaca 3000
acagattctt catgtttgga ccaaatcaac ttgtgatacc atgctcaaag aggcctcaat 3060
tatatttgag tttttaattt ttatggaatt caccccacca gtgcaggctg cctatcagaa 3120
agtggtggct ggtgtggcta atgccctggc ccacaagttt cactaagctc gcttccttgc 3180
tgtccaattt ctattaaagg ttccttggtt ccctaagtcc aactactaaa ctgggggata 3240
ttatgaaggg ccttgagcat ctggattctg cctaataaaa aacatttatt ttcattgcaa 3300
tgatgtattt aaattatttc tgaatatttt actaaaaagg gaatgtggga ggtcagtgca 3360
tttaaaacat aaagaaatga agagctagtt caaaccttgg gaaaatacac tatatcttaa 3420
actccatgaa agaaggtgag gctgcaaaca gctaatgcac attggcaaca gccctgatgc 3480
ctatgcctta ttcatccctc agaaaaggat tcaagtagag gcttgatttg gaggttaaag 3540
tttggctatg ctgtatttta cattacttat tgttttagct gtcctcatga atgtcttttc 3600
actacccatt tgcttatcct gcatctctca gccttgactc cactcagttc tcttgcttag 3660
agataccacc tttcccctga agtgttcctt ccatgtttta cggcgagatg gtttctcctc 3720
gcctggccac tcagccttag ttgtctctgt tgtcttatag aggtctactt gaagaaggaa 3780
aaacaggggg catggtttga ctgtcctgtg agcccttctt ccctgcctcc cccactcaca 3840
gtgacccgga atccctcgac atggcagtct agcactagtg cggccgcaga tctgcttcct 3900
cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 3960
aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 4020
aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 4080
tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 4140
caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 4200
cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 4260
ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 4320
gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 4380
agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 4440
gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 4500
acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 4560
gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 4620
gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 4680
cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 4740
caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 4800
gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 4860
cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 4920
cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 4980
caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 5040
gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 5100
gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 5160
cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 5220
catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 5280
gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 5340
ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 5400
gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 5460
cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 5520
tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 5580
gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 5640
atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 5700
ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 5760
gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 5820
acgt 5824
<210> 2
<211> 8895
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
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 atctgaattc gagatctgcc gccgccatgg 1440
gtgcgagagc gtcagtatta agcgggggag aattagatcg atgggaaaaa attcggttaa 1500
ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag 1560
aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg 1620
gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag 1680
tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag 1740
acaagataga ggaagagcaa aacaaaagta agaaaaaagc acagcaagca gcagctgaca 1800
caggacacag caatcaggtc agccaaaatt accctatagt gcagaacatc caggggcaaa 1860
tggtacatca ggccatatca cctagaactt taaatgcatg ggtaaaagta gtagaagaga 1920
aggctttcag cccagaagtg atacccatgt tttcagcatt atcagaagga gccaccccac 1980
aagatttaaa caccatgcta aacacagtgg ggggacatca agcagccatg caaatgttaa 2040
aagagaccat caatgaggaa gctgcagaat gggatagagt gcatccagtg catgcagggc 2100
ctattgcacc aggccagatg agagaaccaa ggggaagtga catagcagga actactagta 2160
ctagtaccct tcaggaacaa ataggatgga tgacacataa tccacctatc ccagtaggag 2220
aaatctataa aagatggata atcctgggat taaataaaat agtaagaatg tatagcccta 2280
ccagcattct ggacataaga caaggaccaa aggaaccctt tagagactat gtagaccgat 2340
tctataaaac tctaagagcc gagcaagctt cacaagaggt aaaaaattgg atgacagaaa 2400
ccttgttggt ccaaaatgcg aacccagatt gtaagactat tttaaaagca ttgggaccag 2460
gagcgacact agaagaaatg atgacagcat gtcagggagt ggggggaccc ggccataaag 2520
caagagtttt ggctgaagca atgagccaag taacaaatcc agctaccata atgatacaga 2580
aaggcaattt taggaaccaa agaaagactg ttaagtgttt caattgtggc aaagaagggc 2640
acatagccaa aaattgcagg gcccctagga aaaagggctg ttggaaatgt ggaaaggaag 2700
gacaccaaat gaaagattgt actgagagac aggctaattt tttagggaag atctggcctt 2760
cccacaaggg aaggccaggg aattttcttc agagcagacc agagccaaca gccccaccag 2820
aagagagctt caggtttggg gaagagacaa caactccctc tcagaagcag gagccgatag 2880
acaaggaact gtatccttta gcttccctca gatcactctt tggcagcgac ccctcgtcac 2940
aataaagata ggggggcaat taaaggaagc tctattagat acaggagcag atgatacagt 3000
attagaagaa atgaatttgc caggaagatg gaaaccaaaa atgatagggg gaattggagg 3060
ttttatcaaa gtaggacagt atgatcagat actcatagaa atctgtggac ataaagctat 3120
aggtacagta ttagtaggac ctacacctgt caacataatt ggaagaaatc tgttgactca 3180
gattggttgc actttaaatt ttcccattag ccctattgag actgtaccag taaaattaaa 3240
gccaggaatg gatggcccaa aagttaaaca atggccattg acagaagaaa aaataaaagc 3300
attagtagaa atttgtacag agatggaaaa ggaagggaaa atttcaaaaa ttgggcctga 3360
aaatccatac aatactccag tatttgccat aaagaaaaaa gacagtacta aatggagaaa 3420
attagtagat ttcagagaac ttaataagag aactcaagac ttctgggaag ttcaattagg 3480
aataccacat cccgcagggt taaaaaagaa aaaatcagta acagtactgg atgtgggtga 3540
tgcatatttt tcagttccct tagatgaaga cttcaggaag tatactgcat ttaccatacc 3600
tagtataaac aatgagacac cagggattag atatcagtac aatgtgcttc cacagggatg 3660
gaaaggatca ccagcaatat tccaaagtag catgacaaaa atcttagagc cttttagaaa 3720
acaaaatcca gacatagtta tctatcaata catggatgat ttgtatgtag gatctgactt 3780
agaaataggg cagcatagaa caaaaataga ggagctgaga caacatctgt tgaggtgggg 3840
acttaccaca ccagacaaaa aacatcagaa agaacctcca ttcctttgga tgggttatga 3900
actccatcct gataaatgga cagtacagcc tatagtgctg ccagaaaaag acagctggac 3960
tgtcaatgac atacagaagt tagtggggaa attgaattgg gcaagtcaga tttacccagg 4020
gattaaagta aggcaattat gtaaactcct tagaggaacc aaagcactaa cagaagtaat 4080
accactaaca gaagaagcag agctagaact ggcagaaaac agagagattc taaaagaacc 4140
agtacatgga gtgtattatg acccatcaaa agacttaata gcagaaatac agaagcaggg 4200
gcaaggccaa tggacatatc aaatttatca agagccattt aaaaatctga aaacaggaaa 4260
atatgcaaga atgaggggtg cccacactaa tgatgtaaaa caattaacag aggcagtgca 4320
aaaaataacc acagaaagca tagtaatatg gggaaagact cctaaattta aactgcccat 4380
acaaaaggaa acatgggaaa catggtggac agagtattgg caagccacct ggattcctga 4440
gtgggagttt gttaataccc ctcccttagt gaaattatgg taccagttag agaaagaacc 4500
catagtagga gcagaaacct tctatgtaga tggggcagct aacagggaga ctaaattagg 4560
aaaagcagga tatgttacta atagaggaag acaaaaagtt gtcaccctaa ctgacacaac 4620
aaatcagaag actgagttac aagcaattta tctagctttg caggattcgg gattagaagt 4680
aaacatagta acagactcac aatatgcatt aggaatcatt caagcacaac cagatcaaag 4740
tgaatcagag ttagtcaatc aaataataga gcagttaata aaaaaggaaa aggtctatct 4800
ggcatgggta ccagcacaca aaggaattgg aggaaatgaa caagtagata aattagtcag 4860
tgctggaatc aggaaagtac tatttttaga tggaatagat aaggcccaag atgaacatga 4920
gaaatatcac agtaattgga gagcaatggc tagtgatttt aacctgccac ctgtagtagc 4980
aaaagaaata gtagccagct gtgataaatg tcagctaaaa ggagaagcca tgcatggaca 5040
agtagactgt agtccaggaa tatggcaact agtttgtaca catttagaag gaaaagttat 5100
cctggtagca gttcatgtag ccagtggata tatagaagca gaagttattc cagcagaaac 5160
agggcaggaa acagcatatt ttcttttaaa attagcagga agatggccag taaaaacaat 5220
acatactgac aatggcagca atttcaccgg tgctacggtt agggccgcct gttggtgggc 5280
gggaatcaag caggaatttg gaattcccta caatccccaa agtcaaggag tagtagaatc 5340
tatgaataaa gaattaaaga aaattatagg acaggtaaga gatcaggctg aacatcttaa 5400
gacagcagta caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg 5460
gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt 5520
acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc 5580
agtttggaaa ggaccagcaa agctcctctg gaaaggtgaa ggggcagtag taatacaaga 5640
taatagtgac ataaaagtag tgccaagaag aaaagcaaag atcatcaggg attatggaaa 5700
acagatggca ggtgatgatt gtgtggcaag tagacaggat gaggattaac acatggaatt 5760
ccggagcggc cgcaggagct ttgttccttg ggttcttggg agcagcagga agcactatgg 5820
gcgcagcctc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc 5880
agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct 5940
ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac 6000
agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga 6060
atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt 6120
gggacagaga aattaacaat tacacaagct tccgcggaat tcaccccacc agtgcaggct 6180
gcctatcaga aagtggtggc tggtgtggct aatgccctgg cccacaagtt tcactaagct 6240
cgcttccttg ctgtccaatt tctattaaag gttccttggt tccctaagtc caactactaa 6300
actgggggat attatgaagg gccttgagca tctggattct gcctaataaa aaacatttat 6360
tttcattgca atgatgtatt taaattattt ctgaatattt tactaaaaag ggaatgtggg 6420
aggtcagtgc atttaaaaca taaagaaatg aagagctagt tcaaaccttg ggaaaataca 6480
ctatatctta aactccatga aagaaggtga ggctgcaaac agctaatgca cattggcaac 6540
agccctgatg cctatgcctt attcatccct cagaaaagga ttcaagtaga ggcttgattt 6600
ggaggttaaa gtttggctat gctgtatttt acattactta ttgttttagc tgtcctcatg 6660
aatgtctttt cactacccat ttgcttatcc tgcatctctc agccttgact ccactcagtt 6720
ctcttgctta gagataccac ctttcccctg aagtgttcct tccatgtttt acggcgagat 6780
ggtttctcct cgcctggcca ctcagcctta gttgtctctg ttgtcttata gaggtctact 6840
tgaagaagga aaaacagggg gcatggtttg actgtcctgt gagcccttct tccctgcctc 6900
ccccactcac agtgacccgg aatccctcga catggcagtc tagcactagt gcggccgcag 6960
atctgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7020
agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7080
catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7140
tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7200
gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7260
ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7320
cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7380
caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7440
ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7500
taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7560
taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7620
cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7680
tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7740
gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7800
catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7860
atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7920
ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7980
gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8040
agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8100
gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8160
agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8220
catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8280
aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8340
gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8400
taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8460
caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8520
ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8580
ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8640
tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8700
aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8760
actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8820
catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8880
agtgccacct gacgt 8895
<210> 3
<211> 4174
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 60
acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 120
tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 180
ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gaattcgatg 240
tacgggccag atatacgcgt atctgagggg actaggtgtg tttaggcgaa aagcggggct 300
tcggttgtac gcggttagga gtcccctcag gattagtagt ttcgcttttg catagggagg 360
gggaaatgta gtcttatgca atacacttgt agtcttgcaa catggtaacg atgagttagc 420
aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt 480
acgatcgtgc cttattagga aggcaacaga caggtctgac atggattgga cgaaccactg 540
aattccgcat tgcagagata attgtattta agtgcctagc tcgatacaat aaacgccatt 600
tgaccattca ccacattggt gtgcacctcc aagctcgagc tcgtttagtg aaccgtcaga 660
tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca 720
gcctcccctc gaagctagtc gattaggcat ctcctatggc aggaagaagc ggagacagcg 780
acgaagacct cctcaaggca gtcagactca tcaagtttct ctatcaaagc aacccacctc 840
ccaatcccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac 900
agagacagat ccattcgatt agtgaacgga tccttagcac ttatctggga cgatctgcgg 960
agcctgtgcc tcttcagcta ccaccgcttg agagacttac tcttgattgt aacgaggatt 1020
gtggaacttc tgggacgcag ggggtgggaa gccctcaaat attggtggaa tctcctacaa 1080
tattggagtc aggagctaaa gaatagtgct gttagcttgc tcaatgccac agctatagca 1140
gtagctgagg ggacagatag ggttatagaa gtagtacaag aagcttggca ctggccgtcg 1200
ttttacatga tctgagcctg ggagatctct ggctaactag ggaacccact gcttaagcct 1260
caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt 1320
aactagagat caggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc 1380
tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg 1440
cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat 1500
ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg 1560
gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 1620
cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 1680
aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 1740
cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 1800
ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 1860
aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg 1920
ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt 1980
acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc 2040
cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct 2100
tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca 2160
ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg 2220
ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct 2280
atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga 2340
taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc 2400
cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg 2460
aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc 2520
aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact 2580
tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc 2640
ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag 2700
catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat 2760
aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt 2820
ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa 2880
gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc 2940
aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg 3000
gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt 3060
gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca 3120
gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat 3180
gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca 3240
gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg 3300
atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 3360
ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 3420
ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 3480
ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 3540
ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 3600
ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 3660
tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 3720
tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 3780
tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 3840
tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 3900
gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 3960
tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 4020
ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct 4080
gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc 4140
gagcgcagcg agtcagtgag cgaggaagcg gaag 4174
<210> 4
<211> 10704
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 60
ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 120
cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 180
ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 240
gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 300
tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 360
cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 420
attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 480
atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 540
atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 600
tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 660
actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 720
aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 780
gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc 840
gctagcgcta ccggtcgcca ccatggcctc taattttact caatttgtgc ttgtcgataa 900
tggggggacg ggagatgtga ccgttgcccc tagcaatttc gcaaatggcg ttgcagaatg 960
gatctctagc aacagcagaa gccaagcgta caaagtaacg tgttccgttc gccaaagctc 1020
cgcccaaaaa cggaagtata caataaaggt tgaggtgccg aaagtagcca ctcaaacagt 1080
tggtggggta gaattgcccg tagcggcatg gcggtcatat ctcaatatgg aactcactat 1140
cccaatcttc gccacgaata gcgattgtga gctgatagtt aaggctatgc aaggtcttct 1200
caaagatgga aaccctattc catctgctat cgccgccaac agcgggatat acagccagaa 1260
ctatccgatt gtgcagtccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga 1320
cggtaccgcg ggcccgggat ccatggactc gggccgggac ttcctgaccc tgcacggcct 1380
acaggatgat gaggatctac aggcgctgct gaagggcagc cagctcctga aggtgaagtc 1440
cagctcatgg aggagagagc gcttctacaa gttgcaggag gactgcaaga ccatctggca 1500
ggagtcccgc aaggtcatgc ggaccccgga gtcccagctg ttctccatcg aggacattca 1560
ggaggtgcga atggggcacc gcacggaggg tctggagaag ttcgcccgtg atgtgcccga 1620
ggaccgctgc ttctccattg tcttcaagga ccagcgcaat acactagacc tcatcgcccc 1680
atcgccagct gatgcccagc actgggtgct ggggctgcac aagatcatcc accactcagg 1740
ctccatggac cagcgtcaga agctacagca ctggattcac tcctgcttgc gaaaagctga 1800
caaaaacaag gacaacaaga tgagcttcaa ggagctgcag aacttcctga aggagctcaa 1860
catccagcct cgagccgaat tcaccctggc cgccagggcc agcgtgctga gcggcggcga 1920
gctggacagg tgggagaaga tcaggctgag gcccggcggc aagaagaagt ataagctgaa 1980
gcacatcgtg tgggccagca gggagctgga gaggttcgcc gtgaaccccg gcctgctgga 2040
gaccagcgag ggctgcaggc agatcctggg ccagctgcag cccagcctgc agaccggcag 2100
cgaggagctg aggagcctgt acaacaccgt ggccaccctg tactgcgtgc accagaggat 2160
cgagatcaag gacaccaagg aggccctgga caagatcgag gaggagcaga acaagtccaa 2220
gaagaaggcc cagcaggccg ccgccgacac cggccacagc agccaggtga gccagaacta 2280
ccccatcgtg cagaacatcc agggccagat ggtgcaccag gccatcagcc ccaggaccct 2340
gaacgcctgg gtgaaggtgg tggaggagaa ggccttcagc cccgaggtga tccccatgtt 2400
cagcgccctg agcgagggag ccacccccca ggacctgaac accatgctga acaccgtggg 2460
cggccaccag gccgccatgc agatgctgaa ggagaccatc aacgaggagg ccgccgagtg 2520
ggacagggtg caccccgtgc acgccggccc catcgccccc ggccagatga gggagccccg 2580
cggcagcgac atcgccggca ccaccagcac cctgcaggag cagatcggct ggatgaccaa 2640
caaccccccc atccccgtgg gcgaaatcta caagaggtgg atcatcctgg gcctgaacaa 2700
gatcgtgagg atgtacagcc ccaccagcat cctggatatc aggcagggcc ccaaagagcc 2760
cttcagggac tacgtggaca ggttctacaa gaccctgcgc gccgagcagg ccagccagga 2820
ggtgaagaac tggatgaccg agaccctgct ggtgcagaac gccaaccccg actgcaagac 2880
catcctgaag gccctgggac ccgccgccac cctggaggag atgatgaccg cctgccaggg 2940
cgtgggcggc cccggccaca aggccagggt gctggccgag gccatgagcc aggtgaccaa 3000
caccgccacc atcatgatgc agaggggcaa cttcaggaac cagaggaaga tggtgaagtg 3060
cttcaactgc ggcaaggagg gccacaccgc caggaactgc cgcgccccca ggaagaaggg 3120
ctgctggaag tgcggcaagg agggccacca gatgaaggac tgcaccgaga ggcaggctaa 3180
ttttttaggg aagatctggc cttcctacaa gggaaggcca gggaattttc ttcagagcag 3240
accagagcca acagccccac catttcttca gagcagacca gagccaacag ccccaccaga 3300
agagagcttc aggtctgggg tagagacaac aactccccct cagaagcagg agccgataga 3360
caaggaactg tatcctttaa cttccctcag atcactcttt ggcaacgacc cctcgtcaca 3420
ataaagatcg gtggccagct gaaggaggcc ctgctggaca ccggcgccga cgacaccgtg 3480
ctggaggaga tgagcctgcc cggcaggtgg aagcccaaga tgatcggcgg catcggcggc 3540
ttcatcaagg tgaggcagta cgaccagatc ctgatcgaga tctgcggcca caaggccatc 3600
ggcaccgtgc tggtgggacc tacacctgtg aacatcatcg gcaggaacct gctgacccag 3660
atcggctgca ccctgaactt ccccatcagc cccatcgaga ccgtgcccgt gaagctgaag 3720
cccggcatgg acggccctaa ggtgaagcag tggcccctga ccgaggagaa gatcaaggcc 3780
ctggtggaga tctgcaccga gatggagaag gagggcaaga tcagcaagat cggccccgag 3840
aacccctaca acacccccgt gttcgccatc aagaagaagg acagcaccaa gtggaggaag 3900
ctggtggact tcagggagct gaacaagagg acccaggact tctgggaggt gcagctgggc 3960
atcccccacc ccgccggcct gaagaagaag aagagcgtga ccgtgctgga cgtgggcgac 4020
gcctacttca gcgtgcccct ggacgaggac ttcaggaagt ataccgcctt caccatcccc 4080
agcatcaaca acgagacccc cggcatccgc taccagtaca acgtgctgcc ccagggctgg 4140
aagggcagcc ccgccatctt ccagagcagc atgacaaaga tcctggagcc cttcaagaag 4200
cagaaccccg acatcgtgat ctatcagtac atggacgacc tgtacgtggg cagcgacctg 4260
gagatcggcc agcacaggac caagatcgag gagctgaggc agcacctgct gaggtggggc 4320
ctgaccaccc ccgacaagaa gcaccagaag gagcccccat tcctgtggat gggctacgag 4380
ctgcaccccg acaagtggac cgtgcagccc atcgtgctgc ccgagaagga cagctggacc 4440
gtgaacgaca ttcagaagct ggtgggcaag ctgaactggg ccagccagat ctaccccggc 4500
atcaaggtga ggcagctgtg caagctgctg aggggcacaa aggctctgac cgaggtgatc 4560
cccctgaccg aggaggccga gctggagctg gccgagaaca gggagatcct gaaggagccc 4620
gtgcacggcg tgtactacga ccccagcaag gacctgatcg ccgagatcca gaagcagggc 4680
cagggccagt ggacctacca gatctaccag gagcccttca agaacctgaa gaccggcaag 4740
tacgcccgca tgcgcggcgc ccacaccaac gacgtgaagc agctgaccga ggccgtgcag 4800
aagatcacca ccgagagcat cgtgatctgg ggcaagactc ctaagttcaa gctgcccatc 4860
cagaaggaga cctgggagac ctggtggacc gagtactggc aggccacctg gattcccgag 4920
tgggagttcg tgaacacccc tcccctggtg aagctgtggt atcagctgga gaaggagccc 4980
atcgtgggcg ccgagacctt ctacgtggac ggcgccgcca acagggagac caagctgggc 5040
aaggccggct acgtgaccaa caagggccgc cagaaggtgg tgcccctgac caacaccacc 5100
aaccagaaga ccgagctgca ggctatctac ctggccctgc aggactcagg cctggaggtg 5160
aacatcgtga ccgacagcca gtacgccctg ggcatcatcc aggcccagcc cgacaagagc 5220
gagagcgagc tggtgaacca gatcatcgag cagctgatca agaaggagaa ggtgtacctg 5280
gcctgggtgc ccgcccacaa gggcatcggc ggcaacgagc aggtggacaa gctggtgagc 5340
gccggcatca ggaagatcct gttcctggac ggcatcgaca aggcccagga cgagcacgag 5400
aagtaccaca gcaactggag ggctatggct agcgacttca acctgcctcc cgtggtggct 5460
aaggagatcg tggccagctg cgacaagtgc cagctgaagg gcgaggccat gcacggccag 5520
gtggactgca gccccggcat ctggcagctg gtttgcaccc acctggaggg caaggtgatc 5580
ctggtggccg tgcacgtggc ctccggctac atcgaggccg aggtgatccc cgccgagacc 5640
ggccaggaga ccgcctactt cctgctgaag ctggccggcc gctggcccgt gaagaccatc 5700
cacaccgaca acggcagcaa cttcaccagc gccaccgtga aggccgcctg ctggtgggcc 5760
ggcatcaagc aggagttcgg catcccctac aacccccagt ctcagggcgt ggtggagagc 5820
atgaacaagg agctgaagaa gatcatcggc caggtgaggg accaggccga gcacctgaag 5880
accgccgtgc agatggccgt gttcatccac aacttcaaga ggaagggcgg catcggcggc 5940
tacagcgccg gcgagaggat cgtggacatc atcgccaccg acatccagac caaggagctg 6000
cagaagcaga tcaccaagat ccagaacttc agggtgtact acagggacag caggaaccct 6060
ctgtggaagg gccccgccaa gctgctgtgg aagggcgagg gcgccgtggt gatccaggac 6120
aacagcgaca tcaaggtggt gcccaggagg aaggccaaga tcatcaggga ctacggcaag 6180
cagatggccg gcgacgactg cgtggcctcc aggcaggacg aggactgact cgagcatgca 6240
tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac 6300
tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct 6360
ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct 6420
gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg 6480
ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag 6540
aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc 6600
gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcgcgctcc 6660
tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa 6720
tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact 6780
tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt 6840
gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa 6900
ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt 6960
aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag 7020
ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa gcatgcatct 7080
caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 7140
aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 7200
cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 7260
tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt 7320
tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga 7380
tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc 7440
tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 7500
ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 7560
cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc 7620
cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 7680
gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 7740
gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 7800
cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 7860
tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 7920
cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 7980
ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 8040
gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 8100
gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 8160
gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc 8220
gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc 8280
ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 8340
cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat 8400
ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 8460
tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc 8520
tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 8580
ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 8640
tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 8700
ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 8760
gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 8820
gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 8880
ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 8940
ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 9000
cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 9060
ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 9120
tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc 9180
gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 9240
tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 9300
gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 9360
tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag 9420
ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 9480
agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 9540
gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 9600
attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 9660
agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 9720
atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 9780
cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 9840
ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 9900
agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 9960
tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 10020
gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 10080
caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 10140
ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 10200
gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 10260
tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 10320
tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 10380
cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 10440
cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 10500
gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 10560
atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 10620
agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 10680
ccccgaaaag tgccacctga cgtc 10704
<210> 5
<211> 390
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
atggcctcta attttactca atttgtgctt gtcgataatg gggggacggg agatgtgacc 60
gttgccccta gcaatttcgc aaatggcgtt gcagaatgga tctctagcaa cagcagaagc 120
caagcgtaca aagtaacgtg ttccgttcgc caaagctccg cccaaaaacg gaagtataca 180
ataaaggttg aggtgccgaa agtagccact caaacagttg gtggggtaga attgcccgta 240
gcggcatggc ggtcatatct caatatggaa ctcactatcc caatcttcgc cacgaatagc 300
gattgtgagc tgatagttaa ggctatgcaa ggtcttctca aagatggaaa ccctattcca 360
tctgctatcg ccgccaacag cgggatatac 390
<210> 6
<211> 11007
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 60
ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 120
cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 180
ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 240
gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 300
tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 360
cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 420
attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 480
atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 540
atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 600
tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 660
actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 720
aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 780
gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc 840
gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt 900
gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 960
gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 1020
gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 1080
ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 1140
cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 1200
gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 1260
ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 1320
catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 1380
ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 1440
cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 1500
cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 1560
catggacgag ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt 1620
cgacggtacc gcgggcccgg gatccatgga ctcgggccgg gacttcctga ccctgcacgg 1680
cctacaggat gatgaggatc tacaggcgct gctgaagggc agccagctcc tgaaggtgaa 1740
gtccagctca tggaggagag agcgcttcta caagttgcag gaggactgca agaccatctg 1800
gcaggagtcc cgcaaggtca tgcggacccc ggagtcccag ctgttctcca tcgaggacat 1860
tcaggaggtg cgaatggggc accgcacgga gggtctggag aagttcgccc gtgatgtgcc 1920
cgaggaccgc tgcttctcca ttgtcttcaa ggaccagcgc aatacactag acctcatcgc 1980
cccatcgcca gctgatgccc agcactgggt gctggggctg cacaagatca tccaccactc 2040
aggctccatg gaccagcgtc agaagctaca gcactggatt cactcctgct tgcgaaaagc 2100
tgacaaaaac aaggacaaca agatgagctt caaggagctg cagaacttcc tgaaggagct 2160
caacatccag cctcgagccg aattcaccct ggccgccagg gccagcgtgc tgagcggcgg 2220
cgagctggac aggtgggaga agatcaggct gaggcccggc ggcaagaaga agtataagct 2280
gaagcacatc gtgtgggcca gcagggagct ggagaggttc gccgtgaacc ccggcctgct 2340
ggagaccagc gagggctgca ggcagatcct gggccagctg cagcccagcc tgcagaccgg 2400
cagcgaggag ctgaggagcc tgtacaacac cgtggccacc ctgtactgcg tgcaccagag 2460
gatcgagatc aaggacacca aggaggccct ggacaagatc gaggaggagc agaacaagtc 2520
caagaagaag gcccagcagg ccgccgccga caccggccac agcagccagg tgagccagaa 2580
ctaccccatc gtgcagaaca tccagggcca gatggtgcac caggccatca gccccaggac 2640
cctgaacgcc tgggtgaagg tggtggagga gaaggccttc agccccgagg tgatccccat 2700
gttcagcgcc ctgagcgagg gagccacccc ccaggacctg aacaccatgc tgaacaccgt 2760
gggcggccac caggccgcca tgcagatgct gaaggagacc atcaacgagg aggccgccga 2820
gtgggacagg gtgcaccccg tgcacgccgg ccccatcgcc cccggccaga tgagggagcc 2880
ccgcggcagc gacatcgccg gcaccaccag caccctgcag gagcagatcg gctggatgac 2940
caacaacccc cccatccccg tgggcgaaat ctacaagagg tggatcatcc tgggcctgaa 3000
caagatcgtg aggatgtaca gccccaccag catcctggat atcaggcagg gccccaaaga 3060
gcccttcagg gactacgtgg acaggttcta caagaccctg cgcgccgagc aggccagcca 3120
ggaggtgaag aactggatga ccgagaccct gctggtgcag aacgccaacc ccgactgcaa 3180
gaccatcctg aaggccctgg gacccgccgc caccctggag gagatgatga ccgcctgcca 3240
gggcgtgggc ggccccggcc acaaggccag ggtgctggcc gaggccatga gccaggtgac 3300
caacaccgcc accatcatga tgcagagggg caacttcagg aaccagagga agatggtgaa 3360
gtgcttcaac tgcggcaagg agggccacac cgccaggaac tgccgcgccc ccaggaagaa 3420
gggctgctgg aagtgcggca aggagggcca ccagatgaag gactgcaccg agaggcaggc 3480
taatttttta gggaagatct ggccttccta caagggaagg ccagggaatt ttcttcagag 3540
cagaccagag ccaacagccc caccatttct tcagagcaga ccagagccaa cagccccacc 3600
agaagagagc ttcaggtctg gggtagagac aacaactccc cctcagaagc aggagccgat 3660
agacaaggaa ctgtatcctt taacttccct cagatcactc tttggcaacg acccctcgtc 3720
acaataaaga tcggtggcca gctgaaggag gccctgctgg acaccggcgc cgacgacacc 3780
gtgctggagg agatgagcct gcccggcagg tggaagccca agatgatcgg cggcatcggc 3840
ggcttcatca aggtgaggca gtacgaccag atcctgatcg agatctgcgg ccacaaggcc 3900
atcggcaccg tgctggtggg acctacacct gtgaacatca tcggcaggaa cctgctgacc 3960
cagatcggct gcaccctgaa cttccccatc agccccatcg agaccgtgcc cgtgaagctg 4020
aagcccggca tggacggccc taaggtgaag cagtggcccc tgaccgagga gaagatcaag 4080
gccctggtgg agatctgcac cgagatggag aaggagggca agatcagcaa gatcggcccc 4140
gagaacccct acaacacccc cgtgttcgcc atcaagaaga aggacagcac caagtggagg 4200
aagctggtgg acttcaggga gctgaacaag aggacccagg acttctggga ggtgcagctg 4260
ggcatccccc accccgccgg cctgaagaag aagaagagcg tgaccgtgct ggacgtgggc 4320
gacgcctact tcagcgtgcc cctggacgag gacttcagga agtataccgc cttcaccatc 4380
cccagcatca acaacgagac ccccggcatc cgctaccagt acaacgtgct gccccagggc 4440
tggaagggca gccccgccat cttccagagc agcatgacaa agatcctgga gcccttcaag 4500
aagcagaacc ccgacatcgt gatctatcag tacatggacg acctgtacgt gggcagcgac 4560
ctggagatcg gccagcacag gaccaagatc gaggagctga ggcagcacct gctgaggtgg 4620
ggcctgacca cccccgacaa gaagcaccag aaggagcccc cattcctgtg gatgggctac 4680
gagctgcacc ccgacaagtg gaccgtgcag cccatcgtgc tgcccgagaa ggacagctgg 4740
accgtgaacg acattcagaa gctggtgggc aagctgaact gggccagcca gatctacccc 4800
ggcatcaagg tgaggcagct gtgcaagctg ctgaggggca caaaggctct gaccgaggtg 4860
atccccctga ccgaggaggc cgagctggag ctggccgaga acagggagat cctgaaggag 4920
cccgtgcacg gcgtgtacta cgaccccagc aaggacctga tcgccgagat ccagaagcag 4980
ggccagggcc agtggaccta ccagatctac caggagccct tcaagaacct gaagaccggc 5040
aagtacgccc gcatgcgcgg cgcccacacc aacgacgtga agcagctgac cgaggccgtg 5100
cagaagatca ccaccgagag catcgtgatc tggggcaaga ctcctaagtt caagctgccc 5160
atccagaagg agacctggga gacctggtgg accgagtact ggcaggccac ctggattccc 5220
gagtgggagt tcgtgaacac ccctcccctg gtgaagctgt ggtatcagct ggagaaggag 5280
cccatcgtgg gcgccgagac cttctacgtg gacggcgccg ccaacaggga gaccaagctg 5340
ggcaaggccg gctacgtgac caacaagggc cgccagaagg tggtgcccct gaccaacacc 5400
accaaccaga agaccgagct gcaggctatc tacctggccc tgcaggactc aggcctggag 5460
gtgaacatcg tgaccgacag ccagtacgcc ctgggcatca tccaggccca gcccgacaag 5520
agcgagagcg agctggtgaa ccagatcatc gagcagctga tcaagaagga gaaggtgtac 5580
ctggcctggg tgcccgccca caagggcatc ggcggcaacg agcaggtgga caagctggtg 5640
agcgccggca tcaggaagat cctgttcctg gacggcatcg acaaggccca ggacgagcac 5700
gagaagtacc acagcaactg gagggctatg gctagcgact tcaacctgcc tcccgtggtg 5760
gctaaggaga tcgtggccag ctgcgacaag tgccagctga agggcgaggc catgcacggc 5820
caggtggact gcagccccgg catctggcag ctggtttgca cccacctgga gggcaaggtg 5880
atcctggtgg ccgtgcacgt ggcctccggc tacatcgagg ccgaggtgat ccccgccgag 5940
accggccagg agaccgccta cttcctgctg aagctggccg gccgctggcc cgtgaagacc 6000
atccacaccg acaacggcag caacttcacc agcgccaccg tgaaggccgc ctgctggtgg 6060
gccggcatca agcaggagtt cggcatcccc tacaaccccc agtctcaggg cgtggtggag 6120
agcatgaaca aggagctgaa gaagatcatc ggccaggtga gggaccaggc cgagcacctg 6180
aagaccgccg tgcagatggc cgtgttcatc cacaacttca agaggaaggg cggcatcggc 6240
ggctacagcg ccggcgagag gatcgtggac atcatcgcca ccgacatcca gaccaaggag 6300
ctgcagaagc agatcaccaa gatccagaac ttcagggtgt actacaggga cagcaggaac 6360
cctctgtgga agggccccgc caagctgctg tggaagggcg agggcgccgt ggtgatccag 6420
gacaacagcg acatcaaggt ggtgcccagg aggaaggcca agatcatcag ggactacggc 6480
aagcagatgg ccggcgacga ctgcgtggcc tccaggcagg acgaggactg actcgagcat 6540
gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc 6600
gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac 6660
cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg 6720
tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga 6780
ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga 6840
aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc 6900
ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcgcgc 6960
tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 7020
aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 7080
acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 7140
tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 7200
caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg 7260
gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt 7320
cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca 7380
tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 7440
gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 7500
gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 7560
ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt 7620
ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc 7680
tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 7740
ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 7800
ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 7860
gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct 7920
ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 7980
ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 8040
cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 8100
ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 8160
cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 8220
gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 8280
tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 8340
ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 8400
agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 8460
ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 8520
ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 8580
gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 8640
cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 8700
aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 8760
cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 8820
acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 8880
ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 8940
taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 9000
aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 9060
attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 9120
cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 9180
gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 9240
ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 9300
agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 9360
tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 9420
ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag 9480
gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 9540
ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 9600
gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 9660
aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 9720
aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 9780
ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 9840
gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 9900
gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 9960
tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 10020
ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 10080
ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 10140
atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 10200
ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 10260
tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 10320
attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 10380
tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 10440
ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 10500
gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 10560
gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 10620
gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 10680
aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 10740
taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 10800
tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 10860
tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 10920
atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 10980
tttccccgaa aagtgccacc tgacgtc 11007
<210> 7
<211> 19
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
acatgaggat cacccatgt 19
<210> 8
<211> 306
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
cttgcagcac atacatgagg atcacccatg tctgcaggtc gactctagaa aacatgagga 60
tcacccatgt ctgcagtatt cccgggttca ttagatccta aggtacctaa ttgcctagaa 120
aacatgagga tcacccatgt ctgcaggtcg actctagaaa acatgaggat cacccatgtc 180
tgcagtattc ccgggttcat tagatcctaa ggtacctaat tgcctagaaa acatgaggat 240
cacccatgtc tgcaggtcga ctctagaaaa catgaggatc acccatgtgt gcaccttaag 300
cttggc 306
<210> 9
<211> 3897
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
atggactgga cctggatcct gttcctggtg gccgccgcca cccgcgtgca cagctttgtg 60
tttctggtgc tgctgccact ggtgagcagc cagtgtgtga atctgaccac acgcacccag 120
ctgccacccg cctacacaaa tagcttcaca agaggcgtgt actaccctga taaagtgttt 180
aggtctagtg tgctgcactc tacccaggat ctgttcctgc ccttctttag taatgtgacc 240
tggttccacg ccatccatgt gagcggaact aacggcacta agagatttga caacccagtg 300
ctgcctttta acgacggagt gtacttcgcc tccaccgaaa agtctaatat tattagggga 360
tggatcttcg ggacaaccct cgacagtaag acccagagcc tgctgattgt gaataatgct 420
acaaacgtgg tcatcaaggt gtgcgagttc cagttttgta atgatccttt cttgggagtg 480
tactaccaca agaataataa gtcatggatg gagagtgagt ttagggtgta tagttccgcc 540
aataactgta cctttgaata cgtgagtcag ccctttctga tggacctgga aggaaaacag 600
ggcaatttta agaatctgcg cgaatttgtg tttaaaaata tcgacggcta tttcaaaatc 660
tacagcaagc acacccccat caacctggtg agagacctgc cccagggctt cagcgccctg 720
gagcccctgg tggatctccc catcgggatc aatatcacac ggttccagac actgctggcc 780
ctgcacagaa gctatctgac acctggagac tcctcctccg gctggaccgc cggcgcagcc 840
gcatactatg tgggctacct gcagccaagg accttcctgc tgaaatataa cgagaacgga 900
accattacag atgccgtgga ttgtgccctg gatcccctgt ctgagaccaa atgcaccctg 960
aaaagtttta ccgtggagaa gggcatctat cagaccagca atttccgggt gcagcccact 1020
gagtccatcg tccggtttcc taatattacc aacctgtgcc ccttcggcga ggtctttaat 1080
gctacaagat tcgccagcgt gtacgcctgg aatagaaaaa gaatttccaa ctgtgtcgcc 1140
gactattccg tgctgtataa cagtgccagc ttctccacct tcaaatgtta cggagtgagc 1200
cccacaaagc tgaacgacct gtgcttcacc aacgtctacg ccgatagctt cgtgattaga 1260
ggcgacgaag tccggcagat tgctccaggc cagaccggaa agattgccga ttacaactat 1320
aaactccctg atgactttac cggatgcgtg attgcttgga acagtaacaa cctggatagc 1380
aaagtgggcg ggaattataa ctacctgtac cggctcttca gaaaatctaa tttgaaaccc 1440
ttcgagcggg acatcagcac tgagatctac caggccggct ctaccccctg taacggagtg 1500
gagggcttca actgctattt cccactgcag agctatggat tccagcccac taacggcgtc 1560
ggatatcagc catatagggt ggtggtgctg tccttcgaac tcctccacgc ccccgctacc 1620
gtgtgcggcc ctaagaagtc tacaaacctg gttaaaaata agtgcgtgaa ctttaacttt 1680
aacggcctga caggcactgg cgttctgaca gaatctaaca agaagttcct gcccttccag 1740
cagttcggcc gggacatcgc tgacaccacc gacgccgtga gggaccccca gacactggag 1800
atccttgaca tcactccttg ctccttcggc ggcgtgtctg tgatcacacc cggcacaaac 1860
actagtaacc aggtggcagt tctgtaccag gatgtgaact gcactgaggt ccccgttgca 1920
atccacgccg atcagctgac ccccacttgg cgcgtgtact caaccggcag taatgtgttc 1980
cagacccgcg ccggctgcct gataggagcc gagcacgtca ataatagtta cgagtgtgat 2040
atccctattg gtgccggaat ctgtgcttca taccagaccc agaccaattc acccaggcga 2100
gcacgctccg tcgcttccca gagtattatc gcctacacta tgtccctcgg ggcagaaaac 2160
agcgtcgcct atagcaacaa tagcatcgct attccaacaa acttcaccat ctccgtgacc 2220
acagagattc tgcccgtgtc catgactaag acatccgtgg attgcaccat gtatatttgt 2280
ggtgactcta ccgagtgctc taacctcctg ctgcagtacg gctctttctg cacacagctc 2340
aatcgcgcac tgaccgggat cgctgtggag caggacaaga acactcagga agtgttcgca 2400
caggtgaagc agatctataa gacccctccc atcaaagatt tcggaggatt caatttctca 2460
cagatcctgc ccgacccaag caagccatct aaacgctctt tcattgagga tctgctgttc 2520
aataaggtga ccctggctga tgccgggttc atcaagcagt atggcgactg cctcggcgat 2580
atcgctgcca gggacctgat ctgcgcacag aaattcaatg gactgaccgt cctgccacca 2640
ctgctgacag atgagatgat cgcccagtat acttctgccc tgctcgccgg caccatcact 2700
tctgggtgga cctttggcgc cggcgcagcc ctgcagatcc ctttcgccat gcagatggct 2760
taccgcttta atggaatcgg ggtgacccag aacgtgctgt atgagaatca gaaactgatt 2820
gcaaatcagt ttaacagcgc tatcggcaaa atccaggatt ctctgagctc cacagccagc 2880
gccctcggaa agctgcagga tgtcgtgaac cagaatgccc aggcgctgaa taccctggtg 2940
aaacagctca gctctaactt cggcgctatt tcctccgtgc tcaacgatat cctgagcagg 3000
ctggatcctc ccgaagccga agtgcagatt gacagactga tcacaggcag gctccagagc 3060
ctccagacat acgtcaccca gcagctgatt cgggctgccg agatacgggc aagcgctaac 3120
ctggccgcca ctaagatgag cgagtgtgtg ctcggccaga gcaagcgggt ggacttttgc 3180
gggaaaggct atcacctgat gtctttccca cagtccgccc ctcacggggt tgtgttcctg 3240
cacgtgacct acgtgcccgc ccaggagaag aatttcacaa ccgcacccgc catctgccat 3300
gacgggaaag cccactttcc cagggagggc gtgttcgtga gcaatggcac tcactggttt 3360
gtgacccaga gaaatttcta tgagccccag atcatcacca ccgacaatac ctttgtcagt 3420
ggcaactgtg atgtcgtgat tggcatcgtg aacaacactg tgtacgatcc tctgcagccc 3480
gagctggact cctttaaaga ggagctggac aagtatttca aaaaccacac atccccagat 3540
gtggatctgg gcgatatctc cggcatcaac gccagtgtgg tgaatatcca gaaagagatt 3600
gatagactga acgaggtggc taagaacctg aacgaatccc tgattgacct gcaggagctg 3660
ggaaagtacg agcagtatat taagtggcct tggtatatct ggctggggtt catcgctgga 3720
ctgatcgcta ttgtgatggt gaccattatg ctgtgctgca tgactagctg ctgtagttgc 3780
ctgaagggat gctgtagctg cggcagctgc tgcaagtttg atgaggacga ttctgagcct 3840
gtgctgaaag gggtgaagct gcactacacc gattacaaag acgatgacga taagtaa 3897
<210> 10
<211> 4421
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60
cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120
ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180
accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240
attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300
tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360
tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagcttgca tgcctgcagg 420
tcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 480
tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 540
aatgggtgga ctatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 600
caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt 660
acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 720
ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg 780
gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac 840
gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg 900
tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac 960
gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc 1020
tagaggatcc ggtactcgag gaactgaaaa accagaaagt taactggtaa gtttagtctt 1080
tttgtctttt atttcaggtc ccgatccggt ggtggtgcaa atcaaagaac tgctcctcag 1140
tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg 1200
gaattgtacc cgcgggccac catggtgagc aagggcgagg agctgttcac cggggtggtg 1260
cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 1320
ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 1380
ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 1440
cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 1500
gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 1560
aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 1620
gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 1680
atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 1740
gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 1800
gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 1860
gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 1920
atggacgagc tgtacaagta aaattggccg cggggatcca gacatgataa gatacattga 1980
tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg 2040
tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa 2100
ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt cggatcctct 2160
agagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 2220
aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 2280
ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 2340
ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 2400
cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 2460
tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 2520
aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 2580
aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 2640
tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 2700
ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 2760
cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 2820
ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 2880
ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 2940
gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 3000
agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 3060
cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 3120
aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 3180
aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 3240
ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 3300
aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 3360
ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 3420
agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 3480
cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 3540
ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 3600
gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 3660
cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 3720
cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 3780
ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 3840
catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 3900
tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 3960
ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 4020
catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 4080
cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 4140
cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 4200
acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 4260
ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 4320
tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 4380
attaacctat aaaaataggc gtatcacgag gccctttcgt c 4421
<210> 11
<211> 7910
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60
cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120
ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180
accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240
attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300
tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360
tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagcttgca tgcctgcagg 420
tcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 480
tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 540
aatgggtgga ctatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 600
caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt 660
acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 720
ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg 780
gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac 840
gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg 900
tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac 960
gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc 1020
tagaggatcc ggtactcgag gaactgaaaa accagaaagt taactggtaa gtttagtctt 1080
tttgtctttt atttcaggtc ccgatccggt ggtggtgcaa atcaaagaac tgctcctcag 1140
tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg 1200
gaattgtacc cgcggacgcg tgccaccatg gactggacct ggatcctgtt cctggtggcc 1260
gccgccaccc gcgtgcacag ctttgtgttt ctggtgctgc tgccactggt gagcagccag 1320
tgtgtgaatc tgaccacacg cacccagctg ccacccgcct acacaaatag cttcacaaga 1380
ggcgtgtact accctgataa agtgtttagg tctagtgtgc tgcactctac ccaggatctg 1440
ttcctgccct tctttagtaa tgtgacctgg ttccacgcca tccatgtgag cggaactaac 1500
ggcactaaga gatttgacaa cccagtgctg ccttttaacg acggagtgta cttcgcctcc 1560
accgaaaagt ctaatattat taggggatgg atcttcggga caaccctcga cagtaagacc 1620
cagagcctgc tgattgtgaa taatgctaca aacgtggtca tcaaggtgtg cgagttccag 1680
ttttgtaatg atcctttctt gggagtgtac taccacaaga ataataagtc atggatggag 1740
agtgagttta gggtgtatag ttccgccaat aactgtacct ttgaatacgt gagtcagccc 1800
tttctgatgg acctggaagg aaaacagggc aattttaaga atctgcgcga atttgtgttt 1860
aaaaatatcg acggctattt caaaatctac agcaagcaca cccccatcaa cctggtgaga 1920
gacctgcccc agggcttcag cgccctggag cccctggtgg atctccccat cgggatcaat 1980
atcacacggt tccagacact gctggccctg cacagaagct atctgacacc tggagactcc 2040
tcctccggct ggaccgccgg cgcagccgca tactatgtgg gctacctgca gccaaggacc 2100
ttcctgctga aatataacga gaacggaacc attacagatg ccgtggattg tgccctggat 2160
cccctgtctg agaccaaatg caccctgaaa agttttaccg tggagaaggg catctatcag 2220
accagcaatt tccgggtgca gcccactgag tccatcgtcc ggtttcctaa tattaccaac 2280
ctgtgcccct tcggcgaggt ctttaatgct acaagattcg ccagcgtgta cgcctggaat 2340
agaaaaagaa tttccaactg tgtcgccgac tattccgtgc tgtataacag tgccagcttc 2400
tccaccttca aatgttacgg agtgagcccc acaaagctga acgacctgtg cttcaccaac 2460
gtctacgccg atagcttcgt gattagaggc gacgaagtcc ggcagattgc tccaggccag 2520
accggaaaga ttgccgatta caactataaa ctccctgatg actttaccgg atgcgtgatt 2580
gcttggaaca gtaacaacct ggatagcaaa gtgggcggga attataacta cctgtaccgg 2640
ctcttcagaa aatctaattt gaaacccttc gagcgggaca tcagcactga gatctaccag 2700
gccggctcta ccccctgtaa cggagtggag ggcttcaact gctatttccc actgcagagc 2760
tatggattcc agcccactaa cggcgtcgga tatcagccat atagggtggt ggtgctgtcc 2820
ttcgaactcc tccacgcccc cgctaccgtg tgcggcccta agaagtctac aaacctggtt 2880
aaaaataagt gcgtgaactt taactttaac ggcctgacag gcactggcgt tctgacagaa 2940
tctaacaaga agttcctgcc cttccagcag ttcggccggg acatcgctga caccaccgac 3000
gccgtgaggg acccccagac actggagatc cttgacatca ctccttgctc cttcggcggc 3060
gtgtctgtga tcacacccgg cacaaacact agtaaccagg tggcagttct gtaccaggat 3120
gtgaactgca ctgaggtccc cgttgcaatc cacgccgatc agctgacccc cacttggcgc 3180
gtgtactcaa ccggcagtaa tgtgttccag acccgcgccg gctgcctgat aggagccgag 3240
cacgtcaata atagttacga gtgtgatatc cctattggtg ccggaatctg tgcttcatac 3300
cagacccaga ccaattcacc caggcgagca cgctccgtcg cttcccagag tattatcgcc 3360
tacactatgt ccctcggggc agaaaacagc gtcgcctata gcaacaatag catcgctatt 3420
ccaacaaact tcaccatctc cgtgaccaca gagattctgc ccgtgtccat gactaagaca 3480
tccgtggatt gcaccatgta tatttgtggt gactctaccg agtgctctaa cctcctgctg 3540
cagtacggct ctttctgcac acagctcaat cgcgcactga ccgggatcgc tgtggagcag 3600
gacaagaaca ctcaggaagt gttcgcacag gtgaagcaga tctataagac ccctcccatc 3660
aaagatttcg gaggattcaa tttctcacag atcctgcccg acccaagcaa gccatctaaa 3720
cgctctttca ttgaggatct gctgttcaat aaggtgaccc tggctgatgc cgggttcatc 3780
aagcagtatg gcgactgcct cggcgatatc gctgccaggg acctgatctg cgcacagaaa 3840
ttcaatggac tgaccgtcct gccaccactg ctgacagatg agatgatcgc ccagtatact 3900
tctgccctgc tcgccggcac catcacttct gggtggacct ttggcgccgg cgcagccctg 3960
cagatccctt tcgccatgca gatggcttac cgctttaatg gaatcggggt gacccagaac 4020
gtgctgtatg agaatcagaa actgattgca aatcagttta acagcgctat cggcaaaatc 4080
caggattctc tgagctccac agccagcgcc ctcggaaagc tgcaggatgt cgtgaaccag 4140
aatgcccagg cgctgaatac cctggtgaaa cagctcagct ctaacttcgg cgctatttcc 4200
tccgtgctca acgatatcct gagcaggctg gatcctcccg aagccgaagt gcagattgac 4260
agactgatca caggcaggct ccagagcctc cagacatacg tcacccagca gctgattcgg 4320
gctgccgaga tacgggcaag cgctaacctg gccgccacta agatgagcga gtgtgtgctc 4380
ggccagagca agcgggtgga cttttgcggg aaaggctatc acctgatgtc tttcccacag 4440
tccgcccctc acggggttgt gttcctgcac gtgacctacg tgcccgccca ggagaagaat 4500
ttcacaaccg cacccgccat ctgccatgac gggaaagccc actttcccag ggagggcgtg 4560
ttcgtgagca atggcactca ctggtttgtg acccagagaa atttctatga gccccagatc 4620
atcaccaccg acaatacctt tgtcagtggc aactgtgatg tcgtgattgg catcgtgaac 4680
aacactgtgt acgatcctct gcagcccgag ctggactcct ttaaagagga gctggacaag 4740
tatttcaaaa accacacatc cccagatgtg gatctgggcg atatctccgg catcaacgcc 4800
agtgtggtga atatccagaa agagattgat agactgaacg aggtggctaa gaacctgaac 4860
gaatccctga ttgacctgca ggagctggga aagtacgagc agtatattaa gtggccttgg 4920
tatatctggc tggggttcat cgctggactg atcgctattg tgatggtgac cattatgctg 4980
tgctgcatga ctagctgctg tagttgcctg aagggatgct gtagctgcgg cagctgctgc 5040
aagtttgatg aggacgattc tgagcctgtg ctgaaagggg tgaagctgca ctacaccgat 5100
tacaaagacg atgacgataa gtaaaccggt cttgcagcac atacatgagg atcacccatg 5160
tctgcaggtc gactctagaa aacatgagga tcacccatgt ctgcagtatt cccgggttca 5220
ttagatccta aggtacctaa ttgcctagaa aacatgagga tcacccatgt ctgcaggtcg 5280
actctagaaa acatgaggat cacccatgtc tgcagtattc ccgggttcat tagatcctaa 5340
ggtacctaat tgcctagaaa acatgaggat cacccatgtc tgcaggtcga ctctagaaaa 5400
catgaggatc acccatgtgt gcaccttaag cttggcccgc ggggatccag acatgataag 5460
atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg 5520
tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa 5580
caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggttttttc 5640
ggatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt 5700
tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 5760
gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 5820
gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 5880
ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 5940
ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 6000
cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 6060
gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 6120
tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 6180
ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 6240
atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 6300
gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 6360
tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 6420
cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 6480
cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 6540
tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 6600
cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 6660
cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 6720
gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 6780
gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 6840
gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 6900
ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 6960
atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 7020
agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 7080
ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 7140
tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 7200
ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 7260
caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 7320
gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 7380
atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 7440
accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 7500
aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 7560
gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 7620
tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 7680
aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 7740
ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 7800
aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat 7860
tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 7910
<210> 12
<211> 10916
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
aagcttggcc attgcatacg ttgtatccat atcataatat gtacatttat attggctcat 60
gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag taatcaatta 120
cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 180
gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 240
ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 300
ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 360
atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 420
cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 480
acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 540
acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 600
actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 660
gagctcgttt agtgaaccgg ggtctctctg gttagaccag atctgagcct gggagctctc 720
tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 780
agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 840
agtgtggaaa atctctagca gtggcgcccg aacagggacc tgaaagcgaa agggaaacca 900
gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg 960
cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc 1020
gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg 1080
ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa 1140
cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga 1200
cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta 1260
gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac 1320
aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt 1380
cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt 1440
agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag 1500
agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag 1560
cactatgggc gcagcctcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat 1620
agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact 1680
cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa 1740
ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt 1800
gccttggaat gctagttgga gtaataaatc tctggaacag attggaatca cacgacctgg 1860
atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa 1920
tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt 1980
ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata 2040
gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt 2100
aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac 2160
aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta 2220
gtgaacggat ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac 2280
agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa 2340
aaacaaatta caaaaattca aaattttatc gatcacgaga ctagcctcga cgatggtcga 2400
gtaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc 2460
gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg 2520
taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct 2580
agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt 2640
agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg 2700
cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg 2760
aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag 2820
gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc 2880
ctcttcctca tctccgggcc tttcgacctc tagcggacgc gtgccaccat ggactggacc 2940
tggatcctgt tcctggtggc cgccgccacc cgcgtgcaca gctttgtgtt tctggtgctg 3000
ctgccactgg tgagcagcca gtgtgtgaat ctgaccacac gcacccagct gccacccgcc 3060
tacacaaata gcttcacaag aggcgtgtac taccctgata aagtgtttag gtctagtgtg 3120
ctgcactcta cccaggatct gttcctgccc ttctttagta atgtgacctg gttccacgcc 3180
atccatgtga gcggaactaa cggcactaag agatttgaca acccagtgct gccttttaac 3240
gacggagtgt acttcgcctc caccgaaaag tctaatatta ttaggggatg gatcttcggg 3300
acaaccctcg acagtaagac ccagagcctg ctgattgtga ataatgctac aaacgtggtc 3360
atcaaggtgt gcgagttcca gttttgtaat gatcctttct tgggagtgta ctaccacaag 3420
aataataagt catggatgga gagtgagttt agggtgtata gttccgccaa taactgtacc 3480
tttgaatacg tgagtcagcc ctttctgatg gacctggaag gaaaacaggg caattttaag 3540
aatctgcgcg aatttgtgtt taaaaatatc gacggctatt tcaaaatcta cagcaagcac 3600
acccccatca acctggtgag agacctgccc cagggcttca gcgccctgga gcccctggtg 3660
gatctcccca tcgggatcaa tatcacacgg ttccagacac tgctggccct gcacagaagc 3720
tatctgacac ctggagactc ctcctccggc tggaccgccg gcgcagccgc atactatgtg 3780
ggctacctgc agccaaggac cttcctgctg aaatataacg agaacggaac cattacagat 3840
gccgtggatt gtgccctgga tcccctgtct gagaccaaat gcaccctgaa aagttttacc 3900
gtggagaagg gcatctatca gaccagcaat ttccgggtgc agcccactga gtccatcgtc 3960
cggtttccta atattaccaa cctgtgcccc ttcggcgagg tctttaatgc tacaagattc 4020
gccagcgtgt acgcctggaa tagaaaaaga atttccaact gtgtcgccga ctattccgtg 4080
ctgtataaca gtgccagctt ctccaccttc aaatgttacg gagtgagccc cacaaagctg 4140
aacgacctgt gcttcaccaa cgtctacgcc gatagcttcg tgattagagg cgacgaagtc 4200
cggcagattg ctccaggcca gaccggaaag attgccgatt acaactataa actccctgat 4260
gactttaccg gatgcgtgat tgcttggaac agtaacaacc tggatagcaa agtgggcggg 4320
aattataact acctgtaccg gctcttcaga aaatctaatt tgaaaccctt cgagcgggac 4380
atcagcactg agatctacca ggccggctct accccctgta acggagtgga gggcttcaac 4440
tgctatttcc cactgcagag ctatggattc cagcccacta acggcgtcgg atatcagcca 4500
tatagggtgg tggtgctgtc cttcgaactc ctccacgccc ccgctaccgt gtgcggccct 4560
aagaagtcta caaacctggt taaaaataag tgcgtgaact ttaactttaa cggcctgaca 4620
ggcactggcg ttctgacaga atctaacaag aagttcctgc ccttccagca gttcggccgg 4680
gacatcgctg acaccaccga cgccgtgagg gacccccaga cactggagat ccttgacatc 4740
actccttgct ccttcggcgg cgtgtctgtg atcacacccg gcacaaacac tagtaaccag 4800
gtggcagttc tgtaccagga tgtgaactgc actgaggtcc ccgttgcaat ccacgccgat 4860
cagctgaccc ccacttggcg cgtgtactca accggcagta atgtgttcca gacccgcgcc 4920
ggctgcctga taggagccga gcacgtcaat aatagttacg agtgtgatat ccctattggt 4980
gccggaatct gtgcttcata ccagacccag accaattcac ccaggcgagc acgctccgtc 5040
gcttcccaga gtattatcgc ctacactatg tccctcgggg cagaaaacag cgtcgcctat 5100
agcaacaata gcatcgctat tccaacaaac ttcaccatct ccgtgaccac agagattctg 5160
cccgtgtcca tgactaagac atccgtggat tgcaccatgt atatttgtgg tgactctacc 5220
gagtgctcta acctcctgct gcagtacggc tctttctgca cacagctcaa tcgcgcactg 5280
accgggatcg ctgtggagca ggacaagaac actcaggaag tgttcgcaca ggtgaagcag 5340
atctataaga cccctcccat caaagatttc ggaggattca atttctcaca gatcctgccc 5400
gacccaagca agccatctaa acgctctttc attgaggatc tgctgttcaa taaggtgacc 5460
ctggctgatg ccgggttcat caagcagtat ggcgactgcc tcggcgatat cgctgccagg 5520
gacctgatct gcgcacagaa attcaatgga ctgaccgtcc tgccaccact gctgacagat 5580
gagatgatcg cccagtatac ttctgccctg ctcgccggca ccatcacttc tgggtggacc 5640
tttggcgccg gcgcagccct gcagatccct ttcgccatgc agatggctta ccgctttaat 5700
ggaatcgggg tgacccagaa cgtgctgtat gagaatcaga aactgattgc aaatcagttt 5760
aacagcgcta tcggcaaaat ccaggattct ctgagctcca cagccagcgc cctcggaaag 5820
ctgcaggatg tcgtgaacca gaatgcccag gcgctgaata ccctggtgaa acagctcagc 5880
tctaacttcg gcgctatttc ctccgtgctc aacgatatcc tgagcaggct ggatcctccc 5940
gaagccgaag tgcagattga cagactgatc acaggcaggc tccagagcct ccagacatac 6000
gtcacccagc agctgattcg ggctgccgag atacgggcaa gcgctaacct ggccgccact 6060
aagatgagcg agtgtgtgct cggccagagc aagcgggtgg acttttgcgg gaaaggctat 6120
cacctgatgt ctttcccaca gtccgcccct cacggggttg tgttcctgca cgtgacctac 6180
gtgcccgccc aggagaagaa tttcacaacc gcacccgcca tctgccatga cgggaaagcc 6240
cactttccca gggagggcgt gttcgtgagc aatggcactc actggtttgt gacccagaga 6300
aatttctatg agccccagat catcaccacc gacaatacct ttgtcagtgg caactgtgat 6360
gtcgtgattg gcatcgtgaa caacactgtg tacgatcctc tgcagcccga gctggactcc 6420
tttaaagagg agctggacaa gtatttcaaa aaccacacat ccccagatgt ggatctgggc 6480
gatatctccg gcatcaacgc cagtgtggtg aatatccaga aagagattga tagactgaac 6540
gaggtggcta agaacctgaa cgaatccctg attgacctgc aggagctggg aaagtacgag 6600
cagtatatta agtggccttg gtatatctgg ctggggttca tcgctggact gatcgctatt 6660
gtgatggtga ccattatgct gtgctgcatg actagctgct gtagttgcct gaagggatgc 6720
tgtagctgcg gcagctgctg caagtttgat gaggacgatt ctgagcctgt gctgaaaggg 6780
gtgaagctgc actacaccga ttacaaagac gatgacgata agtaactcga gggaattccg 6840
ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 6900
ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 6960
gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 7020
tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 7080
ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 7140
ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 7200
tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc 7260
tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 7320
tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 7380
ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgggaat 7440
tcgagctcgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt 7500
ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt 7560
tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa 7620
ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt 7680
gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg 7740
aaaatctcta gcagcatcta gctagaatta attccgtgta ttctatagtg tcacctaaat 7800
cgtatgtgta tgatacataa ggttatgtat taattgtagc cgcgttctaa cgacaatatg 7860
tacaagccta attgtgtagc atctggctta ctgaagcaga ccctatcatc tctctcgtaa 7920
actgccgtca gagtcggttt ggttggacga accttctgag tttctggtaa cgccgtcccg 7980
cacccggaaa tggtcagcga accaatcagc agggtcatcg ctagcctagg cttttgcgtc 8040
gagacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt 8100
tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc 8160
cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt 8220
tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg 8280
gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 8340
tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 8400
gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 8460
attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 8520
cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 8580
ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 8640
aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa 8700
tttcccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 8760
aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 8820
ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 8880
ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 8940
agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 9000
tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 9060
tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 9120
ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 9180
gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 9240
acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 9300
tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 9360
gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga 9420
actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 9480
aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 9540
cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 9600
tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 9660
cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 9720
tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 9780
ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 9840
ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 9900
cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 9960
aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct 10020
agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 10080
tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 10140
ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 10200
cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 10260
atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 10320
ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 10380
tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 10440
gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 10500
gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 10560
cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 10620
gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat 10680
tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc 10740
aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc 10800
tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca 10860
tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgc 10916
<210> 13
<211> 37
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
tagcaccggt cgccaccatg gcctctaatt ttactca 37
<210> 14
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
tagctccgga ctgcacaatc ggatagttct ggctgtatat cccgctgttg gcgg 54
<210> 15
<211> 21
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
attgacgcaa atgggcggta g 21
<210> 16
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
tagcacgcgt gccaccatgg actggacctg gatcct 36
<210> 17
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 17
ttacttatcg tcatcgtctt 20
<210> 18
<211> 45
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 18
aagacgatga cgataagtaa accggtcttg cagcacatac atgag 45
<210> 19
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 19
tagcccgcgg gccaagctta aggtgcacac 30
<210> 20
<211> 36
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 20
tagcacgcgt gccaccatgg actggacctg gatcct 36
<210> 21
<211> 30
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 21
tagcccgcgg gccaagctta aggtgcacac 30
<210> 22
<211> 20
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 22
gtttattgca gcttataatg 20

Claims (10)

1.一种基于慢病毒外壳修饰和mRNA递送的SARS-CoV-2疫苗，其特征在于，所述SARS-CoV-2疫苗是慢病毒颗粒作为外壳包裹SARS-CoV-2的棘突蛋白Spike的mRNA得到。
2.根据权利要求1所述的SARS-CoV-2疫苗，其特征在于，所述慢病毒颗粒的包膜上装饰有SARS-CoV-2的棘突蛋白Spike；所述棘突蛋白Spike的基因是密码子优化后和氨基酸突变后的基因，序列如SEQ ID NO.9所示。
3.根据权利要求1或2所述的SARS-CoV-2疫苗，其特征在于，表达所述包膜蛋白VSV-G的质粒的序列如SEQ ID NO.1所示。
4.根据权利要求1所述的SARS-CoV-2疫苗，其特征在于，所述棘突蛋白Spike的mRNA由SARS-CoV-2的棘突蛋白Spike基因转录得到；所述棘突蛋白Spike基因是经过密码子优化以及氨基酸突变后的基因，序列如SEQ ID NO.9所示。
5.一种根据权利要求1所述的SARS-CoV-2疫苗的制备方法，其特征在于，通过将疫苗生产质粒转染入病毒生产细胞；收集上清，浓缩，制备而得。
6.根据权利要求5所述的SARS-CoV-2疫苗的制备方法，其特征在于，所述制备方法的具体步骤如下：
S1、将表达VSV-G膜蛋白的质粒、表达含RNA结合蛋白的慢病毒GagPol长链蛋白的质粒、表达含慢病毒GagPol长链蛋白的质粒、表达含RNA结合蛋白所识别的茎环结构和棘突蛋白Spike mRNA的质粒共转染病毒生产细胞；所述茎环结构置于棘突蛋白Spike mRNA的表达框内；所述棘突蛋白Spike mRNA由SARS-CoV-2的棘突蛋白Spike基因转录得到；
S2、收集含病毒颗粒的上清，浓缩，得到含棘突蛋白Spike的mRNA、并在外壳装饰有棘突蛋白Spike的慢病毒颗粒。
7.根据权利要求6所述的SARS-CoV-2疫苗的制备方法，其特征在于，所述表达含RNA结合蛋白的慢病毒GagPol长链蛋白的质粒是通过将RNA结合蛋白整合至第三代慢病毒GagPol长链蛋白的N-端制成，所述RNA结合蛋白为MS2衣壳蛋白；
所述MS2衣壳蛋白的基因序列如SEQ ID NO.5所示，表达含RNA结合蛋白的慢病毒GagPol长链蛋白的质粒序列如SEQ ID NO.4所示。
8.根据权利要求6所述的SARS-CoV-2疫苗的制备方法，其特征在于，所述表达含RNA结合蛋白所识别的茎环结构的棘突蛋白Spike mRNA的质粒是通过将RNA结合蛋白所识别的茎环结构的RNA序列与棘突蛋白Spike的mRNA序列融合而得到，所得质粒的序列如SEQ IDNO.11所示。
9.根据权利要求6所述的SARS-CoV-2疫苗的制备方法，其特征在于，所述含RNA结合蛋白所识别的茎环结构在Spike mRNA的表达框中，茎环结构的RNA序列如SEQ ID NO.8所示。
10.根据权利要求6所述的SARS-CoV-2疫苗的制备方法，其特征在于，所述病毒生产细胞选自293T细胞、293FTX细胞、HEK293细胞；
所述表达含慢病毒GagPol长链蛋白的质粒序列如SEQ ID NO.2所示。
CN202010995678.3A 2020-09-21 2020-09-21 基于慢病毒外壳修饰和mRNA递送的SARS-CoV-2疫苗及其制备方法 Active CN112168958B (zh)

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