CN117858945A - 用于改善产量性状的hect e3泛素连接酶基因的修饰 - Google Patents

用于改善产量性状的hect e3泛素连接酶基因的修饰 Download PDF

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CN117858945A
CN117858945A CN202280056853.XA CN202280056853A CN117858945A CN 117858945 A CN117858945 A CN 117858945A CN 202280056853 A CN202280056853 A CN 202280056853A CN 117858945 A CN117858945 A CN 117858945A
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P·R·玛丽
L·G·马修
H·金
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Pairing Plant Service Co ltd
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Abstract

本发明涉及用于修饰植物中的E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的组合物和方法,任选地以改善产量性状。本发明还涉及使用本发明的方法和组合物产生的具有增加的改善的产量性状的植物。

Description

用于改善产量性状的HECT E3泛素连接酶基因的修饰
关于序列表的电子申请的声明
提供了ASCII文本格式的序列表来代替纸质副本,该序列表根据37C.F.R.§1.821提交,命名为1499.61.WO_ST25.txt,大小为727,911字节,于2022年6月4日生成,并通过EFS-Web提交。此序列表就其公开内容在此通过引用并入说明书中。
优先权声明
此申请根据35U.S.C.§119(e)要求2021年6月24日提交的美国临时申请号63/214,498的权益,其全部内容通过引用并入本文。
技术领域
本发明涉及用于修饰植物中的E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的组合物和方法,任选地以改善产量性状。本发明还涉及使用本发明的方法和组合物产生的具有增加的改善的产量性状的植物。
背景技术
随着需要养活的人口不断增长,不断需要提高农作物产量。目前,包括大豆在内的关键主要农作物产量每年仅增加0.9%-1.6%,这样的产量增加幅度不足以满足粮食生产的未来需要。本发明通过提供用于改善/增强植物(包括大豆、玉米和其他植物物种)的产量性状的新组合物和方法来解决本领域中的这些缺点。
发明内容
本发明的一方面提供了一种植物或其植物部分,其包含编码E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)(HECT E3 UPL)多肽的内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的至少一个非天然突变。
本发明的第二方面提供了一种植物细胞,其包含编辑系统,所述编辑系统包含:(a)CRISPR-Cas效应蛋白;和(b)引导核酸(例如,gRNA、gDNA、crRNA、crDNA、sgRNA、sgDNA),其包含与编码E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)(HECT E3 UPL)多肽的内源靶基因互补的间隔序列。
本发明的第三方面提供了产生/培育无转基因的经编辑的植物的方法,包括:将本发明的植物与无转基因的植物杂交,从而将至少一个非天然突变引入到无转基因的植物中;和选择包含至少一个非天然突变且无转基因的后代植物,从而产生无转基因的经编辑的植物。
本发明的第四方面提供了提供具有一种或多种改善的产量性状(任选地增加的种子大小(例如,种子面积和/或种子重量)和/或种子油含量)的多个植物的方法,该方法包括在生长区域中种植本发明的两个或更多个植物,从而提供与不包含至少一个非天然突变的多个对照植物相比具有一种或多种改善的产量性状的多个植物。
本发明的第五方面提供了在E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)多肽的区域中产生变异的方法,包括:将编辑系统引入到植物细胞中,其中所述编辑系统靶向编码HECT E3 UPL多肽的区域的HECT E3 UPL基因的区域,其中HECT E3 UPL多肽的区域包含(a)与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的序列,(b)与以下的连续氨基酸残基的区域具有至少80%序列同一性的序列:参考SEQ ID NO:78的残基编号,从残基50到残基95、从残基43到残基102、从残基36到残基109、从残基1814到残基1865、从残基1822到残基1857和/或从残基1829到残基1851,参考SEQ ID NO:79的残基编号,从残基48到残基87、从残基41到残基94、从残基34到残基101、从残基1820到残基1870、从残基1827到残基1863和/或从残基1834到残基1856,参考SEQ ID NO:80的残基编号,从残基139到残基194、从残基132到残基201、从残基125到残基208、从残基1838到残基1888、从残基1845到残基1881和/或从残基1852到残基1874,参考SEQ ID NO:81的残基编号,从残基104到残基146、从残基97到残基153、从残基90到残基160、从残基1834到残基1884、从残基1841到残基1877和/或从残基1848到残基1870,参考SEQ ID NO:82的残基编号,从残基1783到残基1833、从残基1790到残基1826和/或从残基1797到残基1818,和/或参考SEQ ID NO:83的残基编号,从残基1776到残基1826、从残基1783到残基1819和/或从残基1790到残基1812;或(c)包含与SEQ ID NO:93的氨基酸序列具有至少90%序列同一性的序列;将HECT E3 UPL基因的区域与所述编辑系统接触,从而将突变引入到植物细胞中的HECT E3 UPL多肽的区域;和在HECT E3 UPL多肽的区域中产生变异。
第六方面提供了用于编辑植物细胞基因组中的特定位点的方法,该方法包括:以位点特异性方式切割植物细胞中内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因内的靶位点,所述内源HECT E3 UPL基因:(a)包含与SEQ ID NO:72-77的核苷酸序列中的任一个具有至少80%序列同一性的序列,(b)编码与SEQ ID NO:78-83中任一个的氨基酸序列具有至少80%序列同一性的区域,(c)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的区域,和/或(d)编码与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域,从而在植物细胞的内源HECT E3 UPL基因中产生编辑并产生包含内源HECT E3UPL基因中的编辑的植物细胞。
第七方面提供了用于制备植物的方法,其包括:(a)使包含内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的植物细胞群体与连接至核酸结合结构域(例如,编辑系统)的核酸酶接触,所述核酸结合结构域结合(a)与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性,(b)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列,(c)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列,和/或(d)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列的序列;(b)从所述群体中选择其中内源HECT E3UPL基因已突变的植物细胞,从而产生包含内源HECT E3 UPL基因中的突变的植物细胞;和(c)将选择的植物细胞生长成植物。
第八方面提供了用于改善植物中的一种或多种产量性状的方法,包括(a)使包含内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的植物细胞与靶向内源HECT E3UPL基因的核酸酶接触,其中所述核酸酶连接至结合内源HECT E3 UPL基因中的靶位点的核酸结合结构域(例如,编辑系统),其中所述内源HECT E3 UPL基因:(i)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列;(ii)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;(iii)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或(iv)编码包含与SEQ IDNO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,以产生包含内源HECT E3UPL基因中的突变的植物细胞;和(b)使植物细胞生长成包含内源HECT E3 UPL基因中的突变的植物,从而产生具有突变的内源HECT E3 UPL基因和一种或多种改善的产量性状(任选地增加的种子大小(例如,种子面积和/或种子重量)和/或种子油含量)的植物。
第九方面提供了用于产生植物或其部分的方法,所述植物或其部分包含至少一个具有突变的内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的细胞,所述方法包括将植物或植物部分中的内源HECT E3 UPL基因中的靶位点与包含切割结构域和核酸结合结构域的核酸酶接触,其中核酸结合结构域结合内源HECT E3 UPL基因中的靶位点,其中内源HECT E3 UPL基因(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列;(b)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;(c)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或(d)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,从而产生包含至少一个具有内源HECT E3 UPL基因中的突变的细胞的植物或其部分。
本发明的第十方面提供了用于产生植物或其部分的方法,所述植物或其部分包含突变的内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因并表现出一种或多种改善的产量性状,所述方法包括将植物或植物部分中的内源HECT E3 UPL基因中的靶位点与包含切割结构域和核酸结合结构域的核酸酶接触,其中核酸结合结构域结合内源HECT E3 UPL基因中的靶位点,其中内源HECT E3 UPL基因:(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列;(b)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;(c)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或(d)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,从而产生包含具有突变的内源HECT E3 UPL基因并表现出一种或多种改善的产量性状(任选地增加的种子大小(例如,种子面积和/或种子重量)和/或增加的种子油含量)的植物或其部分。
第十一方面提供了结合E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的靶位点的引导核酸,该靶位点位于HECT E3 UPL3基因的区域中,所述区域:(a)编码与SEQ IDNO:84-92中任一个的氨基酸序列具有至少80%序列同一性的序列,(b)编码与以下的连续氨基酸残基的区域具有至少80%序列同一性的序列:参考SEQ ID NO:78的残基编号,从残基50到残基95、从残基43到残基102、从残基36到残基109、从残基1814到残基1865、从残基1822到残基1857和/或从残基1829到残基1851,参考SEQ ID NO:79的残基编号,从残基48到残基87、从残基41到残基94、从残基34到残基101、从残基1820到残基1870、从残基1827到残基1863和/或从残基1834到残基1856,参考SEQ ID NO:80的残基编号,从残基139到残基194、从残基132到残基201、从残基125到残基208、从残基1838到残基1888、从残基1845到残基1881和/或从残基1852到残基1874,参考SEQ ID NO:81的残基编号,从残基104到残基146、从残基97到残基153、从残基90到残基160、从残基1834到残基1884、从残基1841到残基1877和/或从残基1848到残基1870,参考SEQ ID NO:82的残基编号,从残基1783到残基1833、从残基1790到残基1826和/或从残基1797到残基1818,和/或参考SEQ ID NO:83的残基编号,从残基1776到残基1826、从残基1783到残基1819和/或从残基1790到残基1812;和/或(c)编码与SEQ ID NO:93的氨基酸序列具有至少90%序列同一性的序列。
在第十二方面,提供了包含本发明的引导核酸和与该引导核酸缔合的CRISPR-Cas效应蛋白的系统。
第十三方面提供了包含与引导核酸缔合的CRISPR-Cas效应蛋白的基因编辑系统,其中引导核酸包含与内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因结合的间隔序列。
在第十四方面,提供了包含引导核酸和包含切割结构域的CRISPR-Cas效应蛋白的复合物,其中引导核酸结合内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的靶位点,其中内源HECT E3 UPL3基因:(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列,(b)编码与SEQ ID NO:78-83中任一个的氨基酸序列具有至少80%序列同一性的序列,(c)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的区域,和/或(d)编码与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域,其中切割结构域切割HECT E3 UPL3基因中的靶标链。
在第十五方面,提供了表达盒,该表达盒包含(a)编码包含切割结构域的CRISPR-Cas效应蛋白的多核苷酸,和(b)结合内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的靶位点的引导核酸,其中所述引导核酸包含间隔序列,所述间隔序列互补于并结合(i)与SEQ ID NO:72-77的核苷酸序列中的任一个具有至少80%序列同一性的序列的一部分;(ii)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列的核酸的一部分;(iii)编码与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的氨基酸序列的核酸的一部分;和/或(iv)编码与SEQ ID NO:93具有至少90%序列同一性的氨基酸序列的核酸的一部分。
在第十六方面,编码E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)多肽的显性失活突变或无效突变的核酸。
在第十七方面,提供了修饰的E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)多肽,所述修饰的HECT E3 UPL多肽包含位于以下位置处的氨基酸残基的突变:参考SEQ ID NO:78的氨基酸位置编号的位置1839,参考SEQ ID NO:79的氨基酸位置编号的位置1844,参考SEQ ID NO:80的氨基酸位置编号的位置1862,参考SEQ ID NO:81的氨基酸位置编号的位置1858,参考SEQ ID NO:82的氨基酸位置编号的位置1807,和/或参考SEQ ID NO:83的氨基酸位置编号的位置1800。
在另外的方面,提供了在植物中产生内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的突变的方法,包括:(a)将基因编辑系统靶向至HECT E3 UPL基因的一部分,其(i)编码对应于参考SEQ ID NO:78的氨基酸位置编号的位置1839、参考SEQ ID NO:79的氨基酸位置编号的位置1844、参考SEQ ID NO:80的氨基酸位置编号的位置1862、参考SEQID NO:81的氨基酸位置编号的位置1858、参考SEQ ID NO:82的氨基酸位置编号的位置1807和/或参考SEQ ID NO:83的氨基酸位置编号的位置1800的氨基酸;(ii)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;(iii)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或(iv)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,并且(b)选择植物,其包含(i)参考SEQ ID NO:78的氨基酸位置编号的位置1839、参考SEQ ID NO:79的氨基酸位置编号的位置1844、参考SEQ ID NO:80的氨基酸位置编号的位置1862、参考SEQ ID NO:81的氨基酸位置编号的位置1858、参考SEQ ID NO:82的氨基酸位置编号的位置1807和/或参考SEQ ID NO:83的氨基酸位置编号的位置1800处的氨基酸残基的取代;(ii)与SEQ IDNO:78-83中的任一个具有至少80%序列同一性的氨基酸序列中的突变;(iii)与SEQ IDNO:84-92中的任一个具有至少80%序列同一性的区域中的突变;和/或(iv)与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域中的突变。
另一方面,提供了在其基因组中包含通过本发明的方法产生的一个或多个突变的E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的植物。
本发明的另一方面提供了一种玉米植物或其植物部分,其包含在至少一个内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的至少一个非天然突变,所述基因具有Zm00001d014920和/或Zm00001d004139的基因识别号(基因ID)。
在另外的方面,提供了大豆植物或其植物部分,其包含在至少一个内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的至少一个非天然突变,所述基因具有Glyma.11G107500、Glyma.12G032500、Glyma.06G003600和/或Glyma.04G004000的基因识别号(基因ID)。
在进一步的方面,提供了引导核酸,其结合E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的靶核酸,该HECT E3 UPL基因具有Zm00001d014920、Zm00001d004139、Glyma.11G107500、Glyma.12G032500、Glyma.06G003600和/或Glyma.04G004000的基因识别号(基因ID)。
还提供了用于制备本发明的植物的多肽、多核苷酸、核酸构建体、表达盒和载体。
本发明的这些和其他方面在下面的本发明的描述中更详细地阐述。
序列简要描述
SEQ ID NO:1-17是可用于本发明的示例性Cas12a氨基酸序列。
SEQ ID NO:18-20是可用于本发明的示例性Cas12a核苷酸序列。
SEQ ID NO:21-22是编码启动子和内含子的示例性调节序列。
SEQ ID NO:23-29是可用于本发明的示例性胞嘧啶脱氨酶序列。
SEQ ID NO:30-40是可用于本发明的示例性腺嘌呤脱氨酶氨基酸序列。
SEQ ID NO:41是可用于本发明的示例性尿嘧啶-DNA糖基化酶抑制剂(UGI)序列。
SEQ ID NO:42-44提供了V型CRISPR-Cas12a核酸酶的前间隔区邻近基序位置的实例。
SEQ ID NO:45-47提供了可用于本发明的示例性肽标签和亲和多肽。
SEQ ID NO:48-58提供了可用于本发明的示例性RNA募集基序和相应的亲和性多肽。
SEQ ID NO:59-60是可用于本发明的示例性Cas9多肽序列。
SEQ ID NO:61-71是可用于本发明的示例性Cas9多核苷酸序列。
SEQ ID NO:72是来自大豆的示例性UPL3a基因组序列。
SEQ ID NO:73是来自大豆的示例性UPL3b基因组序列。
SEQ ID NO:74是来自大豆的示例性UPL3c基因组序列。
SEQ ID NO:75是来自大豆的示例性UPL3d基因组序列。
SEQ ID NO:76是来自玉米的示例性UPL3a基因组序列。
SEQ ID NO:77是来自玉米的示例性UPL3b基因组序列。
SEQ ID NO:78是来自大豆的示例性UPL3a多肽序列。
SEQ ID NO:79是来自大豆的示例性UPL3b多肽序列。
SEQ ID NO:80是来自大豆的示例性UPL3c多肽序列。
SEQ ID NO:81是来自大豆的示例性UPL3d多肽序列。
SEQ ID NO:82是来自玉米的示例性UPL3a多肽序列。
SEQ ID NO:83是来自玉米的示例性UPL3b多肽序列。
SEQ ID NO:84-93是UPL3多肽序列的示例性部分或区域。
SEQ ID NO:94-118是用于本发明的核酸引导物的示例性间隔序列。
SEQ ID NO:119-121是示例性经编辑的大豆序列。
SEQ ID NO:122-125是示例性经编辑的玉米序列。
SEQ ID NO:126和127分别是经编辑的大豆序列SEQ ID NO:120和SEQ ID NO:121的缺失部分。
SEQ ID NO:128是经编辑的玉米序列SEQ ID NO:124的缺失部分。
发明详述
现在将在下文中参考所附实施例来描述本发明,其中示出了本发明的实施方案。该描述并非旨在成为可以实施本发明的所有不同方式或可以添加到本发明的所有特征的详细目录。例如,关于一个实施方案举例说明的特征可以并入其他实施方案中,并且关于特定实施方案举例说明的特征可以从该实施方案中缺失。因此,本发明设想在本发明的一些实施方案中,可以排除或省略本文阐述的任何特征或特征组合。此外,根据本公开内容,对本文提出的各种实施方案的许多变化和添加对于本领域技术人员来说将是明显的,它们不脱离本发明。因此,以下描述旨在举例说明本发明的一些特定实施方案,而不是详尽地指定其所有排列、组合和变化。
除非另有定义,本文使用的所有技术和科学术语与本发明所属领域的普通技术人员通常理解的含义相同。在本文的本发明的描述中使用的术语仅用于描述特定实施方案的目的,并不旨在限制本发明。
本文引用的所有出版物、专利申请、专利和其他参考文献关于与在其中呈现参考文献的句子和/或段落相关的教导通过引用整体并入。
除非上下文另有说明,否则本文所述的本发明的各种特征特别旨在以任何组合形式使用。此外,本发明还设想在本发明的一些实施方案中,可以排除或省略本文阐述的任何特征或特征组合。为了举例说明,如果说明书声明组合物包含组分A、B和C,则特别意在A、B或C中的任何一个或其组合可以单独或以任何组合被省略和弃权。
如在本发明和所附权利要求的描述中使用的,除非上下文另有明确指示,否则单数形式“一个”、“一种”和“该”意在也包括复数形式。
也如本文所用,“和/或”是指并且包括一个或多个相关列出的项目的任何和所有可能的组合,以及在替代(“或”)中解释时组合的缺乏。
如本文所用的术语“约”在提及可测量值诸如量或浓度时,意在涵盖规定值的±10%、±5%、±1%、±0.5%或甚至±0.1%的变化以及规定值。例如,其中X是可测量的值的“约X”意在包括X以及X的±10%、±5%、±1%、±0.5%或甚至±0.1%的变化。此处提供的可测量值的范围可以包括任何其他范围和/或其中的个体值。
如本文所用,诸如“在X和Y之间”和“在约X和Y之间”的短语应当被解释为包括X和Y。如在本文中使用的,诸如“约X和Y之间”的短语意指“约X和约Y之间”并且诸如“从约X至Y”的短语意指“从约X至约Y”。
除非在本文中另有说明,否则本文中数值范围的列举仅旨在用作单独引用落入该范围内的每个单独值的速记方法,并且每个单独的值被并入说明书中,就好像它在本文中单独列举一样。例如,如果公开了范围10至15,则还公开了11、12、13和14。
如本文所用,术语“包括”、“包含”和“含有”指定了所述特征、整数、步骤、操作、元素和/或组件的存在,但不排除一种或多种其他特征、整数、步骤、操作、元素、组件和/或它们的组的存在或添加。
如本文所用,过渡短语“基本上由……组成”是指权利要求的范围应解释为包括权利要求中列举的具体材料或步骤,以及那些不实质影响要求保护的本发明的基本和新颖特征的材料或步骤。因此,当在本发明的权利要求中使用时,术语“基本上由……组成”并不旨在解释为等同于“包括”。
如本文所用,术语“增加”、“增加的”、“增加了”、“增强”、“增强的”、“增强了”和“增强”(及其语法变体)描述了与对照相比至少约5%、10%、15%、20%、25%、50%、75%、100%、150%、200%、300%、400%、500%或更多的升高。例如,包含如本文所述的HECT E3UBL基因中的突变的植物可以表现出改善的产量性状(例如,一种或多种改善的产量性状;例如,任选地增加的种子大小(例如,种子面积和/或种子重量)和/或增加的种子油含量和),其比不包含相同突变的对照植物的产量性状增加至少约5%或更高。对照植物通常是与编辑的植物相同的植物,但对照植物没有经过类似的编辑,因此缺乏突变。对照植物可以是同基因植物和/或野生型植物。因此,对照植物可以是与本文所述的突变渗入其中的主题植物相同的育种系、品种或栽培种,但对照育种系、品种或栽培种不含突变。在一些实施方案中,在相同的生长条件下,例如相同的环境条件(土壤、水合作用、光、热、营养物等)下进行本发明的植物和对照植物之间的比较。
如本文所用,术语“降低”、“降低的”、“降低了”、“减少”、“减小”和“减轻”(及其语法变体)描述例如与对照相比至少约5%、10%、15%、20%、25%、35%、50%、75%、80%、85%、90%、95%、96%、97%、98%、99%、99.5%、99.6%、99.7%、99.8%、99.9%或100%的降低。在特定实施方案中,降低可导致没有或基本上没有(即,微不足道的量,例如,小于约10%或甚至5%)可检测的活性或量。
如本文所用,关于核酸分子和/或核苷酸序列(例如,RNA或DNA)的术语“表达”、“表达的”、“表达了”或“表达”等表示核酸分子和/或核苷酸序列被转录和任选地被翻译。因此,核酸分子和/或核苷酸序列可以表达感兴趣的多肽或例如功能性非翻译RNA。
“异源”或“重组”核苷酸序列是与其引入的宿主细胞不天然相关的核苷酸序列,包括天然存在的核苷酸序列的非天然存在的多个拷贝。“异源”核苷酸/多肽可以源自外来物种,或者如果源自相同物种,则通过有意的人为干预在组成和/或基因组基因座上对其天然形式进行了实质性修饰。
“天然”或“野生型”核酸、核苷酸序列、多肽或氨基酸序列是指天然存在的或内源性核酸、核苷酸序列、多肽或氨基酸序列。在一些情况下,“野生型”核酸是未如本文所述编辑的核酸,并且可以不同于可如本文所述编辑的“内源”基因(例如,突变的内源基因)。在一些情况下,“野生型”核酸(例如,未编辑的)对于其中发现野生型核酸的生物体(例如,转基因生物体)可以是异源的。作为实例,“野生型内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)(HECT E3 UPL)基因”是HECT E3 UPL基因,任选地UPL3a基因、UPL3b基因、UPL3c基因,和/或UPL3d基因,其天然存在于参考生物体(例如植物,例如大豆植物、玉米植物)中或是参考生物体内源的,并且可以进行如本文所述的修饰,之后,这样的修饰的内源基因基因不再是野生型。
如本文所用,术语“杂合的”是指其中不同等位基因位于同源染色体上相应基因座的遗传状态。
如本文所用,术语“纯合的”是指其中相同等位基因位于同源染色体上的相应基因座的遗传状态。
如本文所用,术语“等位基因”是指出现在特定基因座处的两种或更多种不同核苷酸或核苷酸序列之一。
“无效等位基因”是由基因突变引起的非功能性等位基因,该基因突变导致完全缺乏相应蛋白质的产生或产生非功能性蛋白质。
“显性失活突变”是产生改变的基因产物(例如,相对于野生型具有异常功能)的突变,该基因产物不利地影响野生型等位基因或基因产物的功能。例如,“显性失活突变”可能会阻断野生型基因产物的功能。显性失活突变也可称为“反效等位基因突变”。
“半显性突变”是指杂合生物中表型的外显率低于纯合生物中观察到的突变。
“弱功能丧失突变”是导致基因产物与野生型基因产物相比具有部分功能或降低的功能(部分失活)的突变。
“亚形态突变”是导致基因功能部分丧失的突变,其可能通过降低表达(例如,减少蛋白质和/或减少RNA)或降低功能性能(例如,降低活性)而发生,但不是完全丧失功能/活性。“亚形态”等位基因是由基因突变引起的半功能等位基因,该基因突变导致功能在正常效率的1%至99%之间的相应蛋白质的产生。
“超形态突变”是导致基因产物表达增加和/或基因产物活性增加的突变。
“基因座”是基因或标记或等位基因所在的染色体上的位置。在一些实施方案中,基因座可以包含一个或多个核苷酸。
如本文所用,术语“所需等位基因”、“靶等位基因”和/或“感兴趣的等位基因”可互换使用以指代与所需性状相关的等位基因。在一些实施方案中,所需等位基因可与给定性状的增加或减少(相对于对照)或给定性状相关,这取决于所需表型的性质。
当性状与标记相关并且当标记的存在是所需性状或性状形式是否将在包含所述标记的植物/种质中出现和/或以何种程度出现的指示时,所述标记与所述性状“相关”。类似地,当标记与等位基因或染色体区间相关并且当标记的存在是等位基因或染色体区间是否存在于包含标记的植物/种质中的指示时,标记与等位基因或染色体区间“相关”。
如本文所用,术语“回交”和“回交的”是指将后代植物回交至其亲本之一一次或多次(例如,1、2、3、4、5、6、7、8次等)。在回交方案中,“供体”亲本是指具有要渗入的所需基因或基因座的亲本植物。“受体”亲本(使用一次或多次)或“循环”亲本(使用两次或更多次)是指基因或基因座正在渗入的亲本植物。例如,参见Ragot,M.等人,Marker-assistedBackcrossing:A Practical Example,Techniques et Utilisations des MarqueursMoleculaires Les Colloques,Vol.72,pp.45-56(1995);和Openshaw等人,Marker-assisted Selection in Backcross Breeding,Proceedings of the Symposium"Analysis of Molecular Marker Data,"pp.41-43(1994)。最初的杂交产生了F1代。术语“BC1”是指回交双亲的第二次使用,“BC2”是指回交双亲的第三次使用,依此类推。
如本文所用,术语“杂交”或“杂交的”是指配子通过授粉融合以产生后代(例如,细胞、种子或植物)。该术语包括有性杂交(一种植物由另一种植物授粉)和自交(自花授粉,例如,当花粉和胚珠来自同一植物时)。术语“杂交”是指通过授粉融合配子以产生后代的行为。
如本文所用,术语“基因渗入”、“基因渐渗”和“基因渗入的”是指遗传基因座的所需等位基因或所需等位基因组合从一种遗传背景到另一种遗传背景的天然和人工传递。例如,可以通过同一物种的两个亲本之间的有性杂交将特定基因座处的所需等位基因传递给至少一个(例如一个或多个)后代,其中至少一个亲本在其基因组中具有所需等位基因。或者,例如,等位基因的传递可以通过两个供体基因组之间的重组发生,例如在融合的原生质体中,其中至少一个供体原生质体在其基因组中具有所需的等位基因。所需等位基因可以是标记、QTL、转基因等的选定等位基因。包含所需等位基因的后代可以与具有所需遗传背景的品系回交一次或多次(例如,1、2、3、4或更多次),选择所需等位基因,结果是所需等位基因固定在所需的遗传背景中。例如,与在非水胁迫条件下增加的产量相关的标记可以从供体渗入到不包含该标记并且在非水胁迫条件下不表现出增加的产量的回交亲本中。然后可以将所得后代回交一次或多次并进行选择,直到后代具有与回交亲本背景中的非水胁迫条件下的产量增加相关的遗传标记。
“遗传图谱”是对给定物种内一条或多条染色体上基因座之间遗传连锁关系的描述,通常以图解或表格形式描绘。对于每个遗传图谱,基因座之间的距离通过它们之间的重组频率来衡量。可以使用多种标记检测基因座之间的重组。遗传图谱是对群体、使用的标记类型以及不同群体之间每个标记的多态性潜力作图的产物。基因座之间的顺序和遗传距离可以从一个遗传图谱到另一个遗传图谱不同。
如本文所用,术语“基因型”是指个体(或个体群体)在一个或多个遗传基因座处的遗传构成,与可观察和/或可检测和/或表现的性状(表型)形成对比。基因型由个体从其亲本继承的一个或多个已知基因座的等位基因定义。术语基因型可用于指个体在单个基因座、多个基因座处的遗传构成,或更一般地,术语基因型可用于指个体基因组中所有基因的遗传构成。基因型可以间接表征,例如使用标记,和/或通过核酸测序直接表征。
如本文所用,术语“种质”是指来自或来源于个体(例如植物)、个体群体(例如植物品系、品种或科)的遗传物质,或源自品系、品种、物种或培养物的克隆。种质可以是生物体或细胞的一部分,或者可以与生物体或细胞分离。一般来说,种质提供了具有特定的遗传组成的遗传物质,其为生物体或细胞培养物的部分或全部遗传特性提供了基础。如本文所用,种质包括可从其生长新植物的细胞、种子或组织,以及可培养成整株植物的植物部分(例如,叶、茎、芽、根、花粉、细胞等)。.
如本文所用,术语“栽培品种”和“品种”是指通过结构或遗传特征和/或性能可与同一物种内的其他品种区分开来的一组相似植物。
如本文所用,术语“外来的”、“外来品系”和“外来种质”是指非优良的任何植物、品系或种质。一般而言,外来植物/种质不是源自任何已知的优良植物或种质,而是被选择以将一种或多种所需遗传元件引入育种程序中(例如,将新等位基因引入育种程序中)。
如本文所用,植物育种上下文中的术语“杂种”是指作为通过杂交不同品系或品种或物种的植物(包括但不限于两个近交系之间的杂交)产生的遗传不同亲本的后代的植物。
如本文所用,术语“近交”是指基本上纯合的植物或品种。该术语可以指在整个基因组中基本上是纯合的或者就特别感兴趣的基因组的一部分而言基本上是纯合的植物或植物品种。
“单倍型”是在多个基因座处的个体的基因型,即等位基因的组合。通常,定义单倍型的遗传基因座在物理上和遗传上是相连的,即在同一染色体区段上。术语“单倍型”可以指特定基因座处的多态性,例如单个标记基因座,或沿着染色体区段的多个基因座处的多态性。
如本文所用,术语“异源”是指源自外来物种或者如果源自相同物种则通过有意的人为干预在组成和/或基因组基因座上从其天然形式显著修饰的核苷酸/多肽。
其中至少一个(例如,一个或多个,例如,1、2、3或4个或更多个)HECT E3 UBL基因(例如,一个或多个HECT E3 UBL基因;例如,一个或多个HECT E3 UBL3基因)如本文所述修饰(例如,包含如本文所述的修饰)的植物与不包含至少一个HECT E3 UBL基因中的修饰的植物相比可以具有改善的产量性状。如本文所用,“改善的产量性状”是指与生长相关的任何植物性状,例如生物量、产量、氮利用效率(NUE)、花序大小/重量、果实产量、果实质量、果实大小、种子大小(例如,种子面积、种子大小)、种子数、叶组织重量、结瘤数、结瘤质量、结瘤活性、种子头数、分蘖数、分枝数、花量、块茎量、块茎质量、鳞茎质量、种子数、总种子质量、出叶率、分蘖/分枝出现率、出苗率、根的长度、根的数量、根群的大小和/或重量,或它们的任何组合。因此,在一些方面,“改善的产量性状”可以包括但不限于与对照植物或其部分(例如,不包含突变的内源HECT E3 UBL核酸(例如突变的HECT E3 UBL基因)的植物)相比增加的花序产量、增加的果实产量(例如增加的果实数量、重量和/或大小;例如,例如玉米的穗的增加的数量、重量和/或大小)、增加的果实质量、根的增加的数量、大小和/或重量、增加的分生组织大小、增加的种子大小(例如,种子面积和/或种子重量)、增加的生物量、增加的叶大小、增加的氮利用效率、增加的高度、增加的节间数量和/或增加的节间长度。改善的产量性状也可以由本发明的植物的增加的种植密度产生。因此,在一些方面,本发明的植物能够以增加的密度种植(由于内源突变导致的改变的植物结构的结果),这导致与在相同的密度下种植的对照植物相比改善的产量性状。在一些方面,改善的产量性状可以表示为每面积土地生产的谷物数量(例如,每英亩土地的蒲式耳)。在一些实施方案中,改善的产量性状包括但不限于增加的种子大小(例如,种子面积和/或种子重量)和/或增加的种子油含量。
如本文所用,“增加的种子大小”可以指面积增加的种子。在一些实施方案中,与来自对照植物(例如,不包含如本文所述的内源HECT E3 UBL基因中的突变的植物)的种子相比,种子的面积可增加高达约70%(例如,约1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、45、46、47、48、49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64、65、66、67、68、69、70%)。在一些实施方案中,与来自对照植物(例如,不包含如本文所述的内源HECT E3 UBL基因中的突变的植物)的种子相比,种子的重量可增加高达约50%(例如,约1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、45、46、47、48、49或50%)。在一些实施方案中,种子大小的增加可以包括种子面积和种子大小两者的增加。
如本文所用,“增加的种子油含量”是指油/脂质含量与来自对照植物(例如,不包含本文所述的内源HECT E3 UBL基因中的突变的植物)的种子相比增加约30%(例如,约1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29或30%)的种子。在一些实施方案中,具有增加的种子油含量的种子的大小也可以增加(例如,种子面积和/或种子重量增加)。
如本文所用,“对照植物”意指不含有如本文所述的赋予增强/改善的性状(例如,产量性状)或改变的表型的经编辑的一个或多个HECT E3 UBL基因的植物。对照植物用于鉴定和选择如本文所述编辑的并且与对照植物相比具有增强的性状或改变的表型的植物。合适的对照植物可以是用于产生包含突变的一个或多个HECT E3 UBL基因的植物的亲本系的植物,例如,缺乏如本文所述的内源HECT E3UBL基因中的编辑的野生型植物。合适的对照植物还可以是含有赋予其他性状的重组核酸的植物,例如具有增强的除草剂耐受性的转基因植物。在一些情况下,合适的对照植物可以是缺乏如本文所述的突变的HECT E3 UBL基因的杂合或半合转基因植物系的后代,称为负分离子或负等基因系。
增强的性状(例如增加的产量性状)可以是例如与对照植物相比,从种植到成熟的天数减少、茎大小增加、叶子数量增加、营养阶段的植物高度生长速率增加、穗大小增加、每株植物增加的穗干重、每穗增加的籽粒数量、每籽粒增加的重量、每株植物增加的籽粒数量、减少的空穗、延长的谷粒填充期、降低的植物高度、增加的根枝数量、增加的总根长、增加的产量、增加的氮利用效率和增加的水利用效率。改变的表型可以是例如植物高度、生物量、冠层面积、花青素含量、叶绿素含量、施用的水、水含量和水利用效率。
如本文所用,“性状”是植物或特定植物材料或细胞的生理学、形态学、生物化学或物理学特征。在一些情况下,该特征是人眼可见的并且可以机械地测量,例如种子或植物的大小、重量、形状、形式、长度、高度、生长速率和发育阶段,或者可以通过生物化学技术测量,例如检测种子或叶子的蛋白质、淀粉、某些代谢物或油含量,或通过观察代谢或生理过程,例如通过测量对缺水或特定盐或糖浓度的耐受性,或通过测量一个或多个基因的表达水平,例如通过采用Northern分析、RT-PCR、微阵列基因表达测定或报告基因表达系统,或通过农业观察例如高渗胁迫耐受性或产量。然而,可以使用任何技术来测量转基因植物中任何选择的化合物或大分子的量、比较水平或差异。
如本文所用,“增强的性状”是指由本文所述的HECT E3 UBL基因中的突变引起的植物的特征。此类性状包括但不限于增强的农艺性状,其特征在于增强的植物形态、生理学、生长和发育、产量、营养增强、疾病或害虫抗性或环境或化学耐受性。在一些实施方案中,增强的性状/改变的表型可以是例如与对照植物相比,从种植到成熟的天数减少、茎大小增加、叶子数量增加、营养阶段的植物高度生长速率增加、穗大小增加、每株植物增加的穗干重、每穗增加的籽粒数量、每籽粒增加的重量、每株植物增加的籽粒数量、减少的空穗、延长的谷粒填充期、降低的植物高度、增加的根枝数量、增加的总根长、耐旱、增加的水利用效率、耐寒、增加的氮利用效率和增加的产量。在一些实施方案中,性状是在非胁迫条件下增加的产量或在环境胁迫条件下增加的产量。胁迫条件可包括生物胁迫和非生物胁迫,例如干旱、遮荫、真菌病、病毒病、细菌病、昆虫侵扰、线虫侵扰、低温暴露、热暴露、渗透胁迫、减少的氮养分可用性、减少的磷养分可用性和高植物密度。“产量”可以受到许多特性的影响,包括但不限于植物高度、植物生物量、豆荚数量、植物上的豆荚位置、节间数目、豆荚破碎的发生率、谷粒大小、穗大小、穗尖充盈度、籽粒败育、结瘤和固氮效率、养分同化效率、生物和非生物胁迫抗性、碳同化、植物结构、抗倒伏性、种子发芽率、幼苗活力和幼体性状。产量还可能受到以下因素的影响:发芽效率(包括应激条件下的发芽)、生长速率(包括应激条件下的生长速率)、开花时间和持续时间、穗数、穗大小、穗重量、每穗或豆荚的种子数、种子大小、种子成分(淀粉、油、蛋白质)和种子填充物的特征。
本文还使用的术语“性状修饰”涵盖通过在包含如本文所述的内源HECT E3 UBL基因中的突变的植物中相对于不包含该突变的植物(例如野生型植物或负分离子)产生可检测的特征差异来改变天然存在的性状。在一些情况下,可以定量评估性状改变。例如,与对照植物相比,性状修饰可以导致观察到的性状特征或表型的增加或减少。众所周知,经修饰的性状中可能存在自然变异。因此,观察到的性状修饰需要与对照植物相比,植物中性状特征或表型的正态分布和幅度的变化。
本公开内容涉及具有改善的经济上相关的特征的植物,更具体地增加的产量。更具体地,本公开内容涉及包含如本文所述的HECT E3UBL基因(例如HECT E3 UBL3基因)中的突变的植物,其中所述植物与缺乏所述突变的对照植物相比具有增加的产量。在一些实施方案中,如本文所述产生的植物与对照植物相比表现出增加的产量或改善的产量性状成分。在一些实施方案中,本公开内容的植物表现出与产量相关的改善的性状,包括但不限于增加的氮利用效率、增加的氮胁迫耐受性、增加的水利用效率和增加的干旱耐受性,如下文所定义和讨论的。
产量可以被定义为农作物的经济价值的可测量的产出。产量可以在数量和/或质量的范围内定义。产量可以直接取决于几个因素,例如,器官的数量和大小、植物结构(例如分枝数量、植物生物量,例如增加的根生物量、更陡的根角和/或更长的根等)、开花时间和持续时间、谷粒填充期。根结构和发育、光合效率、养分吸收、胁迫耐受性、早期活力、延迟的衰老和功能性持绿表型可能是决定产量的因素。因此,优化上述因素有助于提高作物产量。
本文提及的产量相关性状的增加/改善也可以被认为是指植物的一个或多个部分的生物量(重量)的增加,其可以包括地上和/或地下(可收获)植物部分。具体地,此类可收获部分是种子,并且本公开内容的方法的实施导致相对于合适的对照植物的种子产量具有增加的产量和特别是增加的种子产量的植物。术语植物的“产量”可以涉及该植物的营养生物量(根和/或芽生物量)、生殖器官和/或繁殖体(例如种子)。
本公开内容的植物的增加的产量可以通过多种方式测量,包括测试重量、每株植物的种子数、种子重量、每单位面积的种子数(例如,每英亩的种子或种子的重量)、每英亩的蒲式耳、每英亩的吨或每公顷的公斤。增加的产量可以通过关键生化化合物(例如氮、磷和碳水化合物)的改善的利用,或对环境胁迫例如冷、热、干旱、盐、遮荫、高植物密度以及害虫或病原体攻击的改善的反应来实现。
“增加的产量”可以表现为以下一种或多种:(i)植物的一个或多个部分的增加的植物生物量(重量),特别是植物的地上(可收获)部分,增加的根生物量(增加的根数量、增加的根厚度、增加的根长度)或任何其他可收获部分的增加的生物量;或(ii)增加的早期活力,本文定义为发芽后约三周改善的幼苗地上面积。
“早期活力”是指积极健康的植物生长,尤其是在植物生长的早期阶段,并且可以由于因例如植物更好地适应其环境(例如,优化能源的使用、养分的吸收以及在芽和根之间分配碳份额)而增加的植物适应度引起。例如,早期活力可以是种子在种植后发芽和出苗的能力以及幼苗在出苗后生长和发育的能力的组合。具有早期活力的植物还表现出增加的幼苗存活率和更好的作物定植,这通常导致高度均匀的田地,其中大多数植物基本上同时达到不同的发育阶段,这通常导致增加的产量。因此,早期活力可以通过测量各种因素来确定,例如籽粒重量、发芽率、出苗率、幼苗生长、幼苗高度、根长、根和芽生物量、冠层大小和颜色等。
此外,增加的产量还可以表现为增加的总种子产量,这可能是由于因每株植物和/或单个种子基础上的种子重量的增加所致的种子生物量(种子重量)的增加,每株植物增加的花/圆锥花序数量;增加的豆荚数量;增加的茎节数量;每穗/植物增加的花(“小花”)数量;增加的种子饱满率;增加的饱满种子的数量;增加的种子大小(长度、宽度、面积、周长和/或重量),这也可影响种子的成分;和/或增加的种子体积(这也可影响种子的成分)中的一种或多种引起的。在一个实施方案中,增加的产量可以是增加的种子产量,例如增加的种子重量;增加的饱满种子的数量;和增加的收获指数。
增加的产量也可能导致修饰的结构,或者可能因修饰的植物结构而发生。
增加的产量也可以表现为增加的收获指数,其表示为可收获部分(例如种子)的产量与总生物量的比率。
本公开内容还延伸至植物的可收获部分,例如但不限于种子、叶、果实、花、棉铃、豆荚、长角果、坚果、茎、根茎、块茎和球茎。本公开内容还涉及源自此类植物的可收获部分的产品,例如干颗粒、粉末、油、脂肪和脂肪酸、淀粉或蛋白质。
本公开内容提供了用于增加植物的“产量”或植物或植物部分的“大英亩产量”的方法,植物或植物部分被定义为每单位面积可收获的植物部分,例如每英亩的种子或种子的重量、每英亩的磅、每英亩蒲式耳、每英亩的吨(tones per acre)、每英亩的吨、每公顷的公斤。
如本文所用,“氮利用效率”是指导致施用的每单位氮的植物产量、生物量、活力和生长速率增加的过程。这些过程可以包括植物对氮的吸收、同化、积累、信号传导、传感、再转移(在植物内)和利用。
如本文所用,“增加的氮利用效率”是指当经受与正常或标准条件下相同量的可用/施用的氮时,植物比正常情况更快或更好地生长、发育或生产的能力;植物在经受低于可用/施用的氮的最佳量或在氮限制条件下正常生长、发育或生产或者更快或更好地生长、发育或生产的能力。
如本文所用,“氮限制条件”是指提供低于充分或成功的植物代谢、生长、繁殖成功和/或活力所需的最佳氮量的生长条件或环境。
如本文所用,“增加的氮胁迫耐受性”是指植物在经受低于可用/施加的氮的最佳量时或在氮限制条件下正常生长、发育或生产或者更快或更好地生长、发育或生产的能力。
增加的植物氮利用效率可以在田间转化为在供应更少的氮的同时收获相似数量的产量,或者通过供应最佳/足够量的氮获得增加的产量。增加的氮利用效率可以提高植物氮胁迫耐受性,并且还可以改善作物品质和种子的生化成分,例如蛋白质产量和油产量。术语“增加的氮利用效率”、“增强的氮利用效率”和“氮胁迫耐受性”在本公开内容中可互换使用,是指在氮限制条件下具有提高的生产力的植物。
如本文所用,“水利用效率”是指每单位蒸腾的水蒸气被叶子同化的二氧化碳的量。它构成控制干燥环境中植物生产力的最重要性状。“耐旱性”是指植物适应干燥或干旱条件的程度。植物对缺水的生理反应包括叶子枯萎、叶面积减少、叶子脱落以及通过将养分引导到植物的地下部分来刺激根部生长。通常,植物在开花和种子发育(繁殖阶段)期间更容易受到干旱的影响,因为植物的资源被用来支持根生长。此外,脱落酸(ABA)(一种植物应激激素)诱导叶片气孔(参与气体交换的微孔)关闭,从而减少通过蒸腾作用的水分流失,并降低光合作用速率。这些措施在短期内提高了植物的水利用效率。术语“增加的水利用效率”、“增强的水利用效率”和“增加的耐旱性”在本公开内容中可互换使用,是指在水限制条件下具有改善的生产力的植物。
如本文所用,“增加的水利用效率”是指当经受与正常或标准条件下相同量的可用/施用的水时,植物比正常情况更快或更好地生长、发育或生产的能力;当经受减少量的可用/施用的水(水输入)或在水分胁迫或缺水胁迫条件下时,植物正常生长、发育或生产或者更快或更好地生长、发育或生产的能力。
如本文所用,“增加的耐旱性”是指当经受减少量的可用/施用的水和/或在急性或慢性干旱条件下时,植物正常生长、发育或生产或者比正常情况更快或更好地生长、发育或生产的能力;当经受减少量的可用/施用的水(水输入)或在缺水胁迫条件下或在急性或慢性干旱条件下,植物正常生长、发育或生产的能力。
如本文所用,“干旱胁迫”是指导致缺水并使植物遭受胁迫和/或植物组织损伤和/或负面影响谷物/作物产量的干燥时期(急性或慢性/长期);导致缺水和/或气温升高并使植物遭受胁迫和/或植物组织损伤和/或负面影响谷物/作物产量的干燥时期(急性或慢性/长期)。
如本文所用,“缺水”是指提供低于植物充分/成功生长和发育所需的最佳水量的条件或环境。
如本文所用,“水胁迫”是指提供比植物/作物充分/成功生长和发育所需的水量不适当(更少/不足或更多/过量)的水量的条件或环境,从而使植物遭受胁迫和/或植物组织损伤和/或负面影响谷物/作物产量。
如本文所用,“缺水胁迫”是指提供比植物/作物充分/成功生长和发育所需的水量更少/不足的水量的条件或环境,从而使植物遭受胁迫和/或植物组织损伤和/或负面影响谷物产量。
如本文所用,术语“核酸”、“核酸分子、”“核苷酸序列”和“多核苷酸”是指线性或支链、单链或双链或其混合体的RNA或DNA。该术语还包括RNA/DNA混合体。当合成产生dsRNA时,较少见的碱基如肌苷、5-甲基胞嘧啶、6-甲基腺嘌呤、次黄嘌呤等也可用于反义、dsRNA和核酶配对。例如,含有尿苷和胞苷的C-5丙炔类似物的多核苷酸已被证明以高亲和力结合RNA,并且是基因表达的强效反义抑制剂。还可以进行其他修饰,例如对磷酸二酯骨架的修饰,或RNA的核糖糖基中的2'-羟基。
如本文所用,术语“核苷酸序列”是指核苷酸的杂聚物或这些核苷酸从核酸分子的5’端至3’端的序列,并且包括DNA或RNA分子,包括cDNA、DNA片段或部分、基因组DNA、合成(例如,化学合成)DNA、质粒DNA、mRNA和反义RNA,其中任何一种都可以是单链或双链的。术语“核苷酸序列”、“核酸”、“核酸分子”、“核酸构建体”、“寡核苷酸”和“多核苷酸”在本文中也可互换地用于指核苷酸的杂聚物。本文提供的核酸分子和/或核苷酸序列在本文中从左到右以5’至3’方向呈现,并使用代表美国序列规则、37CFR§§1.821-1.825和世界知识产权组织(WIPO)标准ST.25中所述的核苷酸字符的标准代码来表示。如本文所用,“5’区域”可以是指最接近多核苷酸的5’端的多核苷酸的区域。因此,例如,多核苷酸的5’区域中的元件可以位于从位于多核苷酸5’端的第一核苷酸到位于多核苷酸中间的核苷酸的任何位置。如本文所用,“3’区域”可以是指最接近多核苷酸的3’端的多核苷酸的区域。因此,例如,多核苷酸的3’区域中的元件可以位于从位于多核苷酸3’端的第一核苷酸到位于多核苷酸中间的核苷酸的任何位置。
如本文关于核酸使用的,术语“片段”或“部分”是指相对于参考核酸长度减少(例如减少1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、20、40、50、60、70、80、90、100、110、120、130、140、150、160、170、180、190、200、210、220、230、240、250、260、270、280、290、300、310、320、330、340、350、400、450、500、550、600、650、700、750、800、850或900或更多个核苷酸或其中的任何范围或值)并且包含与参考核酸的相应部分相同或几乎相同(例如,70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%相同)的连续核苷酸的核苷酸序列、基本上由其组成和/或由其组成的核酸。在适当的情况下,这样的核酸片段可以包含在更大的多核苷酸中,该核酸片段是该多核苷酸的组成部分。例如,本发明的引导核酸的重复序列可以包含野生型CRISPR-Cas重复序列(例如,野生型CRISR-Cas重复;例如,来自例如Cas9、Cas12a(Cpf1)、Cas12b、Cas12c(C2c3)、Cas12d(CasY)、Cas12e(CasX)、Cas12g、Cas12h、Cas12i、C2c4、C2c5、C2c8、C2c9、C2c10、Cas14a、Cas14b和/或Cas14c等的CRISPR Cas系统的重复)的“一部分”。
在一些实施方案中,核酸片段可包含编码HECT E3 UPL多肽(例如,HECT E3 UPL3多肽)的核酸的约5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、50、55、60、65、70、75、80、85、90、95、100、110,120、130、140、150、160、170、180、190、200、210、220、230、240、250、260、270、280、285、290、300、310、320、330、340、350、360、370、380、390、395、396、397、398、399、400、410、411、412、413、414、415,420、425、430、435、440、445、450或500个或更多个连续核苷酸或其中的任何范围或值、基本上由其组成或由其组成,任选地HECT E3 UPL基因的片段的长度可以是约5、6、7、8、9或10个连续核苷酸至约300、320、350、375、400或更多个连续核苷酸,可以是约5、6、7、8、9、10、11、12、13、14、15、16、17、18、19或20个连续核苷酸至约30、35、40、45、50、55、60、65、70、75,80、85、90、95或100个连续核苷酸,或约80、85、90、95或100个连续核苷酸至约300、320、350、375、400或更多个连续核苷酸,或其中的任何范围或值。
在一些实施方案中,“序列特异性核酸结合结构域”可以结合编码例如本文所述的HECT E3 UPL多肽的核苷酸序列(例如,DNA、RNA)的一个或多个片段或部分。
如本文所用,关于多肽,术语“片段”或“部分”可以指相对于参考多肽长度减少的多肽,并且其包含与参考多肽的相应部分相同或几乎相同(例如,90%、91%、92%、93%、94%、95%、96%、97%、98%、99%相同)的连续氨基酸的氨基酸序列、基本上由其组成和/或由其组成。在适当的情况下,这样的多肽片段可以包含在其作为其组成部分的较大多肽中。在一些实施方案中,多肽片段可包含参考多肽的至少约2、3、4、5、6、7、8、9、10、11、12、13、14、15、20、25、30、35、40、45、50、55、60、65、70、75、80、85、90、95、100、125、150、175、200、225、250、260、270、280或290个或更多个连续氨基酸、基本上由其组成或由其组成。在一些实施方案中,多肽片段可包含HECT E3 UPL多肽(例如,HECT E3 UPL3多肽)的约10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64、65、66、67、68、69、70、71、72、73、74、75、76、77、78、79、80、81、82、83、84、85、86、87、88、89、90、91、92、93、94、95、96、97、98、99或100个或更多个连续氨基酸残基或其中的任何范围或值(例如,SEQ ID NO:78-83的片段或部分(例如,SEQ ID NO:84-93))、基本上由其组成或由其组成。
在一些实施方案中,这样的缺失可以是无效等位基因、显性失活等位基因、半显性等位基因、超形态突变、弱功能丧失等位基因或亚形态突变,当其包含在植物中时可以导致与不包含所述缺失的植物相比,植物表现出一种或多种改善的产量性状(例如增加的种子大小(例如,种子面积和/或种子重量)和/或增加的种子油含量)。HECT E3 UPL基因(例如,HECT E3 UPL3基因)可以在一个或多于一个位置处进行编辑(并且使用一种或多种不同的编辑工具),从而提供包含一个或多于一个突变的HECT E3 UPL基因。在一些实施方案中,如本文所述突变的HECT E3 UPL多肽可包含一种或多于一种编辑,其可导致具有一个或多于一个氨基酸取代的肽。
在一些实施方案中,涉及核酸的“部分”意指来自基因的至少2、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64、65、66、67、68、69、70、71、72、73、74、75、76、77、78、79、80、81、82、83、84、85、86、87、88、89、90、91、92、93、94、95、96、97、98、99、100、110、120、130、140、150、160、170、180、190、200、210、220、230、240、250、260、270、280、285、290、300、310、320、330、340、350、360、370、380、390、395、396、397、398、399、400、410、411、412、413、414、415、420、425、430、435、440、445、450或500个或更多个连续核苷酸(例如,来自HECT E3 UPL基因的连续核苷酸)。
在一些实施方案中,HECT E3 UPL多肽序列(例如,HECT E3UPL3多肽)的一部分的长度可以是约20至约100或更多个连续氨基酸残基(例如,长度为约20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64、65、66、67、68、69、70、71、72、73、74、75、76、77、78、79、80、81、82、83、84、85、86、87、88、89、90、91、92、93、94、95、96、97、98、99或100或多个连续氨基酸残基(例如,长度为22、51、58、68、71、74或84个连续氨基酸残基)(例如,SEQ ID NO:78-83中任一个的一部分(参见,例如,SEQ ID NO:84-93)。
如本文关于核酸使用的,术语“功能片段”是指编码多肽的功能片段的核酸。
如本文所用,术语“基因”是指能够用于产生mRNA、反义RNA、miRNA、抗microRNA反义寡脱氧核苷酸(AMO)等的核酸分子。基因可能或可能不能够用于产生功能性蛋白或基因产物。基因可以包括编码区和非编码区(例如,内含子、调节元件、启动子、增强子、终止序列和/或5’和3’非翻译区)。基因可以是“分离的”,其是指大体上或基本上不含通常在其自然状态下与核酸相关联的组分的核酸。这些组分包括其他细胞材料、来自重组产生的培养基和/或用于化学合成核酸的各种化学品。
术语“突变”是指点突变(例如,错义或无义,或导致移码的单个碱基对的插入或缺失)、插入、缺失、倒位和/或截短。当突变是氨基酸序列中的一个残基被另一个残基取代,或者是序列中一个或多个残基的缺失或插入时,突变通常通过识别原始残基,然后是该残基在序列中的位置以及新取代的残基的身份来描述。截短可包括在多肽的C端或在多肽的N端的截短。多肽的截短可以是编码该多肽的基因的相应5'末端或3'末端的缺失的结果。当一个或多个碱基对的缺失或插入被引入到基因中时,可能发生移码突变,任选地导致框外突变或框内突变。基因中的移码突变可导致产生与野生型多肽相比更长、更短或相同长度的多肽,这取决于第一个终止密码子何时出现在基因的突变区域之后。作为实例,产生提前终止密码子的框外突变可以产生比野生型多肽更短的多肽,或者在一些实施方案中,该多肽可以不存在/不可检测。在一些实施方案中,突变可以是DNA倒位,任选地具有约10至约2000个连续碱基对的长度的DNA倒位。
如本文所用,术语“互补”或“互补性”是指多核苷酸在允许的盐和温度条件下通过碱基配对的自然结合。例如,序列“A-G-T”(5'至3')与互补序列“T-C-A”(3'至5')结合。两个单链分子之间的互补性可能是“部分的”,其中只有一些核苷酸结合,或者当单链分子之间存在完全互补时,互补可能是完全的。核酸链之间的互补程度对核酸链之间杂交的效率和强度有显著影响。
如本文所用,“互补”可以是指与对照核苷酸序列的100%互补性,或其可以是指与比较核苷酸序列的小于100%的互补性(例如,约70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%等的互补性)。
具有同源性的不同核酸或蛋白质在本文中称为“同源物”。术语同源物包括来自相同和其他物种的同源序列以及来自相同和其他物种的直系同源序列。“同源性”是指两个或更多个核酸和/或氨基酸序列在位置同一性百分比(即序列相似性或同一性)方面的相似性水平。同源性也是指不同核酸或蛋白质之间的相似功能特性的概念。因此,本发明的组合物和方法还包括与本发明的核苷酸序列和多肽序列的同源物。如本文所用,“直系同源物”是指在物种形成过程中从共同祖先基因产生的不同物种中的同源核苷酸序列和/或氨基酸序列。本发明的核苷酸序列的同源物与本发明的所述核苷酸序列具有实质性的序列同一性(例如,至少约70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%、99.5%或100%)。
如本文所用,“序列同一性”是指两个最佳比对的多核苷酸或多肽序列在组分(例如核苷酸或氨基酸)的比对的整个窗口中保持不变的程度。“同一性”可通过已知方法容易计算,包括但不限于以下中描述的那些:Computational Molecular Biology(Lesk,A.M.,ed.)Oxford University Press,New York(1988);Biocomputing:Informatics andGenome Projects(Smith,D.W.,ed.)Academic Press,New York(1993);ComputerAnalysis of Sequence Data,Part I(Griffin,A.M.和Griffin,H.G.,eds.)HumanaPress,New Jersey(1994);Sequence Analysis in Molecular Biology(von Heinje,G.,ed.)Academic Press(1987);和Sequence Analysis Primer(Gribskov,M.和Devereux,J.,eds.)Stockton Press,New York(1991)。
如本文所用,术语“序列同一性百分比”或“同一性百分比”是指当两个序列最佳比对时,与测试(“对象”)多核苷酸分子(或其互补链)相比,参考(“查询”)多核苷酸分子(或其互补链)的线性多核苷酸序列中相同核苷酸的百分比。在一些实施方案中,“序列同一性百分比”可以指与参考多肽相比氨基酸序列中相同氨基酸的百分比。
如本文所用,在两个核酸分子、核苷酸序列或多肽序列的上下文中,短语“基本上相同”或“实质同一性”是指当针对最大对应性进行比较和比对时,具有至少约70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%、99.5%或100%的核苷酸或氨基酸残基同一性的两个或更多个序列或子序列,如使用以下序列比较算法之一或通过目视检查测量的。在本发明的一些实施方案中,实质上的同一性存在于本发明的核苷酸序列的连续核苷酸区域上,该区域的长度为约10个核苷酸至约20个核苷酸、约10个核苷酸至约25个核苷酸、约10个核苷酸至约30个核苷酸、约15个核苷酸至约25个核苷酸、约30个核苷酸至约40个核苷酸、约50个核苷酸至约60个核苷酸、约70个核苷酸至约80个核苷酸、约90个核苷酸至约100个核苷酸、约100个核苷酸至约200个核苷酸、约100个核苷酸至约300个核苷酸、约100个核苷酸至约400个核苷酸、约100个核苷酸至约500个核苷酸、约100个核苷酸至约600个核苷酸、约100个核苷酸至约800个核苷酸、约100个核苷酸至约900个核苷酸或更多个核苷酸,或其中的任何范围,直至序列的全长。在一些实施方案中,核苷酸序列可以在至少约20个核苷酸(例如,约20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、50、60、70或80个核苷酸或更多个)上基本相同。
在本发明的一些实施方案中,实质同一性存在于本发明的多肽的连续氨基酸残基的区域上,所述区域的长度为约3个氨基酸残基至约20个氨基酸残基、约5个氨基酸残基至约25个氨基酸残基、约7个氨基酸残基至约30个氨基酸残基、约10个氨基酸残基至约25个氨基酸残基、约15个氨基酸残基至约30个氨基酸残基、约20个氨基酸残基至约40个氨基酸残基、约25个氨基酸残基至约40个氨基酸残基、约25个氨基酸残基至约50个氨基酸残基、约30个氨基酸残基至约50个氨基酸残基、约40个氨基酸残基至约50个氨基酸残基、约40个氨基酸残基至约70个氨基酸残基、约50个氨基酸残基至约70个氨基酸残基、约60个氨基酸残基至约80个氨基酸残基、约70个氨基酸残基至约80个氨基酸残基、约90个氨基酸残基至约100个氨基酸残基或更多个氨基酸残基,以及其中的任何范围,直至序列的全长。在一些实施方案中,多肽序列可以在至少约8个连续氨基酸残基(例如,长度为约8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64、65、66、67、68、69、70、71、72、73、74、75、76、77、78、79、80、81、82、83、84、85、86、87、88、89、90、91、92、93、94、95、96、97、98、99、100、101、102、103、104、105、106、107、108、109、110、111、112、113、114、115、116、117、118、119、120、130、140、150、175、200、225、250、300、350个或更多个氨基酸或更多个连续氨基酸残基)上彼此基本上相同。在一些实施方案中,两个或更多个HECT E3 UPL多肽可以是相同的或基本上相同的(例如,至少70%至99.9%相同;例如,约70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%、99.5%、99.9%相同或其中的任何范围或值)。
对于序列比较,通常一个序列充当与测试序列进行比较的参考序列。当使用序列比较算法时,将测试序列和参考序列输入计算机,必要时指定子序列坐标,并指定序列算法程序参数。然后,序列比较算法基于指定的程序参数计算测试序列相对于参考序列的序列同一性百分比。
用于对比较窗口进行比对的序列的最佳比对是本领域技术人员熟知的,并且可以通过诸如Smith和Waterman的局部同源算法、Needleman和Wunsch的同源比对算法、Pearson和Lipman的相似性方法的搜索的工具以及可选地通过这些算法的计算机化实现诸如GAP、BESTFIT、FASTA和TFASTA(作为Wisconsin/>(Accelrys Inc.,SanDiego,CA)的一部分提供)来进行。测试序列和参考序列的比对区段的“同一性分数”是指两个比对序列共享的相同组分的数量除以参考序列区段(例如,整个参考序列或参考序列的较小定义部分)中组分的总数。序列同一性百分比表示为同一性分数乘以100。一个或多个多核苷酸序列的比较可以是与全长多核苷酸序列或其一部分,或与更长的多核苷酸序列进行比较。出于本发明的目的,还可以使用BLASTX 2.0版(用于翻译的核苷酸序列)和BLASTN2.0版(用于多核苷酸序列)来确定“同一性百分比”。
当两个序列在严格条件下相互杂交时,也可认为两个核苷酸序列实质上互补。在一些实施方案中,被认为基本上互补的两个核苷酸序列在高度严格的条件下彼此杂交。
“严格杂交条件”和“严格杂交洗涤条件”在核酸杂交实验(如Southern和Northern杂交)的背景下是序列依赖性的,并且在不同的环境参数下是不同的。核酸杂交的广泛指南见Tijssen Laboratory Techniques in Biochemistry and Molecular Biology-Hybridization with Nucleic Acid Probes part I chapter 2"Overview ofprinciples of hybridization and the strategy of nucleic acid probe assays"Elsevier,New York(1993)。通常,在规定的离子强度和pH值下,选择高度严格的杂交和洗涤条件以比特定序列的热熔点(Tm)低约5℃。
Tm是50%靶序列与完全匹配的探针杂交的温度(在规定的离子强度和pH值下)。选择非常严格的条件以等于特定探针的Tm。在Southern或northern印迹中,在过滤器上具有超过100个互补残基的互补核苷酸序列的杂交的严格杂交条件的示例是在42℃下的50%甲酰胺与1mg肝素,杂交过夜进行。高度严格洗涤条件的示例是0.1 5M NaCl,72℃下约15分钟。严格洗涤条件的一个示例是在65℃下0.2×SSC洗涤15分钟(SSC缓冲液的描述见下文的Sambrook)。通常,在高严格洗涤之前先进行低严格洗涤,以去除背景探针信号。例如,超过100个核苷酸的双链体的中等严格洗涤的示例是在45℃下1xSSC持续15分钟。例如,超过100个核苷酸的双链体的低严格性洗涤的示例是在40℃下4-6x SSC持续15分钟。对于短探针(例如,约10至50个核苷酸),严格条件通常涉及在pH 7.0至8.3下小于约1.0M Na离子的盐浓度,通常约0.01至1.0M Na离子浓度(或其他盐),并且温度通常为至少约30℃。加入去稳定剂(例如甲酰胺)也可达到严格条件。一般来说,是在特定杂交测定中对无关探针观察到的信噪比的2x(或更高)的信噪比表明检测到特异性杂交。如果它们编码的蛋白质基本相同,则在严格条件下不相互杂交的核苷酸序列仍然基本相同。例如,当使用遗传密码子允许的最大密码子简并性创建核苷酸序列的拷贝时,可能发生这种情况。
本发明的多核苷酸和/或重组核酸构建体(例如,表达盒和/或载体)可以被密码子优化以用于表达。在一些实施方案中,本发明的编辑系统(例如,包含/编码序列特异性核酸结合结构域(例如,来自多核苷酸引导的核酸内切酶、锌指核酸酶、转录激活因子样效应核酸酶(TALEN)、Argonaute蛋白和/或CRISPR-Cas核酸内切酶(例如,CRISPR-Cas效应蛋白)(例如,I型CRISPR-Cas效应蛋白、II型CRISPR-Cas效应蛋白、III型CRISPR-Cas效应蛋白、IV型CRISPR-Cas效应蛋白、V型CRISPR-Cas效应蛋白或VI型CRISPR-Cas效应蛋白)的序列特异性核酸结合结构域(例如DNA结合结构域))、核酸酶(例如,核酸内切酶(例如,Fok1)、多核苷酸引导的核酸内切酶、CRISPR-Cas核酸内切酶(例如,CRISPR-Cas效应蛋白)、锌指核酸酶和/或转录激活因子样效应核酸酶(TALEN))、脱氨酶蛋白质/结构域(例如,腺嘌呤脱氨酶、胞嘧啶脱氨酶)、编码逆转录酶蛋白质或结构域的多核苷酸、编码5'-3'核酸外切酶多肽的多核苷酸和/或亲和多肽、肽标签等)的多核苷酸、核酸构建体、表达盒和/或载体可以经密码子优化以用于在植物中表达。在一些实施方案中,本发明的密码子优化的核酸、多核苷酸、表达盒和/或载体与没有经过密码子优化的参考核酸、多核苷酸、表达盒和/或载体具有约70%至约99.9%(例如,70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%、99.5%、99.9%或100%)的同一性或更高。
在本文描述的任何实施方案中,本发明的多核苷酸或核酸构建体可以与多种启动子和/或其他调节元件可操作地关联以在植物和/或植物的细胞中表达。因此,在一些实施方案中,本发明的多核苷酸或核酸构建体可以进一步包含与一种或多种核苷酸序列可操作地连接的一种或多种启动子、内含子、增强子和/或终止子。在一些实施方案中,启动子可以与内含子(例如,Ubi1启动子和内含子)可操作地关联。在一些实施方案中,与内含子关联的启动子可以称为“启动子区”(例如,Ubi1启动子和内含子)。
如本文所用,关于多核苷酸的“可操作地连接”或“可操作地关联”是指所示元件在功能上彼此相关,并且通常也是物理相关的。因此,如本文所用,术语“可操作地连接”或“可操作地关联”是指单个核酸分子上功能关联的核苷酸序列。因此,与第二核苷酸序列可操作地连接至的第一核苷酸序列是指当第一核苷酸序列与第二核苷酸序列处于功能关系时的情况。例如,如果启动子影响所述核苷酸序列的转录或表达,则启动子与核苷酸序列可操作地关联。本领域技术人员将理解,控制序列(例如,启动子)不需要与其可操作地连接至的核苷酸序列连续,只要控制序列的功能是指导其表达即可。因此,例如,插入的非翻译但转录的核酸序列可以存在于启动子和核苷酸序列之间,并且启动子仍然可以被认为“可操作地连接”至核苷酸序列。
如本文所用,关于多肽的术语“连接”是指一种多肽与另一种多肽的连接。多肽可以直接(例如,通过肽键)或通过接头与另一多肽(在N-末端或C-末端)连接。
术语“接头”是本领域公认的并且是指化学基团,或连接两个分子或部分(例如融合蛋白的两个结构域)的分子,例如核酸结合多肽或结构域和肽标签,和/或与肽标签结合的逆转录酶和亲和多肽;或DNA核酸内切酶多肽或结构域和肽标签和/或与肽标签结合的逆转录酶和亲和多肽。接头可以由单个连接分子组成或可以包含多于一个连接分子。在一些实施方案中,接头可以是有机分子、基团、聚合物或化学部分,例如二价有机部分。在一些实施方案中,接头可以是氨基酸或者它可以是肽。在一些实施方案中,接头是肽。
在一些实施方案中,可用于本发明的肽接头长度可为约2至约100个或更多个氨基酸,例如约2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64、65、66、67、68、69、70、71、72、73、74、75、76、77、78、79、80、81、82、83、84、85、86、87、88、89、90、91、92、93、94、95、96、97、98、99、100个或更多个氨基酸(例如,约2至约40、约2至约50、约2至约60、约4至约40、约4至约50、约4至约60、约5至约40、约5至约50、约5至约60、约9至约40、约9至约50、约9至约60、约10至约40、约10至约50、约10至约60、或约2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25个氨基酸至约26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64、65、66、67、68、69、70、71、72、73、74、75、76、77、78、79、80、81、82、83、84、85、86、87、88、89、90、91、92、93、94、95、96、97、98、99、100或更多个氨基酸的长度(例如,约105、110、115、120、130、140、150或更多个氨基酸的长度)。在一些实施方案中,肽接头可以是GS接头。
如本文所用,关于多核苷酸的术语“连接的”或“融合的”是指一个多核苷酸与另一个多核苷酸的连接。在一些实施方案中,两个或更多个多核苷酸分子可以通过接头连接,接头可以是有机分子、基团、聚合物或化学部分,例如二价有机部分。多核苷酸可以通过共价或非共价键合或结合,包括例如Watson-Crick碱基配对,或通过一个或多个连接核苷酸连接或融合到另一个多核苷酸(在5'末端或3'末端)。在一些实施方案中,可以将某种结构的多核苷酸基序插入另一个多核苷酸序列中(例如,引导RNA中发夹结构的延伸)。在一些实施方案中,连接核苷酸可以是天然存在的核苷酸。在一些实施方案中,连接核苷酸可以是非天然存在的核苷酸。
“启动子”是控制或调节与启动子可操作地关联的核苷酸序列(例如,编码序列)的转录的核苷酸序列。由启动子控制或调节的编码序列可以编码多肽和/或功能性RNA。通常,“启动子”是指包含RNA聚合酶II结合位点并指导转录起始的核苷酸序列。通常,启动子位于相对于相应编码序列的编码区起点的5'或上游。启动子可以包含其他作为基因表达调节因子的元件;例如,启动子区。这些包括TATA盒共有序列,并且通常是CAAT盒共有序列(Breathnach和Chambon,(1981)Annu.Rev.Biochem.50:349)。在植物中,CAAT盒可以被AGGA盒替代(Messing等人,(1983),Genetic Engineering of Plants,T.Kosuge,C.Meredith和A.Hollaender(eds.),Plenum Press,pp.211-227)。
可用于本发明的启动子可以包括例如组成型、诱导型、时间调节的、发育调节的、化学调节的、组织优选的和/或组织特异性的启动子,用于制备重组核酸分子,例如“合成核酸构建体”或“蛋白质-RNA复合物”。这些不同类型的启动子是本领域已知的。
启动子的选择可能取决于表达的时间和空间要求而不同,并且也可能基于要转化的宿主细胞而不同。许多不同生物的启动子在本领域中是众所周知的。基于本领域现有的广泛知识,可以为感兴趣的特定宿主生物选择合适的启动子。因此,例如,关于在模式生物中高度组成型表达的基因的上游的启动子了解很多,并且可以在其他系统中适当地访问和实施此类知识。
在一些实施方案中,在植物中起作用的启动子可以与本发明的构建体一起使用。可用于在植物中驱动表达的启动子的非限制性实例包括RubisCo小亚基基因1的启动子(PrbcS1)、肌动蛋白基因的启动子(Pactin)、硝酸还原酶基因的启动子(Pnr)和重复碳酸酐酶基因1的启动子(Pdca1)(参见Walker等人,Plant Cell Rep.23:727-735(2005);Li等人,Gene 403:132-142(2007);Li等人,Mol Biol.Rep.37:1143-1154(2010))。PrbcS1和Pactin是组成型启动子,Pnr和Pdca1是诱导型启动子。Pnr由硝酸盐诱导并由铵抑制(Li等人,Gene403:132-142(2007)),Pdca1由盐诱导(Li等人,Mol Biol.Rep.37:1143-1154(2010))。在一些实施方案中,可用于本发明的启动子是RNA聚合酶II(Pol II)启动子。在一些实施方案中,来自玉米的U6启动子或7SL启动子可用于本发明的构建体。在一些实施方案中,来自玉米的U6c启动子和/或7SL启动子可用于驱动引导核酸的表达。在一些实施方案中,来自大豆的U6c启动子、U6i启动子和/或7SL启动子可用于本发明的构建体。在一些实施方案中,来自大豆的U6c启动子、U6i启动子和/或7SL启动子可用于驱动引导核酸的表达。
可用于植物的组成型启动子的实例包括但不限于:cestrum病毒启动子(cmp)(美国专利号7,166,770)、水稻肌动蛋白1启动子(Wang等人(1992)Mol.Cell.Biol.12:3399-3406;以及美国专利号5,641,876)、CaMV 35S启动子(Odell等人(1985)Nature 313:810-812)、CaMV 19S启动子(Lawton等人(1987)Plant Mol.Biol.9:315-324)、nos启动子(Ebert等人(1987)Proc.Natl.Acad.Sci USA 84:5745-5749)、Adh启动子(Walker等人(1987)Proc.Natl.Acad.Sci.USA 84:6624-6629)、蔗糖合酶启动子(Yang&Russell(1990)Proc.Natl.Acad.Sci.USA 87:4144-4148)和泛素启动子。源自泛素的组成型启动子在许多细胞类型中积累。泛素启动子已经从几种植物物种(例如向日葵(Binet等人,1991.PlantScience 79:87-94)、玉米(Christensen等人,1989.Plant Molec.Biol.12:619-632)和拟南芥(Norris等人1993.Plant Molec.Biol.21:895-906))中克隆出来用于转基因植物。玉米泛素启动子(UbiP)已在转基因单子叶植物系统中开发,其序列和构建用于单子叶植物转化的载体在专利公开EP 0 342 926中公开。泛素启动子适用于本发明的核苷酸序列在转基因植物,尤其是单子叶植物中的表达。此外,McElroy等人(Mol.Gen.Genet.231:150-160(1991))描述的启动子表达盒可以容易地被修饰以表达本发明的核苷酸序列并且特别适用于单子叶植物宿主。
在一些实施方案中,组织特异性/组织优选启动子可用于在植物细胞中表达异源多核苷酸。组织特异性或优选表达模式包括但不限于绿色组织特异性或优选、根特异性或优选、茎特异性或优选、花特异性或优选或花粉特异性或优选。适合在绿色组织中表达的启动子包括许多调节参与光合作用的基因,其中许多已从单子叶植物和双子叶植物中克隆。在一个实施方案中,可用于本发明的启动子是来自磷酸烯醇羧化酶基因的玉米PEPC启动子(Hudspeth&Grula,Plant Molec.Biol.12:579-589(1989))。组织特异性启动子的非限制性实例包括与编码种子贮藏蛋白(例如β-伴大豆球蛋白、cruciferin、napin和菜豆蛋白)、玉米醇溶蛋白或油体蛋白(例如油质蛋白)或涉及脂肪酸生物合成的蛋白质(包括酰基载体蛋白、硬脂酰-ACP去饱和酶和脂肪酸去饱和酶(fad2-1))的基因以及在胚发育过程中表达的其他核酸(例如Bce4,参见例如Kridl等人(1991)Seed Sci.Res.1:209-219;以及EP专利号255378)相关的那些启动子。可用于在植物,特别是玉米中表达本发明的核苷酸序列的组织特异性或组织优选启动子包括但不限于直接在根、髓、叶或花粉中表达的那些。此类启动子公开于例如WO93/07278中,其通过引用整体并入本文。可用于本发明的组织特异性或组织优选启动子的其他非限制性实例是美国专利6,040,504中公开的棉花rubisco启动子;美国专利5,604,121中公开的水稻蔗糖合酶启动子;de Framond(FEBS290:103-106(1991);Ciba-Geigy的EP 0 452 269)描述的根特异性启动子;在美国专利5,625,136(Ciba-Geigy)中描述的茎特异性启动子,其驱动玉米trpA基因的表达;WO01/73087中公开的certrum黄卷叶病毒启动子;和花粉特异性或优选启动子,包括但不限于来自水稻的ProOsLPS10和ProOsLPS11(Nguyen等人,Plant Biotechnol.Reports 9(5):297-306(2015)),来自玉米的ZmSTK2_USP(Wang等人Genome 60(6):485-495(2017)),来自番茄的LAT52和LAT59(Twell等人Development 109(3):705-713(1990)),Zm13(美国专利号10,421,972),来自拟南芥的PLA2-δ启动子(美国专利号7,141,424)和/或来自玉米的ZmC5启动子(国际PCT公开号WO1999/042587。
植物组织特异性/组织优选启动子的其他例子包括但不限于根毛特异性顺式元件(RHE)(Kim等人,The Plant Cell 18:2958-2970(2006)),根特异性启动子RCc3(Jeong等人,Plant Physiol.153:185-197(2010))和RB7(美国专利号5459252),凝集素启动子(Lindstrom等人(1990)Der.Genet.11:160-167;和Vodkin(1983)Prog.Clin.Biol.Res.138:87-98),玉米醇脱氢酶1启动子(Dennis等人(1984)NucleicAcids Res.12:3983-4000),S-腺苷-L-甲硫氨酸合成酶(SAMS)(Vander Mijnsbrugge等人(1996)Plant and Cell Physiology,37(8):1108-1115),玉米光收获复合启动子(Bansal等人(1992)Proc.Natl.Acad.Sci.USA 89:3654-3658),玉米热休克蛋白启动子(O'Dell等人(1985)EMBO J.5:451-458;和Rochester等人(1986)EMBO J.5:451-458),豌豆小亚基RuBP羧化酶启动子(Cashmore,"Nuclear genes encoding the small subunit ofribulose-l,5-bisphosphate carboxylase"pp.29-39于:Genetic Engineering ofPlants(Hollaender ed.,Plenum Press 1983;和Poulsen等人(1986)Mol.Gen.Genet.205:193-200),Ti质粒甘露碱合酶启动子(Langridge等人(1989)Proc.Natl.Acad.Sci.USA 86:3219-3223),Ti质粒胭脂碱合酶启动子(Langridge等人(1989),同上),矮牵牛查尔酮异构酶启动子(van Tunen等人(1988)EMBO J.7:1257-1263),豆甘氨酸富集蛋白1启动子(Keller等人(1989)Genes Dev.3:1639-1646),截短的CaMV 35S启动子(O'Dell等人(1985)Nature 313:810-812),马铃薯patatin启动子(Wenzler等人(1989)Plant Mol.Biol.13:347-354),根细胞启动子(Yamamoto等人(1990)Nucleic Acids Res.18:7449),玉米玉米醇溶蛋白启动子(Kriz等人(1987)Mol.Gen.Genet.207:90-98;Langridge等人(1983)Cell34:1015-1022;Reina等人(1990)Nucleic Acids Res.18:6425;Reina等人(1990)NucleicAcids Res.18:7449;和Wandelt等人(1989)Nucleic Acids Res.17:2354),球蛋白-1启动子(Belanger等人(1991)Genetics 129:863-872),α-微管蛋白cab启动子(Sullivan等人(1989)Mol.Gen.Genet.215:431-440),PEPCase启动子(Hudspeth&Grula(1989)PlantMol.Biol.12:579-589),R基因复合物相关启动子(Chandler等人(1989)Plant Cell 1:1175-1183)和查耳酮合酶启动子(Franken等人(1991)EMBO J.10:2605-2612)。
可用于种子特异性表达的是豌豆球蛋白启动子(Czako等人(1992)Mol.Gen.Genet.235:33-40;以及美国专利号5,625,136中公开的种子特异性启动子。可用于在成熟叶中的表达的启动子是在衰老开始时被转换的那些,例如来自拟南芥的SAG启动子(Gan等人(1995)Science 270:1986-1988)。
此外,可以使用在叶绿体中起作用的启动子。此类启动子的非限制性实例包括噬菌体T3基因9 5'UTR和美国专利号7,579,516中公开的其他启动子。可用于本发明的其他启动子包括但不限于S-E9小亚基RuBP羧化酶启动子和Kunitz胰蛋白酶抑制剂基因启动子(Kti3)。
可用于本发明的其他调节元件包括但不限于内含子、增强子、终止序列和/或5'和3'非翻译区。
可用于本发明的内含子可以是在植物中鉴定和分离的内含子,然后插入到表达盒中以用于植物的转化。如本领域技术人员将理解的,内含子可包含自我切除所需的序列,并以符合阅读框的方式并入核酸构建体/表达盒中。内含子可用作间隔区以分隔一个核酸构建体中的多个蛋白质编码序列,或者内含子可用于一个蛋白质编码序列内以例如稳定mRNA。如果它们在蛋白质编码序列中使用,它们将被插入“框内”并包含切除位点。内含子也可以与启动子关联以改善或修饰表达。例如,可用于本发明的启动子/内含子组合包括但不限于玉米Ubi1启动子和内含子的组合(参见例如SEQ ID NO:21和SEQ ID NO:22)。
可用于本发明的内含子的非限制性实例包括来自ADHI基因(例如,Adh1-S内含子1、2和6)、泛素基因(Ubi1)、RuBisCO小亚基(rbcS)基因、RuBisCO大亚基(rbcL)基因、肌动蛋白基因(例如,肌动蛋白-1内含子)、丙酮酸脱氢酶激酶基因(pdk)、硝酸还原酶基因(nr)、重复碳酸酐酶基因1(Tdca1)、psbA基因、atpA基因的内含子或其任意组合。
在一些实施方案中,本发明的多核苷酸和/或核酸构建体可以是“表达盒”或可以包含在表达盒内。如本文所用,“表达盒”是指重组核酸分子,其包含例如一种或多种本发明的多核苷酸(例如,编码序列特异性核酸结合结构域的多核苷酸、编码脱氨酶蛋白或结构域的多核苷酸,编码逆转录酶蛋白质或结构域的多核苷酸、编码5'-3'外切核酸酶多肽或结构域的多核苷酸、引导核酸和/或逆转录酶(RT)模板),其中一种或多种多核苷酸与一个或多个控制序列(例如,启动子、终止子等)可操作地关联。因此,在一些实施方案中,可以提供一种或多种表达盒,其被设计为表达例如本发明的核酸构建体(例如,编码序列特异性核酸结合结构域的多核苷酸、编码核酸酶多肽/结构域的多核苷酸、编码脱氨酶蛋白质/结构域的多核苷酸、编码逆转录酶蛋白质/结构域的多核苷酸、编码5'-3'外切核酸酶多肽/结构域的多核苷酸、编码肽标签的多核苷酸和/或编码亲和多肽的多核苷酸等,或包含引导核酸、延伸的引导核酸和/或RT模板等)。当本发明的表达盒包含多于一个多核苷酸时,多核苷酸可以与驱动所有多核苷酸表达的单个启动子可操作地连接,或者多核苷酸可以与一个或多个单独的启动子(例如三个多核苷酸可以由一个、两个或三个任意组合的启动子驱动)可操作地连接。当使用两个或更多个单独的启动子时,启动子可以是相同的启动子或者它们可以是不同的启动子。因此,编码序列特异性核酸结合结构域的多核苷酸、编码核酸酶蛋白/结构域的多核苷酸、编码CRISPR-Cas效应蛋白/结构域的多核苷酸、编码脱氨酶蛋白/结构域的多核苷酸、编码逆转录酶多肽/结构域(例如,RNA依赖性DNA聚合酶)的多核苷酸和/或编码5'-3'外切核酸酶多肽/结构域的多核苷酸、引导核酸、延伸的引导核酸和/或当包含在单个表达盒中时可以各自可操作地连接到单个启动子的RT模板,或任何组合的单独的启动子。
包含本发明的核酸构建体的表达盒可以是嵌合的,这意味着它的至少一种(例如一种或多种)成分相对于它的至少一种其他成分是异源的(例如,来自宿主生物的启动子可操作地连接到在宿主生物体中表达的感兴趣的多核苷酸,其中感兴趣的多核苷酸来自与宿主不同的生物体或通常不与该启动子关联)。表达盒也可以是天然存在的但已经以可用于异源表达的重组形式获得的表达盒。
表达盒可以任选地包括在所选宿主细胞中起作用的转录和/或翻译终止区(即终止区)和/或增强子区。多种转录终止子和增强子是本领域已知的并且可用于表达盒。转录终止子负责转录的终止和正确的mRNA多聚腺苷酸化。终止区和/或增强子区对于转录起始区可以是天然的,对于例如编码序列特异性核酸结合蛋白的基因、编码核酸酶的基因、编码逆转录酶的基因、编码脱氨酶的基因等可以是天然的,或者对于宿主细胞可以是天然的,或者对于另一来源(例如,对于例如启动子、对于编码序列特异性核酸结合蛋白的基因、编码核酸酶的基因、编码逆转录酶的基因、编码脱氨酶的基因等,或对于宿主细胞或其任何组合来说是外来的或异源的)可以是天然的。
本发明的表达盒还可包括编码可选择标记的多核苷酸,其可用于选择转化的宿主细胞。如本文所用,“可选择标记”是指多核苷酸序列,其在表达时赋予表达该标记的宿主细胞不同的表型,从而允许将此类转化的细胞与不具有该标记的细胞区分开来。这样的多核苷酸序列可以编码可选择的或可筛选的标记,这取决于标记是否赋予可以通过化学手段(例如通过使用选择剂(例如抗生素等))选择的性状,或者取决于是否标记只是一种可以通过观察或测试来识别的性状,例如通过筛选(例如,荧光)。合适的可选择标记的许多例子是本领域已知的并且可以用于本文所述的表达盒中。
除了表达盒之外,本文所述的核酸分子/构建体和多核苷酸序列可以与载体结合使用。术语“载体”是指用于将核酸(或多种核酸)转移、递送或引入细胞中的组合物。载体包含含有待转移、递送或引入的核苷酸序列的核酸构建体(例如表达盒)。用于宿主生物转化的载体在本领域是众所周知的。一般类型的载体的非限制性实例包括双链或单链线性或环状形式的病毒载体、质粒载体、噬菌体载体、噬菌粒载体、粘粒载体、fosmid载体、噬菌体、人工染色体、小环或农杆菌双元载体,其可以或者可能无法自我传播或移动。在一些实施方案中,病毒载体可以包括但不限于逆转录病毒、慢病毒、腺病毒、腺相关病毒或单纯疱疹病毒载体。本文定义的载体可以通过整合到细胞基因组中或存在于染色体外(例如具有复制起点的自主复制质粒)来转化原核或真核宿主。另外包括穿梭载体,其意指能够在两种不同宿主生物(其可以选自放线菌和相关物种、细菌和真核生物(例如高等植物、哺乳动物、酵母或真菌细胞))中(天然或通过设计)复制的DNA载体。在一些实施方案中,载体中的核酸合适启动子或其他调节元件的控制下并且可操作地与其连接以用于宿主细胞中转录。该载体可以是在多个宿主中起作用的双功能表达载体。在基因组DNA的情况下,这可以包含其自身的启动子和/或其他调节元件,而在cDNA的情况下,这可以在合适的启动子和/或其他调节元件的控制下以用于在宿主细胞中表达。因此,本发明的核酸或多核苷酸和/或包含其的表达盒可以包含在如本文所述和本领域已知的载体中。
如本文所用,“接触”、“接触的”、“接触了”及其语法变体是指将所需反应的组分放在适合进行所需反应(例如,转化、转录控制、基因组编辑、切口和/或切割)的条件下。例如,靶核酸可以与序列特异性核酸结合蛋白(例如,多核苷酸引导的核酸内切酶、CRISPR-Cas核酸内切酶(例如,CRISPR-Cas效应蛋白)、锌指核酸酶、转录激活因子样效应核酸酶(TALEN)和/或Argonaute蛋白))和脱氨酶或编码其的核酸构建体在表达序列特异性核酸结合蛋白、逆转录酶和/或脱氨酶并且序列特异性核酸结合蛋白与靶核酸结合的条件下接触,逆转录酶和/或脱氨酶可以与序列特异性核酸结合蛋白融合或募集到序列特异性核酸结合蛋白上(例如,通过与序列特异性核酸结合蛋白融合的肽标签和与逆转录酶和/或脱氨酶融合的亲和标签),因此脱氨酶和/或逆转录酶位于靶核酸附近,从而修饰靶核酸。可以使用利用其他蛋白质-蛋白质相互作用的募集逆转录酶和/或脱氨酶的其他方法,RNA-蛋白质相互作用和化学相互作用也可以用于蛋白质-蛋白质和蛋白质-核酸募集。
如本文所用,关于靶核酸的“修饰”或“修饰的”包括编辑(例如突变)、共价修饰、交换/取代核酸/核苷酸碱基、缺失、切割、切口和/或改变靶核酸的转录控制。在一些实施方案中,修饰可以包括任何类型的一个或多个单碱基改变(SNP)。
在感兴趣的多核苷酸的上下文中,“引入”、“引入了”、“引入的”(及其语法变体)是指将感兴趣的核苷酸序列(例如,多核苷酸、RT模板、核酸构建体和/或引导核酸)以使得核苷酸序列能够进入细胞内部的方式呈递给植物、其植物部分或其细胞。
术语“转化”或“转染”可以互换使用,并且如本文所用,是指将异源核酸引入细胞。细胞的转化可以是稳定的或瞬时的。因此,在一些实施方案中,宿主细胞或宿主生物体(例如植物)可以用本发明的多核苷酸/核酸分子稳定转化。在一些实施方案中,宿主细胞或宿主生物体可以用本发明的多核苷酸/核酸分子瞬时转化。
在多核苷酸的上下文中,“瞬时转化”是指多核苷酸被引入细胞中并且不整合到细胞的基因组中。
在将多核苷酸引入细胞中的上下文中,“稳定地引入”或“稳定地引入的”意指引入的多核苷酸被稳定地掺入细胞的基因组中,因此细胞被多核苷酸稳定地转化。
如本文所用,“稳定转化”或“稳定转化的”是指将核酸分子引入细胞中并整合到细胞的基因组中。因此,整合的核酸分子能够被其后代,更具体地,被多个连续世代的后代遗传。如本文所用,“基因组”包括核基因组和质体基因组,因此包括将核酸整合到例如叶绿体或线粒体基因组中。如本文所用,稳定转化还可以指在染色体外维持的转基因,例如,作为微型染色体或质粒。
瞬时转化可以通过例如酶联免疫吸附测定(ELISA)或蛋白质印迹来检测,其可以检测由引入生物体的一种或多种转基因编码的肽或多肽的存在。细胞的稳定转化可以通过例如细胞的基因组DNA与核酸序列的Southern印迹杂交测定来检测,所述核酸序列与引入生物体(例如植物)中的转基因的核苷酸序列特异性杂交。细胞的稳定转化可以通过例如细胞的RNA与核酸序列的Northern印迹杂交测定来检测,所述核酸序列与引入宿主生物的转基因的核苷酸序列特异性杂交。细胞的稳定转化也可以通过例如聚合酶链式反应(PCR)或本领域熟知的其他扩增反应来检测,使用与转基因的靶序列杂交的特异性引物序列,导致转基因序列的扩增,其可以根据标准方法检测。转化也可以通过本领域公知的直接测序和/或杂交方案检测。
因此,在一些实施方案中,本发明的核苷酸序列、多核苷酸、核酸构建体和/或表达盒可以瞬时表达和/或它们可以稳定地掺入宿主生物的基因组中。因此,在一些实施方案中,本发明的核酸构建体(例如,一种或多种包含用于如本文所述进行编辑的多核苷酸的表达盒)可以与引导核酸一起瞬时引入细胞中,因此,没有DNA保持在细胞中。
可以通过本领域技术人员已知的任何方法将本发明的核酸构建体引入到植物细胞中。转化方法的非限制性实例包括通过细菌介导的核酸递送(例如,通过农杆菌)、病毒介导的核酸递送、碳化硅或核酸晶须介导的核酸递送、脂质体介导的核酸递送、显微注射、微粒轰击、磷酸钙介导的转化、环糊精介导的转化、电穿孔、纳米颗粒介导的转化、超声处理、渗透、PEG介导的核酸摄取以及导致将核酸引入植物细胞的任何其他电学、化学、物理(机械)和/或生物机制的转化,包括其任何组合。用于转化真核生物和原核生物的程序在本领域中是众所周知的并且是常规的并且在整个文献中都有描述(参见,例如,Jiang等人,2013.Nat.Biotechnol.31:233-239;Ran等人,Nature Protocols 8:2281-2308(2013))。本领域已知的各种植物转化方法的一般指南包括Miki等人("Procedures for IntroducingForeign DNA into Plants"Methods in Plant Molecular Biology and Biotechnology,Glick,B.R.和Thompson,J.E.,Eds.(CRC Press,Inc.,Boca Raton,1993),第67-88页)和Rakowoczy-Trojanowska(Cell.Mol.Biol.Lett.7:849-858(2002))。
在本发明的一些实施方案中,细胞的转化可以包括核转化。在其他实施方案中,细胞的转化可以包括质体转化(例如,叶绿体转化)。在更进一步的实施方案中,可以通过常规育种技术将本发明的核酸引入细胞中。在一些实施方案中,可以通过农杆菌转化将一种或多种多核苷酸、表达盒和/或载体引入到植物细胞中。
因此,可以以本领域公知的许多方式将多核苷酸引入植物、植物部分、植物细胞中。本发明的方法不依赖于将一个或多个核苷酸序列引入植物的特定方法,仅依赖于它们能够进入细胞内部。在要引入多于多核苷酸的情况下,它们可以组装为单个核酸构建体的一部分,或作为单独的核酸构建体,并且可以位于相同或不同的核酸构建体上。因此,可以在单个转化事件中或在单独的转化事件中将多核苷酸引入感兴趣的细胞,或者,作为育种方案的一部分,可以将多核苷酸掺入植物中。
泛素连接酶是负责识别靶蛋白并促进底物降解的细胞机制的一部分。它们是导致底物蛋白泛素化并通过26s蛋白酶体降解它们的关键途径中高度多样化的关键调节因子(Chen和Hellmann,Molecular Plant,6(5):1388-1404(2013))。泛素连接酶参与多个关键过程,包括植物发育、非生物和生物胁迫(Shu和Yang,Plant Cell Physiol.58(9):1461-1476(2017)),You等人,Cell Host&Microbe 20:758-769(2016))。Miller等人(The PlantCell 31:2370-2385(2019))从一组甘蓝型油菜(Brassica napus)品种中鉴定出了BnaUPL3.C03的启动子区域中的遗传变异,其可以影响种子大小、脂质含量和最终作物产量。这些作者鉴定了一种机制,其中编码HECT E3泛素连接酶的BnaUPL3.C03的减少的表达导致LEAFY COTYLEDON 2(LEC2)(一种控制种子成熟的转录因子)的升高的水平。这延长了种子成熟过程,导致增加的种子大小(例如种子面积、种子重量)和脂质含量(例如含油量)。Miller等人进一步发现拟南芥功能丧失upl3突变体(upl3-4和upl3-5)具有比野生型大约10%的种子,并且种子脂质含量增加12%。相比之下,使用35S启动子(35S:3HA-UPL3转基因)过度表达AtUPL3降低种子大小和脂质含量。有趣的是,芜菁(Brassica rapa)(甘蓝型油菜的A基因组供体)中UPL3基因的突变体比各自的分离野生型具有更大的种子,因此表明UPL3在调节拟南芥和芸苔属物种的种子大小方面的潜在作用。
因此,如本文所述,编辑技术用于靶向植物中的内源HECT E3UPL基因(例如,HECTE3 UPL3基因;例如,HECT E3 UPL3a、HECT E3 UPL3b、HECT E3 UPL3c、HECT E3 UPL3d),以提供具有/表现出一种或更改善的产量性状(任选地增加的种子大小(例如,种子面积和/或种子重量)和/或增加的种子油含量)的植物。可用于产生具有一种或多种改善的产量性状的植物的突变包括例如取代、缺失、插入和/或倒位。在一些方面,由编辑技术产生的突变可以是点突变。在一些方面,如本文所述产生的突变可以是显性失活突变、半显性突变、无效突变、超形态突变、亚形态突变或弱功能丧失突变,任选地无效突变或显性失活突变。
在一些实施方案中,本发明提供了一种植物或其植物部分,其包含编码HECT E3UPL多肽的内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的至少一个(例如,一个或多个)非天然突变。在一些实施方案中,所述至少一个非天然突变提供显性失活突变、半显性突变、无效突变、超形态突变、亚形态突变和/或弱功能丧失突变,任选地,其中至少一个非天然突变提供无效突变或显性失活突变。
在一些实施方案中,提供了植物细胞,该植物细胞包含编辑系统,该编辑系统包含:(a)CRISPR-Cas效应蛋白;和(b)引导核酸(例如,gRNA、gDNA、crRNA、crDNA、sgRNA、sgDNA),其包含与编码HECT E3 UPL蛋白的内源靶基因互补的间隔序列。编辑系统可用于在编码HECT E3 UPL蛋白的内源靶基因中产生突变。在一些实施方案中,突变是非天然突变。在一些实施方案中,编辑系统的引导核酸可以包含SEQ ID NO:94-118(例如SEQ ID NO:94(PWsp1159)、SEQ ID NO:95(PWsp1160)、SEQ ID NO:96(PWsp1161)、SEQ ID NO:97(PWsp1162)、SEQ ID NO:98(PWsp1236)、SEQ ID NO:99(PWsp1237)、SEQ ID NO:100(PWsp1238)、SEQ ID NO:101(PWsp1239)、SEQ ID NO:102(PWsp1384)、SEQ ID NO:103(PWsp1385)、SEQ ID NO:104(PWsp1386)、SEQ ID NO:105(PWsp1387)、SEQ ID NO:106(PWsp1388)、SEQ ID NO:107(PWsp1389)、SEQ ID NO:108(PWsp1390)、SEQ ID NO:109(PWsp1391)、SEQ ID NO:110(PWsp1392)、SEQ ID NO:111(PWsp1393)、SEQ ID NO:112(PWsp1394)、SEQ ID NO:113(PWsp1397)、SEQ ID NO:114(PWsp1398)、SEQ ID NO:115(PWsp1399)、SEQ ID NO:116(PWsp1340)、SEQ ID NO:117(PWsp1401)和/或SEQ ID NO:118(PWsp1402))中任一个的核苷酸序列(间隔序列,例如,一个或多个间隔区)。
植物、其植物部分或植物细胞的HECT E3 UPL基因中的突变可以是任何类型的突变,包括碱基取代、碱基缺失、碱基插入和/或倒位。在一些实施方案中,非天然突变可包含对A、T、G或C的碱基取代,任选地,碱基取代可以是从G至C。在一些实施方案中,非天然突变可以是至少一个碱基对(例如,1个碱基对至约5个碱基对;例如,1、2、3、4、5个连续碱基对)的缺失或至少一个碱基对(例如,1个碱基对至约5个碱基对;例如,1、2、3、4、5个连续碱基对)的插入,任选地框外或框内缺失或插入。在一些实施方案中,非天然突变可以是至少5个碱基对至约100个碱基对或更多(例如,5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64、65、66、67、68、69、70、71、72、73、74、75、76、77、78、79、80、81、82、83、84、85、86、87、88、89、90、91、92、93、94、95、96、97、98、99、100或更多个连续碱基对)的缺失,任选地框外或框内缺失。在一些实施方案中,非天然突变可以是HECT E3 UPL基因的约10至约2000个连续碱基对的倒位。
在一些实施方案中,HECT E3 UPL基因的突变位于编码HECT E3 UPL多肽的内源HECT E3 UPL基因的一部分内(例如,编码区(外显子))。作为实例,突变可以在编码HECT E3UPL多肽的活性/催化位点的HECT E3 UPL基因的区域中,任选地,其中突变导致HECT E3UPL多肽与野生型成熟HECT E3 UPL肽相比具有氨基酸取代(例如,Cys>Ser)。在一些实施方案中,催化位点变体可以通过产生框内缺失或框内插入来获得。在一些实施方案中,HECT E3UPL基因中的突变是导致例如提前终止密码子的框外缺失或框外插入。在一些实施方案中,内源HECT E3 UPL3基因中的至少一个非天然突变可以导致所编码的HECT E3 UPL3多肽的N端区域中的突变。在一些实施方案中,编码HECT E3 UPL多肽的HECT E3 UPL基因区域中的突变可以是至少10至约2000个连续核苷酸的倒位(例如,来自外显子或其部分的连续核苷酸的倒位),任选地,至少10至约15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64、65、66、67、68、69、70、71、72、73、74、75、76、77、78、79、80、81、82、83、84、85、86、87、88、89、90、91、92、93、94、95、96、97、98、99、100、110、120、130、140、150、160、170、180、190或200个或更多个连续碱基对的倒位。
可用于在HECT E3 UPL基因中产生这些和其他突变的编辑工具的类型包括任何碱基编辑器或切割器,其使用与如本文所述的HECT E3 UPL基因的一部分具有至少80%互补性的间隔区被引导至靶位点。
在一些实施方案中,HECT E3 UPL基因(例如,HECT E3 UPL3基因)的突变位于编码HECT E3 UPL多肽(例如,HECT E3 UPL3多肽)的N端区域的内源HECT E3 UPL基因的一部分内,其中HECT E3UPL多肽的N端区域包含例如(a)与SEQ ID NO:84-87中的任一个具有至少80%序列同一性的序列,和/或(b)与以下的连续氨基酸残基区域具有至少80%序列同一性的氨基酸序列:参考SEQ ID NO:78的残基编号,从残基50到残基95、从残基43到残基102和/或从残基36到残基109,参考SEQ ID NO:79的残基编号,从残基48到残基87、从残基41到残基94和/或从残基34到残基101,参考SEQ ID NO:80的残基编号,从残基139到残基194、从残基132到残基201和/或从残基125到残基208,和/或参考SEQ ID NO:81的残基编号,从残基104到残基146、从残基97到残基153和/或从残基90到残基160。
在一些实施方案中,突变可以位于由内源HECT E3 UPL基因(例如,HECT E3 UPL3基因)编码的HECT E3 UPL多肽(例如,HECT E3 UPL3多肽)的活性/催化位点中,任选地,其中活性位点位于HECT E3 UPL多肽的区域中,该区域包含:(a)与SEQ ID NO:88-93中的任一个具有至少80%同一性的氨基酸序列和/或(b)与以下的连续氨基酸残基区域具有至少80%序列同一性的氨基酸序列:参考SEQ ID NO:78的残基编号,从残基1814到残基1865、从残基1822到残基1857和/或从残基1829到残基1851,参考SEQ ID NO:79的残基编号,从残基1820到残基1870、从残基1827到残基1863和/或从残基1834到残基1856,参考SEQ ID NO:80的残基编号,从残基1838到残基1888、从残基1845到残基1881和/或从残基1852到残基1874,参考SEQ ID NO:81的残基编号,从残基1834到残基1884、从残基1841到残基1877和/或从残基1848到残基1870,参考SEQ ID NO:82的残基编号,从残基1783到残基1833、从残基1790到残基1826和/或从残基1797到残基1818,或参考SEQ ID NO:83的残基编号、从残基1776到残基1826、从残基1783到残基1819和/或从残基1790到残基1812。
在一些实施方案中,HECT E3 UPL基因(例如,HECT E3 UPL3基因)的活性/催化位点中的突变可以导致取代的氨基酸残基,任选地其中氨基酸残基的取代位于:参考SEQ IDNO:78的氨基酸位置编号的位置1839,参考SEQ ID NO:79的氨基酸位置编号的位置1844,参考SEQ ID NO:80的氨基酸位置编号的位置1862,参考SEQ ID NO:81的氨基酸位置编号的位置1858,参考SEQ ID NO:82的氨基酸位置编号的位置1807,或者参考SEQ ID NO:83的氨基酸位置编号的位置1800。在一些实施方案中,取代可以是半胱氨酸至丝氨酸。
可用于本发明的内源HECT E3 UPL基因(例如,内源靶基因)编码E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)蛋白,任选地HECT E3 UPL3蛋白(例如,HECT E3 UPL3a、HECT E3 UPL3b、HECT E3UPL3c、HECT E3 UPL3d)。在一些实施方案中,内源HECT E3 UPL基因(例如,靶基因;例如,HECT E3 UPL3基因)可包含与SEQ ID NO:72-77的核苷酸序列中的任一个具有至少80%序列同一性的序列,和/或可以编码这样的多肽:(a)包含与SEQ IDNO:78-83的氨基酸序列中的任一个具有至少80%序列同一性的序列,(b)包含与SEQ IDNO:84-92的氨基酸序列中的任一个具有至少80%序列同一性的区域,和/或(c)包含与SEQID NO:93具有至少90%同一性的区域。
HECT E3 UPL基因中的示例性非天然突变可以提供突变的HECT E3 UPL基因(例如,突变的HECT E3 UPL3基因)和/或可以编码突变的HECT E3 UPL蛋白(例如,突变的HECTE3 UPL3蛋白)。
在一些实施方案中,在内源HECT E3 UPL(例如,HECT E3 UPL3)基因中包含至少一个(例如,一个或多个)非天然突变的植物(例如,玉米植物、大豆植物)与缺乏所述至少一个突变的植物(例如,同基因植物(例如,野生型未经编辑的植物或无效分离子)相比表现出一种或多种)改善的产量性状。在一些实施方案中,一种或多种改善的产量性状可以包括,但不限于,更高的产量(蒲式耳/英亩)、增加的籽粒行数,任选地其中穗长度基本上没有减少、增加的籽粒数、增加的籽粒大小、增加的穗长度、减少的分蘖数、减少的雄穗分枝数、每个节增加的豆荚数、每株植物增加的豆荚数、增加的种子重量和/或增加的种子油含量。在一些实施方案中,包含内源HECT E3 UPL基因中的至少一个突变的植物与缺乏相同HECT E3 UPL突变的植物相比,表现出一种或多种改善的产量性状的表型,任选地增加的种子大小(例如面积、重量)和/或增加的种子油含量。在一些实施方案中,植物可以从包含如本文所述的HECT E3 UPL基因中的突变的本发明的植物部分和/或植物细胞再生,其中再生的植物包含内源HECT E3 UPL基因中的突变和与缺乏HECT E3 UPL基因中的相同突变的植物相比改善的一种或多种产量性状的表型。
如本文所用,术语“穗长度基本上没有减少”是指由于如本文所述的一个或多个HECT E3 UPL基因中的一个或多个突变而具有增加的籽粒行数的穗的长度,其中与缺乏相同HECT E3 UPL基因中的相同突变的植物的穗相比穗长度减少不超过约5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24,25%、26%、27%、28%、29%或30%。
在一些实施方案中,提供了植物细胞,该植物细胞包含内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因内的至少一个(例如,一个或多个)非天然突变,其中所述突变是使用编辑系统引入的取代、插入、缺失或倒位,所述编辑系统包含结合内源HECT E3 UPL基因中的靶位点的核酸结合结构域,任选地其中内源HECT E3 UPL基因是内源HECT E3UPL3基因(例如,HECT E3 UPL3a基因、HECT E3 UPL3b基因、HECT E3 UPL3c基因、HECT E3UPL3d基因)。在一些实施方案中,取代、插入、缺失或倒位导致例如显性失活等位基因或无效等位基因。在一些实施方案中,至少一个非天然突变是点突变。在一些实施方案中,HECTE3 UPL基因内的至少一个非天然突变是插入和/或缺失,任选地,至少一个非天然突变是框外插入或缺失或框内插入或缺失,任选地其中框外插入或缺失导致提前终止密码子。在一些实施方案中,HECT E3 UPL基因中的靶位点在HECT E3 UPL基因的区域内,该区域(a)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的序列,(b)编码与以下的连续氨基酸残基的区域具有至少80%序列同一性的序列:参考SEQ ID NO:78的残基编号,从残基50到残基95、从残基43到残基102、从残基36到残基109、从残基1814到残基1865、从残基1822到残基1857和/或从残基1829到残基1851,参考SEQ ID NO:79的残基编号,从残基48到残基87、从残基41到残基94、从残基34到残基101、从残基1820到残基1870、从残基1827到残基1863和/或从残基1834到残基1856,参考SEQ ID NO:80的残基编号,从残基139到残基194、从残基132到残基201、从残基125到残基208、从残基1838到残基1888、从残基1845到残基1881和/或从残基1852到残基1874,参考SEQ ID NO:81的残基编号,从残基104到残基146、从残基97到残基153、从残基90到残基160、从残基1834到残基1884、从残基1841到残基1877和/或从残基1848到残基1870,参考SEQ ID NO:82的残基编号,从残基1783到残基1833、从残基1790到残基1826和/或从残基1797到残基1818,和/或参考SEQ ID NO:83的残基编号,从残基1776到残基1826、从残基1783到残基1819和/或从残基1790到残基1812;和/或(c)编码与SEQ ID NO:93的氨基酸序列具有至少90%序列同一性的序列。在一些实施方案中,在通过编辑系统切割后进行突变,所述编辑系统包含核酸酶和核酸结合结构域,所述核酸结合结构域结合与编码SEQ ID NO:84-92中任一个的序列具有至少80%序列同一性或与编码SEQ ID NO:93的序列具有至少90%的序列同一性的序列内的靶位点,并且HECTE3 UPL基因内的至少一个非天然突变是在核酸酶的切割后产生的。在一些实施方案中,至少一个非天然突变是位于以下位置处的经修饰的氨基酸残基:参考SEQ ID NO:78的氨基酸位置编号的位置1839,参考SEQ ID NO:79的氨基酸位置编号的位置1844,参考SEQ ID NO:80的氨基酸位置编号的位置1862,参考SEQ ID NO:81的氨基酸位置编号的位置1858,参考SEQ ID NO:82的氨基酸位置编号的位置1807,或者参考SEQ ID NO:83的氨基酸位置编号的位置1800。在一些实施方案中,HECT E3 UPL基因是HECT E3 UPL3基因,任选地HECT E3UPL3a基因、HECT E3 UPL3b基因、HECT E3 UPL3c基因、HECT E3 UPL3d基因。在一些实施方案中,植物细胞再生为包含至少一个非天然突变的植物,并且该突变导致显性失活等位基因或无效等位基因。在一些实施方案中,从植物细胞再生的植物当与不包含/缺乏等位基因的野生型植物(例如,同基因野生型植物)相比时,表现出至少一种(一种或多种)改善的产量性状的表型,任选地其中所述一种或多种改善的产量性状包括与缺乏所述至少一个非天然突变的对照植物相比更高的产量(蒲式耳/英亩)、增加的籽粒行数,任选地其中穗长度基本上没有减少、增加的籽粒数量、增加的籽粒大小(例如,增加的面积和/或增加的重量)、增加的穗长度、减少的分蘖数、减少的雄穗分枝数、每个节增加的豆荚数、每株植物增加的豆荚数、增加的种子重量、增加的种子面积和/或增加的种子油含量。在一些实施方案中,从植物细胞再生的植物表现出具有增加的种子重量/大小和/或增加的种子油含量的种子表型。
在一些实施方案中,提供了产生/培育无转基因的经编辑的植物(例如,玉米植物、大豆植物)的方法,该方法包括:使本发明的植物(例如,包含一个或多个HECT E3 UPL基因中的一个或多个突变(例如,非天然突变)并且具有增加的籽粒行数的植物)与无转基因的植物杂交,从而将突变引入到无转基因的植物中;和选择包含突变且无转基因的后代植物,从而产生无转基因的经编辑的植物。
本文还提供了提供具有一种或多种改善的产量性状的多个植物(例如,玉米植物、大豆植物)的方法,该方法包括在生长区域(例如,田地(例如,耕地、农田)、生长室、温室、休闲区、草坪和/或路边等)中种植本发明的两个或更多个植物(例如,2、3、4、5、6、7、8、9、10或更多个植物,其包含一个或多个HECT E3 UPL基因、任选地一个或多个HECT E3 UPL多肽中的一个或多个突变(例如,非天然突变)并且具有一种或多种改善的产量性状),从而提供与没有突变的多个对照植物相比具有一种或多种改善的产量性状的多个植物。
本发明进一步提供了在E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)多肽的区域中产生变异的方法,包括:将编辑系统引入到植物细胞中,其中该编辑系统靶向编码HECTE3 UPL多肽的区域的HECT E3 UPL基因的区域,其中HECT E3 UPL多肽的区域包含(a)与SEQID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的序列,(b)与以下的连续氨基酸残基的区域具有至少80%序列同一性的序列:参考SEQ ID NO:78的残基编号,从残基50到残基95、从残基43到残基102、从残基36到残基109、从残基1814到残基1865、从残基1822到残基1857和/或从残基1829到残基1851,参考SEQ ID NO:79的残基编号,从残基48到残基87、从残基41到残基94、从残基34到残基101、从残基1820到残基1870、从残基1827到残基1863和/或从残基1834到残基1856,参考SEQ ID NO:80的残基编号,从残基139到残基194、从残基132到残基201、从残基125到残基208、从残基1838到残基1888、从残基1845到残基1881和/或从残基1852到残基1874,参考SEQ ID NO:81的残基编号,从残基104到残基146、从残基97到残基153、从残基90到残基160、从残基1834到残基1884、从残基1841到残基1877和/或从残基1848到残基1870,参考SEQ ID NO:82的残基编号,从残基1783到残基1833、从残基1790到残基1826和/或从残基1797到残基1818,和/或参考SEQ ID NO:83的残基编号,从残基1776到残基1826、从残基1783到残基1819和/或从残基1790到残基1812;或(c)包含与SEQ ID NO:93的氨基酸序列具有至少90%序列同一性的序列;将HECT E3 UPL基因的区域与所述编辑系统接触,从而将突变引入到植物细胞中的HECT E3 UPL多肽的区域;和在HECT E3 UPL多肽的区域中产生变异。在一些实施方案中,可以在通过编辑系统切割后进行突变,所述编辑系统包含核酸酶和核酸结合结构域,所述核酸结合结构域结合与SEQID NO:72-77的核苷酸序列中的任一个具有至少80%序列同一性的序列内的靶位点。在一些实施方案中,使植物细胞中的HECT E3 UPL基因的区域与所述编辑系统接触产生在其基因组中包含编辑的内源HECT E3 UPL基因的植物细胞,该方法还包括(a)从该植物细胞再生植物;(b)使植物自交以产生后代植物(E1);(c)针对一种或多种改善的产量性状测定(b)的后代植物;和(d)选择表现出一种或多种改善的产量性状的后代植物,以产生与对照植物相比表现出一种或多种改善的产量性状的所选择的后代植物。在一些实施方案中,该方法可以进一步包括(e)使(d)的选择的后代植物自交以产生后代植物(E2);(f)针对一种或多种改善的产量性状测定(e)的后代植物;和(g)选择表现出一种或多种改善的产量性状的后代植物,以产生与对照植物相比表现出一种或多种改善的产量性状的所选择的后代植物,任选地重复(e)至(g)一次或多次。在一些实施方案中,HECT E3 UPL基因是HECT E3 UPL3基因,任选地HECT E3 UPL3a基因、HECT E3 UPL3b基因、HECT E3 UPL3c基因、HECT E3 UPL3d基因。
在一些实施方案中,通过本发明的方法产生的突变的HECT E3 UPL基因可包含与SEQ ID NO:119-125中的任一个具有至少90%序列同一性的序列。在一些实施方案中,植物(例如,大豆植物、玉米植物)可以包含一个或多个(例如,1、2、3、4、5、6或更多个)本文所述的突变HECT E3 UPL基因,包括但不包括限于,与SEQ ID NO:119-125具有至少90%序列同一性的突变的HECT E3 UPL基因。作为实例,如本文所述编辑的大豆植物可包含(1)SEQ IDNO:119、(2)SEQ ID NO:120、(3)SEQ ID NO:121、(4)SEQ ID NO:120,和/或SEQ ID NO:121,或如本文所述编辑的玉米植物可以包含(1)SEQ ID NO:122,(2)SEQ ID NO:123,(3)SEQ IDNO:124,和/或(4)SEQ ID NO:125,任选地其中对于任何给定等位基因处的一个或多个突变,经编辑的植物可以是杂合的或纯合的,或其组合。在一些实施方案中,植物可以是杂合的并且在其基因组中的特定基因座处的HECT E3 UPL基因的一个等位基因中包含突变,并且在相同基因的第二拷贝中的相同基因座处是野生型。在一些实施方案中,在特定的HECTE3 UPL基因座中,植物可以在特定HECT E3 UPL基因的每个等位基因处包含不同的突变,或者可以在每个等位基因处包含相同的突变。
在一些实施方案中,提供了用于编辑植物细胞基因组中的特定位点的方法,该方法包括:以位点特异性方式切割植物细胞中内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因内的靶位点,所述内源HECT E3 UPL基因:(a)包含与SEQ ID NO:72-77的核苷酸序列中的任一个具有至少80%序列同一性的序列,(b)编码与SEQ ID NO:78-83中任一个的氨基酸序列具有至少80%序列同一性的序列,(c)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的区域,和/或(d)编码与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域,从而在植物细胞的内源HECT E3 UPL基因中产生编辑,并且产生包含内源HECT E3 UPL基因中的编辑的植物细胞,任选地其中HECT E3 UPL基因是HECT E3UPL3基因,任选地HECT E3 UPL3a基因、HECT E3 UPL3b基因、HECT E3 UPL3c基因和/或HECTE3 UPL3d基因。在一些实施方案中,编辑导致与SEQ ID NO:119-125的核苷酸序列中的任一个具有至少90%序列同一性的突变的HECT E3 UPL基因。
在一些实施方案中,编辑导致非天然突变,包括但不限于缺失、取代、插入或倒位,其中编辑可以导致无效突变、显性失活突变、半显性突变、超形态突变、亚形态突变或弱功能丧失突变,任选地无效突变或显性失活突变。在一些实施方案中,编辑可以是G至C的核苷酸取代(G>C)。在一些实施方案中,编辑导致HECT E3 UPL蛋白的编码区中氨基酸的变化。例如,编辑可导致位于以下位置处的氨基酸残基的变异:参考SEQ ID NO:78的氨基酸位置编号的位置1839,参考SEQ ID NO:79的氨基酸位置编号的位置1844,参考SEQ ID NO:80的氨基酸位置编号的位置1862,参考SEQ ID NO:81的氨基酸位置编号的位置1858,参考SEQ IDNO:82的氨基酸位置编号的位置1807,或者参考SEQ ID NO:83的氨基酸位置编号的位置1800。在一些实施方案中,编辑产生HECT E3 UPL多肽中半胱氨酸(C)至丝氨酸(S)的氨基酸取代。
在一些实施方案中,编辑方法还可包括从包含内源HECT E3 UPL基因(例如,内源HECT E3 UPL3基因)中的编辑的植物细胞再生植物,从而产生包含其内源HECT E3 UPL基因中的编辑并且与缺乏编辑的对照植物相比具有一种或多种改善的产量性状的表型的植物,任选地其中编辑的HECT E3 UPL基因包含与SEQ ID NO:119-125中的任一个具有至少90%序列同一性的序列。
在一些实施方案中,提供了用于制备植物的方法,其包括:(a)使包含内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的植物细胞群体与连接至核酸结合结构域(例如,编辑系统)的核酸酶接触,所述核酸结合结构域结合(a)与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性,(b)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列,(c)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列,和/或(d)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列的序列;(b)从所述群体中选择其中内源HECT E3 UPL基因已突变的植物细胞,从而产生包含内源HECT E3 UPL基因中的突变的植物细胞;和(c)将选择的植物细胞生长成植物,任选地其中HECT E3 UPL基因是HECT E3 UPL3基因,任选地HECT E3 UPL3a基因、HECT E3 UPL3b基因、HECT E3 UPL3c基因、HECT E3 UPL3d基因。
在一些实施方案中,提供了改善植物中的一种或多种产量性状的方法,该方法包括(a)使包含内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的植物细胞与靶向内源HECT E3 UPL基因的核酸酶接触,其中所述核酸酶连接至结合内源HECT E3 UPL基因中的靶位点的核酸结合结构域(例如,编辑系统),其中所述内源HECT E3 UPL基因:(i)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列;(ii)编码与SEQID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;(iii)编码包含与SEQ IDNO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或(iv)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,以产生包含内源HECT E3 UPL基因中的突变的植物细胞;(b)使植物细胞生长成包含内源HECT E3 UPL基因中的突变的植物,从而产生具有突变的内源HECT E3 UPL基因和一种或多种改善的产量性状的植物,任选地其中HECT E3 UPL基因是HECT E3 UPL3基因,任选地HECT E3 UPL3a基因、HECT E3 UPL3b基因、HECT E3 UPL3c基因、HECT E3 UPL3d基因。
在一些实施方案中,提供了用于产生包含至少一个具有突变的内源HECT E3 UPL基因的细胞的植物或其部分的方法,该方法包括使植物或植物部分中的内源HECT E3 UPL基因中的靶位点与包含切割结构域和核酸结合结构域的核酸酶接触,其中核酸结合结构域结合内源HECT E3 UPL基因中的靶位点,其中内源HECT E3 UPL基因(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列;(b)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;(c)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或(d)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,从而产生包含至少一个具有内源HECT E3 UPL基因中的突变的细胞的植物或其部分,任选地其中HECT E3 UPL基因是HECTE3 UPL3基因,任选地HECT E3 UPL3a基因、HECT E3 UPL3b基因、HECT E3 UPL3c基因、HECTE3 UPL3d基因。
本文还提供了用于产生包含突变的内源HECT E3 UPL基因并表现出一种或多种改善的产量性状的植物或其部分的方法,该方法包括使植物或植物部分中的内源HECT E3UPL基因中的靶位点与包含切割结构域和核酸结合结构域的核酸酶,其中所述核酸结合结构域结合内源HECT E3 UPL基因中的靶位点,其中所述内源HECT E3 UPL基因:(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列;(b)编码与SEQID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;(c)编码包含与SEQ IDNO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或(d)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,从而产生包含具有突变的内源HECT E3 UPL基因并表现出一种或多种改善的产量性状的植物或其部分,任选地其中一种或多种改善的产量性状包括与缺乏所述至少一个非天然突变的对照植物相比更高的产量(蒲式耳/英亩)、增加的籽粒行数,任选地其中穗长度基本上没有减少、增加的籽粒数量、增加的籽粒大小、增加的穗长度、减少的分蘖数、减少的雄穗分枝数、每个节增加的豆荚数、每株植物增加的豆荚数和/或增加的种子重量。在一些实施方案中改善的产量性状可以包括但不限于增加的种子大小(例如,面积、重量)和/或增加的种子油含量。在一些实施方案中,HECT E3 UPL基因是HECT E3 UPL3基因,任选地HECT E3 UPL3a基因、HECT E3UPL3b基因、HECT E3 UPL3c基因、HECT E3 UPL3d基因。在一些实施方案中,如本文所述编辑的植物或其部分包含与SEQ ID NO:119-125的核苷酸序列中的任一个具有至少90%序列同一性的突变的内源HECT E3 UPL基因。
在一些实施方案中,核酸酶可以切割内源HECT E3 UPL基因(例如,内源HECT E3UPL3基因),从而将突变引入到内源HECT E3 UPL基因中。可用于本发明的核酸酶可以是可用于编辑/修饰靶核酸的任何核酸酶。此类核酸酶包括但不限于锌指核酸酶、转录激活因子样效应核酸酶(TALEN)、核酸内切酶(例如,Fok1)和/或CRISPR-Cas效应蛋白。同样,可用于本发明的任何核酸结合结构域可以是可用于编辑/修饰靶核酸的任何DNA结合结构域或RNA结合结构域。此类核酸结合结构域包括但不限于锌指、转录激活因子样DNA结合结构域(TAL)、argonaute和/或CRISPR-Cas效应子DNA结合结构域。
在一些实施方案中,核酸结合结构域(例如,DNA结合结构域)包含在核酸结合多肽中。如本文所用,“核酸结合蛋白”或“核酸结合多肽”是指以位点和/或序列特异性方式结合和/或能够结合核酸的多肽。在一些实施方案中,核酸结合多肽可以是序列特异性核酸结合多肽(例如,序列特异性DNA结合结构域),例如但不限于来自例如多核苷酸引导的核酸内切酶、CRISPR-Cas效应蛋白(例如,CRISPR-Cas核酸内切酶)、锌指核酸酶、转录激活因子样效应核酸酶(TALEN)和/或Argonaute蛋白的序列特异性结合多肽和/或结构域。在一些实施方案中,核酸结合多肽包含切割多肽(例如,核酸酶多肽和/或结构域),例如但不限于核酸内切酶(例如,Fok1)、多核苷酸引导的核酸内切酶、CRISPR-Cas核酸内切酶、锌指核酸酶和/或转录激活因子样效应物核酸酶(TALEN)。在一些实施方案中,核酸结合多肽与一种或多种核酸分子缔合和/或能够与一种或多种核酸分子缔合(例如,形成复合物)(例如,与本文所述的引导核酸形成复合物),所述一种或多种核酸分子可以引导或指导核酸结合多肽至与一个或多个核酸分子(或其部分或区域)互补的特定靶核苷酸序列(例如,基因组的基因座),从而引起核酸结合多肽与特定靶位点处的核苷酸序列结合。在一些实施方案中,核酸结合多肽是本文所述的CRISPR-Cas效应蛋白。在一些实施方案中,为了简单起见,特指CRISPR-Cas效应蛋白,但也可以使用本文所述的核酸结合多肽。在一些实施方案中,本发明的多核苷酸和/或核酸构建体可以是“表达盒”或可以包含在表达盒内。
在一些实施方案中,提供了编辑植物或植物部分中的内源HECT E3 UPL基因的方法,该方法包括使植物或植物部分中的HECT E3 UPL基因中的靶位点与包含胞嘧啶脱氨酶和与HECT E3 UPL基因中的靶位点结合的核酸结合结构域的胞嘧啶碱基编辑系统接触,所述HECT E3 UPL基因,其中所述内源HECT E3 UPL基因:(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列;(b)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;(c)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或(d)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,从而编辑植物或其部分中的内源HECT E3UPL基因并产生包含在内源HECT E3 UPL基因中具有突变的至少一个细胞的植物或其部分。
在一些实施方案中,提供了编辑植物或植物部分中的内源HECT E3 UPL基因的方法,该方法包括使植物或植物部分中的HECT E3 UPL基因中的靶位点与包含腺苷脱氨酶和与HECT E3 UPL基因中的靶位点结合的核酸结合结构域的腺苷碱基编辑系统接触,所述HECT E3 UPL基因(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列;(b)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;(c)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或(d)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,从而编辑植物或其部分中的内源HECT E3 UPL基因并产生包含在内源HECT E3 UPL基因中具有突变的至少一个细胞的植物或其部分。
在一些实施方案中,提供了检测突变型HECT E3 UPL基因(内源HECT E3 UPL基因中的突变)的方法,该方法包括检测植物基因组中的编码包含活性/催化位点中的突变的HECT E3 UPL多肽的内源HECT E3 UPL基因,任选地其中活性位点在HECT E3 UPL3多肽的区域中,该区域包含:(a)与SEQ ID NO:88-93中的任一个具有至少80%同一性的氨基酸序列和/或(b)与以下的连续氨基酸残基区域具有至少80%序列同一性的氨基酸序列:参考SEQID NO:78的残基编号,从残基1814到残基1865、从残基1822到残基1857和/或从残基1829到残基1851,参考SEQ ID NO:79的残基编号,从残基1820到残基1870、从残基1827到残基1863和/或从残基1834到残基1856,参考SEQ ID NO:80的残基编号,从残基1838到残基1888、从残基1845到残基1881和/或从残基1852到残基1874,参考SEQ ID NO:81的残基编号,从残基1834到残基1884、从残基1841到残基1877和/或从残基1848到残基1870,参考SEQ ID NO:82的残基编号,从残基1783到残基1833、从残基1790到残基1826和/或从残基1797到残基1818,或参考SEQ ID NO:83的残基编号、从残基1776到残基1826、从残基1783到残基1819和/或从残基1790到残基1812。在一些实施方案中,提供了检测突变型HECT E3 UPL基因(内源HECT E3 UPL基因中的突变)的方法,该方法包括检测植物基因组中的编码SEQ ID NO:78-83的氨基酸序列的核酸,其中SEQ ID NO:78-83的氨基酸序列包含位于以下位置处的一个或多个氨基酸残基的取代:参考SEQ ID NO:78的氨基酸位置编号的位置1839,参考SEQID NO:79的氨基酸位置编号的位置1844,参考SEQ ID NO:80的氨基酸位置编号的位置1862,参考SEQ ID NO:81的氨基酸位置编号的位置1858,参考SEQ ID NO:82的氨基酸位置编号的位置1807,或者参考SEQ ID NO:83的氨基酸位置编号的位置1800。在一些实施方案中,突变是G至C的核苷酸取代(G>C)的结果。在一些实施方案中,突变的HECT E3 UPL肽包含至少一个包含C取代S的突变。
在一些实施方案中,提供了检测突变型HECT E3 UPL基因(内源HECT E3 UPL基因;例如内源HECT E3 UPL3基因中的突变)的方法,该方法包括检测植物基因组中的包含框内或框外插入缺失的HECT E3 UPL基因,任选地其中框外插入缺失导致提前终止密码子。在一些实施方案中,检测突变型HECT E3 UPL基因的方法包括检测与SEQ ID NO:119-125中的任一个具有至少90%序列同一性的核苷酸序列。
在一些实施方案中,本发明提供了产生包含内源HECT E3 UPL基因(例如,HECT E3UPL3基因,例如HECT E3 UPL3a、HECT E3 UPL3b、HECT E3 UPL3c、HECT E3 UPL3d)中的突变和至少一种感兴趣的多核苷酸的植物的方法,该方法包括将包含内源HECT E3 UPL基因中的至少一个突变的本发明的植物(第一植物)与包含至少一种感兴趣的多核苷酸的第二植物杂交以产生后代植物;和选择包含HECT E3 UPL基因中的至少一个突变和至少一种感兴趣的多核苷酸的后代植物,从而产生包含内源HECT E3 UPL基因中的突变和至少一种感兴趣的多核苷酸的植物。
本发明还提供了产生包含内源HECT E3 UPL基因中的突变和至少一种感兴趣的多核苷酸的植物的方法,该方法包括将至少一种感兴趣的多核苷酸引入到包含HECT E3 UPL基因中的至少一个突变的本发明的植物中,从而产生包含HECT E3 UPL基因中的至少一个突变和至少一种感兴趣的多核苷酸的植物。在一些实施方案中,植物是玉米植物。在一些实施方案中,植物是大豆植物。
感兴趣的多核苷酸可以是能够赋予所需的表型或以其他方式修饰植物的表型或基因型的任何多核苷酸。在一些实施方案中,感兴趣的多核苷酸可以是赋予除草剂耐受性、昆虫抗性、线虫抗性、抗病性、增加的产量、增加的养分利用效率或非生物胁迫抗性的多核苷酸。
可用于本发明的HECT E3 UPL基因包括其中本文所述的突变可以赋予包含该突变的植物或其部分中一种或多种产量性状的改善的任何HECT E3 UPL基因。在一些实施方案中,用于本发明的HECT E3 UPL基因编码HECT E3 UPL3多肽,任选地HECT E3 UPL3a多肽、HECT E3 UPL3b多肽、HECT E3 UPL3c多肽和/或HECT E3 UPL3d多肽。在一些实施方案中,内源HECT E3 UPL多肽包含与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的序列的氨基酸序列,包含与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的区域,和/或包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域。
在一些实施方案中,植物中内源HECT E3 UPL基因中的至少一个非天然突变可以是碱基取代、碱基缺失和/或碱基插入或倒位。在一些实施方案中,植物中内源HECT E3 UPL基因中的至少一个非天然突变可以是导致无效突变、显性失活突变、半显性突变、超形态突变、亚形态突变或弱功能丧失突变(任选地无效突变或显性失活突变)以及与缺乏编辑/突变的对照植物相比具有一种或多种改善的产量性状的表型的植物的取代、缺失、插入和/或倒位,任选地其中改善的产量性状可以包括但不限于增加的种子大小(例如,种子面积和/或种子重量)和/或增加的种子油含量。例如,突变可以是一个或多个氨基酸残基(例如,HECT E3 UPL3多肽的1、2、3、4、5、6、7、8、9或10个氨基酸)的取代、缺失和/或插入,或突变可以是编码HECT E3 UPL3多肽的基因中的至少1个核苷酸至约15个核苷酸(例如,约1、2、3、4、5、6、7、8、9、10、11、12、13、14或15个核苷酸,或其中的任何范围或值)的碱基取代、碱基缺失和/或碱基插入,或者可以是至少10个连续核苷酸至约2000个连续核苷酸(例如,约10、15、20、25、30、35、40、45、50、55、60、65、70、75、80、85、90、95、100、110、120、130、140、150、160、170、180、190、200、210、220、230、240、250、260、270、280、290、300、350、400、450、500、550、600、650、700,750、800、850、900、950、1000、1100、1110、1120、1130、1140、1150、1160、1170、1180、1190、1200、1210、1220、1230、1240、1250、1260、1270、1280、1290、1300、1350、1400、1450、1500、1550、1600、1650、1700、1750、1800、1850、1900、1950或2000个或更多个连续核苷酸,或其中的任何范围或值)的DNA倒位。在一些实施方案中,至少一个非天然突变可以是对A、T、G或C的碱基取代。在一些实施方案中,至少一个非天然突变可以是例如从G到C的碱基取代(G>C)。突变可以是点突变。在一些实施方案中,突变导致位于以下位置处的取代的氨基酸残基:参考SEQ ID NO:78的氨基酸位置编号的位置1839,参考SEQ ID NO:79的氨基酸位置编号的位置1844,参考SEQ ID NO:80的氨基酸位置编号的位置1862,参考SEQ IDNO:81的氨基酸位置编号的位置1858,参考SEQ ID NO:82的氨基酸位置编号的位置1807,或者参考SEQ ID NO:83的氨基酸位置编号的位置1800。
在一些实施方案中,内源HECT E3 UPL基因中的突变可在通过编辑系统切割后产生,所述编辑系统包含核酸酶和结合靶核酸(例如,HECT E3 UPL基因)内的靶位点的核酸结合结构域,所述靶核酸包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列,任选地其中靶位点位于HECT E3 UPL基因的区域中:该区域(a)编码与SEQID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的序列,(b)编码与以下的连续氨基酸残基的区域具有至少80%序列同一性的序列:参考SEQ ID NO:78的残基编号,从残基50到残基95、从残基43到残基102、从残基36到残基109、从残基1814到残基1865、从残基1822到残基1857和/或从残基1829到残基1851,参考SEQ ID NO:79的残基编号,从残基48到残基87、从残基41到残基94、从残基34到残基101、从残基1820到残基1870、从残基1827到残基1863和/或从残基1834到残基1856,参考SEQ ID NO:80的残基编号,从残基139到残基194、从残基132到残基201、从残基125到残基208、从残基1838到残基1888、从残基1845到残基1881和/或从残基1852到残基1874,参考SEQ ID NO:81的残基编号,从残基104到残基146、从残基97到残基153、从残基90到残基160、从残基1834到残基1884、从残基1841到残基1877和/或从残基1848到残基1870,参考SEQ ID NO:82的残基编号,从残基1783到残基1833、从残基1790到残基1826和/或从残基1797到残基1818,和/或参考SEQ ID NO:83的残基编号,从残基1776到残基1826、从残基1783到残基1819和/或从残基1790到残基1812;和/或(c)编码与SEQ ID NO:93的氨基酸序列具有至少90%序列同一性的序列。
进一步提供了结合E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的靶位点的引导核酸(例如,gRNA、gDNA、crRNA、crDNA),该靶位点位于HECT E3 UPL3基因的区域中:所述区域(a)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的序列,(b)编码与以下的连续氨基酸残基的区域具有至少80%序列同一性的序列:参考SEQID NO:78的残基编号,从残基50到残基95、从残基43到残基102、从残基36到残基109、从残基1814到残基1865、从残基1822到残基1857和/或从残基1829到残基1851,参考SEQ ID NO:79的残基编号,从残基48到残基87、从残基41到残基94、从残基34到残基101、从残基1820到残基1870、从残基1827到残基1863和/或从残基1834到残基1856,参考SEQ ID NO:80的残基编号,从残基139到残基194、从残基132到残基201、从残基125到残基208、从残基1838到残基1888、从残基1845到残基1881和/或从残基1852到残基1874,参考SEQ ID NO:81的残基编号,从残基104到残基146、从残基97到残基153、从残基90到残基160、从残基1834到残基1884、从残基1841到残基1877和/或从残基1848到残基1870,参考SEQ ID NO:82的残基编号,从残基1783到残基1833、从残基1790到残基1826和/或从残基1797到残基1818,和/或参考SEQ ID NO:83的残基编号,从残基1776到残基1826、从残基1783到残基1819和/或从残基1790到残基1812;和/或(c)编码与SEQ ID NO:93的氨基酸序列具有至少90%序列同一性的序列。在一些实施方案中,引导核酸包含具有SEQ ID NO:94-118中任一个的核苷酸序列的间隔区。
在一些实施方案中,提供了一种玉米植物或其植物部分,其包含在至少一个内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的至少一个非天然突变,该基因具有Zm00001d014920和/或Zm00001d004139的基因识别号(基因ID),任选地其中具有至少一个非天然突变的HECT E3 UPL基因包含与SEQ ID NO:122-125中的任一个具有至少90%序列同一性的核苷酸序列。
在一些实施方案中,提供了一种大豆植物或其植物部分,其包含在至少一个内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的至少一个非天然突变,该基因具有Glyma.11G107500、Glyma.12G032500、Glyma.06G003600和/或Glyma.04G004000的基因识别号(基因ID),任选地其中具有至少一个非天然突变的HECT E3 UPL基因包含与SEQ ID NO:119-121中的任一个具有至少90%序列同一性的核苷酸序列。
在一些实施方案中,提供了引导核酸,其结合E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的靶核酸,所述基因具有Zm00001d014920、Zm00001d004139、Glyma.11G107500、Glyma.12G032500、Glyma.06G003600和/或Glyma.04G004000的基因识别号(基因ID)。
在一些实施方案中,提供了一种系统,其包含引导核酸和CRISPR-Cas效应蛋白,所述引导核酸包含具有SEQ ID NO:94-118中任一个的核苷酸序列的间隔区(例如,一个或多个间隔区),所述CRISPR-Cas效应蛋白与引导核酸缔合。在一些实施方案中,该系统还可以包含与引导核酸和CRISPR-Cas效应蛋白缔合的tracr核酸,任选地其中tracr核酸和引导核酸共价连接。如本文所用,“与引导核酸缔合的CRISPR-Cas效应蛋白”是指在CRISPR-Cas效应蛋白和引导核酸之间形成的复合物,以将CRISPR-Cas效应蛋白引导至基因中的靶位点。
本发明还提供了基因编辑系统,其包含与引导核酸缔合的CRISPR-Cas效应蛋白,并且引导核酸包含结合E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的间隔序列,任选地其中HECT E3 UPL基因(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列,(b)编码与SEQ ID NO:78-83中任一个的氨基酸序列具有至少80%序列同一性的序列,(c)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的区域,和/或(d)编码与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域。在一些实施方案中,引导核酸的间隔序列可以包含SEQ ID NO:94-118中任一个的核苷酸序列。在一些实施方案中,基因编辑系统还可包含与引导核酸和CRISPR-Cas效应蛋白缔合的tracr核酸,任选地其中tracr核酸和引导核酸共价连接。
本发明还提供了包含含有切割结构域的CRISPR-Cas效应蛋白和引导核酸的复合物,其中所述引导核酸结合内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的靶位点,其中内源HECT E3 UPL3基因:(a)包含与SEQ ID NO:72-77的核苷酸序列中的任一个具有至少80%序列同一性的序列,(b)编码与SEQ ID NO:78-83中任一个的氨基酸序列具有至少80%序列同一性的序列,(c)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的区域,和/或(d)编码与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域,其中切割结构域切割HECT E3 UPL3基因中的靶标链。
在一些实施方案中,提供了表达盒,其包含(a)编码包含切割结构域的CRISPR-Cas效应蛋白的多核苷酸和(b)结合内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的靶位点的引导核酸,其中引导核酸包含间隔序列,所述间隔序列互补于并结合:(i)与SEQID NO:72-77的核苷酸序列中的任一个具有至少80%序列同一性的序列的一部分;(i)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列的核酸的一部分;(iii)编码与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的氨基酸序列的核酸的一部分;和/或(iv)编码与SEQ ID NO:93具有至少90%序列同一性的氨基酸序列的核酸的一部分。
本文还提供了包含突变的HECT E3 UPL基因的核酸,当其存在于植物或植物部分中时,导致与不缺乏突变的植物或植物部分相比包含一种或多种改善的产量性状的表型的植物,其中改善的产量性状可包括但不限于增加的种子大小和/或增加的种子油含量,任选地其中突变是显性失活突变或无效突变,任选地其中突变的HECT E3 UPL基因可与SEQ IDNO:119-125中的任一个包含至少90%的序列同一性。
本发明的核酸构建体(例如,包含序列特异性核酸结合结构域(例如,序列特异性DNA结合结构域)、CRISPR-Cas效应结构域、脱氨酶结构域、逆转录酶(RT)、RT模板和/或引导核酸等的构建体)和包含其的表达盒/载体可以用作本发明的编辑系统,用于修饰靶核酸(例如,内源HECT E3 UPL基因)和/或其表达。
包含当如本文所述进行修饰时能够赋予至少一种改善的产量性状的内源HECT E3UPL基因(例如,HECT E3 UPL3基因,例如HECT E3 UPL3a基因、HECT E3 UPL3b基因、HECT E3UPL3c基因、HECT E3 UPL3d基因)的任何植物可以如本文所述进行修饰(例如,突变,例如碱基编辑、切割、切口等)(例如,使用本发明的多肽、多核苷酸、RNP、核酸构建体、表达盒和/或载体)以改善植物中的一种或多种产量性状。表现出改善的产量性状的植物可以与缺乏突变的内源HECT E3 UPL基因的植物或其部分相比表现出产量性状的约5%至约100%(例如,约5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64、65、66、67、68、69、70、71、72、73、74、75、76、77、78、79、80、81、82、83、84、85、86、87、88、89、90、91、92、93、94、95、96、97、98、99或100%或更多或其中的任何范围或值;例如,约5%至约10%、约5%至约15%、约5%至约20%、约10%至约50%、约10%至约80%、约10%至约90%、约10%至约100%、约20%至约50%、约20%至约80%、约20%至约90%、约20%至约100%、约30%至约50%、约30%至约80%、约30%至约90%、约30%至约100%、约50%至约100%、约75%至约100%或更多以及其中的任何范围或值)的改善。
可用于本发明的编辑系统可以是现在已知或以后开发的任何位点特异性(序列特异性)基因组编辑系统,该系统可以以靶标特异性方式引入突变。例如,编辑系统(例如,位点特异性或序列特异性编辑系统)可以包括但不限于CRISPR-Cas编辑系统、大范围核酸酶编辑系统、锌指核酸酶(ZFN)编辑系统、转录激活因子样效应核酸酶(TALEN)编辑系统、碱基编辑系统和/或引导编辑系统,其各自可以包含一种或多种多肽和/或一种或多种多核苷酸,当其在细胞中表达为系统时,可以以序列特异性方式修饰(突变)靶核酸。在一些实施方案中,编辑系统(例如,位点特异性或序列特异性编辑系统)可以包含一种或多种多核苷酸和/或一种或多种多肽,包括但不限于核酸结合结构域(DNA结合结构域)、核酸酶,和/或其他多肽,和/或多核苷酸。
在一些实施方案中,编辑系统可以包含一个或多个序列特异性核酸结合结构域(DNA结合结构域),其可以来自例如多核苷酸引导的核酸内切酶、CRISPR-Cas核酸内切酶(例如,CRISPR-Cas效应蛋白)、锌指核酸酶、转录激活因子样效应核酸酶(TALEN)和/或Argonaute蛋白。在一些实施方案中,编辑系统可以包含一个或多个切割结构域(例如,核酸酶),包括但不限于核酸内切酶(例如,Fok1)、多核苷酸引导的核酸内切酶、CRISPR-Cas核酸内切酶(例如,CRISPR-Cas效应蛋白)、锌指核酸酶和/或转录激活因子样效应核酸酶(TALEN)。在一些实施方案中,编辑系统可以包含一种或多种多肽,其包括但不限于脱氨酶(例如,胞嘧啶脱氨酶、腺嘌呤脱氨酶)、逆转录酶、Dna2多肽和/或5’瓣核酸内切酶(FEN)。在一些实施方案中,编辑系统可包含一种或多种多核苷酸,其包括但不限于CRISPR阵列(CRISPR引导)核酸、延伸的引导核酸和/或逆转录酶模板。
在一些实施方案中,修饰或编辑E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的方法可以包括使靶核酸(例如,编码HECT E3UPL多肽的核酸)与融合至脱氨酶结构域(例如,腺嘌呤脱氨酶和/或胞嘧啶脱氨酶)的碱基编辑融合蛋白(例如,序列特异性DNA结合蛋白(例如,CRISPR-Cas效应蛋白或结构域))和引导核酸接触,其中引导核酸能够将碱基编辑融合蛋白引导/靶向至靶核酸,从而编辑靶核酸内的基因座。在一些实施方案中,碱基编辑融合蛋白和引导核酸可以包含在一个或多个表达盒中。在一些实施方案中,靶核酸可以与碱基编辑融合蛋白和包含引导核酸的表达盒接触。在一些实施方案中,序列特异性核酸结合融合蛋白和引导物可以作为核糖核蛋白(RNP)提供。在一些实施方案中,细胞可以与多于一种碱基编辑融合蛋白和/或可以靶向细胞中的一种或多种靶核酸的一种或多种引导核酸接触。
在一些实施方案中,修饰或编辑E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的方法可以包括使靶核酸(例如,编码HECT E3 UPL多肽(例如HECT E3 UPL3多肽;例如,HECT E3 UPL3a、HECT E3 UPL3b、HECT E3 UPL3c、HECT E3 UPL3d)的核酸)与融合至肽标签的序列特异性核酸结合融合蛋白(例如,序列特异性DNA结合蛋白(例如,CRISPR-Cas效应蛋白或结构域))、包含融合至能够结合肽标签的亲和多肽的脱氨酶结构域(例如,腺嘌呤脱氨酶和/或胞嘧啶脱氨酶)的脱氨酶融合蛋白和引导核酸接触,其中引导核酸能够将序列特异性核酸结合融合蛋白引导/靶向至靶核酸,并且序列特异性核酸结合融合蛋白能够通过肽标签-亲和多肽相互作用将脱氨酶融合蛋白募集至靶核酸,从而编辑靶核酸内的基因座。在一些实施方案中,序列特异性核酸结合融合蛋白可与结合肽标签的亲和多肽融合,并且脱氨酶可与肽标签融合,从而将脱氨酶募集至序列特异性核酸结合融合蛋白和靶核酸。在一些实施方案中,序列特异性结合融合蛋白、脱氨酶融合蛋白和引导核酸可以包含在一个或多个表达盒中。在一些实施方案中,靶核酸可以与序列特异性结合融合蛋白、脱氨酶融合蛋白和包含引导核酸的表达盒接触。在一些实施方案中,序列特异性核酸结合融合蛋白、脱氨酶融合蛋白和引导物可以作为核糖核蛋白(RNP)提供。
在一些实施方案中,诸如引导编辑的方法可用于在内源HECT E3UPL基因中产生突变。在引导编辑中,RNA依赖性DNA聚合酶(逆转录酶,RT)和逆转录酶模板(RT模板)与赋予以序列特异性方式识别和结合靶标的能力并且还可能导致靶标内含有PAM的链的切口的序列特异性核酸结合域组合使用。核酸结合结构域可以是CRISPR-Cas效应蛋白,并且在这种情况下,CRISPR阵列或引导RNA可以是延伸的引导物,其包含含有引物结合位点(PSB)的延伸部分和待掺入基因组(模板)的编辑。与碱基编辑类似,引物编辑可以利用招募蛋白质用于编辑靶位点的各种方法,这些方法包括所选择的基因组编辑过程中使用的蛋白质和核酸之间的非共价和共价相互作用。
如本文所用,“CRISPR-Cas效应蛋白”是裂解或剪切核酸、结合核酸(例如,靶核酸和/或引导核酸)和/或鉴定、识别或结合本文定义的引导核酸的蛋白质或多肽或其结构域。在一些实施方案中,CRISPR-Cas效应蛋白可以是酶(例如,核酸酶、核酸内切酶、切口酶等)或其部分和/或可以起到酶的作用。在一些实施方案中,CRISPR-Cas效应蛋白是指CRISPR-Cas核酸酶多肽或其结构域,其包含核酸酶活性或其中核酸酶活性已降低或消除,和/或包含切口酶活性或其中切口酶已减少或消除,和/或包含单链DNA切割活性(ss DNAse活性)或其中ss DNAse活性已降低或消除,和/或包含自加工RNAse活性或其中自加工RNAse活性已被减少或消除。CRISPR-Cas效应蛋白可以结合靶核酸。
在一些实施方案中,序列特异性核酸结合结构域可以是CRISPR-Cas效应蛋白。在一些实施方案中,CRISPR-Cas效应蛋白可以来自I型CRISPR-Cas系统、II型CRISPR-Cas系统、III型CRISPR-Cas系统、IV型CRISPR-Cas系统、V型CRISPR-Cas系统或VI型CRISPR-Cas系统。在一些实施方案中,本发明的CRISPR-Cas效应蛋白可以来自II型CRISPR-Cas系统或V型CRISPR-Cas系统。在一些实施方案中,CRISPR-Cas效应蛋白可以是II型CRISPR-Cas效应蛋白,例如Cas9效应蛋白。在一些实施方案中,CRISPR-Cas效应蛋白可以是V型CRISPR-Cas效应蛋白,例如Cas12效应蛋白。
在一些实施方案中,CRISPR-Cas效应蛋白可以包括但不限于Cas9、C2c1、C2c3、Cas12a(也称为Cpf1)、Cas12b、Cas12c、Cas12d、Cas12e、Cas13a、Cas13b、Cas13c、Cas13d、Casl、CaslB、Cas2、Cas3、Cas3'、Cas3"、Cas4、Cas5、Cas6、Cas7、Cas8、Cas9(也称为Csnl和Csx12)、Cas10、Csyl、Csy2、Csy3、Csel、Cse2、Cscl、Csc2、Csa5、Csn2、Csm2、Csm3、Csm4、Csm5、Csm6、Cmrl、Cmr3、Cmr4、Cmr5、Cmr6、Csbl、Csb2、Csb3、Csxl7、Csxl4、Csx10、Csx16、CsaX、Csx3、Csxl、Csxl5、Csfl、Cs、Csf2、Csf3、Csf4(dinG)和/或Csf5核酸酶,任选其中CRISPR-Cas效应蛋白可以是Cas9、Cas12a(Cpf1)、Cas12b、Cas12c(C2c3)、Cas12d(CasY)、Cas12e(CasX)、Cas12g、Cas12h、Cas12i、C2c4、C2c5、C2c8、C2c9、C2c10、Cas14a、Cas14b和/或Cas14c效应蛋白。
在一些实施方案中,可用于本发明的CRISPR-Cas效应蛋白可以在其核酸酶活性位点(例如,RuvC、HNH,例如,Cas12a核酸酶结构域的RuvC位点;例如,Cas9核酸酶结构域的RuvC位点和/或HNH位点)中包含突变。在其核酸酶活性位点具有突变并因此不再包含核酸酶活性的CRISPR-Cas效应蛋白通常被称为“死”,例如dCas。在一些实施方案中,与没有突变的相同CRISPR-Cas效应蛋白(例如切口酶,例如Cas9切口酶,Cas12a切口酶)相比,在其核酸酶活性位点具有突变的CRISPR-Cas效应蛋白结构域或多肽可能具有受损的活性或降低的活性。
可用于本发明的CRISPR Cas9效应蛋白或CRISPR Cas9效应结构域可以是任何已知的或后来鉴定的Cas9核酸酶。在一些实施方案中,CRISPR Cas9多肽可以是来自例如链球菌属的几个种(例如,化脓性链球菌(S.pyogenes)、嗜热链球菌(S.thermophilus))、乳杆菌属的几个种(Lactobacillus spp.)、双歧杆菌属的几个种(Bifidobacterium spp.)、坎德勒氏菌属的几个种(Kandleria spp.)、明串珠菌属的几个种(Leuconostoc spp.)、酒球菌属的几个种(Oenococcus spp.)、片球菌属的几个种(Pediococcus spp.)、魏斯氏菌属的几个种(Weissella spp.)和/或奥森氏菌属的几个种(Olsenella spp.)的Cas9多肽。示例性Cas9序列包括但不限于SEQ ID NO:59和SEQ ID NO:60的氨基酸序列或SEQ ID NO:61-71的核苷酸序列。
在一些实施方案中,CRISPR-Cas效应蛋白可以是源自化脓性链球菌并且识别PAM序列基序NGG、NAG、NGA的Cas9多肽(Mali等人,Science 2013;339(6121):823-826)。在一些实施方案中,CRISPR-Cas效应蛋白可以是源自嗜热链球菌并且识别PAM序列基序NGGNG和/或NNAGAAW(W=A或T)的Cas9多肽(参见例如Horvath等人,Science,2010;327(5962):167-170,和Deveau等人,J Bacteriol 2008;190(4):1390-1400)。在一些实施方案中,CRISPR-Cas效应蛋白可以是源自变形链球菌并且识别PAM序列基序NGG和/或NAAR(R=A或G)的Cas9多肽(参见例如Deveau等人,J BACTERIOL 2008;190(4):1390-1400)。在一些实施方案中,CRISPR-Cas效应蛋白可以是源自金黄色链球菌并识别PAM序列基序NNGRR(R=A或G)的Cas9多肽。在一些实施方案中,CRISPR-Cas效应蛋白可以是源自金黄色葡萄球菌的Cas9蛋白,其识别PAM序列基序N GRRT(R=A或G)。在一些实施方案中,CRISPR-Cas效应蛋白可以是源自金黄色葡萄球菌的Cas9多肽,其识别PAM序列基序N GRRV(R=A或G)。在一些实施方案中,CRISPR-Cas效应蛋白可以是源自脑膜炎奈瑟菌并且识别PAM序列基序N GATT或N GCTT(R=A或G,V=A、G或C)的Cas9多肽(参见,例如,Hou等人,PNAS2013,1-6)。在上述实施方案中,N可以是任何核苷酸残基,例如A、G、C或T中的任何一个。在一些实施方案中,CRISPR-Cas效应蛋白可以是源自沙氏纤毛菌(Leptotrichia shahii)的Cas13a蛋白,其识别单个3'A、U或C的前间隔区侧翼序列(PFS)(或RNA PAM(rPAM))序列基序,其可能位于靶核酸内。
在一些实施方案中,CRISPR-Cas效应蛋白可以源自Cas12a,其是V型聚集规则间隔短回文重复序列(CRISPR)-Cas核酸酶,参见例如SEQ ID NO:1-20)。Cas12a在几个方面与更知名的II型CRISPR Cas9核酸酶不同。例如,Cas9识别富含G的前间隔区邻近基序(PAM),它位于其引导RNA(gRNA、sgRNA、crRNA、crDNA、CRISPR阵列)结合位点(前间隔区、靶核酸、靶DNA)的3'(3'-NGG),而Cas12a识别位于靶核酸的5'的富含T的PAM(5'-TTN,5'-TTTN。事实上,其中Cas9和Cas12a结合其引导RNA的方向关于其N和C末端是几乎相反的。此外,Cas12a酶使用单引导RNA(gRNA、CRISPR阵列、crRNA),而不是在天然Cas9系统中发现的双引导RNA(sgRNA(例如crRNA和tracrRNA)),并且Cas12a加工其自身的gRNA。此外,Cas12a核酸酶活性产生交错的DNA双链断裂,而不是由Cas9核酸酶活性产生的平末端,并且Cas12a依赖于单个RuvC结构域来切割两条DNA链,而Cas9利用HNH结构域和RuvC结构域用于切割。
可用于本发明的CRISPR Cas12a效应蛋白/结构域可以是任何已知的或后来鉴定的Cas12a多肽(以前称为Cpf1)(参见例如美国专利号9,790,490,其关于Cpf1(Cas12a)序列的公开内容通过引用并入本文)。术语“Cas12a”、“Cas12a多肽”或“Cas12a结构域”是指包含Cas12a多肽或其片段的RNA引导的核酸酶,其包含Cas12a的引导核酸结合结构域和/或Cas12a的活性、非活性或部分活性DNA切割结构域。在一些实施方案中,可用于本发明的Cas12a可包含核酸酶活性位点(例如,Cas12a结构域的RuvC位点)中的突变。在其核酸酶活性位点中具有突变并因此不再包含核酸酶活性的Cas12a结构域或Cas12a多肽通常被称为死Cas12a(例如,dCas12a)。在一些实施方案中,在其核酸酶活性位点中具有突变的Cas12a结构域或Cas12a多肽可能具有受损的活性,例如,可能具有切口酶活性。
可用于碱基编辑的任何脱氨酶结构域/多肽可用于本发明。在一些实施方案中,脱氨酶结构域可以是胞嘧啶脱氨酶结构域或腺嘌呤脱氨酶结构域。可用于本发明的胞嘧啶脱氨酶(或胞苷脱氨酶)可以是来自任何生物体的任何已知或后来鉴定的胞嘧啶脱氨酶(参见例如美国专利号10,167,457和Thuronyi等人,Nat.Biotechnol.37:1070-1079(2019),其各自关于其胞嘧啶脱氨酶的公开内容通过引用并入本文)。胞嘧啶脱氨酶可分别催化胞苷或脱氧胞苷水解脱氨为尿苷或脱氧尿苷。因此,在一些实施方案中,可用于本发明的脱氨酶或脱氨酶结构域可以是胞苷脱氨酶结构域,其催化胞嘧啶水解脱氨为尿嘧啶。在一些实施方案中,胞嘧啶脱氨酶可以是天然存在的胞嘧啶脱氨酶的变体,包括但不限于灵长类动物(例如人、猴、黑猩猩、大猩猩)、狗、牛、大鼠或小鼠。因此,在一些实施方案中,可用于本发明的胞嘧啶脱氨酶可以与野生型胞嘧啶脱氨酶约70%至约100%相同(例如,与天然存在的胞嘧啶脱氨酶约70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%相同,以及其中的任何范围或值)。
在一些实施方案中,可用于本发明的胞嘧啶脱氨酶可以是载脂蛋白B mRNA-编辑复合物(APOBEC)家族脱氨酶。在一些实施方案中,胞嘧啶脱氨酶可以是APOBEC1脱氨酶、APOBEC2脱氨酶、APOBEC3A脱氨酶、APOBEC3B脱氨酶、APOBEC3C脱氨酶、APOBEC3D脱氨酶、APOBEC3F脱氨酶、APOBEC3G脱氨酶、APOBEC3H脱氨酶、APOBEC4脱氨酶、人活化诱导脱氨酶(hAID)、rAPOBEC1、FERNY和/或CDA1,任选地pmCDA1、atCDA1(例如,At2g19570),以及它们的进化形式(例如,SEQ ID NO:27、SEQ ID NO:28或SEQ ID NO:29)。在一些实施方案中,胞嘧啶脱氨酶可以是具有SEQ ID NO:23的氨基酸序列的APOBEC1脱氨酶。在一些实施方案中,胞嘧啶脱氨酶可以是具有SEQ ID NO:24的氨基酸序列的APOBEC3A脱氨酶。在一些实施方案中,胞嘧啶脱氨酶可以是CDA1脱氨酶,任选地是具有SEQ ID NO:25的氨基酸序列的CDA1。在一些实施方案中,胞嘧啶脱氨酶可以是FERNY脱氨酶,任选地是具有SEQ ID NO:26的氨基酸序列的FERNY。在一些实施方案中,可用于本发明的胞嘧啶脱氨酶可以与天然存在的胞嘧啶脱氨酶(例如进化的脱氨酶)的氨基酸序列约70%至约100%相同的(例如,70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%、99.5%或100%相同)。在一些实施方案中,可用于本发明的胞嘧啶脱氨酶可与SEQ ID NO:23、SEQ IDNO:24、SEQ ID NO:25或SEQ ID NO:26的氨基酸序列约70%至约99.5%相同(例如,约70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或99.5%相同)(例如,与SEQ ID NO:23、SEQ ID NO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28或SEQ ID NO:29的氨基酸序列至少80%、至少85%、至少90%、至少92%、至少95%、至少96%、至少97%、至少98%、至少99%、或至少99.5%相同)。在一些实施方案中,可以对编码胞嘧啶脱氨酶的多核苷酸进行密码子优化以用于在植物中表达,并且密码子优化的多肽可以与参考多核苷酸约70%至99.5%相同。
在一些实施方案中,本发明的核酸构建体可以进一步编码尿嘧啶糖基化酶抑制剂(UGI)(例如,尿嘧啶-DNA糖基化酶抑制剂)多肽/结构域。因此,在一些实施方案中,编码CRISPR-Cas效应蛋白和胞嘧啶脱氨酶结构域的核酸构建体(例如,编码包含与胞嘧啶脱氨酶结构域融合的CRISPR-Cas效应蛋白结构域的融合蛋白,和/或与肽标签或能够结合肽标签的亲和多肽融合的CRISPR-Cas效应蛋白结构域和/或与肽标签或能够结合肽标签的亲和多肽融合的脱氨酶蛋白结构域)可以进一步编码尿嘧啶-DNA糖基化酶抑制剂(UGI),任选地其中UGI可以被密码子优化以用于在植物中表达。在一些实施方案中,本发明提供了包含CRISPR-Cas效应多肽、脱氨酶结构域和UGI的融合蛋白和/或一种或多种编码它们的多核苷酸,任选地其中所述一种或多种多核苷酸可以被密码子优化以用于在植物中表达。在一些实施方案中,本发明提供了融合蛋白,其中CRISPR-Cas效应多肽、脱氨酶结构域和UGI可以与本文所述的肽标签和亲和多肽的任何组合融合,从而将脱氨酶结构域和UGI募集到CRISPR-Cas效应多肽和靶核酸。在一些实施方案中,引导核酸可以与募集RNA基序连接,并且脱氨酶结构域和/或UGI中的一个或多个可以与能够与募集RNA基序相互作用的亲和多肽融合,从而将脱氨酶结构域和UGI募集到靶核酸。
可用于本发明的“尿嘧啶糖基化酶抑制剂”可以是能够抑制尿嘧啶-DNA糖基化酶碱基切除修复酶的任何蛋白质。在一些实施方案中,UGI结构域包含野生型UGI或其片段。在一些实施方案中,可用于本发明的UGI结构域可以与天然存在的UGI结构域的氨基酸序列约70%至约100%相同(例如,70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%、99.5%或100%相同以及其中的任何范围或值)。在一些实施方案中,UGI结构域可包含SEQ ID NO:41的氨基酸序列或与SEQ ID NO:41的氨基酸序列具有约70%至约99.5%序列同一性的多肽(例如,与SEQ ID NO:41的氨基酸序列具有至少80%,至少85%、至少90%、至少92%、至少95%、至少96%、至少97%、至少98%、至少99%或至少99.5%的序列同一性)。例如,在一些实施方案中,UGI结构域可以包含SEQ ID NO:41的氨基酸序列的片段,其与SEQ ID NO:41的氨基酸序列的连续核苷酸的一部分(例如,10、15、20、25、30、35、40、45、50、55、60、65、70、75、80个连续核苷酸;例如,约10、15、20、25、30、35、40、45至约50、55、60、65、70、75、80个连续核苷酸)100%相同。在一些实施方案中,UGI结构域可以是与已知UGI具有约70%至约99.5%序列同一性(例如,70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%、99.5%序列同一性,以及其中的任何范围或值)的已知UGI(例如,SEQ ID NO:41)的变体。在一些实施方案中,编码UGI的多核苷酸可以针对在植物(例如植物)中表达进行密码子优化,并且密码子优化的多肽可以与参考多核苷酸具有约70%至约99.5%的同一性。
可用于本发明的腺嘌呤脱氨酶(或腺苷脱氨酶)可以是来自任何生物体的任何已知的或以后鉴定的腺嘌呤脱氨酶(参见例如美国专利号10,113,163,其关于其腺嘌呤脱氨酶的公开内容通过引用并入本文)。腺嘌呤脱氨酶可以催化腺嘌呤或腺苷的水解脱氨。在一些实施方案中,腺嘌呤脱氨酶可分别催化腺苷或脱氧腺苷水解脱氨为肌苷或脱氧肌苷。在一些实施方案中,腺苷脱氨酶可以催化DNA中腺嘌呤或腺苷的水解脱氨。在一些实施方案中,由本发明的核酸构建体编码的腺嘌呤脱氨酶可以在靶核酸的有义链(例如“+”;模板)中产生A→G转换或在靶核酸的反义链(例如,“-”,互补的)中产生T→C转换。
在一些实施方案中,腺苷脱氨酶可以是天然存在的腺嘌呤脱氨酶的变体。因此,在一些实施方案中,腺苷脱氨酶可以与野生型腺嘌呤脱氨酶具有约70%至100%的同一性(例如,与天然存在的腺嘌呤脱氨酶约70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或100%相同,以及其中的任何范围或值)。在一些实施方案中,一种或多种脱氨酶在自然界中不存在并且可以被称为工程化的、突变的或进化的腺苷脱氨酶。因此,例如,工程化的、突变的或进化的腺嘌呤脱氨酶多肽或腺嘌呤脱氨酶结构域可以与天然存在的腺嘌呤脱氨酶多肽/结构域具有约70%至99.9%的同一性(例如,与天然存在的腺嘌呤脱氨酶多肽或腺嘌呤脱氨酶结构域约70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%、99.1%、99.2%、99.3%、99.4%、99.5%、99.6%、99.7%、99.8%或99.9%相同,以及其中的任何范围或值)。在一些实施方案中,腺苷脱氨酶可以来自细菌(例如,大肠杆菌、金黄色葡萄球菌、流感嗜血杆菌、新月柄杆菌(Caulobacter crescentus)等)。在一些实施方案中,编码腺嘌呤脱氨酶多肽/结构域的多核苷酸可以进行密码子优化以用于在植物中表达。
在一些实施方案中,腺嘌呤脱氨酶结构域可以是野生型tRNA特异性腺苷脱氨酶结构域,例如tRNA特异性腺苷脱氨酶(TadA)和/或突变/进化的腺苷脱氨酶结构域,例如突变/进化的tRNA特异性腺苷脱氨酶结构域(TadA*)。在一些实施方案中,TadA结构域可以来自大肠杆菌。在一些实施方案中,TadA可以被修饰,例如截短,相对于全长TadA缺失一个或多个N端和/或C端氨基酸(例如,相对于全长TadA,可能缺失1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、6、17、18、19或20个N端和/或C端氨基酸残基。在一些实施方案中,TadA多肽或TadA结构域不包含N-末端甲硫氨酸。在一些实施方案中,野生型大肠杆菌TadA包含SEQ ID NO:30的氨基酸序列。在一些实施方案中,突变/进化的大肠杆菌TadA*包含SEQ ID NO:31-40(例如,SEQ ID NO:31、32、33、34、35、36、37、38、39或40)的氨基酸序列。在一些实施方案中,编码TadA/TadA*的多核苷酸可以进行密码子优化以用于在植物中表达。
胞嘧啶脱氨酶催化胞嘧啶脱氨并产生胸苷(通过尿嘧啶中间体),导致基因组中互补链中的C到T转换或G到A转换。因此,在一些实施方案中,由本发明的多核苷酸编码的胞嘧啶脱氨酶在靶核酸的有义链(例如“+”;模板)中产生C→T转换或在靶核酸的反义链(例如,"-",互补的)中产生G→A转换。
在一些实施方案中,由本发明的核酸构建体编码的腺嘌呤脱氨酶在靶核酸的有义链(例如“+”;模板)中产生A→G转换或在靶核酸的反义链(例如,"-",互补的)中产生T→C转换。
本发明的编码包含序列特异性核酸结合蛋白和胞嘧啶脱氨酶多肽的碱基编辑器的核酸构建体,以及编码其的核酸构建体/表达盒/载体,可以与引导核酸组合使用以用于修饰靶标核酸包括但不限于在靶核酸(包括但不限于质粒序列)中产生C→T或G→A突变;在编码序列中产生C→T或G→A突变以改变氨基酸同一性;在编码序列中产生C→T或G→A突变以产生终止密码子;在编码序列中产生C→T或G→A突变以破坏起始密码子;在基因组DNA中产生点突变以破坏功能;和/或在基因组DNA中产生点突变以破坏剪接点。
本发明的编码包含序列特异性核酸结合蛋白和腺嘌呤脱氨酶多肽的碱基编辑器的核酸构建体,以及编码其的表达盒和/或载体,可以与引导核酸组合使用以用于修饰靶核酸,包括但不限于在靶核酸(包括但不限于质粒序列)中产生A→G或T→C突变;在编码序列中产生A→G或T→C突变以改变氨基酸同一性;在编码序列中产生A→G或T→C突变以产生终止密码子;在编码序列中产生A→G或T→C突变以破坏起始密码子;在基因组DNA中产生点突变以破坏功能;和/或在基因组DNA中产生点突变以破坏剪接点。
包含CRISPR-Cas效应蛋白或其融合蛋白的本发明的核酸构建体可以与引导RNA(gRNA、CRISPR阵列、CRISPR RNA、crRNA)组合使用,该引导RNA被设计成与编码的CRISPR-Cas效应蛋白或结构域一起发挥作用以修饰靶核酸。可用于本发明的引导核酸包含至少一个间隔序列和至少一个重复序列。引导核酸能够与由本发明的核酸构建体编码和表达的CRISPR-Cas核酸酶结构域形成复合物,并且间隔序列能够与靶核酸杂交,从而引导复合物(例如,CRISPR-Cas效应融合蛋白(例如,与脱氨酶结构域融合的CRISPR-Cas效应结构域和/或与肽标签或亲和多肽融合以募集脱氨酶结构域和任选地UGI的CRISPR-Cas效应结构域)至靶核酸,其中靶核酸可以被脱氨酶结构域修饰(例如,切割或编辑)或调节(例如,调节转录)。
例如,编码与胞嘧啶脱氨酶结构域(例如,融合蛋白)连接的Cas9结构域的核酸构建体可以与Cas9引导核酸组合使用以修饰靶核酸,其中融合蛋白的胞嘧啶脱氨酶结构域使靶核酸中的胞嘧啶碱基脱氨基,从而编辑靶核酸。在另一个实例中,编码与腺嘌呤脱氨酶结构域(例如,融合蛋白)连接的Cas9结构域的核酸构建体可与Cas9引导核酸组合使用以修饰靶核酸,其中融合蛋白的腺嘌呤脱氨酶结构域使靶核酸中的腺苷碱基脱氨基,从而编辑靶核酸。
同样,编码与胞嘧啶脱氨酶结构域或腺嘌呤脱氨酶结构域连接的Cas12a结构域(或其他选择的CRISPR-Cas核酸酶,例如C2c1、C2c3、Cas12b、Cas12c、Cas12d、Cas12e、Cas13a、Cas13b、Cas13c、Cas13d、Cas1、Cas1B、Cas2、Cas3、Cas3'、Cas3"、Cas4、Cas5、Cas6、Cas7、Cas8、Cas9(也称为Csnl和Csx12)、Cas10、Csyl、Csy2、Csy3、Csel、Cse2、Cscl、Csc2、Csa5、Csn2、Csm2、Csm3、Csm4、Csm5、Csm6、Cmrl、Cmr3、Cmr4、Cmr5、Cmr6、Csbl、Csb2、Csb3、Csxl7、Csxl4、Csx10、Csx16、CsaX、Csx3、Csxl、Csxl5、Csfl、Csf2、Csf3、Csf4(dinG)和/或Csf5)(例如,融合蛋白)可与Cas12a引导核酸(或其他选择的CRISPR-Cas核酸酶的引导核酸)组合使用以修饰靶核酸,其中融合蛋白的胞嘧啶脱氨酶结构域或腺嘌呤脱氨酶结构域使靶核酸中的胞嘧啶碱基脱氨基,从而编辑靶核酸。
本文所用的“引导核酸”、“引导RNA”、“gRNA”、“CRISPR RNA/DNA”、“crRNA”或“crDNA”是指包含至少一个与靶DNA(例如,前间隔区)互补(和杂交)的间隔序列和至少一个重复序列(例如,V型Cas12a CRISPR-Cas系统的重复序列,或其片段或部分;II型Cas9CRISPR-Cas系统的重复序列,或其片段;V型C2c1 CRISPR Cas系统的重复序列,或其片段;例如C2c3、Cas12a(也称为Cpf1)、Cas12b、Cas12c、Cas12d、Cas12e、Cas13a、Cas13b、Cas13c、Cas13d、Casl、CaslB、Cas2、Cas3、Cas3'、Cas3"、Cas4、Cas5、Cas6、Cas7、Cas8、Cas9(也称为Csnl和Csx12)、Cas10、Csyl、Csy2、Csy3、Csel、Cse2、Cscl、Csc2、Csa5、Csn2、Csm2、Csm3、Csm4、Csm5、Csm6、Cmrl、Cmr3、Cmr4、Cmr5、Cmr6、Csbl、Csb2、Csb3、Csxl7、Csxl4、Csx10、Csx16、CsaX、Csx3、Csxl、Csxl5、Csfl、Csf2、Csf3、Csf4(dinG)和/或Csf5的CRISPR-Cas系统的重复序列,或其片段)的核酸,其中重复序列可以连接到间隔序列的5'端和/或3'端。本发明的gRNA的设计可以基于I型、II型、III型、IV型、V型或VI型CRISPR-Cas系统。
在一些实施方案中,Cas12a gRNA可以从5'到3'包含重复序列(全长或其部分(“柄”);例如,假结样结构)和间隔序列。
在一些实施方案中,引导核酸可以包含多于一个重复序列-间隔序列(例如,2、3、4、5、6、7、8、9、10个或更多个重复序列-间隔序列)(例如,重复序列-间隔序列-重复序列,例如,重复序列-间隔序列-重复序列-间隔序列-重复序列-间隔序列-重复序列-间隔序列-重复序列-间隔序列等)。本发明的引导核酸是合成的、人造的并且在自然界中不存在。gRNA可以很长,并且可以用作适配体(如在MS2募集策略中)或拉开(hanging off)间隔区上的其他RNA结构。
如本文所用,“重复序列”是指例如野生型CRISPR Cas基因座(例如,Cas9基因座、Cas12a基因座、C2c1基因座等)的任何重复序列或与本发明的核酸构建体编码的CRISPR-Cas效应蛋白一起起作用的合成crRNA。可用于本发明的重复序列可以是CRISPR-Cas基因座的任何已知或以后鉴定的重复序列(例如,I型、II型、III型、IV型、V型或VI型),或者它可以是设计用于I、II、III、IV、V或VI型CRISPR-Cas系统的合成的重复序列。重复序列可以包含发夹结构和/或茎环结构。在一些实施方案中,重复序列可以在其5'端形成假结样结构(即,“柄”)。因此,在一些实施方案中,重复序列可以与来自野生型I型、CRISPR-Cas基因座、II型CRISPR-Cas基因座、III型CRISPR-Cas基因座、IV型CRISPR-Cas基因座、V型CRISPR-Cas基因座和/或VI型CRISPR-Cas基因座的重复序列相同或基本相同。来自野生型CRISPR-Cas基因座的重复序列可以通过已建立的算法来确定,例如使用通过CRISPRdb提供的CRISPRfinder(参见Grissa等人Nucleic Acids Res.35(Web Server issue):W52-7)。在一些实施方案中,重复序列或其部分在其3'末端连接到间隔序列的5'末端,从而形成重复间隔序列(例如,引导核酸、引导RNA/DNA、crRNA、crDNA)。
在一些实施方案中,重复序列包含、基本上由或由至少10个核苷酸组成,这取决于特定的重复序列以及包含重复序列的引导核酸是加工的还是未加工的(例如,约10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50至100个或更多个核苷酸,或其中的任何范围或值)。在一些实施方案中,重复序列包含约10至约20、约10至约30、约10至约45、约10至约50、约15至约30、约15至约40、约15至约45、约15至约50、约20至约30、约20至约40、约20至约50、约30至约40、约40至约80、约50至约100或更多个核苷酸、基本上由其组成或由其组成。
与间隔序列的5'末端连接的重复序列可以包含重复序列的一部分(例如,野生型重复序列的5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35个或更多个连续核苷酸)。在一些实施方案中,与间隔序列的5'末端连接的重复序列的一部分的长度可以是约5至约10个连续核苷酸(例如,约5、6、7、8、9、10个核苷酸)并且与野生型CRISPR Cas重复核苷酸序列的相同区域(例如,5'端)具有至少90%的序列同一性(例如,至少约90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或更多(例如,99.1、99.2、99.3、99.4、99.5、99.6、99.7、99.8、99.9或100%))。在一些实施方案中,重复序列的一部分可以在其5'端包含假结样结构(例如,“柄”)。
如本文所用,“间隔序列”是与靶核酸(例如,靶DNA)(例如,前间隔区)(例如,(a)包含与SEQ ID NO:72-77中的任一个具有至少80%序列同一性的连续核苷酸的序列;(b)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的连续氨基酸的序列;(c)包含编码与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的连续氨基酸的序列的区域;或(d)包含编码与SEQ ID NO:93具有至少90%序列同一性的连续氨基酸的序列的区域的序列的连续核苷酸的一部分)互补的核苷酸序列。在一些实施方案中,间隔序列(例如,一个或多个间隔区)可包括但不限于SEQ ID NO:94-118中任一个的核苷酸序列。间隔序列可以与靶核酸完全互补或基本上互补(例如,至少约70%互补(例如,约70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或更多(例如,99.1、99.2、99.3、99.4、99.5、99.6、99.7、99.8、99.9或100%))。因此,在一些实施方案中,与靶核酸相比,间隔序列可以具有一个、两个、三个、四个或五个错配,这些错配可以是连续的或不连续的。在一些实施方案中,间隔序列可以与靶核酸具有70%的互补性。在其他实施方案中,间隔核苷酸序列可以与靶核酸具有80%的互补性。在其他实施方案中,间隔核苷酸序列可以与靶核酸(前间隔区)具有85%、90%、95%、96%、97%、98%、99%或99.5%的互补性等。在一些实施方案中,间隔序列与靶核酸100%互补。间隔序列可具有约15个核苷酸至约30个核苷酸(例如,15、16、17、18、19、20、21、22、23、24、25、26、27、28、29或30个核苷酸或其中的任何范围或值)的长度。因此,在一些实施方案中,间隔序列可在长度为至少约15个核苷酸至约30个核苷酸的靶核酸(例如,前间隔区)的区域上具有完全互补性或基本互补性。在一些实施方案中,间隔区的长度为约20个核苷酸。在一些实施方案中,间隔区的长度为约21、22或23个核苷酸。
在一些实施方案中,引导核酸的间隔序列的5'区域可以与靶DNA相同,而间隔区的3'区域可以与靶DNA基本上互补(例如,V型CRISPR-Cas),或引导核酸的间隔序列的3'区域可以与靶DNA相同,而间隔区的5'区域可以与靶DNA基本上互补(例如,II型CRISPR-Cas),并且因此,间隔序列与靶DNA的总体互补性可能低于100%。因此,例如,在V型CRISPR-Cas系统的引导物中,例如20个核苷酸的间隔序列的5'区域(即种子区域)中的前1、2、3、4、5、6、7、8、9、10个核苷酸可以与靶DNA 100%互补,而间隔序列3’区域中的剩余核苷酸与靶DNA基本上互补(例如,至少约70%互补)。在一些实施方案中,间隔序列的5'末端的前1至8个核苷酸(例如,前1、2、3、4、5、6、7、8个核苷酸和其中的任何范围)可以与靶DNA 100%互补,而间隔序列3'区域中的剩余核苷酸与靶DNA基本上互补(例如,至少约50%互补(例如,50%、55%、60%、65%、70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或更多))。
作为另一个例子,在II型CRISPR-Cas系统的引导物中,例如20个核苷酸的间隔序列的3'区域(即种子区域)中的前1、2、3、4、5、6、7、8、9、10个核苷酸可以与靶DNA 100%互补,而间隔序列的5'区域中的剩余核苷酸与靶DNA基本上互补(例如,至少约70%互补)。在一些实施方案中,间隔序列的3'端的前1至10个核苷酸(例如,前1、2、3、4、5、6、7、8、9、10个核苷酸,以及其中的任何范围)可以与靶DNA 100%互补,而间隔序列的5'区域的剩余核苷酸与靶DNA基本上互补(例如,至少约50%互补(例如,至少约50%、55%、60%、65%、70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或更多或其中的任何范围或值))。
在一些实施方案中,间隔区的种子区可以是约8至约10个核苷酸的长度、约5至约6个核苷酸的长度或约6个核苷酸的长度。
如本文所用,“靶核酸”、“靶DNA”、“靶核苷酸序列”、“靶区域”或“基因组中的靶区域”是指植物基因组中与本发明的引导核酸中的间隔序列完全互补(100%互补)或基本互补(例如,至少70%互补(例如,70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%或更多))的区域。可用于CRISPR-Cas系统的靶区域可以紧邻生物体基因组(例如植物基因组)中的PAM序列的3'(例如,对于V型CRISPR-Cas系统)或紧邻其5'(例如,对于II型CRISPR-Cas系统)定位。靶区域可以选自紧邻PAM序列定位的至少15个连续核苷酸(例如,16、17、18、19、20、21、22、23、24、25、26、27、28、29、30个核苷酸等)的任何区域。
“前间隔序列”是指靶双链DNA,并且特别是指与CRISPR重复序列-间隔序列(例如,引导核酸、CRISPR阵列、crRNA)完全或基本上互补(和杂交)的靶DNA部分(例如,或基因组中的靶区域)。
在V型CRISPR-Cas(例如,Cas12a)系统和II型CRISPR-Cas(Cas9)系统的情况下,前间隔序列的两侧是(例如,紧邻)前间隔区邻近基序(PAM)。对于IV型CRISPR-Cas系统,PAM位于非靶标链的5'端和靶标链的3'端(例如,见下文)。
在II型CRISPR-Cas(例如Cas9)系统的情况下,PAM紧邻靶区域的3'定位。I型CRISPR-Cas系统的PAM位于靶标链的5'。没有已知的用于III型CRISPR-Cas系统的PAM。Makarova等人描述了CRISPR系统的所有类别、类型和亚型的命名法(Nature ReviewsMicrobiology 13:722-736(2015))。R.Barrangou(Genome Biol.16:247(2015))描述了引导物结构和PAM。
典型的Cas12a PAM富含T。在一些实施方案中,典型的Cas12aPAM序列可以是5'-TTN、5'-TTTN或5'-TTTV。在一些实施方案中,典型的Cas9(例如,化脓性链球菌)PAM可以是5'-NGG-3'。在一些实施方案中,可以使用非典型PAM,但效率可能较低。
本领域技术人员可以通过已建立的实验和计算方法确定另外的PAM序列。因此,例如,实验方法包括靶向侧接所有可能的核苷酸序列的序列并识别不经历靶向的序列成员,例如通过靶质粒DNA的转化(Esvelt等人,2013.Nat.Methods 10:1116-1121;Jiang等人,2013.Nat.Biotechnol.31:233-239)。在一些方面,计算方法可以包括对天然间隔区进行BLAST搜索以鉴定噬菌体或质粒中的原始靶DNA序列,并比对这些序列以确定与靶序列相邻的保守序列(Briner和Barrangou.2014.Appl.Environ.Microbiol.80:994-1001;Mojica等人2009.Microbiology 155:733-740)。
在一些实施方案中,本发明提供了包含本发明的核酸构建体(例如,本发明的编辑系统的一种或多种组分)的表达盒和/或载体。在一些实施方案中,可以提供包含本发明的核酸构建体和/或一种或多种引导核酸的表达盒和/或载体。在一些实施方案中,编码碱基编辑器(例如,包含CRISPR-Cas效应蛋白和脱氨酶结构域的构建体(例如,融合蛋白))的本发明的核酸构建体或用于碱基编辑的组分(例如,与肽标签或亲和多肽融合的CRISPR-Cas效应蛋白、与肽标签或亲和多肽融合的脱氨酶结构域和/或与肽标签或亲和多肽融合的UGI)可以包含在相同的表达盒或载体上或在与包含一种或多种引导核酸的表达盒或载体分开的表达盒或载体上。当编码碱基编辑器的核酸构建体或用于碱基编辑的组分包含在与包含引导核酸的表达盒或载体不同的表达盒或载体上时,靶核酸可以彼此以任何顺序接触(例如,与其一起提供)编码碱基编辑器或用于碱基编辑的组分的表达盒或载体,例如,在包含引导核酸的表达盒之前、同时或之后提供(例如,与靶核酸接触)。
本发明的融合蛋白可以包含与如本领域已知的肽标签或与肽标签相互作用的亲和多肽融合的序列特异性核酸结合结构域(序列特异性DNA结合结构域)、CRISPR-Cas多肽和/或脱氨酶结构域,用于募集脱氨酶到靶核酸。募集方法还可以包括与RNA募集基序连接的引导核酸和与能够与RNA募集基序相互作用的亲和多肽融合的脱氨酶,从而将脱氨酶募集至靶核酸。或者,化学相互作用可用于将多肽(例如脱氨酶)募集到靶核酸。
可用于本发明的肽标签(例如表位)可以包括但不限于GCN4肽标签(例如Sun-Tag)、c-Myc亲和标签、HA亲和标签、His亲和标签、S亲和标签、甲硫氨酸-His亲和标签、RGD-His亲和标签、FLAG八肽、strep标签或strep标签II、V5标签和/或VSV-G表位。可以与多肽连接并且存在可以与另一多肽连接的相应亲和多肽的任何表位可用于本发明作为肽标签。在一些实施方案中,肽标签可包含1或2个或更多个拷贝的肽标签(例如,重复单元、多聚化表位(例如,串联重复))(例如,1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25或更多个重复单元)。在一些实施方案中,与肽标签相互作用/结合的亲和多肽可以是抗体。在一些实施方案中,抗体可以是scFv抗体。在一些实施方案中,与肽标签结合的亲和多肽可以是合成的(例如,进化用于亲和相互作用),包括,但不限于,affibody、anticalin、monobody和/或DARPin(参见,例如,Sha等人,Protein Sci.26(5):910-924(2017));Gilbreth(Curr Opin Struc Biol 22(4):413-420(2013)),美国专利号9,982,053,其各自关于affibody、anticalins、monobody和/或DARPin的教导内容通过引用整体并入本文。示例性肽标签序列及其亲和多肽包括但不限于SEQ ID NO:45-47的氨基酸序列。
在一些实施方案中,引导核酸可与RNA募集基序连接,待募集的多肽(例如脱氨酶)可与结合RNA募集基序的亲和多肽融合,其中引导物与靶核酸结合和RNA募集基序与亲和多肽结合,从而将多肽募集到引导物并使靶核酸与多肽(例如脱氨酶)接触。在一些实施方案中,可以将两种或更多种多肽募集到引导核酸,从而使靶核酸与两种或更多种多肽(例如,脱氨酶)接触。示例性RNA募集基序及其亲和多肽包括但不限于SEQ ID NO:48-58的序列。
在一些实施方案中,与亲和多肽融合的多肽可以是逆转录酶并且引导核酸可以是与RNA募集基序连接的延伸的引导核酸。在一些实施方案中,RNA募集基序可以位于延伸的引导核酸的延伸部分的3'末端(例如,5'-3',重复序列-间隔区-延伸部分(RT模板-引物结合位点)-RNA募集基序)。在一些实施方案中,RNA募集基序可以嵌入延伸部分中。
在本发明的一些实施方案中,延伸的引导RNA和/或引导RNA可以与一个或两个或更多个RNA募集基序(例如,1、2、3、4、5、6、7、8、9、10个或更多个基序;例如,至少10至约25个基序)连接,任选地其中两个或更多个RNA募集基序可以是相同的RNA募集基序或不同的RNA募集基序。在一些实施方案中,RNA募集基序和相应的亲和多肽可以包括但不限于端粒酶Ku结合基序(例如,Ku结合发夹)和相应的亲和多肽Ku(例如,Ku异二聚体),端粒酶Sm7结合基序和相应的亲和多肽Sm7、MS2噬菌体操纵子茎环和相应的亲和多肽MS2外壳蛋白(MCP)、PP7噬菌体操纵子茎环和相应的亲和多肽PP7外壳蛋白(PCP)、SfMu噬菌体Com茎环和相应的亲和多肽Com RNA结合蛋白、PUF结合位点(PBS)和亲和多肽Pumilio/fem-3mRNA结合因子(PUF),和/或合成的RNA适配体和适配体配体作为相应的亲和多肽。在一些实施方案中,RNA募集基序和相应的亲和多肽可以是MS2噬菌体操纵子茎环和亲和多肽MS2外壳蛋白(MCP)。在一些实施方案中,RNA募集基序和相应的亲和多肽可以是PUF结合位点(PBS)和亲和多肽Pumilio/fem-3m RNA结合因子(PUF)。
在一些实施方案中,用于募集多肽和核酸的组分可以是通过化学相互作用起作用的那些组分,其可以包括但不限于雷帕霉素诱导的FRB-FKBP二聚化;生物素-链霉亲和素;SNAP标签;Halo标签;CLIP标签;化合物诱导的DmrA-DmrC异二聚体;双功能配体(例如,两种蛋白质结合化学物质一起的融合体;例如二氢叶酸还原酶(DHFR)。
在一些实施方案中,经优化以用于在植物中表达的本发明的核酸构建体、表达盒或载体可以与包含相同多核苷酸但尚未针对在植物中的表达进行密码子优化的核酸构建体、表达盒或载体约70%至100%相同(例如,约70%、71%、72%、73%、74%、75%、76%、77%、78%、79%、80%、81%、82%、83%、84%、85%、86%、87%、88%、89%、90%、91%、92%、93%、94%、95%、96%、97%、98%、99%、99.5%或100%)。
本文还提供了包含本发明的一种或多种多核苷酸、引导核酸、核酸构建体、表达盒或载体的细胞。
本发明的核酸构建体(例如,包含序列特异性DNA结合结构域、CRISPR-Cas效应结构域、脱氨酶结构域、逆转录酶(RT)、RT模板和/或引导核酸等的构建体)和包含它们的表达盒/载体可以用作本发明的编辑系统用于修饰靶核酸和/或其表达。
任何植物或植物部分(或植物分组,例如属或更高级分类)的靶核酸可以使用本发明的多肽、多核苷酸、核糖核蛋白(RNP)、核酸构建体、表达盒和/或载体修饰(例如,突变,例如碱基编辑、切割、切口等),包括被子植物、裸子植物、单子叶植物、双子叶植物、C3、C4、CAM植物、苔藓植物、蕨类植物和/或拟蕨植物、微藻和/或大型藻类。可如本文所述修饰的植物和/或植物部分可以是任何植物物种/品种/栽培品种的植物和/或植物部分。在一些实施方案中,可如本文所述修饰的植物是单子叶植物。在一些实施方案中,可如本文所述修饰的植物是双子叶植物。
如本文所用,术语“植物部分”包括但不限于生殖组织(例如,花瓣、萼片、雄蕊、雌蕊、花托、花药、花粉、花、果实、花蕾、胚珠、种子、胚、坚果、仁、穗、穗轴和壳);营养组织(例如,叶柄、茎、根、根毛、根尖、髓、胚芽鞘、茎、芽、枝、树皮、顶端分生组织、腋芽、子叶、下胚轴和叶);维管组织(例如韧皮部和木质部);特化细胞,例如表皮细胞、薄壁细胞、胆囊细胞、厚壁细胞、气孔、保卫细胞、角质层、叶肉细胞;愈伤组织;和插条。术语“植物部分”还包括植物细胞,包括在植物和/或植物部分、植物原生质体、植物组织、植物器官、植物细胞组织培养物、植物愈伤组织、植物团块等中完整的植物细胞。如本文所用,“芽”是指地上部分,包括叶和茎。如本文所用,术语“组织培养物”包括组织、细胞、原生质体和愈伤组织的培养物。
如本文所用,“植物细胞”是指植物的结构和生理单位,其通常包括细胞壁,但也包括原生质体。本发明的植物细胞可以是分离的单细胞的形式,或者可以是培养的细胞,或者可以是更高组织单元例如植物组织(包括愈伤组织)或植物器官的一部分。在一些实施方案中,植物细胞可以是藻类细胞。“原生质体”是没有细胞壁或仅具有部分细胞壁的分离的植物细胞。因此,在本发明的一些实施方案中,包含本发明的核酸分子和/或核苷酸序列的转基因细胞是任何植物或植物部分的细胞,包括但不限于根细胞、叶细胞、组织培养细胞、种子细胞、花细胞、果实细胞、花粉细胞等。在本发明的一些方面,植物部分可以是植物种质。在一些方面,植物细胞可以是不再生成植物的非繁殖植物细胞。
“植物细胞培养物”是指植物单元(例如原生质体、细胞培养细胞、植物组织中的细胞、花粉、花粉管、胚珠、胚囊、合子和处于不同发育阶段的胚)的培养物。
如本文所用,“植物器官”是植物的独特且明显的结构化和分化的部分,例如根、茎、叶、花蕾或胚。
如本文所用,“植物组织”是指组织成结构和功能单元的一组植物细胞。包括植物中或培养物中的任何植物组织。该术语包括但不限于整株植物、植物器官、植物种子、组织培养物和组织成结构和/或功能单元的任何植物细胞群。该术语与以上列出的或本定义所包含的任何特定类型的植物组织结合或不结合使用时,并不旨在排除任何其他类型的植物组织。
在本发明的一些实施方案中,提供了转基因组织培养物或转基因植物细胞培养物,其中转基因组织或细胞培养物包含本发明的核酸分子/核苷酸序列。在一些实施方案中,可以通过使转基因植物与非转基因植物育种并在后代中选择包含所需基因编辑而不是用于产生该编辑的转基因的植物,可以从由转基因组织或细胞发育而来的植物中消除转基因。
包含E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因(例如内源HECT E3 UPL3基因,例如内源HECT E3 UPL3a基因、HECT E3 UPL3b基因、HECT E3 UPL3c基因、HECT E3UPL3d基因)的任何植物)可以如本文所述进行修饰以改善一种或多种产量性状。可以如本文所述进行修饰的植物的非限制性实例可以包括但不限于草坪草(例如,早熟禾、翦股颖、黑麦草、羊茅)、羽苇草、丛生毛草、芒草、芦竹、柳枝稷、蔬菜作物,包括洋蓟、大头菜、芝麻菜、韭葱、芦笋、莴苣(例如头菜、叶菜、生菜)、黄肉芋、香瓜(例如甜瓜、西瓜、克伦肖瓜、白兰瓜、哈密瓜)、芸苔属作物(例如球芽甘蓝、卷心菜、花椰菜、西兰花、羽衣甘蓝、无头甘蓝、大白菜、白菜)、刺菜蓟、胡萝卜、纳帕白菜(napa)、秋葵、洋葱、芹菜、欧芹、鹰嘴豆、防风草、菊苣、辣椒、土豆、葫芦科植物(例如嫩葫芦、黄瓜、西葫芦、南瓜、番瓜、白兰瓜、西瓜、哈密瓜)、萝卜、干洋葱、芜菁甘蓝、茄子、婆罗门参、阔叶菊苣、青葱、苦苣、大蒜、菠菜、绿色洋葱、南瓜、绿叶蔬菜、甜菜(糖甜菜和饲料用甜菜)、甜薯、君达菜、辣根、西红柿、红萝卜和香料;水果作物,例如苹果、杏、樱桃、油桃、桃子、梨、李子、西梅、樱桃、榅桲、无花果、坚果(例如栗子、山核桃、开心果、榛子、开心果、花生、核桃、澳洲坚果、杏仁等)、柑橘(例如克莱门氏小柑橘、金橘、橙子、葡萄柚、橘子、柑橘(mandarin)、柠檬、酸橙等)、蓝莓、黑树莓、波森莓、酸果蔓、醋栗果、醋栗、罗甘莓、树莓、草莓、黑莓、葡萄(酒和佐餐)、牛油果、香蕉、猕猴桃、柿子、石榴、菠萝、热带水果、梨果、香瓜、芒果、木瓜和荔枝,大田作物如三叶草、苜蓿、梯牧草、月见草、草甸泡沫(meadow foam)、玉米/玉蜀黎(田间玉米、甜玉米、爆米花)、啤酒花、荷荷巴油、荞麦、红花、藜麦、小麦、大米、大麦、黑麦、小米、高粱、燕麦、小黑麦、高粱、烟草、木棉、豆科植物(豆类(例如,绿色和干燥的)、扁豆、豌豆、大豆)、油料植物(油菜(rape)、油菜(canola)、芥末、罂粟、橄榄、向日葵、椰子、蓖麻油植物、可可豆、花生、油棕)、浮萍、拟南芥、纤维植物(棉花、亚麻、大麻(hemp)、黄麻)、大麻(Cannabis)(例如Cannabis sativa、印度大麻(Cannabis indica)和粗大麻(Cannabis ruderalis))、樟科植物(肉桂、樟脑)或植物诸如咖啡、甘蔗、茶和天然橡胶植物;和/或花坛植物,例如开花植物、仙人掌、肉质植物和/或观赏植物(例如,玫瑰、郁金香、紫罗兰),以及树木,例如林木(阔叶树和常绿树,例如如针叶树;例如榆树、白蜡树、橡树、枫树、冷杉、云杉、雪松、松树、桦树、柏树、桉树、柳树)以及灌木和其他苗木。在一些实施方案中,本发明的核酸构建体和/或编码其的表达盒和/或载体可用于修饰玉米、大豆、小麦、油菜、水稻、番茄、胡椒或向日葵。
在一些实施方案中,可如本文所述修饰的植物可包括但不限于玉米、大豆、油菜、小麦、水稻、棉花、甘蔗、甜菜、大麦、燕麦、苜蓿、向日葵、红花、油棕榈、芝麻、椰子、烟草、马铃薯、甜薯、木薯、咖啡、苹果、李子、杏、桃、樱桃、梨、无花果、香蕉、柑橘、可可、鳄梨、橄榄、杏仁、核桃、草莓、西瓜、胡椒、葡萄、番茄、黄瓜或芸苔属植物物种(例如,甘蓝型油菜(B.napus)、甘蓝(B.oleracea)、芜菁(B.rapa)、芥菜(B.juncea)和/或黑芥(B.nigra))。在一些实施方案中,可如本文所述修饰的植物是双子叶植物。在一些实施方案中,可如本文所述修饰的植物是单子叶植物。在一些实施方案中,可以如本文所述进行修饰的植物是玉米(即,玉蜀黍(Zea mays))。在一些实施方案中,可以如本文所述进行修饰的植物是大豆(即,Glycine max)。
因此,根据本发明要优先处理的植物或植物栽培品种包括通过遗传修饰接受遗传物质的所有植物,所述遗传物质赋予这些植物特别有利的有用特性(“性状”)。此类特性的实例是更好的植物生长、活力、胁迫耐受性、站立性、抗倒伏性、养分吸收、植物营养和/或产量,特别是改善的生长、对高温或低温的增加的耐受性、对干旱或水分水平或土壤盐分的增加的耐受性、增强的开花性能、更容易收获、加速成熟、更高产量、更高质量和/或更高营养价值的收获产品、更好的储存寿命和/或收获产品的可加工性。
这样的特性的进一步和特别强调的例子是针对动物和微生物害虫(例如针对昆虫、蛛形纲动物、线虫、螨虫、蛞蝓和蜗牛,例如针对植物中形成的毒素)的增加的抗性。在编码赋予针对此类动物和微生物害虫特别是昆虫的耐受性特性的蛋白质的DNA序列中,将特别提及来自苏云金芽孢杆菌的编码文献中广泛描述且为本领域技术人员熟知的Bt蛋白质的遗传物质。还将提及从细菌如发光杆菌(Photorhabdus)中提取的蛋白质(WO97/17432和WO98/08932)。具体而言,将提及Bt Cry或VIP蛋白,其包括Cry1A、CryIAb、CryIAc、CryIIA、CryIIIA、CryIIIB2、Cry9c、Cry2Ab、Cry3Bb和CryIF蛋白或其毒性片段,以及它们的杂合体或组合,尤其是CrylF蛋白或衍生自CrylF蛋白的杂合体(例如杂合CrylA-CrylF蛋白或其毒性片段)、CrylA型蛋白或其毒性片段,优选CrylAc蛋白或衍生自CrylAc蛋白的杂合体(例如杂合CrylAb-CrylAc蛋白)或CrylAb或Bt2蛋白或其毒性片段、Cry2Ae、Cry2Af或Cry2Ag蛋白或其毒性片段、CrylA.105蛋白或其毒性片段、VIP3Aa19蛋白、VIP3Aa20蛋白、在COT202或COT203棉花事件中产生的VIP3A蛋白、如Estruch等人(1996),Proc Natl Acad Sci USA.28;93(11):5389-94所述的VIP3Aa蛋白或其毒性片段、如WO2001/47952中所述的Cry蛋白、来自异短杆菌属(Xenorhabdus)(如WO98/50427中所述)、沙雷氏菌属(Serratia)(特别是来自嗜虫沙雷氏菌(S.entomophila))或发光杆菌属物种菌株的杀虫蛋白,例如来自发光杆菌属的Tc-蛋白质,如WO98/08932中所述。本文还包括了这些蛋白质中的任何一种的与上述任何序列在一些氨基酸(1-10,优选1-5)中不同的任何变体或突变体,特别是它们的毒性片段的序列,或融合至转运肽,例如质体转运肽或另一种蛋白质或肽。
这种性质的另一个和特别强调的例子是赋予对一种或多种除草剂例如咪唑啉酮、磺酰脲、草甘膦或草丁膦的耐受性。在编码赋予转化的植物细胞和植物对某些除草剂的耐受性特性的蛋白质(即感兴趣的多核苷酸)的DNA序列中,将特别提及WO2009/152359中描述的bar或PAT基因或天蓝色链霉菌基因,其赋予对草铵膦除草剂的耐受性,编码合适的EPSPS(5-烯醇丙酮酸莽草酸-3-磷酸-合酶)的基因,其赋予对以EPSPS作为靶标的除草剂(尤其是诸如草甘膦及其盐的除草剂)的耐受性,编码草甘膦-n-乙酰转移酶的基因,或编码草甘膦氧化还原酶的基因。其他合适的除草剂耐受性状包括至少一种ALS(乙酰乳酸合酶)抑制剂(例如WO2007/024782)、突变的拟南芥ALS/AHAS基因(例如美国专利6,855,533)、编码赋予对2,4-D(2,4-二氯苯氧乙酸)的耐受性的2,4-D-单加氧酶的基因,和编码赋予对麦草畏(3,6-二氯-2-甲氧基苯甲酸)的耐受性的麦草畏单加氧酶的基因。
此类特性的进一步实例包括但不限于由于例如系统性获得性抗性(SAR)、系统素、植物抗毒素、诱导子以及抗性基因和相应表达的蛋白质和毒素而对植物病原真菌、细菌和/或病毒的增加的抗性。
可以根据本发明处理的转基因植物或植物栽培品种中特别有用的转基因事件包括事件531/PV-GHBK04(棉花,昆虫控制,描述于WO2002/040677),事件1143-14A(棉花,昆虫控制,未保藏,描述于WO2006/128569);事件1143-51B(棉花,昆虫控制,未保藏,描述于WO2006/128570);事件1445(棉花、除草剂耐受性、未保藏,描述于US-A2002-120964或WO2002/034946);事件17053(水稻,除草剂耐受性,保藏为PTA-9843,描述于WO2010/117737);事件17314(水稻,除草剂耐受性,保藏为PTA-9844,描述于WO2010/117735);事件281-24-236(棉花,昆虫控制-除草剂耐受性,保藏为PTA-6233,描述于WO2005/103266或US-A2005-216969);事件3006-210-23(棉花,昆虫控制-除草剂耐受性,保藏为PTA-6233,描述于US-A2007-143876或WO2005/103266);事件3272(玉米,质量性状,保藏为PTA-9972,描述于WO2006/098952或US-A2006-230473);事件33391(小麦,除草剂耐受性,保藏为PTA-2347,描述于WO2002/027004),事件40416(玉米,昆虫控制-除草剂耐受性,保藏为ATCC PTA-11508,描述于WO11/075593);事件43A47(玉米,昆虫控制-除草剂耐受性,保藏为ATCC PTA-11509,描述于WO2011/075595);事件5307(玉米,昆虫控制,保藏为ATCC PTA-9561,描述于WO2010/077816);事件ASR-368(常绿草,除草剂耐受性,保藏为ATCC PTA-4816,描述于US-A2006-162007或WO2004/053062);事件B16(玉米,除草剂耐受性,未保藏,描述于US-A2003-126634);事件BPS-CV127-9(大豆,除草剂耐受性,保藏为NCIMB No.41603,描述于WO2010/080829);事件BLR1(油菜,雄性不育的恢复,保藏为NCIMB 41193,描述于WO2005/074671),事件CE43-67B(棉花,昆虫控制,保藏为DSM ACC2724,描述于US-A2009-217423或WO2006/128573);事件CE44-69D(棉花,昆虫控制,未保藏,描述于US-A2010-0024077);事件CE44-69D(棉花,昆虫控制,未保藏,描述于WO2006/128571);事件CE46-02A(棉花,昆虫控制,未保藏,描述于WO2006/128572);事件COT102(棉花,昆虫控制,未保藏,描述于US-A2006-130175或WO2004/039986);事件COT202(棉花,昆虫控制,未保藏,描述于US-A2007-067868或WO2005/054479);事件COT203(棉花,昆虫控制,未保藏,描述于WO2005/054480);事件DAS21606-3/1606(大豆,除草剂耐受性,保藏为PTA-11028,描述于WO2012/033794),事件DAS40278(玉米,除草剂耐受性,保藏为ATCC PTA-10244,描述于WO2011/022469);事件DAS-44406-6/pDAB8264.44.06.l(大豆,除草剂耐受性,保藏为PTA-11336,描述于WO2012/075426),事件DAS-14536-7/pDAB8291.45.36.2(大豆,除草剂耐受性,保藏为PTA-11335,描述于WO2012/075429),事件DAS-59122-7(玉米,昆虫控制-除草剂耐受性,保藏为ATCC PTA11384,描述于US-A2006-070139);事件DAS-59132(玉米,昆虫控制-除草剂耐受性,未保藏,描述于WO2009/100188);事件DAS68416(大豆,除草剂耐受性,保藏为ATCC PTA-10442,描述于WO2011/066384或WO2011/066360);事件DP-098140-6(玉米,除草剂耐受性,保藏为ATCCPTA-8296,描述于US-A2009-137395或WO08/112019);事件DP-305423-1(大豆,质量性状,未保藏,描述于US-A2008-312082或WO2008/054747);事件DP-32138-1(玉米,杂交系统,保藏为ATCC PTA-9158,描述于US-A2009-0210970或WO2009/103049);事件DP-356043-5(大豆,除草剂耐受性,保藏为ATCC PTA-8287,描述于US-A2010-0184079或WO2008/002872);事件EE-I(茄子,昆虫控制,未保藏,描述于WO07/091277);事件Fil17(玉米,除草剂耐受性,保藏为ATCC209031,描述于US-A2006-059581或WO98/044140);事件FG72(大豆,除草剂耐受性,保藏为PTA-11041,描述于WO2011/063413),事件GA21(玉米,除草剂耐受性,保藏为ATCC209033,描述于US-A2005-086719或WO98/044140);事件GG25(玉米,除草剂耐受性,保藏为ATCC209032,描述于US-A2005-188434或WO98/044140);事件GHB119(棉花,昆虫控制-除草剂耐受性,保藏为ATCC PTA-8398,描述于WO2008/151780);事件GHB614(棉花,除草剂耐受性,保藏为ATCC PTA-6878,描述于US-A2010-050282或W02007/017186);事件GJ11(玉米,除草剂耐受性,保藏为ATCC209030,描述于US-A2005-188434或WO98/044140);事件GMRZ13(甜菜,病毒抗性,保藏为NCIMB-41601,描述于WO2010/076212);事件H7-l(甜菜,除草剂耐受性,保藏为NCIMB41158或NCIMB41159,描述于US-A2004-172669或WO2004/074492);事件JOPLINl(小麦,抗病性,未保藏,描述于US-A2008-064032);事件LL27(大豆,除草剂耐受性,保藏为NCIMB41658,描述于WO2006/108674或US-A2008-320616);事件LL55(大豆,除草剂耐受性,保藏为NCIMB41660,描述于WO2006/108675或US-A2008-196127);事件LLcotton25(棉花,除草剂耐受性,保藏为ATCC PTA-3343,描述于WO2003/013224或US-A2003-097687);事件LLRICE06(水稻,除草剂耐受性,保藏为ATCC203353,描述于US6,468,747或WO2000/026345);事件LLRice62(水稻,除草剂耐受性,保藏为ATCC203352,描述于WO2000/026345),事件LLRICE601(水稻,除草剂耐受性,保藏为ATCC PTA-2600,描述于US-A2008-2289060或WO2000/026356);事件LY038(玉米,质量性状,保藏为ATCC PTA-5623,描述于US-A2007-028322或WO2005/061720);事件MIR162(玉米,昆虫控制,保藏为PTA-8166,描述于US-A2009-300784或WO2007/142840);事件MIR604(玉米,昆虫控制,未保藏,描述于US-A2008-167456或WO2005/103301);事件MON15985(棉花,昆虫控制,保藏为ATCC PTA-2516,描述于US-A2004-250317或WO2002/100163);事件MON810(玉米,昆虫控制,未保藏,描述于US-A2002-102582);事件MON863(玉米,昆虫控制,以ATCC PTA-2605保藏,描述于WO2004/011601或US-A2006-095986);事件MON87427(玉米,授粉对照,保藏为ATCC PTA-7899,描述于WO2011/062904);事件MON87460(玉米,胁迫耐受性,保藏为ATCC PTA-8910,描述于WO2009/111263或US-A2011-0138504);事件MON87701(大豆,昆虫控制,保藏为ATCCPTA-8194,描述于US-A2009-130071或WO2009/064652);事件MON87705(大豆,质量性状-除草剂耐受性,保藏为ATCC PTA-9241,描述于US-A2010-0080887或WO2010/037016);事件MON87708(大豆,除草剂耐受性,保藏为ATCC PTA-9670,描述于WO2011/034704);事件MON87712(大豆,产量,保藏为PTA-10296,描述于WO2012/051199),事件MON87754(大豆,质量性状,保藏为ATCC PTA-9385,描述于WO2010/024976);事件MON87769(大豆,质量性状,保藏为ATCC PTA-8911,描述于US-A2011-0067141或WO2009/102873);事件MON88017(玉米,昆虫控制-除草剂耐受性,保藏为ATCC PTA-5582,描述于US-A2008-028482或WO2005/059103);事件MON88913(棉花,除草剂耐受性,保藏为ATCC PTA-4854,描述于WO2004/072235或US-A2006-059590);事件MON88302(油菜,除草剂耐受性,保藏为PTA-10955,描述于WO2011/153186),事件MON88701(棉花,除草剂耐受性,保藏为PTA-11754,描述于WO2012/134808),事件MON89034(玉米,昆虫控制,保藏为ATCC PTA-7455描述于WO07/140256或US-A2008-260932);事件MON89788(大豆,除草剂耐受性,保藏为ATCC PTA-6708,描述于US-A2006-282915或WO2006/130436);事件MSl 1(油菜,授粉控制-除草剂耐受性,保藏为ATCCPTA-850或PTA-2485,描述于WO2001/031042);事件MS8(油菜,授粉控制-除草剂耐受性,保藏为ATCC PTA-730,描述于WO2001/041558或US-A2003-188347);事件NK603(玉米,除草剂耐受性,保藏为ATCC PTA-2478,描述于US-A2007-292854);事件PE-7(水稻,昆虫控制,未保藏,描述于WO2008/114282);事件RF3(油菜,授粉控制-除草剂耐受性,保藏为ATCC PTA-730,描述于WO2001/041558或US-A2003-188347);事件RT73(油菜,除草剂耐受性,未保藏,描述于WO2002/036831或US-A2008-070260);事件SYHT0H2/SYN-000H2-5(大豆,除草剂耐受性,保藏为PTA-11226,描述于WO2012/082548),事件T227-1(甜菜,除草剂耐受性,未保藏,描述于WO2002/44407或US-A2009-265817);事件T25(玉米、除草剂耐受性、未保藏,描述于US-A2001-029014或WO2001/051654);事件T304-40(棉花,昆虫控制-除草剂耐受性,保藏为ATCC PTA-8171,描述于US-A2010-077501或WO2008/122406);事件T342-142(棉花,昆虫控制,未保藏,描述于WO2006/128568);事件TC1507(玉米,昆虫控制-除草剂耐受性,未保藏,描述于US-A2005-039226或WO2004/099447);事件VIP1034(玉米,昆虫控制-除草剂耐受性,保藏为ATCC PTA-3925,描述于WO2003/052073),事件32316(玉米,昆虫控制-除草剂耐受性,保藏为PTA-11507,描述于WO2011/084632),事件4114(玉米,昆虫控制-除草剂耐受性,保藏为PTA-11506,描述于WO2011/084621),事件EE-GM3/FG72(大豆,除草剂耐受性,ATCC登录号PTA-11041)任选与事件EE-GM1叠加/LL27或事件EE-GM2/LL55(WO2011/063413A2),事件DAS-68416-4(大豆,除草剂耐受性,ATCC登录号PTA-10442,WO2011/066360A1),事件DAS-68416-4(大豆,除草剂耐受性,ATCC登录号PTA-10442,WO2011/066384A1),事件DP-040416-8(玉米,昆虫控制,ATCC登录号PTA-11508,WO2011/075593A1),事件DP-043A47-3(玉米,昆虫对照,ATCC登录号PTA-11509,WO2011/075595A1),事件DP-004114-3(玉米,昆虫对照,ATCC登记号PTA-11506,WO2011/084621A1),事件DP-032316-8(玉米,昆虫控制,ATCC登录号PTA-11507,WO2011/084632A1),事件MON-88302-9(油菜,除草剂耐受性,ATCC登录号PTA-10955,WO2011/153186A1),事件DAS-21606-3(大豆,除草剂耐受性,ATCC登录号PTA-11028,WO2012/033794A2),事件MON-87712-4(大豆,质量性状,ATCC登录号PTA-10296,WO2012/051199A2),事件DAS-44406-6(大豆,叠加除草剂耐受性,ATCC登录号PTA-11336,WO2012/075426A1),事件DAS-14536-7(大豆,叠加除草剂耐受性,ATCC登录号PTA-11335,WO2012/075429A1),事件SYN-000H2-5(大豆,除草剂耐受性,ATCC登录号PTA-11226,WO2012/082548A2),事件DP-061061-7(油菜,除草剂耐受性,无可用的保藏号,WO2012071039A1),事件DP-073496-4(油菜,除草剂耐受性,无保藏号,US2012131692),事件8264.44.06.1(大豆,叠加除草剂耐受性,登录号PTA-11336,WO2012075426A2),事件8291.45.36.2(大豆,叠加除草剂耐受性,登录号PTA-11335,WO2012075429A2),事件SYHT0H2(大豆,ATCC登录号PTA-11226,WO2012/082548A2),事件MON88701(棉花ATCC登录号PTA-11754,WO2012/134808A1),事件KK179-2(苜蓿,ATCC登录号PTA-11833,WO2013/003558A1),事件pDAB8264.42.32.1(大豆,堆叠除草剂耐受性,ATCC登录号PTA-11993,WO2013/010094A1),事件MZDT09Y(玉米,ATCC登录号PTA-13025,WO2013/012775A1)。
赋予所需性状的基因/事件(例如,感兴趣的多核苷酸)也可以彼此组合存在于转基因植物中。转基因植物的实例包括但不限于农作物,例如谷物(小麦、水稻、黑小麦、大麦、黑麦、燕麦)、玉米、大豆、马铃薯、甜菜、甘蔗、西红柿、豌豆和其他类型的蔬菜、棉花、烟草、油菜以及水果植物(包括水果苹果、梨、柑橘类水果和葡萄),特别强调玉米、大豆、小麦、水稻、马铃薯、棉花、甘蔗、烟草和油菜。特别强调的特性是植物对昆虫、蛛形纲动物、线虫和蛞蝓和蜗牛的增加的抗性,以及植物对一种或多种除草剂的增加的抗性。
可根据本发明优选处理的此类植物、植物部分或植物种子的可商购实例包括以RIBROUNDUP/>VT DOUBLE/>VT TRIPLE/>BOLLGARD/>ROUNDUP READY 2/> ROUNDUP/>2XTENDTM,INTACTA RR2/>VISTIVE/>和/或XTENDFLEXTM商品名销售或分销的商业产品,例如植物种子。
现在将参考以下实施例描述本发明。应当理解,这些实施方案并非旨在将权利要求的范围限制于本发明,而是旨在作为某些实施方案的示例。本领域技术人员想到的示例性方法的任何变化都旨在落入本发明的范围内。
实施例
实施例1.编辑策略
设计策略以在11g107500(SEQ ID NO:72)和12g032500(SEQ ID NO:73)的大豆HECT E3泛素连接酶基因中产生敲除或敲低编辑。为了产生一系列等位基因,设计了包含与大豆HECT E3泛素连接酶基因内的靶标互补的间隔区(SEQ ID NO:94-97)(参见表1)的多个CRISPR-Cas引导核酸,并将其置于各种构建体中。
对携带HECT E3泛素连接酶基因中的编辑的品系进行筛选,那些显示大约10%的测序读数具有在靶向的基因中的编辑的品系将被推进到下一代。
包含不同间隔区的其他引导构建体(参见例如表1)被设计用于工程化靶向的编辑以生成其他大豆HECT E3泛素连接酶基因的敲除或敲低版本。
表1.用于靶向HECT E3泛素连接酶基因的示例性间隔区
实施例2.HECT E3泛素连接酶基因中的敲除编辑的表征
开发含有间隔区pWsp1159(SEQ ID NO:94)、pWsp1161(SEQ ID NO:96)、pWsp1160(SEQ ID NO:95)和pWsp1162(SEQ ID NO:97)的编辑构建体pWISE2921来产生11g107500(SEQ ID NO:72)和12g032500(SEQ ID NO:73)的HECT E3基因中的敲除编辑。
鉴定出两种大豆植物CE50904和CE50894,其在靶向的HECT E3基因11g107500(SEQID NO:72)和12g032500(SEQ ID NO:73)之一或两者中含有一系列编辑。将这些大豆植物转移到温室中结籽,并收集每株植物产生的E1种子。在E1代中分离HECT E3基因的经编辑的等位基因范围。E1代被种植在温室中,并允许植物结籽以产生E2种子群体。E2种子在温室中发芽,并对所得植物群体针对HECT E3基因的经编辑的等位基因范围以及改变的种子表型例如种子大小和产量进行分析。
实施例3.HECT E3基因12g032500的敲除编辑的表征
E0植物CE50894产生了E1植物CE75549。允许CE75549结籽,产生E2群体,将其种植用于如下所述的表型分析。对CE75549中经编辑的HECT E3基因12g032500的分析表明,该基因对于5bp缺失是纯合的,如表2所示。
如实施例2中所述,E0植物CE50904产生E1植物CE75626。允许CE75626结籽,产生E2群体,将其种植用于如下所述的表型分析。对CE75626中经编辑的HECT E3基因12g032500的分析表明,该基因对于10bp缺失是纯合的,如表2所示。
对源自CE75549的二十七个E2群体和源自CE75626的十五个E2群体针对种子表型(例如种子长度、种子面积和100粒种子重量)进行评估。通过对收集的种子进行成像并直接测量种子图像的长度和面积来确定种子长度和种子面积。从E2植物收集的种子均未表现出改变的种子表型。这些观察表明,单独的12g032500的这些特定编辑不足以产生所需的改变的种子表型。
表2.HECT E3基因12g032500的编辑的等位基因
实施例4.HECT E3基因11g107500的敲除编辑的表征
E0植物CE50904产生了E1植物CE75600。CE75600对于11g107500(SEQ ID NO:72)中的11bp缺失是纯合的,并且对于12g032500(SEQ ID NO:73)的实施例3中描述的相同10bp缺失也是纯合的。E0植物CE50904还产生E1植物CE75640,其被确定为对于11g107500(SEQ IDNO:72)中的11bp缺失是纯合的。经编辑的等位基因信息总结于下表3中。
E1植物CE75600和CE75640在温室中生长并使其结出E2种子,E2种子也种植在温室中。对E2植物的产量性状进行评估,并且还允许结出E3代种子。还评估了E3种子的改变的种子表型。通过对种子成像并测量种子面积和种子长度来收集一系列植物种子表型。与野生型种子相比,源自CE75640的E2群体的E3种子显示出种子面积的统计上显著的增加,以及100粒种子重量的增加。在源自CE75600的E2群体中没有观察到改变的种子表型,这表明12g032500中的编辑干扰了11g107500的经编辑的等位基因。这些观察结果表明,HECT E3基因中的敲除编辑可以改变大豆的种子结构。
表3.实施例4中的编辑的等位基因
实施例5:玉米中的催化位点变异
设计了同时编辑玉米HECT E3a基因Zm00001d004139和HECT E3b基因Zm00001d014920两者的催化位点的策略。使用农杆菌将含有间隔区PWsp1401(CCTAGTGTCATGACTTGCGCCAA SEQ ID NO:117)和PWsp1402(CCTAGTGTGATGACGTGCGCCAA SEQID NO:118)的CRISPR-Cas编辑构建体引入到干燥的切除的玉米胚中。转化的组织在体外在抗生素选择的情况下维持以再生阳性转化体,其被再生成为植物。通过下一代测序针对靶基因中的编辑筛选再生的植物,并且将那些显示约10%的测序读数具有在被靶向的基因中的编辑的植物转移至温室以结籽。从再生的植物(E0代)中收集组织用于DNA提取,并采用随后的分子筛选来鉴定HECT E3靶基因中的编辑。
表4.来自实施例5的编辑的等位基因
允许几株E0植物结籽以产生E1种子,并在个体植物水平上测定100粒种子重量。如表4所示,所有编辑均显示100粒种子重量的增加。这些观察结果表明,玉米HECT E3基因的催化位点中的编辑可以导致增加的种子大小。
表5.由经编辑的玉米植物产生的种子的100粒种子重量的测定。
“WT”是指对照同基因植物。
前述内容是对本发明的说明并且不应被解释为对其进行限制。本发明由所附权利要求限定,权利要求的等同物也包括在其中。
序列表
<110> 成对植物服务股份有限公司
MARRI, Pradeep R.
MATHEW, Lolita G.
KIM, HaeJin
<120> 用于改善产量性状的HECT E3泛素连接酶基因的修饰
<130> 1499.61.WO
<150> US 63/214,498
<151> 2021-06-24
<160> 128
<170> PatentIn version 3.5
<210> 1
<211> 1228
<212> PRT
<213> 未知
<220>
<223> 毛螺菌科物种(Lachnospiraceae sp.)
<400> 1
Met Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr
1 5 10 15
Leu Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp
20 25 30
Asn Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys
35 40 45
Gly Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp
50 55 60
Val Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu
65 70 75 80
Phe Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn
85 90 95
Leu Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn
100 105 110
Glu Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu
115 120 125
Pro Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe
130 135 140
Asn Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn
145 150 155 160
Met Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile
165 170 175
Asn Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys
180 185 190
Val Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys
195 200 205
Ile Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe
210 215 220
Phe Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile
225 230 235 240
Ile Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn
245 250 255
Glu Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys
260 265 270
Phe Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser
275 280 285
Phe Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe
290 295 300
Arg Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys
305 310 315 320
Leu Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile
325 330 335
Phe Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe
340 345 350
Gly Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp
355 360 365
Ile His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp
370 375 380
Arg Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu
385 390 395 400
Gln Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu
405 410 415
Ile Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser
420 425 430
Glu Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys
435 440 445
Asn Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys
450 455 460
Ser Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr
465 470 475 480
Asn Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile
485 490 495
Leu Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr
500 505 510
Gln Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro
515 520 525
Gln Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala
530 535 540
Thr Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys
545 550 555 560
Lys Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly
565 570 575
Asn Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met
580 585 590
Leu Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro
595 600 605
Ser Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly
610 615 620
Asp Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys
625 630 635 640
Asp Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn
645 650 655
Phe Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu
660 665 670
Val Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys
675 680 685
Glu Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile
690 695 700
Tyr Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His
705 710 715 720
Thr Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile
725 730 735
Arg Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys
740 745 750
Lys Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys
755 760 765
Asn Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr
770 775 780
Lys Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile
785 790 795 800
Ala Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu Val
805 810 815
Arg Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile Asp
820 825 830
Arg Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys Gly
835 840 845
Asn Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe Asn
850 855 860
Gly Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys Glu
865 870 875 880
Lys Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn Ile
885 890 895
Lys Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile Cys
900 905 910
Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu Asn
915 920 925
Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val Tyr Gln
930 935 940
Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met Val Asp Lys
945 950 955 960
Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys Gly Tyr Gln Ile
965 970 975
Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser Thr Gln Asn Gly Phe
980 985 990
Ile Phe Tyr Ile Pro Ala Trp Leu Thr Ser Lys Ile Asp Pro Ser Thr
995 1000 1005
Gly Phe Val Asn Leu Leu Lys Thr Lys Tyr Thr Ser Ile Ala Asp
1010 1015 1020
Ser Lys Lys Phe Ile Ser Ser Phe Asp Arg Ile Met Tyr Val Pro
1025 1030 1035
Glu Glu Asp Leu Phe Glu Phe Ala Leu Asp Tyr Lys Asn Phe Ser
1040 1045 1050
Arg Thr Asp Ala Asp Tyr Ile Lys Lys Trp Lys Leu Tyr Ser Tyr
1055 1060 1065
Gly Asn Arg Ile Arg Ile Phe Arg Asn Pro Lys Lys Asn Asn Val
1070 1075 1080
Phe Asp Trp Glu Glu Val Cys Leu Thr Ser Ala Tyr Lys Glu Leu
1085 1090 1095
Phe Asn Lys Tyr Gly Ile Asn Tyr Gln Gln Gly Asp Ile Arg Ala
1100 1105 1110
Leu Leu Cys Glu Gln Ser Asp Lys Ala Phe Tyr Ser Ser Phe Met
1115 1120 1125
Ala Leu Met Ser Leu Met Leu Gln Met Arg Asn Ser Ile Thr Gly
1130 1135 1140
Arg Thr Asp Val Asp Phe Leu Ile Ser Pro Val Lys Asn Ser Asp
1145 1150 1155
Gly Ile Phe Tyr Asp Ser Arg Asn Tyr Glu Ala Gln Glu Asn Ala
1160 1165 1170
Ile Leu Pro Lys Asn Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala
1175 1180 1185
Arg Lys Val Leu Trp Ala Ile Gly Gln Phe Lys Lys Ala Glu Asp
1190 1195 1200
Glu Lys Leu Asp Lys Val Lys Ile Ala Ile Ser Asn Lys Glu Trp
1205 1210 1215
Leu Glu Tyr Ala Gln Thr Ser Val Lys His
1220 1225
<210> 2
<211> 1307
<212> PRT
<213> 氨基酸球菌属物种(Acidaminococcus sp.)
<400> 2
Met Thr Gln Phe Glu Gly Phe Thr Asn Leu Tyr Gln Val Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Lys His Ile Gln
20 25 30
Glu Gln Gly Phe Ile Glu Glu Asp Lys Ala Arg Asn Asp His Tyr Lys
35 40 45
Glu Leu Lys Pro Ile Ile Asp Arg Ile Tyr Lys Thr Tyr Ala Asp Gln
50 55 60
Cys Leu Gln Leu Val Gln Leu Asp Trp Glu Asn Leu Ser Ala Ala Ile
65 70 75 80
Asp Ser Tyr Arg Lys Glu Lys Thr Glu Glu Thr Arg Asn Ala Leu Ile
85 90 95
Glu Glu Gln Ala Thr Tyr Arg Asn Ala Ile His Asp Tyr Phe Ile Gly
100 105 110
Arg Thr Asp Asn Leu Thr Asp Ala Ile Asn Lys Arg His Ala Glu Ile
115 120 125
Tyr Lys Gly Leu Phe Lys Ala Glu Leu Phe Asn Gly Lys Val Leu Lys
130 135 140
Gln Leu Gly Thr Val Thr Thr Thr Glu His Glu Asn Ala Leu Leu Arg
145 150 155 160
Ser Phe Asp Lys Phe Thr Thr Tyr Phe Ser Gly Phe Tyr Glu Asn Arg
165 170 175
Lys Asn Val Phe Ser Ala Glu Asp Ile Ser Thr Ala Ile Pro His Arg
180 185 190
Ile Val Gln Asp Asn Phe Pro Lys Phe Lys Glu Asn Cys His Ile Phe
195 200 205
Thr Arg Leu Ile Thr Ala Val Pro Ser Leu Arg Glu His Phe Glu Asn
210 215 220
Val Lys Lys Ala Ile Gly Ile Phe Val Ser Thr Ser Ile Glu Glu Val
225 230 235 240
Phe Ser Phe Pro Phe Tyr Asn Gln Leu Leu Thr Gln Thr Gln Ile Asp
245 250 255
Leu Tyr Asn Gln Leu Leu Gly Gly Ile Ser Arg Glu Ala Gly Thr Glu
260 265 270
Lys Ile Lys Gly Leu Asn Glu Val Leu Asn Leu Ala Ile Gln Lys Asn
275 280 285
Asp Glu Thr Ala His Ile Ile Ala Ser Leu Pro His Arg Phe Ile Pro
290 295 300
Leu Phe Lys Gln Ile Leu Ser Asp Arg Asn Thr Leu Ser Phe Ile Leu
305 310 315 320
Glu Glu Phe Lys Ser Asp Glu Glu Val Ile Gln Ser Phe Cys Lys Tyr
325 330 335
Lys Thr Leu Leu Arg Asn Glu Asn Val Leu Glu Thr Ala Glu Ala Leu
340 345 350
Phe Asn Glu Leu Asn Ser Ile Asp Leu Thr His Ile Phe Ile Ser His
355 360 365
Lys Lys Leu Glu Thr Ile Ser Ser Ala Leu Cys Asp His Trp Asp Thr
370 375 380
Leu Arg Asn Ala Leu Tyr Glu Arg Arg Ile Ser Glu Leu Thr Gly Lys
385 390 395 400
Ile Thr Lys Ser Ala Lys Glu Lys Val Gln Arg Ser Leu Lys His Glu
405 410 415
Asp Ile Asn Leu Gln Glu Ile Ile Ser Ala Ala Gly Lys Glu Leu Ser
420 425 430
Glu Ala Phe Lys Gln Lys Thr Ser Glu Ile Leu Ser His Ala His Ala
435 440 445
Ala Leu Asp Gln Pro Leu Pro Thr Thr Leu Lys Lys Gln Glu Glu Lys
450 455 460
Glu Ile Leu Lys Ser Gln Leu Asp Ser Leu Leu Gly Leu Tyr His Leu
465 470 475 480
Leu Asp Trp Phe Ala Val Asp Glu Ser Asn Glu Val Asp Pro Glu Phe
485 490 495
Ser Ala Arg Leu Thr Gly Ile Lys Leu Glu Met Glu Pro Ser Leu Ser
500 505 510
Phe Tyr Asn Lys Ala Arg Asn Tyr Ala Thr Lys Lys Pro Tyr Ser Val
515 520 525
Glu Lys Phe Lys Leu Asn Phe Gln Met Pro Thr Leu Ala Ser Gly Trp
530 535 540
Asp Val Asn Lys Glu Lys Asn Asn Gly Ala Ile Leu Phe Val Lys Asn
545 550 555 560
Gly Leu Tyr Tyr Leu Gly Ile Met Pro Lys Gln Lys Gly Arg Tyr Lys
565 570 575
Ala Leu Ser Phe Glu Pro Thr Glu Lys Thr Ser Glu Gly Phe Asp Lys
580 585 590
Met Tyr Tyr Asp Tyr Phe Pro Asp Ala Ala Lys Met Ile Pro Lys Cys
595 600 605
Ser Thr Gln Leu Lys Ala Val Thr Ala His Phe Gln Thr His Thr Thr
610 615 620
Pro Ile Leu Leu Ser Asn Asn Phe Ile Glu Pro Leu Glu Ile Thr Lys
625 630 635 640
Glu Ile Tyr Asp Leu Asn Asn Pro Glu Lys Glu Pro Lys Lys Phe Gln
645 650 655
Thr Ala Tyr Ala Lys Lys Thr Gly Asp Gln Lys Gly Tyr Arg Glu Ala
660 665 670
Leu Cys Lys Trp Ile Asp Phe Thr Arg Asp Phe Leu Ser Lys Tyr Thr
675 680 685
Lys Thr Thr Ser Ile Asp Leu Ser Ser Leu Arg Pro Ser Ser Gln Tyr
690 695 700
Lys Asp Leu Gly Glu Tyr Tyr Ala Glu Leu Asn Pro Leu Leu Tyr His
705 710 715 720
Ile Ser Phe Gln Arg Ile Ala Glu Lys Glu Ile Met Asp Ala Val Glu
725 730 735
Thr Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Lys
740 745 750
Gly His His Gly Lys Pro Asn Leu His Thr Leu Tyr Trp Thr Gly Leu
755 760 765
Phe Ser Pro Glu Asn Leu Ala Lys Thr Ser Ile Lys Leu Asn Gly Gln
770 775 780
Ala Glu Leu Phe Tyr Arg Pro Lys Ser Arg Met Lys Arg Met Ala His
785 790 795 800
Arg Leu Gly Glu Lys Met Leu Asn Lys Lys Leu Lys Asp Gln Lys Thr
805 810 815
Pro Ile Pro Asp Thr Leu Tyr Gln Glu Leu Tyr Asp Tyr Val Asn His
820 825 830
Arg Leu Ser His Asp Leu Ser Asp Glu Ala Arg Ala Leu Leu Pro Asn
835 840 845
Val Ile Thr Lys Glu Val Ser His Glu Ile Ile Lys Asp Arg Arg Phe
850 855 860
Thr Ser Asp Lys Phe Phe Phe His Val Pro Ile Thr Leu Asn Tyr Gln
865 870 875 880
Ala Ala Asn Ser Pro Ser Lys Phe Asn Gln Arg Val Asn Ala Tyr Leu
885 890 895
Lys Glu His Pro Glu Thr Pro Ile Ile Gly Ile Asp Arg Gly Glu Arg
900 905 910
Asn Leu Ile Tyr Ile Thr Val Ile Asp Ser Thr Gly Lys Ile Leu Glu
915 920 925
Gln Arg Ser Leu Asn Thr Ile Gln Gln Phe Asp Tyr Gln Lys Lys Leu
930 935 940
Asp Asn Arg Glu Lys Glu Arg Val Ala Ala Arg Gln Ala Trp Ser Val
945 950 955 960
Val Gly Thr Ile Lys Asp Leu Lys Gln Gly Tyr Leu Ser Gln Val Ile
965 970 975
His Glu Ile Val Asp Leu Met Ile His Tyr Gln Ala Val Val Val Leu
980 985 990
Glu Asn Leu Asn Phe Gly Phe Lys Ser Lys Arg Thr Gly Ile Ala Glu
995 1000 1005
Lys Ala Val Tyr Gln Gln Phe Glu Lys Met Leu Ile Asp Lys Leu
1010 1015 1020
Asn Cys Leu Val Leu Lys Asp Tyr Pro Ala Glu Lys Val Gly Gly
1025 1030 1035
Val Leu Asn Pro Tyr Gln Leu Thr Asp Gln Phe Thr Ser Phe Ala
1040 1045 1050
Lys Met Gly Thr Gln Ser Gly Phe Leu Phe Tyr Val Pro Ala Pro
1055 1060 1065
Tyr Thr Ser Lys Ile Asp Pro Leu Thr Gly Phe Val Asp Pro Phe
1070 1075 1080
Val Trp Lys Thr Ile Lys Asn His Glu Ser Arg Lys His Phe Leu
1085 1090 1095
Glu Gly Phe Asp Phe Leu His Tyr Asp Val Lys Thr Gly Asp Phe
1100 1105 1110
Ile Leu His Phe Lys Met Asn Arg Asn Leu Ser Phe Gln Arg Gly
1115 1120 1125
Leu Pro Gly Phe Met Pro Ala Trp Asp Ile Val Phe Glu Lys Asn
1130 1135 1140
Glu Thr Gln Phe Asp Ala Lys Gly Thr Pro Phe Ile Ala Gly Lys
1145 1150 1155
Arg Ile Val Pro Val Ile Glu Asn His Arg Phe Thr Gly Arg Tyr
1160 1165 1170
Arg Asp Leu Tyr Pro Ala Asn Glu Leu Ile Ala Leu Leu Glu Glu
1175 1180 1185
Lys Gly Ile Val Phe Arg Asp Gly Ser Asn Ile Leu Pro Lys Leu
1190 1195 1200
Leu Glu Asn Asp Asp Ser His Ala Ile Asp Thr Met Val Ala Leu
1205 1210 1215
Ile Arg Ser Val Leu Gln Met Arg Asn Ser Asn Ala Ala Thr Gly
1220 1225 1230
Glu Asp Tyr Ile Asn Ser Pro Val Arg Asp Leu Asn Gly Val Cys
1235 1240 1245
Phe Asp Ser Arg Phe Gln Asn Pro Glu Trp Pro Met Asp Ala Asp
1250 1255 1260
Ala Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Gln Leu Leu Leu
1265 1270 1275
Asn His Leu Lys Glu Ser Lys Asp Leu Lys Leu Gln Asn Gly Ile
1280 1285 1290
Ser Asn Gln Asp Trp Leu Ala Tyr Ile Gln Glu Leu Arg Asn
1295 1300 1305
<210> 3
<211> 1241
<212> PRT
<213> Butyrivibrio proteoclasticus
<400> 3
Met Leu Leu Tyr Glu Asn Tyr Thr Lys Arg Asn Gln Ile Thr Lys Ser
1 5 10 15
Leu Arg Leu Glu Leu Arg Pro Gln Gly Lys Thr Leu Arg Asn Ile Lys
20 25 30
Glu Leu Asn Leu Leu Glu Gln Asp Lys Ala Ile Tyr Ala Leu Leu Glu
35 40 45
Arg Leu Lys Pro Val Ile Asp Glu Gly Ile Lys Asp Ile Ala Arg Asp
50 55 60
Thr Leu Lys Asn Cys Glu Leu Ser Phe Glu Lys Leu Tyr Glu His Phe
65 70 75 80
Leu Ser Gly Asp Lys Lys Ala Tyr Ala Lys Glu Ser Glu Arg Leu Lys
85 90 95
Lys Glu Ile Val Lys Thr Leu Ile Lys Asn Leu Pro Glu Gly Ile Gly
100 105 110
Lys Ile Ser Glu Ile Asn Ser Ala Lys Tyr Leu Asn Gly Val Leu Tyr
115 120 125
Asp Phe Ile Asp Lys Thr His Lys Asp Ser Glu Glu Lys Gln Asn Ile
130 135 140
Leu Ser Asp Ile Leu Glu Thr Lys Gly Tyr Leu Ala Leu Phe Ser Lys
145 150 155 160
Phe Leu Thr Ser Arg Ile Thr Thr Leu Glu Gln Ser Met Pro Lys Arg
165 170 175
Val Ile Glu Asn Phe Glu Ile Tyr Ala Ala Asn Ile Pro Lys Met Gln
180 185 190
Asp Ala Leu Glu Arg Gly Ala Val Ser Phe Ala Ile Glu Tyr Glu Ser
195 200 205
Ile Cys Ser Val Asp Tyr Tyr Asn Gln Ile Leu Ser Gln Glu Asp Ile
210 215 220
Asp Ser Tyr Asn Arg Leu Ile Ser Gly Ile Met Asp Glu Asp Gly Ala
225 230 235 240
Lys Glu Lys Gly Ile Asn Gln Thr Ile Ser Glu Lys Asn Ile Lys Ile
245 250 255
Lys Ser Glu His Leu Glu Glu Lys Pro Phe Arg Ile Leu Lys Gln Leu
260 265 270
His Lys Gln Ile Leu Glu Glu Arg Glu Lys Ala Phe Thr Ile Asp His
275 280 285
Ile Asp Ser Asp Glu Glu Val Val Gln Val Thr Lys Glu Ala Phe Glu
290 295 300
Gln Thr Lys Glu Gln Trp Glu Asn Ile Lys Lys Ile Asn Gly Phe Tyr
305 310 315 320
Ala Lys Asp Pro Gly Asp Ile Thr Leu Phe Ile Val Val Gly Pro Asn
325 330 335
Gln Thr His Val Leu Ser Gln Leu Ile Tyr Gly Glu His Asp Arg Ile
340 345 350
Arg Leu Leu Leu Glu Glu Tyr Glu Lys Asn Thr Leu Glu Val Leu Pro
355 360 365
Arg Arg Thr Lys Ser Glu Asp Ala Arg Tyr Asp Lys Phe Val Asn Ala
370 375 380
Val Pro Lys Lys Val Ala Lys Glu Ser His Thr Phe Asp Gly Leu Gln
385 390 395 400
Lys Met Thr Gly Asp Asp Arg Leu Phe Ile Leu Tyr Arg Asp Glu Leu
405 410 415
Ala Arg Asn Tyr Met Arg Ile Lys Glu Ala Tyr Gly Thr Phe Glu Arg
420 425 430
Asp Ile Leu Lys Ser Arg Arg Gly Ile Lys Gly Asn Arg Asp Val Gln
435 440 445
Glu Ser Leu Val Ser Phe Tyr Asp Glu Leu Thr Lys Phe Arg Ser Ala
450 455 460
Leu Arg Ile Ile Asn Ser Gly Asn Asp Glu Lys Ala Asp Pro Ile Phe
465 470 475 480
Tyr Asn Thr Phe Asp Gly Ile Phe Glu Lys Ala Asn Arg Thr Tyr Lys
485 490 495
Ala Glu Asn Leu Cys Arg Asn Tyr Val Thr Lys Ser Pro Ala Asp Asp
500 505 510
Ala Arg Ile Met Ala Ser Cys Leu Gly Thr Pro Ala Arg Leu Arg Thr
515 520 525
His Trp Trp Asn Gly Glu Glu Asn Phe Ala Ile Asn Asp Val Ala Met
530 535 540
Ile Arg Arg Gly Asp Glu Tyr Tyr Tyr Phe Val Leu Thr Pro Asp Val
545 550 555 560
Lys Pro Val Asp Leu Lys Thr Lys Asp Glu Thr Asp Ala Gln Ile Phe
565 570 575
Val Gln Arg Lys Gly Ala Lys Ser Phe Leu Gly Leu Pro Lys Ala Leu
580 585 590
Phe Lys Cys Ile Leu Glu Pro Tyr Phe Glu Ser Pro Glu His Lys Asn
595 600 605
Asp Lys Asn Cys Val Ile Glu Glu Tyr Val Ser Lys Pro Leu Thr Ile
610 615 620
Asp Arg Arg Ala Tyr Asp Ile Phe Lys Asn Gly Thr Phe Lys Lys Thr
625 630 635 640
Asn Ile Gly Ile Asp Gly Leu Thr Glu Glu Lys Phe Lys Asp Asp Cys
645 650 655
Arg Tyr Leu Ile Asp Val Tyr Lys Glu Phe Ile Ala Val Tyr Thr Arg
660 665 670
Tyr Ser Cys Phe Asn Met Ser Gly Leu Lys Arg Ala Asp Glu Tyr Asn
675 680 685
Asp Ile Gly Glu Phe Phe Ser Asp Val Asp Thr Arg Leu Cys Thr Met
690 695 700
Glu Trp Ile Pro Val Ser Phe Glu Arg Ile Asn Asp Met Val Asp Lys
705 710 715 720
Lys Glu Gly Leu Leu Phe Leu Val Arg Ser Met Phe Leu Tyr Asn Arg
725 730 735
Pro Arg Lys Pro Tyr Glu Arg Thr Phe Ile Gln Leu Phe Ser Asp Ser
740 745 750
Asn Met Glu His Thr Ser Met Leu Leu Asn Ser Arg Ala Met Ile Gln
755 760 765
Tyr Arg Ala Ala Ser Leu Pro Arg Arg Val Thr His Lys Lys Gly Ser
770 775 780
Ile Leu Val Ala Leu Arg Asp Ser Asn Gly Glu His Ile Pro Met His
785 790 795 800
Ile Arg Glu Ala Ile Tyr Lys Met Lys Asn Asn Phe Asp Ile Ser Ser
805 810 815
Glu Asp Phe Ile Met Ala Lys Ala Tyr Leu Ala Glu His Asp Val Ala
820 825 830
Ile Lys Lys Ala Asn Glu Asp Ile Ile Arg Asn Arg Arg Tyr Thr Glu
835 840 845
Asp Lys Phe Phe Leu Ser Leu Ser Tyr Thr Lys Asn Ala Asp Ile Ser
850 855 860
Ala Arg Thr Leu Asp Tyr Ile Asn Asp Lys Val Glu Glu Asp Thr Gln
865 870 875 880
Asp Ser Arg Met Ala Val Ile Val Thr Arg Asn Leu Lys Asp Leu Thr
885 890 895
Tyr Val Ala Val Val Asp Glu Lys Asn Asn Val Leu Glu Glu Lys Ser
900 905 910
Leu Asn Glu Ile Asp Gly Val Asn Tyr Arg Glu Leu Leu Lys Glu Arg
915 920 925
Thr Lys Ile Lys Tyr His Asp Lys Thr Arg Leu Trp Gln Tyr Asp Val
930 935 940
Ser Ser Lys Gly Leu Lys Glu Ala Tyr Val Glu Leu Ala Val Thr Gln
945 950 955 960
Ile Ser Lys Leu Ala Thr Lys Tyr Asn Ala Val Val Val Val Glu Ser
965 970 975
Met Ser Ser Thr Phe Lys Asp Lys Phe Ser Phe Leu Asp Glu Gln Ile
980 985 990
Phe Lys Ala Phe Glu Ala Arg Leu Cys Ala Arg Met Ser Asp Leu Ser
995 1000 1005
Phe Asn Thr Ile Lys Glu Gly Glu Ala Gly Ser Ile Ser Asn Pro
1010 1015 1020
Ile Gln Val Ser Asn Asn Asn Gly Asn Ser Tyr Gln Asp Gly Val
1025 1030 1035
Ile Tyr Phe Leu Asn Asn Ala Tyr Thr Arg Thr Leu Cys Pro Asp
1040 1045 1050
Thr Gly Phe Val Asp Val Phe Asp Lys Thr Arg Leu Ile Thr Met
1055 1060 1065
Gln Ser Lys Arg Gln Phe Phe Ala Lys Met Lys Asp Ile Arg Ile
1070 1075 1080
Asp Asp Gly Glu Met Leu Phe Thr Phe Asn Leu Glu Glu Tyr Pro
1085 1090 1095
Thr Lys Arg Leu Leu Asp Arg Lys Glu Trp Thr Val Lys Ile Ala
1100 1105 1110
Gly Asp Gly Ser Tyr Phe Asp Lys Asp Lys Gly Glu Tyr Val Tyr
1115 1120 1125
Val Asn Asp Ile Val Arg Glu Gln Ile Ile Pro Ala Leu Leu Glu
1130 1135 1140
Asp Lys Ala Val Phe Asp Gly Asn Met Ala Glu Lys Phe Leu Asp
1145 1150 1155
Lys Thr Ala Ile Ser Gly Lys Ser Val Glu Leu Ile Tyr Lys Trp
1160 1165 1170
Phe Ala Asn Ala Leu Tyr Gly Ile Ile Thr Lys Lys Asp Gly Glu
1175 1180 1185
Lys Ile Tyr Arg Ser Pro Ile Thr Gly Thr Glu Ile Asp Val Ser
1190 1195 1200
Lys Asn Thr Thr Tyr Asn Phe Gly Lys Lys Phe Met Phe Lys Gln
1205 1210 1215
Glu Tyr Arg Gly Asp Gly Asp Phe Leu Asp Ala Phe Leu Asn Tyr
1220 1225 1230
Met Gln Ala Gln Asp Ile Ala Val
1235 1240
<210> 4
<211> 1238
<212> PRT
<213> Candidatus Methanoplasma termitum
<400> 4
Met Asn Asn Tyr Asp Glu Phe Thr Lys Leu Tyr Pro Ile Gln Lys Thr
1 5 10 15
Ile Arg Phe Glu Leu Lys Pro Gln Gly Arg Thr Met Glu His Leu Glu
20 25 30
Thr Phe Asn Phe Phe Glu Glu Asp Arg Asp Arg Ala Glu Lys Tyr Lys
35 40 45
Ile Leu Lys Glu Ala Ile Asp Glu Tyr His Lys Lys Phe Ile Asp Glu
50 55 60
His Leu Thr Asn Met Ser Leu Asp Trp Asn Ser Leu Lys Gln Ile Ser
65 70 75 80
Glu Lys Tyr Tyr Lys Ser Arg Glu Glu Lys Asp Lys Lys Val Phe Leu
85 90 95
Ser Glu Gln Lys Arg Met Arg Gln Glu Ile Val Ser Glu Phe Lys Lys
100 105 110
Asp Asp Arg Phe Lys Asp Leu Phe Ser Lys Lys Leu Phe Ser Glu Leu
115 120 125
Leu Lys Glu Glu Ile Tyr Lys Lys Gly Asn His Gln Glu Ile Asp Ala
130 135 140
Leu Lys Ser Phe Asp Lys Phe Ser Gly Tyr Phe Ile Gly Leu His Glu
145 150 155 160
Asn Arg Lys Asn Met Tyr Ser Asp Gly Asp Glu Ile Thr Ala Ile Ser
165 170 175
Asn Arg Ile Val Asn Glu Asn Phe Pro Lys Phe Leu Asp Asn Leu Gln
180 185 190
Lys Tyr Gln Glu Ala Arg Lys Lys Tyr Pro Glu Trp Ile Ile Lys Ala
195 200 205
Glu Ser Ala Leu Val Ala His Asn Ile Lys Met Asp Ile Val Phe Ser
210 215 220
Leu Glu Tyr Phe Asn Lys Val Leu Asn Gln Glu Gly Ile Gln Arg Tyr
225 230 235 240
Asn Leu Ala Leu Gly Gly Tyr Val Thr Lys Ser Gly Glu Lys Met Met
245 250 255
Gly Leu Asn Asp Ala Leu Asn Leu Ala His Gln Ser Glu Lys Ser Ser
260 265 270
Lys Gly Arg Ile His Met Thr Pro Leu Phe Lys Gln Ile Leu Ser Glu
275 280 285
Lys Glu Ser Phe Ser Tyr Ile Pro Asp Val Phe Thr Glu Asp Ser Gln
290 295 300
Leu Leu Pro Ser Ile Gly Gly Phe Phe Ala Gln Ile Glu Asn Asp Lys
305 310 315 320
Asp Gly Asn Ile Phe Asp Arg Ala Leu Glu Leu Ile Ser Ser Tyr Ala
325 330 335
Glu Tyr Asp Thr Glu Arg Ile Tyr Ile Arg Gln Ala Asp Ile Asn Arg
340 345 350
Val Ser Asn Val Ile Phe Gly Glu Trp Gly Thr Leu Gly Gly Leu Met
355 360 365
Arg Glu Tyr Lys Ala Asp Ser Ile Asn Asp Ile Asn Leu Glu Arg Thr
370 375 380
Cys Lys Lys Val Asp Lys Trp Leu Asp Ser Lys Glu Phe Ala Leu Ser
385 390 395 400
Asp Val Leu Glu Ala Ile Asp Arg Thr Gly Asn Asn Asp Ala Phe Asn
405 410 415
Glu Tyr Ile Ser Lys Met Arg Thr Ala Arg Glu Lys Ile Asp Ala Ala
420 425 430
Arg Lys Glu Met Lys Phe Ile Ser Glu Lys Ile Ser Gly Asp Glu Glu
435 440 445
Ser Ile His Ile Ile Lys Thr Leu Leu Asp Ser Val Gln Gln Phe Leu
450 455 460
His Phe Phe Asn Leu Phe Lys Ala Arg Gln Asp Ile Pro Leu Asp Gly
465 470 475 480
Ala Phe Tyr Ala Glu Phe Asp Glu Val His Ser Lys Leu Phe Ala Ile
485 490 495
Val Pro Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr Lys Asn Asn Leu
500 505 510
Asn Thr Lys Lys Ile Lys Leu Asn Phe Lys Asn Pro Thr Leu Ala Asn
515 520 525
Gly Trp Asp Gln Asn Lys Val Tyr Asp Tyr Ala Ser Leu Ile Phe Leu
530 535 540
Arg Asp Gly Asn Tyr Tyr Leu Gly Ile Ile Asn Pro Lys Arg Lys Lys
545 550 555 560
Asn Ile Lys Phe Glu Gln Gly Ser Gly Asn Gly Pro Phe Tyr Arg Lys
565 570 575
Met Val Tyr Lys Gln Ile Pro Gly Pro Asn Lys Asn Leu Arg Pro Val
580 585 590
Phe Leu Thr Ser Thr Lys Gly Lys Lys Glu Tyr Lys Pro Ser Lys Glu
595 600 605
Ile Ile Glu Gly Tyr Glu Ala Asp Lys His Ile Arg Gly Asp Lys Phe
610 615 620
Asp Leu Asp Phe Cys His Lys Leu Ile Asp Phe Phe Lys Glu Ser Ile
625 630 635 640
Glu Lys His Lys Asp Trp Ser Lys Phe Asn Phe Tyr Phe Ser Pro Thr
645 650 655
Glu Ser Tyr Gly Asp Ile Ser Glu Phe Tyr Leu Asp Val Glu Lys Gln
660 665 670
Gly Tyr Arg Met His Phe Glu Asn Ile Ser Ala Glu Thr Ile Asp Glu
675 680 685
Tyr Val Glu Lys Gly Asp Leu Phe Leu Phe Gln Ile Tyr Asn Lys Asp
690 695 700
Phe Val Lys Ala Ala Thr Gly Lys Lys Asp Met His Thr Ile Tyr Trp
705 710 715 720
Asn Ala Ala Phe Ser Pro Glu Asn Leu Gln Asp Val Val Val Lys Leu
725 730 735
Asn Gly Glu Ala Glu Leu Phe Tyr Arg Asp Lys Ser Asp Ile Lys Glu
740 745 750
Ile Val His Arg Glu Gly Glu Ile Leu Val Asn Arg Thr Tyr Asn Gly
755 760 765
Arg Thr Pro Val Pro Asp Lys Ile His Lys Lys Leu Thr Asp Tyr His
770 775 780
Asn Gly Arg Thr Lys Asp Leu Gly Glu Ala Lys Glu Tyr Leu Asp Lys
785 790 795 800
Val Arg Tyr Phe Lys Ala His Tyr Asp Ile Thr Lys Asp Arg Arg Tyr
805 810 815
Leu Asn Asp Lys Ile Tyr Phe His Val Pro Leu Thr Leu Asn Phe Lys
820 825 830
Ala Asn Gly Lys Lys Asn Leu Asn Lys Met Val Ile Glu Lys Phe Leu
835 840 845
Ser Asp Glu Lys Ala His Ile Ile Gly Ile Asp Arg Gly Glu Arg Asn
850 855 860
Leu Leu Tyr Tyr Ser Ile Ile Asp Arg Ser Gly Lys Ile Ile Asp Gln
865 870 875 880
Gln Ser Leu Asn Val Ile Asp Gly Phe Asp Tyr Arg Glu Lys Leu Asn
885 890 895
Gln Arg Glu Ile Glu Met Lys Asp Ala Arg Gln Ser Trp Asn Ala Ile
900 905 910
Gly Lys Ile Lys Asp Leu Lys Glu Gly Tyr Leu Ser Lys Ala Val His
915 920 925
Glu Ile Thr Lys Met Ala Ile Gln Tyr Asn Ala Ile Val Val Met Glu
930 935 940
Glu Leu Asn Tyr Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln
945 950 955 960
Ile Tyr Gln Lys Phe Glu Asn Met Leu Ile Asp Lys Met Asn Tyr Leu
965 970 975
Val Phe Lys Asp Ala Pro Asp Glu Ser Pro Gly Gly Val Leu Asn Ala
980 985 990
Tyr Gln Leu Thr Asn Pro Leu Glu Ser Phe Ala Lys Leu Gly Lys Gln
995 1000 1005
Thr Gly Ile Leu Phe Tyr Val Pro Ala Ala Tyr Thr Ser Lys Ile
1010 1015 1020
Asp Pro Thr Thr Gly Phe Val Asn Leu Phe Asn Thr Ser Ser Lys
1025 1030 1035
Thr Asn Ala Gln Glu Arg Lys Glu Phe Leu Gln Lys Phe Glu Ser
1040 1045 1050
Ile Ser Tyr Ser Ala Lys Asp Gly Gly Ile Phe Ala Phe Ala Phe
1055 1060 1065
Asp Tyr Arg Lys Phe Gly Thr Ser Lys Thr Asp His Lys Asn Val
1070 1075 1080
Trp Thr Ala Tyr Thr Asn Gly Glu Arg Met Arg Tyr Ile Lys Glu
1085 1090 1095
Lys Lys Arg Asn Glu Leu Phe Asp Pro Ser Lys Glu Ile Lys Glu
1100 1105 1110
Ala Leu Thr Ser Ser Gly Ile Lys Tyr Asp Gly Gly Gln Asn Ile
1115 1120 1125
Leu Pro Asp Ile Leu Arg Ser Asn Asn Asn Gly Leu Ile Tyr Thr
1130 1135 1140
Met Tyr Ser Ser Phe Ile Ala Ala Ile Gln Met Arg Val Tyr Asp
1145 1150 1155
Gly Lys Glu Asp Tyr Ile Ile Ser Pro Ile Lys Asn Ser Lys Gly
1160 1165 1170
Glu Phe Phe Arg Thr Asp Pro Lys Arg Arg Glu Leu Pro Ile Asp
1175 1180 1185
Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala Leu Arg Gly Glu Leu
1190 1195 1200
Thr Met Arg Ala Ile Ala Glu Lys Phe Asp Pro Asp Ser Glu Lys
1205 1210 1215
Met Ala Lys Leu Glu Leu Lys His Lys Asp Trp Phe Glu Phe Met
1220 1225 1230
Gln Thr Arg Gly Asp
1235
<210> 5
<211> 1281
<212> PRT
<213> 挑剔真杆菌(Eubacterium eligens)
<400> 5
Met Asn Gly Asn Arg Ser Ile Val Tyr Arg Glu Phe Val Gly Val Ile
1 5 10 15
Pro Val Ala Lys Thr Leu Arg Asn Glu Leu Arg Pro Val Gly His Thr
20 25 30
Gln Glu His Ile Ile Gln Asn Gly Leu Ile Gln Glu Asp Glu Leu Arg
35 40 45
Gln Glu Lys Ser Thr Glu Leu Lys Asn Ile Met Asp Asp Tyr Tyr Arg
50 55 60
Glu Tyr Ile Asp Lys Ser Leu Ser Gly Val Thr Asp Leu Asp Phe Thr
65 70 75 80
Leu Leu Phe Glu Leu Met Asn Leu Val Gln Ser Ser Pro Ser Lys Asp
85 90 95
Asn Lys Lys Ala Leu Glu Lys Glu Gln Ser Lys Met Arg Glu Gln Ile
100 105 110
Cys Thr His Leu Gln Ser Asp Ser Asn Tyr Lys Asn Ile Phe Asn Ala
115 120 125
Lys Leu Leu Lys Glu Ile Leu Pro Asp Phe Ile Lys Asn Tyr Asn Gln
130 135 140
Tyr Asp Val Lys Asp Lys Ala Gly Lys Leu Glu Thr Leu Ala Leu Phe
145 150 155 160
Asn Gly Phe Ser Thr Tyr Phe Thr Asp Phe Phe Glu Lys Arg Lys Asn
165 170 175
Val Phe Thr Lys Glu Ala Val Ser Thr Ser Ile Ala Tyr Arg Ile Val
180 185 190
His Glu Asn Ser Leu Ile Phe Leu Ala Asn Met Thr Ser Tyr Lys Lys
195 200 205
Ile Ser Glu Lys Ala Leu Asp Glu Ile Glu Val Ile Glu Lys Asn Asn
210 215 220
Gln Asp Lys Met Gly Asp Trp Glu Leu Asn Gln Ile Phe Asn Pro Asp
225 230 235 240
Phe Tyr Asn Met Val Leu Ile Gln Ser Gly Ile Asp Phe Tyr Asn Glu
245 250 255
Ile Cys Gly Val Val Asn Ala His Met Asn Leu Tyr Cys Gln Gln Thr
260 265 270
Lys Asn Asn Tyr Asn Leu Phe Lys Met Arg Lys Leu His Lys Gln Ile
275 280 285
Leu Ala Tyr Thr Ser Thr Ser Phe Glu Val Pro Lys Met Phe Glu Asp
290 295 300
Asp Met Ser Val Tyr Asn Ala Val Asn Ala Phe Ile Asp Glu Thr Glu
305 310 315 320
Lys Gly Asn Ile Ile Gly Lys Leu Lys Asp Ile Val Asn Lys Tyr Asp
325 330 335
Glu Leu Asp Glu Lys Arg Ile Tyr Ile Ser Lys Asp Phe Tyr Glu Thr
340 345 350
Leu Ser Cys Phe Met Ser Gly Asn Trp Asn Leu Ile Thr Gly Cys Val
355 360 365
Glu Asn Phe Tyr Asp Glu Asn Ile His Ala Lys Gly Lys Ser Lys Glu
370 375 380
Glu Lys Val Lys Lys Ala Val Lys Glu Asp Lys Tyr Lys Ser Ile Asn
385 390 395 400
Asp Val Asn Asp Leu Val Glu Lys Tyr Ile Asp Glu Lys Glu Arg Asn
405 410 415
Glu Phe Lys Asn Ser Asn Ala Lys Gln Tyr Ile Arg Glu Ile Ser Asn
420 425 430
Ile Ile Thr Asp Thr Glu Thr Ala His Leu Glu Tyr Asp Asp His Ile
435 440 445
Ser Leu Ile Glu Ser Glu Glu Lys Ala Asp Glu Met Lys Lys Arg Leu
450 455 460
Asp Met Tyr Met Asn Met Tyr His Trp Ala Lys Ala Phe Ile Val Asp
465 470 475 480
Glu Val Leu Asp Arg Asp Glu Met Phe Tyr Ser Asp Ile Asp Asp Ile
485 490 495
Tyr Asn Ile Leu Glu Asn Ile Val Pro Leu Tyr Asn Arg Val Arg Asn
500 505 510
Tyr Val Thr Gln Lys Pro Tyr Asn Ser Lys Lys Ile Lys Leu Asn Phe
515 520 525
Gln Ser Pro Thr Leu Ala Asn Gly Trp Ser Gln Ser Lys Glu Phe Asp
530 535 540
Asn Asn Ala Ile Ile Leu Ile Arg Asp Asn Lys Tyr Tyr Leu Ala Ile
545 550 555 560
Phe Asn Ala Lys Asn Lys Pro Asp Lys Lys Ile Ile Gln Gly Asn Ser
565 570 575
Asp Lys Lys Asn Asp Asn Asp Tyr Lys Lys Met Val Tyr Asn Leu Leu
580 585 590
Pro Gly Ala Asn Lys Met Leu Pro Lys Val Phe Leu Ser Lys Lys Gly
595 600 605
Ile Glu Thr Phe Lys Pro Ser Asp Tyr Ile Ile Ser Gly Tyr Asn Ala
610 615 620
His Lys His Ile Lys Thr Ser Glu Asn Phe Asp Ile Ser Phe Cys Arg
625 630 635 640
Asp Leu Ile Asp Tyr Phe Lys Asn Ser Ile Glu Lys His Ala Glu Trp
645 650 655
Arg Lys Tyr Glu Phe Lys Phe Ser Ala Thr Asp Ser Tyr Ser Asp Ile
660 665 670
Ser Glu Phe Tyr Arg Glu Val Glu Met Gln Gly Tyr Arg Ile Asp Trp
675 680 685
Thr Tyr Ile Ser Glu Ala Asp Ile Asn Lys Leu Asp Glu Glu Gly Lys
690 695 700
Ile Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Glu Asn Ser Thr
705 710 715 720
Gly Lys Glu Asn Leu His Thr Met Tyr Phe Lys Asn Ile Phe Ser Glu
725 730 735
Glu Asn Leu Asp Lys Ile Ile Lys Leu Asn Gly Gln Ala Glu Leu Phe
740 745 750
Tyr Arg Arg Ala Ser Val Lys Asn Pro Val Lys His Lys Lys Asp Ser
755 760 765
Val Leu Val Asn Lys Thr Tyr Lys Asn Gln Leu Asp Asn Gly Asp Val
770 775 780
Val Arg Ile Pro Ile Pro Asp Asp Ile Tyr Asn Glu Ile Tyr Lys Met
785 790 795 800
Tyr Asn Gly Tyr Ile Lys Glu Ser Asp Leu Ser Glu Ala Ala Lys Glu
805 810 815
Tyr Leu Asp Lys Val Glu Val Arg Thr Ala Gln Lys Asp Ile Val Lys
820 825 830
Asp Tyr Arg Tyr Thr Val Asp Lys Tyr Phe Ile His Thr Pro Ile Thr
835 840 845
Ile Asn Tyr Lys Val Thr Ala Arg Asn Asn Val Asn Asp Met Val Val
850 855 860
Lys Tyr Ile Ala Gln Asn Asp Asp Ile His Val Ile Gly Ile Asp Arg
865 870 875 880
Gly Glu Arg Asn Leu Ile Tyr Ile Ser Val Ile Asp Ser His Gly Asn
885 890 895
Ile Val Lys Gln Lys Ser Tyr Asn Ile Leu Asn Asn Tyr Asp Tyr Lys
900 905 910
Lys Lys Leu Val Glu Lys Glu Lys Thr Arg Glu Tyr Ala Arg Lys Asn
915 920 925
Trp Lys Ser Ile Gly Asn Ile Lys Glu Leu Lys Glu Gly Tyr Ile Ser
930 935 940
Gly Val Val His Glu Ile Ala Met Leu Ile Val Glu Tyr Asn Ala Ile
945 950 955 960
Ile Ala Met Glu Asp Leu Asn Tyr Gly Phe Lys Arg Gly Arg Phe Lys
965 970 975
Val Glu Arg Gln Val Tyr Gln Lys Phe Glu Ser Met Leu Ile Asn Lys
980 985 990
Leu Asn Tyr Phe Ala Ser Lys Glu Lys Ser Val Asp Glu Pro Gly Gly
995 1000 1005
Leu Leu Lys Gly Tyr Gln Leu Thr Tyr Val Pro Asp Asn Ile Lys
1010 1015 1020
Asn Leu Gly Lys Gln Cys Gly Val Ile Phe Tyr Val Pro Ala Ala
1025 1030 1035
Phe Thr Ser Lys Ile Asp Pro Ser Thr Gly Phe Ile Ser Ala Phe
1040 1045 1050
Asn Phe Lys Ser Ile Ser Thr Asn Ala Ser Arg Lys Gln Phe Phe
1055 1060 1065
Met Gln Phe Asp Glu Ile Arg Tyr Cys Ala Glu Lys Asp Met Phe
1070 1075 1080
Ser Phe Gly Phe Asp Tyr Asn Asn Phe Asp Thr Tyr Asn Ile Thr
1085 1090 1095
Met Gly Lys Thr Gln Trp Thr Val Tyr Thr Asn Gly Glu Arg Leu
1100 1105 1110
Gln Ser Glu Phe Asn Asn Ala Arg Arg Thr Gly Lys Thr Lys Ser
1115 1120 1125
Ile Asn Leu Thr Glu Thr Ile Lys Leu Leu Leu Glu Asp Asn Glu
1130 1135 1140
Ile Asn Tyr Ala Asp Gly His Asp Ile Arg Ile Asp Met Glu Lys
1145 1150 1155
Met Asp Glu Asp Lys Lys Ser Glu Phe Phe Ala Gln Leu Leu Ser
1160 1165 1170
Leu Tyr Lys Leu Thr Val Gln Met Arg Asn Ser Tyr Thr Glu Ala
1175 1180 1185
Glu Glu Gln Glu Asn Gly Ile Ser Tyr Asp Lys Ile Ile Ser Pro
1190 1195 1200
Val Ile Asn Asp Glu Gly Glu Phe Phe Asp Ser Asp Asn Tyr Lys
1205 1210 1215
Glu Ser Asp Asp Lys Glu Cys Lys Met Pro Lys Asp Ala Asp Ala
1220 1225 1230
Asn Gly Ala Tyr Cys Ile Ala Leu Lys Gly Leu Tyr Glu Val Leu
1235 1240 1245
Lys Ile Lys Ser Glu Trp Thr Glu Asp Gly Phe Asp Arg Asn Cys
1250 1255 1260
Leu Lys Leu Pro His Ala Glu Trp Leu Asp Phe Ile Gln Asn Lys
1265 1270 1275
Arg Tyr Glu
1280
<210> 6
<211> 1300
<212> PRT
<213> 新凶手弗朗西丝氏菌(Francisella novicida)
<400> 6
Met Ser Ile Tyr Gln Glu Phe Val Asn Lys Tyr Ser Leu Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Glu Asn Ile Lys
20 25 30
Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys
35 40 45
Lys Ala Lys Gln Ile Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu
50 55 60
Ile Leu Ser Ser Val Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser
65 70 75 80
Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys
85 90 95
Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys Lys Gln Ile Ser Glu Tyr
100 105 110
Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile
115 120 125
Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln
130 135 140
Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr
145 150 155 160
Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr
165 170 175
Thr Tyr Phe Lys Gly Phe His Glu Asn Arg Lys Val Asn Tyr Ser Ser
180 185 190
Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu
195 200 205
Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr Glu Ser Leu Lys Asp Lys
210 215 220
Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu
225 230 235 240
Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg
245 250 255
Val Phe Ser Leu Asp Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr
260 265 270
Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn Thr Ile Ile Gly Gly Lys
275 280 285
Phe Val Asn Gly Glu Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile
290 295 300
Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys Thr Leu Lys Lys Tyr Lys
305 310 315 320
Met Ser Val Leu Phe Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser
325 330 335
Phe Val Ile Asp Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met
340 345 350
Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr Val Glu Glu Lys
355 360 365
Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln
370 375 380
Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr
385 390 395 400
Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala
405 410 415
Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala Pro Lys Asn Leu Asp Asn
420 425 430
Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala
435 440 445
Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn
450 455 460
Lys His Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Leu Ala
465 470 475 480
Asn Phe Ala Ala Ile Pro Met Ile Phe Asp Glu Ile Ala Gln Asn Lys
485 490 495
Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys
500 505 510
Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp Val Lys Ala Ile Lys Asp
515 520 525
Leu Leu Asp Gln Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His
530 535 540
Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His
545 550 555 560
Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val
565 570 575
Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser
580 585 590
Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly
595 600 605
Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys
610 615 620
Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile
625 630 635 640
Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys
645 650 655
Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val
660 665 670
Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr Asn Pro Ser Glu Asp Ile
675 680 685
Leu Arg Ile Arg Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln
690 695 700
Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe
705 710 715 720
Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys His Pro Glu Trp Lys Asp
725 730 735
Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg Tyr Asn Ser Ile Asp Glu
740 745 750
Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn
755 760 765
Ile Ser Glu Ser Tyr Ile Asp Ser Val Val Asn Gln Gly Lys Leu Tyr
770 775 780
Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Ala Tyr Ser Lys Gly Arg
785 790 795 800
Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn
805 810 815
Leu Gln Asp Val Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr
820 825 830
Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala
835 840 845
Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys Lys Glu Ser Val Phe Glu
850 855 860
Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe
865 870 875 880
His Cys Pro Ile Thr Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe
885 890 895
Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His
900 905 910
Ile Leu Ser Ile Asp Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu
915 920 925
Val Asp Gly Lys Gly Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile
930 935 940
Gly Asn Asp Arg Met Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile
945 950 955 960
Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn
965 970 975
Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile
980 985 990
Ala Lys Leu Val Ile Glu Tyr Asn Ala Ile Val Val Phe Glu Asp Leu
995 1000 1005
Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Val
1010 1015 1020
Tyr Gln Lys Leu Glu Lys Met Leu Ile Glu Lys Leu Asn Tyr Leu
1025 1030 1035
Val Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly Val Leu Arg
1040 1045 1050
Ala Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys Met Gly
1055 1060 1065
Lys Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr Ser
1070 1075 1080
Lys Ile Cys Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys
1085 1090 1095
Tyr Glu Ser Val Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp
1100 1105 1110
Lys Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe
1115 1120 1125
Asp Tyr Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly Lys Trp Thr
1130 1135 1140
Ile Ala Ser Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn Ser Asp
1145 1150 1155
Lys Asn His Asn Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys Glu
1160 1165 1170
Leu Glu Lys Leu Leu Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly
1175 1180 1185
Glu Cys Ile Lys Ala Ala Ile Cys Gly Glu Ser Asp Lys Lys Phe
1190 1195 1200
Phe Ala Lys Leu Thr Ser Val Leu Asn Thr Ile Leu Gln Met Arg
1205 1210 1215
Asn Ser Lys Thr Gly Thr Glu Leu Asp Tyr Leu Ile Ser Pro Val
1220 1225 1230
Ala Asp Val Asn Gly Asn Phe Phe Asp Ser Arg Gln Ala Pro Lys
1235 1240 1245
Asn Met Pro Gln Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Gly
1250 1255 1260
Leu Lys Gly Leu Met Leu Leu Gly Arg Ile Lys Asn Asn Gln Glu
1265 1270 1275
Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu Tyr Phe Glu
1280 1285 1290
Phe Val Gln Asn Arg Asn Asn
1295 1300
<210> 7
<211> 1206
<212> PRT
<213> 未知
<220>
<223> 毛螺菌科物种
<400> 7
Met Tyr Tyr Glu Ser Leu Thr Lys Gln Tyr Pro Val Ser Lys Thr Ile
1 5 10 15
Arg Asn Glu Leu Ile Pro Ile Gly Lys Thr Leu Asp Asn Ile Arg Gln
20 25 30
Asn Asn Ile Leu Glu Ser Asp Val Lys Arg Lys Gln Asn Tyr Glu His
35 40 45
Val Lys Gly Ile Leu Asp Glu Tyr His Lys Gln Leu Ile Asn Glu Ala
50 55 60
Leu Asp Asn Cys Thr Leu Pro Ser Leu Lys Ile Ala Ala Glu Ile Tyr
65 70 75 80
Leu Lys Asn Gln Lys Glu Val Ser Asp Arg Glu Asp Phe Asn Lys Thr
85 90 95
Gln Asp Leu Leu Arg Lys Glu Val Val Glu Lys Leu Lys Ala His Glu
100 105 110
Asn Phe Thr Lys Ile Gly Lys Lys Asp Ile Leu Asp Leu Leu Glu Lys
115 120 125
Leu Pro Ser Ile Ser Glu Asp Asp Tyr Asn Ala Leu Glu Ser Phe Arg
130 135 140
Asn Phe Tyr Thr Tyr Phe Thr Ser Tyr Asn Lys Val Arg Glu Asn Leu
145 150 155 160
Tyr Ser Asp Lys Glu Lys Ser Ser Thr Val Ala Tyr Arg Leu Ile Asn
165 170 175
Glu Asn Phe Pro Lys Phe Leu Asp Asn Val Lys Ser Tyr Arg Phe Val
180 185 190
Lys Thr Ala Gly Ile Leu Ala Asp Gly Leu Gly Glu Glu Glu Gln Asp
195 200 205
Ser Leu Phe Ile Val Glu Thr Phe Asn Lys Thr Leu Thr Gln Asp Gly
210 215 220
Ile Asp Thr Tyr Asn Ser Gln Val Gly Lys Ile Asn Ser Ser Ile Asn
225 230 235 240
Leu Tyr Asn Gln Lys Asn Gln Lys Ala Asn Gly Phe Arg Lys Ile Pro
245 250 255
Lys Met Lys Met Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Glu Ser
260 265 270
Phe Ile Asp Glu Phe Gln Ser Asp Glu Val Leu Ile Asp Asn Val Glu
275 280 285
Ser Tyr Gly Ser Val Leu Ile Glu Ser Leu Lys Ser Ser Lys Val Ser
290 295 300
Ala Phe Phe Asp Ala Leu Arg Glu Ser Lys Gly Lys Asn Val Tyr Val
305 310 315 320
Lys Asn Asp Leu Ala Lys Thr Ala Met Ser Val Ile Val Phe Glu Asn
325 330 335
Trp Arg Thr Phe Asp Asp Leu Leu Asn Gln Glu Tyr Asp Leu Ala Asn
340 345 350
Glu Asn Lys Lys Lys Asp Asp Lys Tyr Phe Glu Lys Arg Gln Lys Glu
355 360 365
Leu Lys Lys Asn Lys Ser Tyr Ser Leu Glu His Leu Cys Asn Leu Ser
370 375 380
Glu Asp Ser Cys Asn Leu Ile Glu Asn Tyr Ile His Gln Ile Ser Asp
385 390 395 400
Asp Ile Glu Asn Ile Ile Ile Asn Asn Glu Thr Phe Leu Arg Ile Val
405 410 415
Ile Asn Glu His Asp Arg Ser Arg Lys Leu Ala Lys Asn Arg Lys Ala
420 425 430
Val Lys Ala Ile Lys Asp Phe Leu Asp Ser Ile Lys Val Leu Glu Arg
435 440 445
Glu Leu Lys Leu Ile Asn Ser Ser Gly Gln Glu Leu Glu Lys Asp Leu
450 455 460
Ile Val Tyr Ser Ala His Glu Glu Leu Leu Val Glu Leu Lys Gln Val
465 470 475 480
Asp Ser Leu Tyr Asn Met Thr Arg Asn Tyr Leu Thr Lys Lys Pro Phe
485 490 495
Ser Thr Glu Lys Val Lys Leu Asn Phe Asn Arg Ser Thr Leu Leu Asn
500 505 510
Gly Trp Asp Arg Asn Lys Glu Thr Asp Asn Leu Gly Val Leu Leu Leu
515 520 525
Lys Asp Gly Lys Tyr Tyr Leu Gly Ile Met Asn Thr Ser Ala Asn Lys
530 535 540
Ala Phe Val Asn Pro Pro Val Ala Lys Thr Glu Lys Val Phe Lys Lys
545 550 555 560
Val Asp Tyr Lys Leu Leu Pro Val Pro Asn Gln Met Leu Pro Lys Val
565 570 575
Phe Phe Ala Lys Ser Asn Ile Asp Phe Tyr Asn Pro Ser Ser Glu Ile
580 585 590
Tyr Ser Asn Tyr Lys Lys Gly Thr His Lys Lys Gly Asn Met Phe Ser
595 600 605
Leu Glu Asp Cys His Asn Leu Ile Asp Phe Phe Lys Glu Ser Ile Ser
610 615 620
Lys His Glu Asp Trp Ser Lys Phe Gly Phe Lys Phe Asp Thr Gln Ala
625 630 635 640
Ser Tyr Asn Asp Ile Ser Glu Phe Tyr Arg Glu Val Glu Lys Gln Gly
645 650 655
Tyr Lys Leu Thr Tyr Thr Asp Ile Asp Glu Thr Tyr Ile Asn Asp Leu
660 665 670
Ile Glu Arg Asn Glu Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe
675 680 685
Ser Met Tyr Ser Lys Gly Lys Leu Asn Leu His Thr Leu Tyr Phe Met
690 695 700
Met Leu Phe Asp Gln Arg Asn Ile Asp Asp Val Val Tyr Lys Leu Asn
705 710 715 720
Gly Glu Ala Glu Val Phe Tyr Arg Pro Ala Ser Ile Ser Glu Asp Glu
725 730 735
Leu Ile Ile His Lys Ala Gly Glu Glu Ile Lys Asn Lys Asn Pro Asn
740 745 750
Arg Ala Arg Thr Lys Glu Thr Ser Thr Phe Ser Tyr Asp Ile Val Lys
755 760 765
Asp Lys Arg Tyr Ser Lys Asp Lys Phe Thr Leu His Ile Pro Ile Thr
770 775 780
Met Asn Phe Gly Val Asp Glu Val Lys Arg Phe Asn Asp Ala Val Asn
785 790 795 800
Ser Ala Ile Arg Ile Asp Glu Asn Val Asn Val Ile Gly Ile Asp Arg
805 810 815
Gly Glu Arg Asn Leu Leu Tyr Val Val Val Ile Asp Ser Lys Gly Asn
820 825 830
Ile Leu Glu Gln Ile Ser Leu Asn Ser Ile Ile Asn Lys Glu Tyr Asp
835 840 845
Ile Glu Thr Asp Tyr His Ala Leu Leu Asp Glu Arg Glu Gly Gly Arg
850 855 860
Asp Lys Ala Arg Lys Asp Trp Asn Thr Val Glu Asn Ile Arg Asp Leu
865 870 875 880
Lys Ala Gly Leu Tyr Leu Gln Val Val Asn Val Val Ala Lys Leu Val
885 890 895
Leu Lys Tyr Asn Ala Ile Ile Cys Leu Glu Asp Leu Asn Phe Gly Phe
900 905 910
Lys Arg Gly Arg Gln Lys Val Glu Lys Gln Val Tyr Gln Lys Phe Glu
915 920 925
Lys Met Leu Ile Asp Lys Leu Asn Tyr Leu Val Ile Asp Lys Ser Arg
930 935 940
Glu Gln Thr Ser Pro Lys Glu Leu Gly Gly Ala Leu Asn Ala Leu Gln
945 950 955 960
Leu Thr Ser Lys Phe Lys Ser Phe Lys Glu Leu Gly Lys Gln Ser Gly
965 970 975
Val Ile Tyr Tyr Val Pro Ala Tyr Leu Thr Ser Lys Ile Asp Pro Thr
980 985 990
Thr Gly Phe Ala Asn Leu Phe Tyr Met Lys Cys Glu Asn Val Glu Lys
995 1000 1005
Ser Lys Arg Phe Phe Asp Gly Phe Asp Phe Ile Arg Phe Asn Ala
1010 1015 1020
Leu Glu Asn Val Phe Glu Phe Gly Phe Asp Tyr Arg Ser Phe Thr
1025 1030 1035
Gln Arg Ala Cys Gly Ile Asn Ser Lys Trp Thr Val Cys Thr Asn
1040 1045 1050
Gly Glu Arg Ile Ile Lys Tyr Arg Asn Pro Asp Lys Asn Asn Met
1055 1060 1065
Phe Asp Glu Lys Val Val Val Val Thr Asp Glu Met Lys Asn Leu
1070 1075 1080
Phe Glu Gln Tyr Lys Ile Pro Tyr Glu Asp Gly Arg Asn Val Lys
1085 1090 1095
Asp Met Ile Ile Ser Asn Glu Glu Ala Glu Phe Tyr Arg Arg Leu
1100 1105 1110
Tyr Arg Leu Leu Gln Gln Thr Leu Gln Met Arg Asn Ser Thr Ser
1115 1120 1125
Asp Gly Thr Arg Asp Tyr Ile Ile Ser Pro Val Lys Asn Lys Arg
1130 1135 1140
Glu Ala Tyr Phe Asn Ser Glu Leu Ser Asp Gly Ser Val Pro Lys
1145 1150 1155
Asp Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala Arg Lys Gly Leu
1160 1165 1170
Trp Val Leu Glu Gln Ile Arg Gln Lys Ser Glu Gly Glu Lys Ile
1175 1180 1185
Asn Leu Ala Met Thr Asn Ala Glu Trp Leu Glu Tyr Ala Gln Thr
1190 1195 1200
His Leu Leu
1205
<210> 8
<211> 1233
<212> PRT
<213> 未知
<220>
<223> 毛螺菌科物种
<400> 8
Met Asp Tyr Gly Asn Gly Gln Phe Glu Arg Arg Ala Pro Leu Thr Lys
1 5 10 15
Thr Ile Thr Leu Arg Leu Lys Pro Ile Gly Glu Thr Arg Glu Thr Ile
20 25 30
Arg Glu Gln Lys Leu Leu Glu Gln Asp Ala Ala Phe Arg Lys Leu Val
35 40 45
Glu Thr Val Thr Pro Ile Val Asp Asp Cys Ile Arg Lys Ile Ala Asp
50 55 60
Asn Ala Leu Cys His Phe Gly Thr Glu Tyr Asp Phe Ser Cys Leu Gly
65 70 75 80
Asn Ala Ile Ser Lys Asn Asp Ser Lys Ala Ile Lys Lys Glu Thr Glu
85 90 95
Lys Val Glu Lys Leu Leu Ala Lys Val Leu Thr Glu Asn Leu Pro Asp
100 105 110
Gly Leu Arg Lys Val Asn Asp Ile Asn Ser Ala Ala Phe Ile Gln Asp
115 120 125
Thr Leu Thr Ser Phe Val Gln Asp Asp Ala Asp Lys Arg Val Leu Ile
130 135 140
Gln Glu Leu Lys Gly Lys Thr Val Leu Met Gln Arg Phe Leu Thr Thr
145 150 155 160
Arg Ile Thr Ala Leu Thr Val Trp Leu Pro Asp Arg Val Phe Glu Asn
165 170 175
Phe Asn Ile Phe Ile Glu Asn Ala Glu Lys Met Arg Ile Leu Leu Asp
180 185 190
Ser Pro Leu Asn Glu Lys Ile Met Lys Phe Asp Pro Asp Ala Glu Gln
195 200 205
Tyr Ala Ser Leu Glu Phe Tyr Gly Gln Cys Leu Ser Gln Lys Asp Ile
210 215 220
Asp Ser Tyr Asn Leu Ile Ile Ser Gly Ile Tyr Ala Asp Asp Glu Val
225 230 235 240
Lys Asn Pro Gly Ile Asn Glu Ile Val Lys Glu Tyr Asn Gln Gln Ile
245 250 255
Arg Gly Asp Lys Asp Glu Ser Pro Leu Pro Lys Leu Lys Lys Leu His
260 265 270
Lys Gln Ile Leu Met Pro Val Glu Lys Ala Phe Phe Val Arg Val Leu
275 280 285
Ser Asn Asp Ser Asp Ala Arg Ser Ile Leu Glu Lys Ile Leu Lys Asp
290 295 300
Thr Glu Met Leu Pro Ser Lys Ile Ile Glu Ala Met Lys Glu Ala Asp
305 310 315 320
Ala Gly Asp Ile Ala Val Tyr Gly Ser Arg Leu His Glu Leu Ser His
325 330 335
Val Ile Tyr Gly Asp His Gly Lys Leu Ser Gln Ile Ile Tyr Asp Lys
340 345 350
Glu Ser Lys Arg Ile Ser Glu Leu Met Glu Thr Leu Ser Pro Lys Glu
355 360 365
Arg Lys Glu Ser Lys Lys Arg Leu Glu Gly Leu Glu Glu His Ile Arg
370 375 380
Lys Ser Thr Tyr Thr Phe Asp Glu Leu Asn Arg Tyr Ala Glu Lys Asn
385 390 395 400
Val Met Ala Ala Tyr Ile Ala Ala Val Glu Glu Ser Cys Ala Glu Ile
405 410 415
Met Arg Lys Glu Lys Asp Leu Arg Thr Leu Leu Ser Lys Glu Asp Val
420 425 430
Lys Ile Arg Gly Asn Arg His Asn Thr Leu Ile Val Lys Asn Tyr Phe
435 440 445
Asn Ala Trp Thr Val Phe Arg Asn Leu Ile Arg Ile Leu Arg Arg Lys
450 455 460
Ser Glu Ala Glu Ile Asp Ser Asp Phe Tyr Asp Val Leu Asp Asp Ser
465 470 475 480
Val Glu Val Leu Ser Leu Thr Tyr Lys Gly Glu Asn Leu Cys Arg Ser
485 490 495
Tyr Ile Thr Lys Lys Ile Gly Ser Asp Leu Lys Pro Glu Ile Ala Thr
500 505 510
Tyr Gly Ser Ala Leu Arg Pro Asn Ser Arg Trp Trp Ser Pro Gly Glu
515 520 525
Lys Phe Asn Val Lys Phe His Thr Ile Val Arg Arg Asp Gly Arg Leu
530 535 540
Tyr Tyr Phe Ile Leu Pro Lys Gly Ala Lys Pro Val Glu Leu Glu Asp
545 550 555 560
Met Asp Gly Asp Ile Glu Cys Leu Gln Met Arg Lys Ile Pro Asn Pro
565 570 575
Thr Ile Phe Leu Pro Lys Leu Val Phe Lys Asp Pro Glu Ala Phe Phe
580 585 590
Arg Asp Asn Pro Glu Ala Asp Glu Phe Val Phe Leu Ser Gly Met Lys
595 600 605
Ala Pro Val Thr Ile Thr Arg Glu Thr Tyr Glu Ala Tyr Arg Tyr Lys
610 615 620
Leu Tyr Thr Val Gly Lys Leu Arg Asp Gly Glu Val Ser Glu Glu Glu
625 630 635 640
Tyr Lys Arg Ala Leu Leu Gln Val Leu Thr Ala Tyr Lys Glu Phe Leu
645 650 655
Glu Asn Arg Met Ile Tyr Ala Asp Leu Asn Phe Gly Phe Lys Asp Leu
660 665 670
Glu Glu Tyr Lys Asp Ser Ser Glu Phe Ile Lys Gln Val Glu Thr His
675 680 685
Asn Thr Phe Met Cys Trp Ala Lys Val Ser Ser Ser Gln Leu Asp Asp
690 695 700
Leu Val Lys Ser Gly Asn Gly Leu Leu Phe Glu Ile Trp Ser Glu Arg
705 710 715 720
Leu Glu Ser Tyr Tyr Lys Tyr Gly Asn Glu Lys Val Leu Arg Gly Tyr
725 730 735
Glu Gly Val Leu Leu Ser Ile Leu Lys Asp Glu Asn Leu Val Ser Met
740 745 750
Arg Thr Leu Leu Asn Ser Arg Pro Met Leu Val Tyr Arg Pro Lys Glu
755 760 765
Ser Ser Lys Pro Met Val Val His Arg Asp Gly Ser Arg Val Val Asp
770 775 780
Arg Phe Asp Lys Asp Gly Lys Tyr Ile Pro Pro Glu Val His Asp Glu
785 790 795 800
Leu Tyr Arg Phe Phe Asn Asn Leu Leu Ile Lys Glu Lys Leu Gly Glu
805 810 815
Lys Ala Arg Lys Ile Leu Asp Asn Lys Lys Val Lys Val Lys Val Leu
820 825 830
Glu Ser Glu Arg Val Lys Trp Ser Lys Phe Tyr Asp Glu Gln Phe Ala
835 840 845
Val Thr Phe Ser Val Lys Lys Asn Ala Asp Cys Leu Asp Thr Thr Lys
850 855 860
Asp Leu Asn Ala Glu Val Met Glu Gln Tyr Ser Glu Ser Asn Arg Leu
865 870 875 880
Ile Leu Ile Arg Asn Thr Thr Asp Ile Leu Tyr Tyr Leu Val Leu Asp
885 890 895
Lys Asn Gly Lys Val Leu Lys Gln Arg Ser Leu Asn Ile Ile Asn Asp
900 905 910
Gly Ala Arg Asp Val Asp Trp Lys Glu Arg Phe Arg Gln Val Thr Lys
915 920 925
Asp Arg Asn Glu Gly Tyr Asn Glu Trp Asp Tyr Ser Arg Thr Ser Asn
930 935 940
Asp Leu Lys Glu Val Tyr Leu Asn Tyr Ala Leu Lys Glu Ile Ala Glu
945 950 955 960
Ala Val Ile Glu Tyr Asn Ala Ile Leu Ile Ile Glu Lys Met Ser Asn
965 970 975
Ala Phe Lys Asp Lys Tyr Ser Phe Leu Asp Asp Val Thr Phe Lys Gly
980 985 990
Phe Glu Thr Lys Lys Leu Ala Lys Leu Ser Asp Leu His Phe Arg Gly
995 1000 1005
Ile Lys Asp Gly Glu Pro Cys Ser Phe Thr Asn Pro Leu Gln Leu
1010 1015 1020
Cys Gln Asn Asp Ser Asn Lys Ile Leu Gln Asp Gly Val Ile Phe
1025 1030 1035
Met Val Pro Asn Ser Met Thr Arg Ser Leu Asp Pro Asp Thr Gly
1040 1045 1050
Phe Ile Phe Ala Ile Asn Asp His Asn Ile Arg Thr Lys Lys Ala
1055 1060 1065
Lys Leu Asn Phe Leu Ser Lys Phe Asp Gln Leu Lys Val Ser Ser
1070 1075 1080
Glu Gly Cys Leu Ile Met Lys Tyr Ser Gly Asp Ser Leu Pro Thr
1085 1090 1095
His Asn Thr Asp Asn Arg Val Trp Asn Cys Cys Cys Asn His Pro
1100 1105 1110
Ile Thr Asn Tyr Asp Arg Glu Thr Lys Lys Val Glu Phe Ile Glu
1115 1120 1125
Glu Pro Val Glu Glu Leu Ser Arg Val Leu Glu Glu Asn Gly Ile
1130 1135 1140
Glu Thr Asp Thr Glu Leu Asn Lys Leu Asn Glu Arg Glu Asn Val
1145 1150 1155
Pro Gly Lys Val Val Asp Ala Ile Tyr Ser Leu Val Leu Asn Tyr
1160 1165 1170
Leu Arg Gly Thr Val Ser Gly Val Ala Gly Gln Arg Ala Val Tyr
1175 1180 1185
Tyr Ser Pro Val Thr Gly Lys Lys Tyr Asp Ile Ser Phe Ile Gln
1190 1195 1200
Ala Met Asn Leu Asn Arg Lys Cys Asp Tyr Tyr Arg Ile Gly Ser
1205 1210 1215
Lys Glu Arg Gly Glu Trp Thr Asp Phe Val Ala Gln Leu Ile Asn
1220 1225 1230
<210> 9
<211> 1227
<212> PRT
<213> 未知
<220>
<223> 毛螺菌科物种
<400> 9
Met Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr
1 5 10 15
Leu Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp
20 25 30
Asn Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys
35 40 45
Gly Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp
50 55 60
Val Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu
65 70 75 80
Phe Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn
85 90 95
Leu Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn
100 105 110
Glu Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu
115 120 125
Pro Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe
130 135 140
Asn Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn
145 150 155 160
Met Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile
165 170 175
Asn Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys
180 185 190
Val Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys
195 200 205
Ile Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe
210 215 220
Phe Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile
225 230 235 240
Ile Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn
245 250 255
Glu Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys
260 265 270
Phe Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser
275 280 285
Phe Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe
290 295 300
Arg Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys
305 310 315 320
Leu Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile
325 330 335
Phe Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe
340 345 350
Gly Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp
355 360 365
Ile His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp
370 375 380
Arg Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu
385 390 395 400
Gln Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu
405 410 415
Ile Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser
420 425 430
Glu Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys
435 440 445
Asn Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys
450 455 460
Ser Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr
465 470 475 480
Asn Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile
485 490 495
Leu Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr
500 505 510
Gln Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro
515 520 525
Gln Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala
530 535 540
Thr Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys
545 550 555 560
Lys Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly
565 570 575
Asn Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met
580 585 590
Leu Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro
595 600 605
Ser Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly
610 615 620
Asp Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys
625 630 635 640
Asp Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn
645 650 655
Phe Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu
660 665 670
Val Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys
675 680 685
Glu Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile
690 695 700
Tyr Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His
705 710 715 720
Thr Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile
725 730 735
Arg Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys
740 745 750
Lys Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys
755 760 765
Asn Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr
770 775 780
Lys Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile
785 790 795 800
Ala Asn Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu
805 810 815
Val Arg Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile
820 825 830
Asp Arg Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys
835 840 845
Gly Asn Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe
850 855 860
Asn Gly Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys
865 870 875 880
Glu Lys Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn
885 890 895
Ile Lys Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile
900 905 910
Cys Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu
915 920 925
Asn Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val Tyr
930 935 940
Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met Val Asp
945 950 955 960
Lys Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys Gly Tyr Gln
965 970 975
Ile Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser Thr Gln Asn Gly
980 985 990
Phe Ile Phe Tyr Ile Pro Ala Trp Leu Thr Ser Lys Ile Asp Pro Ser
995 1000 1005
Thr Gly Phe Val Asn Leu Leu Lys Thr Lys Tyr Thr Ser Ile Ala
1010 1015 1020
Asp Lys Lys Phe Ile Ser Ser Phe Asp Arg Ile Met Tyr Val Pro
1025 1030 1035
Glu Glu Asp Leu Phe Glu Phe Ala Leu Asp Tyr Lys Asn Phe Ser
1040 1045 1050
Arg Thr Asp Ala Asp Tyr Ile Lys Lys Trp Lys Leu Tyr Ser Tyr
1055 1060 1065
Gly Asn Arg Ile Arg Ile Phe Arg Asn Pro Lys Lys Asn Asn Val
1070 1075 1080
Phe Asp Trp Glu Glu Val Cys Leu Thr Ser Ala Tyr Lys Glu Leu
1085 1090 1095
Phe Asn Lys Tyr Gly Ile Asn Tyr Gln Gln Gly Asp Ile Arg Ala
1100 1105 1110
Leu Leu Cys Glu Gln Ser Asp Lys Ala Phe Tyr Ser Ser Phe Met
1115 1120 1125
Ala Leu Met Ser Leu Met Leu Gln Met Arg Asn Ser Ile Thr Gly
1130 1135 1140
Arg Thr Asp Val Asp Phe Leu Ile Ser Pro Val Lys Asn Ser Asp
1145 1150 1155
Gly Ile Phe Tyr Asp Ser Arg Asn Tyr Glu Ala Gln Glu Asn Ala
1160 1165 1170
Ile Leu Pro Lys Asn Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala
1175 1180 1185
Arg Lys Val Leu Trp Ala Ile Gly Gln Phe Lys Lys Ala Glu Asp
1190 1195 1200
Glu Lys Leu Asp Lys Val Lys Ile Ala Ser Asn Lys Glu Trp Leu
1205 1210 1215
Glu Tyr Ala Gln Thr Ser Val Lys His
1220 1225
<210> 10
<211> 1264
<212> PRT
<213> Leptospira inadai
<400> 10
Met Glu Asp Tyr Ser Gly Phe Val Asn Ile Tyr Ser Ile Gln Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr Leu Glu His Ile Glu
20 25 30
Lys Lys Gly Phe Leu Lys Lys Asp Lys Ile Arg Ala Glu Asp Tyr Lys
35 40 45
Ala Val Lys Lys Ile Ile Asp Lys Tyr His Arg Ala Tyr Ile Glu Glu
50 55 60
Val Phe Asp Ser Val Leu His Gln Lys Lys Lys Lys Asp Lys Thr Arg
65 70 75 80
Phe Ser Thr Gln Phe Ile Lys Glu Ile Lys Glu Phe Ser Glu Leu Tyr
85 90 95
Tyr Lys Thr Glu Lys Asn Ile Pro Asp Lys Glu Arg Leu Glu Ala Leu
100 105 110
Ser Glu Lys Leu Arg Lys Met Leu Val Gly Ala Phe Lys Gly Glu Phe
115 120 125
Ser Glu Glu Val Ala Glu Lys Tyr Asn Lys Asn Leu Phe Ser Lys Glu
130 135 140
Leu Ile Arg Asn Glu Ile Glu Lys Phe Cys Glu Thr Asp Glu Glu Arg
145 150 155 160
Lys Gln Val Ser Asn Phe Lys Ser Phe Thr Thr Tyr Phe Thr Gly Phe
165 170 175
His Ser Asn Arg Gln Asn Ile Tyr Ser Asp Glu Lys Lys Ser Thr Ala
180 185 190
Ile Gly Tyr Arg Ile Ile His Gln Asn Leu Pro Lys Phe Leu Asp Asn
195 200 205
Leu Lys Ile Ile Glu Ser Ile Gln Arg Arg Phe Lys Asp Phe Pro Trp
210 215 220
Ser Asp Leu Lys Lys Asn Leu Lys Lys Ile Asp Lys Asn Ile Lys Leu
225 230 235 240
Thr Glu Tyr Phe Ser Ile Asp Gly Phe Val Asn Val Leu Asn Gln Lys
245 250 255
Gly Ile Asp Ala Tyr Asn Thr Ile Leu Gly Gly Lys Ser Glu Glu Ser
260 265 270
Gly Glu Lys Ile Gln Gly Leu Asn Glu Tyr Ile Asn Leu Tyr Arg Gln
275 280 285
Lys Asn Asn Ile Asp Arg Lys Asn Pro Leu Asn Val Lys Ile Leu Phe
290 295 300
Lys Gln Ile Leu Gly Asp Arg Glu Thr Lys Ser Phe Ile Pro Glu Ala
305 310 315 320
Phe Pro Asp Asp Gln Ser Val Leu Asn Ser Ile Thr Glu Phe Ala Lys
325 330 335
Tyr Leu Lys Leu Asp Lys Lys Lys Lys Ser Ile Ile Ala Glu Leu Lys
340 345 350
Lys Phe Leu Ser Ser Phe Asn Arg Tyr Glu Leu Asp Gly Ile Tyr Leu
355 360 365
Ala Asn Asp Asn Ser Leu Ala Ser Ile Ser Thr Phe Leu Phe Asp Asp
370 375 380
Trp Ser Phe Ile Lys Lys Ser Val Ser Phe Lys Tyr Asp Glu Ser Val
385 390 395 400
Gly Asp Pro Lys Lys Lys Ile Lys Ser Pro Leu Lys Tyr Glu Lys Glu
405 410 415
Lys Glu Lys Trp Leu Lys Gln Lys Tyr Tyr Thr Ile Ser Phe Leu Asn
420 425 430
Asp Ala Ile Glu Ser Tyr Ser Lys Ser Gln Asp Glu Lys Arg Val Lys
435 440 445
Ile Arg Leu Glu Ala Tyr Phe Ala Glu Phe Lys Ser Lys Asp Asp Ala
450 455 460
Lys Lys Gln Phe Asp Leu Leu Glu Arg Ile Glu Glu Ala Tyr Ala Ile
465 470 475 480
Val Glu Pro Leu Leu Gly Ala Glu Tyr Pro Arg Asp Arg Asn Leu Lys
485 490 495
Ala Asp Lys Lys Glu Val Gly Lys Ile Lys Asp Phe Leu Asp Ser Ile
500 505 510
Lys Ser Leu Gln Phe Phe Leu Lys Pro Leu Leu Ser Ala Glu Ile Phe
515 520 525
Asp Glu Lys Asp Leu Gly Phe Tyr Asn Gln Leu Glu Gly Tyr Tyr Glu
530 535 540
Glu Ile Asp Ile Ser Gly His Leu Tyr Asn Lys Val Arg Asn Tyr Leu
545 550 555 560
Thr Gly Lys Ile Tyr Ser Lys Glu Lys Phe Lys Leu Asn Phe Glu Asn
565 570 575
Ser Thr Leu Leu Lys Gly Trp Asp Glu Asn Arg Glu Val Ala Asn Leu
580 585 590
Cys Val Ile Phe Arg Glu Asp Gln Lys Tyr Tyr Leu Gly Val Met Asp
595 600 605
Lys Glu Asn Asn Thr Ile Leu Ser Asp Ile Pro Lys Val Lys Pro Asn
610 615 620
Glu Leu Phe Tyr Glu Lys Met Val Tyr Lys Leu Ile Pro Thr Pro His
625 630 635 640
Met Gln Leu Pro Arg Ile Ile Phe Ser Ser Asp Asn Leu Ser Ile Tyr
645 650 655
Asn Pro Ser Lys Ser Ile Leu Lys Ile Arg Glu Ala Lys Ser Phe Lys
660 665 670
Glu Gly Lys Asn Phe Lys Leu Lys Asp Cys His Lys Phe Ile Asp Phe
675 680 685
Tyr Lys Glu Ser Ile Ser Lys Asn Glu Asp Trp Ser Arg Phe Asp Phe
690 695 700
Lys Phe Ser Lys Thr Ser Ser Tyr Glu Asn Ile Ser Glu Phe Tyr Arg
705 710 715 720
Glu Val Glu Arg Gln Gly Tyr Asn Leu Asp Phe Lys Lys Val Ser Lys
725 730 735
Phe Tyr Ile Asp Ser Leu Val Glu Asp Gly Lys Leu Tyr Leu Phe Gln
740 745 750
Ile Tyr Asn Lys Asp Phe Ser Ile Phe Ser Lys Gly Lys Pro Asn Leu
755 760 765
His Thr Ile Tyr Phe Arg Ser Leu Phe Ser Lys Glu Asn Leu Lys Asp
770 775 780
Val Cys Leu Lys Leu Asn Gly Glu Ala Glu Met Phe Phe Arg Lys Lys
785 790 795 800
Ser Ile Asn Tyr Asp Glu Lys Lys Lys Arg Glu Gly His His Pro Glu
805 810 815
Leu Phe Glu Lys Leu Lys Tyr Pro Ile Leu Lys Asp Lys Arg Tyr Ser
820 825 830
Glu Asp Lys Phe Gln Phe His Leu Pro Ile Ser Leu Asn Phe Lys Ser
835 840 845
Lys Glu Arg Leu Asn Phe Asn Leu Lys Val Asn Glu Phe Leu Lys Arg
850 855 860
Asn Lys Asp Ile Asn Ile Ile Gly Ile Asp Arg Gly Glu Arg Asn Leu
865 870 875 880
Leu Tyr Leu Val Met Ile Asn Gln Lys Gly Glu Ile Leu Lys Gln Thr
885 890 895
Leu Leu Asp Ser Met Gln Ser Gly Lys Gly Arg Pro Glu Ile Asn Tyr
900 905 910
Lys Glu Lys Leu Gln Glu Lys Glu Ile Glu Arg Asp Lys Ala Arg Lys
915 920 925
Ser Trp Gly Thr Val Glu Asn Ile Lys Glu Leu Lys Glu Gly Tyr Leu
930 935 940
Ser Ile Val Ile His Gln Ile Ser Lys Leu Met Val Glu Asn Asn Ala
945 950 955 960
Ile Val Val Leu Glu Asp Leu Asn Ile Gly Phe Lys Arg Gly Arg Gln
965 970 975
Lys Val Glu Arg Gln Val Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp
980 985 990
Lys Leu Asn Phe Leu Val Phe Lys Glu Asn Lys Pro Thr Glu Pro Gly
995 1000 1005
Gly Val Leu Lys Ala Tyr Gln Leu Thr Asp Glu Phe Gln Ser Phe
1010 1015 1020
Glu Lys Leu Ser Lys Gln Thr Gly Phe Leu Phe Tyr Val Pro Ser
1025 1030 1035
Trp Asn Thr Ser Lys Ile Asp Pro Arg Thr Gly Phe Ile Asp Phe
1040 1045 1050
Leu His Pro Ala Tyr Glu Asn Ile Glu Lys Ala Lys Gln Trp Ile
1055 1060 1065
Asn Lys Phe Asp Ser Ile Arg Phe Asn Ser Lys Met Asp Trp Phe
1070 1075 1080
Glu Phe Thr Ala Asp Thr Arg Lys Phe Ser Glu Asn Leu Met Leu
1085 1090 1095
Gly Lys Asn Arg Val Trp Val Ile Cys Thr Thr Asn Val Glu Arg
1100 1105 1110
Tyr Phe Thr Ser Lys Thr Ala Asn Ser Ser Ile Gln Tyr Asn Ser
1115 1120 1125
Ile Gln Ile Thr Glu Lys Leu Lys Glu Leu Phe Val Asp Ile Pro
1130 1135 1140
Phe Ser Asn Gly Gln Asp Leu Lys Pro Glu Ile Leu Arg Lys Asn
1145 1150 1155
Asp Ala Val Phe Phe Lys Ser Leu Leu Phe Tyr Ile Lys Thr Thr
1160 1165 1170
Leu Ser Leu Arg Gln Asn Asn Gly Lys Lys Gly Glu Glu Glu Lys
1175 1180 1185
Asp Phe Ile Leu Ser Pro Val Val Asp Ser Lys Gly Arg Phe Phe
1190 1195 1200
Asn Ser Leu Glu Ala Ser Asp Asp Glu Pro Lys Asp Ala Asp Ala
1205 1210 1215
Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Leu Met Asn Leu Leu
1220 1225 1230
Val Leu Asn Glu Thr Lys Glu Glu Asn Leu Ser Arg Pro Lys Trp
1235 1240 1245
Lys Ile Lys Asn Lys Asp Trp Leu Glu Phe Val Trp Glu Arg Asn
1250 1255 1260
Arg
<210> 11
<211> 1373
<212> PRT
<213> Moraxella bovoculi
<400> 11
Met Leu Phe Gln Asp Phe Thr His Leu Tyr Pro Leu Ser Lys Thr Val
1 5 10 15
Arg Phe Glu Leu Phe Ile Asp Arg Thr Leu Glu His Ile His Ala Lys
20 25 30
Asn Phe Leu Ser Gln Asp Glu Thr Met Ala Asp Met His Gln Lys Val
35 40 45
Lys Val Ile Leu Asp Asp Tyr His Arg Asp Phe Ile Ala Asp Met Met
50 55 60
Gly Glu Val Lys Leu Thr Lys Leu Ala Glu Phe Tyr Asp Val Tyr Leu
65 70 75 80
Lys Phe Arg Lys Asn Pro Lys Asp Asp Glu Leu Gln Lys Ala Gln Leu
85 90 95
Lys Asp Leu Gln Ala Val Leu Arg Lys Glu Ile Val Lys Pro Ile Gly
100 105 110
Asn Gly Gly Lys Tyr Lys Ala Gly Tyr Asp Arg Leu Phe Gly Ala Lys
115 120 125
Leu Phe Lys Asp Gly Lys Glu Leu Gly Asp Leu Ala Lys Phe Val Ile
130 135 140
Ala Gln Glu Gly Glu Ser Ser Pro Lys Leu Ala His Leu Ala His Phe
145 150 155 160
Glu Lys Phe Ser Thr Tyr Phe Thr Gly Phe His Asp Asn Arg Lys Asn
165 170 175
Met Tyr Ser Asp Glu Asp Lys His Thr Ala Ile Ala Tyr Arg Leu Ile
180 185 190
His Glu Asn Leu Pro Arg Phe Ile Asp Asn Leu Gln Ile Leu Thr Thr
195 200 205
Ile Lys Gln Lys His Ser Ala Leu Tyr Asp Gln Ile Ile Asn Glu Leu
210 215 220
Thr Ala Ser Gly Leu Asp Val Ser Leu Ala Ser His Leu Asp Gly Tyr
225 230 235 240
His Lys Leu Leu Thr Gln Glu Gly Ile Thr Ala Tyr Asn Thr Leu Leu
245 250 255
Gly Gly Ile Ser Gly Glu Ala Gly Ser Pro Lys Ile Gln Gly Ile Asn
260 265 270
Glu Leu Ile Asn Ser His His Asn Gln His Cys His Lys Ser Glu Arg
275 280 285
Ile Ala Lys Leu Arg Pro Leu His Lys Gln Ile Leu Ser Asp Gly Met
290 295 300
Ser Val Ser Phe Leu Pro Ser Lys Phe Ala Asp Asp Ser Glu Met Cys
305 310 315 320
Gln Ala Val Asn Glu Phe Tyr Arg His Tyr Ala Asp Val Phe Ala Lys
325 330 335
Val Gln Ser Leu Phe Asp Gly Phe Asp Asp His Gln Lys Asp Gly Ile
340 345 350
Tyr Val Glu His Lys Asn Leu Asn Glu Leu Ser Lys Gln Ala Phe Gly
355 360 365
Asp Phe Ala Leu Leu Gly Arg Val Leu Asp Gly Tyr Tyr Val Asp Val
370 375 380
Val Asn Pro Glu Phe Asn Glu Arg Phe Ala Lys Ala Lys Thr Asp Asn
385 390 395 400
Ala Lys Ala Lys Leu Thr Lys Glu Lys Asp Lys Phe Ile Lys Gly Val
405 410 415
His Ser Leu Ala Ser Leu Glu Gln Ala Ile Glu His Tyr Thr Ala Arg
420 425 430
His Asp Asp Glu Ser Val Gln Ala Gly Lys Leu Gly Gln Tyr Phe Lys
435 440 445
His Gly Leu Ala Gly Val Asp Asn Pro Ile Gln Lys Ile His Asn Asn
450 455 460
His Ser Thr Ile Lys Gly Phe Leu Glu Arg Glu Arg Pro Ala Gly Glu
465 470 475 480
Arg Ala Leu Pro Lys Ile Lys Ser Gly Lys Asn Pro Glu Met Thr Gln
485 490 495
Leu Arg Gln Leu Lys Glu Leu Leu Asp Asn Ala Leu Asn Val Ala His
500 505 510
Phe Ala Lys Leu Leu Thr Thr Lys Thr Thr Leu Asp Asn Gln Asp Gly
515 520 525
Asn Phe Tyr Gly Glu Phe Gly Val Leu Tyr Asp Glu Leu Ala Lys Ile
530 535 540
Pro Thr Leu Tyr Asn Lys Val Arg Asp Tyr Leu Ser Gln Lys Pro Phe
545 550 555 560
Ser Thr Glu Lys Tyr Lys Leu Asn Phe Gly Asn Pro Thr Leu Leu Asn
565 570 575
Gly Trp Asp Leu Asn Lys Glu Lys Asp Asn Phe Gly Val Ile Leu Gln
580 585 590
Lys Asp Gly Cys Tyr Tyr Leu Ala Leu Leu Asp Lys Ala His Lys Lys
595 600 605
Val Phe Asp Asn Ala Pro Asn Thr Gly Lys Ser Ile Tyr Gln Lys Met
610 615 620
Ile Tyr Lys Tyr Leu Glu Val Arg Lys Gln Phe Pro Lys Val Phe Phe
625 630 635 640
Ser Lys Glu Ala Ile Ala Ile Asn Tyr His Pro Ser Lys Glu Leu Val
645 650 655
Glu Ile Lys Asp Lys Gly Arg Gln Arg Ser Asp Asp Glu Arg Leu Lys
660 665 670
Leu Tyr Arg Phe Ile Leu Glu Cys Leu Lys Ile His Pro Lys Tyr Asp
675 680 685
Lys Lys Phe Glu Gly Ala Ile Gly Asp Ile Gln Leu Phe Lys Lys Asp
690 695 700
Lys Lys Gly Arg Glu Val Pro Ile Ser Glu Lys Asp Leu Phe Lys Asp
705 710 715 720
Ile Asn Gly Ile Phe Ser Ser Lys Pro Lys Leu Glu Met Glu Asp Phe
725 730 735
Phe Ile Gly Glu Phe Lys Arg Tyr Asn Pro Ser Gln Asp Leu Val Asp
740 745 750
Gln Tyr Asn Ile Tyr Lys Lys Ile Asp Ser Asn Asp Asn Arg Lys Lys
755 760 765
Glu Asn Phe Tyr Asn Asn His Pro Lys Phe Lys Lys Asp Leu Val Arg
770 775 780
Tyr Tyr Tyr Glu Ser Met Cys Lys His Glu Glu Trp Glu Glu Ser Phe
785 790 795 800
Glu Phe Ser Lys Lys Leu Gln Asp Ile Gly Cys Tyr Val Asp Val Asn
805 810 815
Glu Leu Phe Thr Glu Ile Glu Thr Arg Arg Leu Asn Tyr Lys Ile Ser
820 825 830
Phe Cys Asn Ile Asn Ala Asp Tyr Ile Asp Glu Leu Val Glu Gln Gly
835 840 845
Gln Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Pro Lys Ala
850 855 860
His Gly Lys Pro Asn Leu His Thr Leu Tyr Phe Lys Ala Leu Phe Ser
865 870 875 880
Glu Asp Asn Leu Ala Asp Pro Ile Tyr Lys Leu Asn Gly Glu Ala Gln
885 890 895
Ile Phe Tyr Arg Lys Ala Ser Leu Asp Met Asn Glu Thr Thr Ile His
900 905 910
Arg Ala Gly Glu Val Leu Glu Asn Lys Asn Pro Asp Asn Pro Lys Lys
915 920 925
Arg Gln Phe Val Tyr Asp Ile Ile Lys Asp Lys Arg Tyr Thr Gln Lys
930 935 940
Asp Phe Met Leu His Val Pro Ile Thr Met Asn Phe Gly Val Gln Gly
945 950 955 960
Met Thr Ile Lys Glu Phe Asn Lys Lys Val Asn Gln Ser Ile Gln Gln
965 970 975
Tyr Asp Glu Val Asn Val Ile Gly Ile Asp Arg Gly Glu Arg His Leu
980 985 990
Leu Tyr Leu Thr Val Ile Asn Ser Lys Gly Glu Ile Leu Glu Gln Cys
995 1000 1005
Ser Leu Asn Asp Ile Thr Thr Ala Ser Ala Asn Gly Thr Gln Met
1010 1015 1020
Thr Thr Pro Tyr His Lys Ile Leu Asp Lys Arg Glu Ile Glu Arg
1025 1030 1035
Leu Asn Ala Arg Val Gly Trp Gly Glu Ile Glu Thr Ile Lys Glu
1040 1045 1050
Leu Lys Ser Gly Tyr Leu Ser His Val Val His Gln Ile Ser Gln
1055 1060 1065
Leu Met Leu Lys Tyr Asn Ala Ile Val Val Leu Glu Asp Leu Asn
1070 1075 1080
Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Ile Tyr
1085 1090 1095
Gln Asn Phe Glu Asn Ala Leu Ile Lys Lys Leu Asn His Leu Val
1100 1105 1110
Leu Lys Asp Lys Ala Asp Asp Glu Ile Gly Ser Tyr Lys Asn Ala
1115 1120 1125
Leu Gln Leu Thr Asn Asn Phe Thr Asp Leu Lys Ser Ile Gly Lys
1130 1135 1140
Gln Thr Gly Phe Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Lys
1145 1150 1155
Ile Asp Pro Glu Thr Gly Phe Val Asp Leu Leu Lys Pro Arg Tyr
1160 1165 1170
Glu Asn Ile Gln Ala Ser Gln Ala Phe Phe Gly Lys Phe Asp Lys
1175 1180 1185
Ile Cys Tyr Asn Ala Asp Lys Asp Tyr Phe Glu Phe His Ile Asp
1190 1195 1200
Tyr Ala Lys Phe Thr Asp Lys Ala Lys Asn Ser Arg Gln Ile Trp
1205 1210 1215
Thr Ile Cys Ser His Gly Asp Lys Arg Tyr Val Tyr Asp Lys Thr
1220 1225 1230
Ala Asn Gln Asn Lys Gly Ala Ala Lys Gly Ile Asn Val Asn Asp
1235 1240 1245
Ile Leu Lys Ser Leu Phe Ala Arg His His Ile Asn Glu Lys Gln
1250 1255 1260
Pro Asn Leu Val Met Asp Ile Cys Gln Asn Asn Asp Lys Glu Phe
1265 1270 1275
His Lys Ser Leu Met Tyr Leu Leu Lys Thr Leu Leu Ala Leu Arg
1280 1285 1290
Tyr Ser Asn Ala Ser Ser Asp Glu Asp Phe Ile Leu Ser Pro Val
1295 1300 1305
Ala Asn Asp Glu Gly Val Phe Phe Asn Ser Ala Leu Ala Asp Asp
1310 1315 1320
Thr Gln Pro Gln Asn Ala Asp Ala Asn Gly Ala Tyr His Ile Ala
1325 1330 1335
Leu Lys Gly Leu Trp Leu Leu Asn Glu Leu Lys Asn Ser Asp Asp
1340 1345 1350
Leu Asn Lys Val Lys Leu Ala Ile Asp Asn Gln Thr Trp Leu Asn
1355 1360 1365
Phe Ala Gln Asn Arg
1370
<210> 12
<211> 1352
<212> PRT
<213> 未知
<220>
<223> Parcubacteria细菌
<400> 12
Met Glu Asn Ile Phe Asp Gln Phe Ile Gly Lys Tyr Ser Leu Ser Lys
1 5 10 15
Thr Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr Glu Asp Phe Leu
20 25 30
Lys Ile Asn Lys Val Phe Glu Lys Asp Gln Thr Ile Asp Asp Ser Tyr
35 40 45
Asn Gln Ala Lys Phe Tyr Phe Asp Ser Leu His Gln Lys Phe Ile Asp
50 55 60
Ala Ala Leu Ala Ser Asp Lys Thr Ser Glu Leu Ser Phe Gln Asn Phe
65 70 75 80
Ala Asp Val Leu Glu Lys Gln Asn Lys Ile Ile Leu Asp Lys Lys Arg
85 90 95
Glu Met Gly Ala Leu Arg Lys Arg Asp Lys Asn Ala Val Gly Ile Asp
100 105 110
Arg Leu Gln Lys Glu Ile Asn Asp Ala Glu Asp Ile Ile Gln Lys Glu
115 120 125
Lys Glu Lys Ile Tyr Lys Asp Val Arg Thr Leu Phe Asp Asn Glu Ala
130 135 140
Glu Ser Trp Lys Thr Tyr Tyr Gln Glu Arg Glu Val Asp Gly Lys Lys
145 150 155 160
Ile Thr Glu Ser Lys Ala Asp Leu Lys Gln Lys Gly Ala Asp Phe Leu
165 170 175
Thr Ala Ala Gly Ile Leu Lys Val Leu Lys Tyr Glu Phe Pro Glu Glu
180 185 190
Lys Glu Lys Glu Phe Gln Ala Lys Asn Gln Pro Ser Leu Phe Val Glu
195 200 205
Glu Lys Glu Asn Pro Gly Gln Lys Arg Tyr Ile Phe Asp Ser Phe Asp
210 215 220
Lys Phe Ala Gly Tyr Leu Thr Lys Phe Gln Gln Thr Lys Lys Asn Leu
225 230 235 240
Tyr Ala Ala Asp Gly Thr Ser Thr Ala Val Ala Thr Arg Ile Ala Asp
245 250 255
Asn Phe Ile Ile Phe His Gln Asn Thr Lys Val Phe Arg Asp Lys Tyr
260 265 270
Lys Asn Asn His Thr Asp Leu Gly Phe Asp Glu Glu Asn Ile Phe Glu
275 280 285
Ile Glu Arg Tyr Lys Asn Cys Leu Leu Gln Arg Glu Ile Glu His Ile
290 295 300
Lys Asn Glu Asn Ser Tyr Asn Lys Ile Ile Gly Arg Ile Asn Lys Lys
305 310 315 320
Ile Lys Glu Tyr Arg Asp Gln Lys Ala Lys Asp Thr Lys Leu Thr Lys
325 330 335
Ser Asp Phe Pro Phe Phe Lys Asn Leu Asp Lys Gln Ile Leu Gly Glu
340 345 350
Val Glu Lys Glu Lys Gln Leu Ile Glu Lys Thr Arg Glu Lys Thr Glu
355 360 365
Glu Asp Val Leu Ile Glu Arg Phe Lys Glu Phe Ile Glu Asn Asn Glu
370 375 380
Glu Arg Phe Thr Ala Ala Lys Lys Leu Met Asn Ala Phe Cys Asn Gly
385 390 395 400
Glu Phe Glu Ser Glu Tyr Glu Gly Ile Tyr Leu Lys Asn Lys Ala Ile
405 410 415
Asn Thr Ile Ser Arg Arg Trp Phe Val Ser Asp Arg Asp Phe Glu Leu
420 425 430
Lys Leu Pro Gln Gln Lys Ser Lys Asn Lys Ser Glu Lys Asn Glu Pro
435 440 445
Lys Val Lys Lys Phe Ile Ser Ile Ala Glu Ile Lys Asn Ala Val Glu
450 455 460
Glu Leu Asp Gly Asp Ile Phe Lys Ala Val Phe Tyr Asp Lys Lys Ile
465 470 475 480
Ile Ala Gln Gly Gly Ser Lys Leu Glu Gln Phe Leu Val Ile Trp Lys
485 490 495
Tyr Glu Phe Glu Tyr Leu Phe Arg Asp Ile Glu Arg Glu Asn Gly Glu
500 505 510
Lys Leu Leu Gly Tyr Asp Ser Cys Leu Lys Ile Ala Lys Gln Leu Gly
515 520 525
Ile Phe Pro Gln Glu Lys Glu Ala Arg Glu Lys Ala Thr Ala Val Ile
530 535 540
Lys Asn Tyr Ala Asp Ala Gly Leu Gly Ile Phe Gln Met Met Lys Tyr
545 550 555 560
Phe Ser Leu Asp Asp Lys Asp Arg Lys Asn Thr Pro Gly Gln Leu Ser
565 570 575
Thr Asn Phe Tyr Ala Glu Tyr Asp Gly Tyr Tyr Lys Asp Phe Glu Phe
580 585 590
Ile Lys Tyr Tyr Asn Glu Phe Arg Asn Phe Ile Thr Lys Lys Pro Phe
595 600 605
Asp Glu Asp Lys Ile Lys Leu Asn Phe Glu Asn Gly Ala Leu Leu Lys
610 615 620
Gly Trp Asp Glu Asn Lys Glu Tyr Asp Phe Met Gly Val Ile Leu Lys
625 630 635 640
Lys Glu Gly Arg Leu Tyr Leu Gly Ile Met His Lys Asn His Arg Lys
645 650 655
Leu Phe Gln Ser Met Gly Asn Ala Lys Gly Asp Asn Ala Asn Arg Tyr
660 665 670
Gln Lys Met Ile Tyr Lys Gln Ile Ala Asp Ala Ser Lys Asp Val Pro
675 680 685
Arg Leu Leu Leu Thr Ser Lys Lys Ala Met Glu Lys Phe Lys Pro Ser
690 695 700
Gln Glu Ile Leu Arg Ile Lys Lys Glu Lys Thr Phe Lys Arg Glu Ser
705 710 715 720
Lys Asn Phe Ser Leu Arg Asp Leu His Ala Leu Ile Glu Tyr Tyr Arg
725 730 735
Asn Cys Ile Pro Gln Tyr Ser Asn Trp Ser Phe Tyr Asp Phe Gln Phe
740 745 750
Gln Asp Thr Gly Lys Tyr Gln Asn Ile Lys Glu Phe Thr Asp Asp Val
755 760 765
Gln Lys Tyr Gly Tyr Lys Ile Ser Phe Arg Asp Ile Asp Asp Glu Tyr
770 775 780
Ile Asn Gln Ala Leu Asn Glu Gly Lys Met Tyr Leu Phe Glu Val Val
785 790 795 800
Asn Lys Asp Ile Tyr Asn Thr Lys Asn Gly Ser Lys Asn Leu His Thr
805 810 815
Leu Tyr Phe Glu His Ile Leu Ser Ala Glu Asn Leu Asn Asp Pro Val
820 825 830
Phe Lys Leu Ser Gly Met Ala Glu Ile Phe Gln Arg Gln Pro Ser Val
835 840 845
Asn Glu Arg Glu Lys Ile Thr Thr Gln Lys Asn Gln Cys Ile Leu Asp
850 855 860
Lys Gly Asp Arg Ala Tyr Lys Tyr Arg Arg Tyr Thr Glu Lys Lys Ile
865 870 875 880
Met Phe His Met Ser Leu Val Leu Asn Thr Gly Lys Gly Glu Ile Lys
885 890 895
Gln Val Gln Phe Asn Lys Ile Ile Asn Gln Arg Ile Ser Ser Ser Asp
900 905 910
Asn Glu Met Arg Val Asn Val Ile Gly Ile Asp Arg Gly Glu Lys Asn
915 920 925
Leu Leu Tyr Tyr Ser Val Val Lys Gln Asn Gly Glu Ile Ile Glu Gln
930 935 940
Ala Ser Leu Asn Glu Ile Asn Gly Val Asn Tyr Arg Asp Lys Leu Ile
945 950 955 960
Glu Arg Glu Lys Glu Arg Leu Lys Asn Arg Gln Ser Trp Lys Pro Val
965 970 975
Val Lys Ile Lys Asp Leu Lys Lys Gly Tyr Ile Ser His Val Ile His
980 985 990
Lys Ile Cys Gln Leu Ile Glu Lys Tyr Ser Ala Ile Val Val Leu Glu
995 1000 1005
Asp Leu Asn Met Arg Phe Lys Gln Ile Arg Gly Gly Ile Glu Arg
1010 1015 1020
Ser Val Tyr Gln Gln Phe Glu Lys Ala Leu Ile Asp Lys Leu Gly
1025 1030 1035
Tyr Leu Val Phe Lys Asp Asn Arg Asp Leu Arg Ala Pro Gly Gly
1040 1045 1050
Val Leu Asn Gly Tyr Gln Leu Ser Ala Pro Phe Val Ser Phe Glu
1055 1060 1065
Lys Met Arg Lys Gln Thr Gly Ile Leu Phe Tyr Thr Gln Ala Glu
1070 1075 1080
Tyr Thr Ser Lys Thr Asp Pro Ile Thr Gly Phe Arg Lys Asn Val
1085 1090 1095
Tyr Ile Ser Asn Ser Ala Ser Leu Asp Lys Ile Lys Glu Ala Val
1100 1105 1110
Lys Lys Phe Asp Ala Ile Gly Trp Asp Gly Lys Glu Gln Ser Tyr
1115 1120 1125
Phe Phe Lys Tyr Asn Pro Tyr Asn Leu Ala Asp Glu Lys Tyr Lys
1130 1135 1140
Asn Ser Thr Val Ser Lys Glu Trp Ala Ile Phe Ala Ser Ala Pro
1145 1150 1155
Arg Ile Arg Arg Gln Lys Gly Glu Asp Gly Tyr Trp Lys Tyr Asp
1160 1165 1170
Arg Val Lys Val Asn Glu Glu Phe Glu Lys Leu Leu Lys Val Trp
1175 1180 1185
Asn Phe Val Asn Pro Lys Ala Thr Asp Ile Lys Gln Glu Ile Ile
1190 1195 1200
Lys Lys Ile Lys Ala Gly Asp Leu Gln Gly Glu Lys Glu Leu Asp
1205 1210 1215
Gly Arg Leu Arg Asn Phe Trp His Ser Phe Ile Tyr Leu Phe Asn
1220 1225 1230
Leu Val Leu Glu Leu Arg Asn Ser Phe Ser Leu Gln Ile Lys Ile
1235 1240 1245
Lys Ala Gly Glu Val Ile Ala Val Asp Glu Gly Val Asp Phe Ile
1250 1255 1260
Ala Ser Pro Val Lys Pro Phe Phe Thr Thr Pro Asn Pro Tyr Ile
1265 1270 1275
Pro Ser Asn Leu Cys Trp Leu Ala Val Glu Asn Ala Asp Ala Asn
1280 1285 1290
Gly Ala Tyr Asn Ile Ala Arg Lys Gly Val Met Ile Leu Lys Lys
1295 1300 1305
Ile Arg Glu His Ala Lys Lys Asp Pro Glu Phe Lys Lys Leu Pro
1310 1315 1320
Asn Leu Phe Ile Ser Asn Ala Glu Trp Asp Glu Ala Ala Arg Asp
1325 1330 1335
Trp Gly Lys Tyr Ala Gly Thr Thr Ala Leu Asn Leu Asp His
1340 1345 1350
<210> 13
<211> 1260
<212> PRT
<213> Porphyromonas crevioricanis
<400> 13
Met Asp Ser Leu Lys Asp Phe Thr Asn Leu Tyr Pro Val Ser Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr Leu Glu Asn Ile Glu
20 25 30
Lys Ala Gly Ile Leu Lys Glu Asp Glu His Arg Ala Glu Ser Tyr Arg
35 40 45
Arg Val Lys Lys Ile Ile Asp Thr Tyr His Lys Val Phe Ile Asp Ser
50 55 60
Ser Leu Glu Asn Met Ala Lys Met Gly Ile Glu Asn Glu Ile Lys Ala
65 70 75 80
Met Leu Gln Ser Phe Cys Glu Leu Tyr Lys Lys Asp His Arg Thr Glu
85 90 95
Gly Glu Asp Lys Ala Leu Asp Lys Ile Arg Ala Val Leu Arg Gly Leu
100 105 110
Ile Val Gly Ala Phe Thr Gly Val Cys Gly Arg Arg Glu Asn Thr Val
115 120 125
Gln Asn Glu Lys Tyr Glu Ser Leu Phe Lys Glu Lys Leu Ile Lys Glu
130 135 140
Ile Leu Pro Asp Phe Val Leu Ser Thr Glu Ala Glu Ser Leu Pro Phe
145 150 155 160
Ser Val Glu Glu Ala Thr Arg Ser Leu Lys Glu Phe Asp Ser Phe Thr
165 170 175
Ser Tyr Phe Ala Gly Phe Tyr Glu Asn Arg Lys Asn Ile Tyr Ser Thr
180 185 190
Lys Pro Gln Ser Thr Ala Ile Ala Tyr Arg Leu Ile His Glu Asn Leu
195 200 205
Pro Lys Phe Ile Asp Asn Ile Leu Val Phe Gln Lys Ile Lys Glu Pro
210 215 220
Ile Ala Lys Glu Leu Glu His Ile Arg Ala Asp Phe Ser Ala Gly Gly
225 230 235 240
Tyr Ile Lys Lys Asp Glu Arg Leu Glu Asp Ile Phe Ser Leu Asn Tyr
245 250 255
Tyr Ile His Val Leu Ser Gln Ala Gly Ile Glu Lys Tyr Asn Ala Leu
260 265 270
Ile Gly Lys Ile Val Thr Glu Gly Asp Gly Glu Met Lys Gly Leu Asn
275 280 285
Glu His Ile Asn Leu Tyr Asn Gln Gln Arg Gly Arg Glu Asp Arg Leu
290 295 300
Pro Leu Phe Arg Pro Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Gln
305 310 315 320
Leu Ser Tyr Leu Pro Glu Ser Phe Glu Lys Asp Glu Glu Leu Leu Arg
325 330 335
Ala Leu Lys Glu Phe Tyr Asp His Ile Ala Glu Asp Ile Leu Gly Arg
340 345 350
Thr Gln Gln Leu Met Thr Ser Ile Ser Glu Tyr Asp Leu Ser Arg Ile
355 360 365
Tyr Val Arg Asn Asp Ser Gln Leu Thr Asp Ile Ser Lys Lys Met Leu
370 375 380
Gly Asp Trp Asn Ala Ile Tyr Met Ala Arg Glu Arg Ala Tyr Asp His
385 390 395 400
Glu Gln Ala Pro Lys Arg Ile Thr Ala Lys Tyr Glu Arg Asp Arg Ile
405 410 415
Lys Ala Leu Lys Gly Glu Glu Ser Ile Ser Leu Ala Asn Leu Asn Ser
420 425 430
Cys Ile Ala Phe Leu Asp Asn Val Arg Asp Cys Arg Val Asp Thr Tyr
435 440 445
Leu Ser Thr Leu Gly Gln Lys Glu Gly Pro His Gly Leu Ser Asn Leu
450 455 460
Val Glu Asn Val Phe Ala Ser Tyr His Glu Ala Glu Gln Leu Leu Ser
465 470 475 480
Phe Pro Tyr Pro Glu Glu Asn Asn Leu Ile Gln Asp Lys Asp Asn Val
485 490 495
Val Leu Ile Lys Asn Leu Leu Asp Asn Ile Ser Asp Leu Gln Arg Phe
500 505 510
Leu Lys Pro Leu Trp Gly Met Gly Asp Glu Pro Asp Lys Asp Glu Arg
515 520 525
Phe Tyr Gly Glu Tyr Asn Tyr Ile Arg Gly Ala Leu Asp Gln Val Ile
530 535 540
Pro Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr Arg Lys Pro Tyr Ser
545 550 555 560
Thr Arg Lys Val Lys Leu Asn Phe Gly Asn Ser Gln Leu Leu Ser Gly
565 570 575
Trp Asp Arg Asn Lys Glu Lys Asp Asn Ser Cys Val Ile Leu Arg Lys
580 585 590
Gly Gln Asn Phe Tyr Leu Ala Ile Met Asn Asn Arg His Lys Arg Ser
595 600 605
Phe Glu Asn Lys Met Leu Pro Glu Tyr Lys Glu Gly Glu Pro Tyr Phe
610 615 620
Glu Lys Met Asp Tyr Lys Phe Leu Pro Asp Pro Asn Lys Met Leu Pro
625 630 635 640
Lys Val Phe Leu Ser Lys Lys Gly Ile Glu Ile Tyr Lys Pro Ser Pro
645 650 655
Lys Leu Leu Glu Gln Tyr Gly His Gly Thr His Lys Lys Gly Asp Thr
660 665 670
Phe Ser Met Asp Asp Leu His Glu Leu Ile Asp Phe Phe Lys His Ser
675 680 685
Ile Glu Ala His Glu Asp Trp Lys Gln Phe Gly Phe Lys Phe Ser Asp
690 695 700
Thr Ala Thr Tyr Glu Asn Val Ser Ser Phe Tyr Arg Glu Val Glu Asp
705 710 715 720
Gln Gly Tyr Lys Leu Ser Phe Arg Lys Val Ser Glu Ser Tyr Val Tyr
725 730 735
Ser Leu Ile Asp Gln Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys
740 745 750
Asp Phe Ser Pro Cys Ser Lys Gly Thr Pro Asn Leu His Thr Leu Tyr
755 760 765
Trp Arg Met Leu Phe Asp Glu Arg Asn Leu Ala Asp Val Ile Tyr Lys
770 775 780
Leu Asp Gly Lys Ala Glu Ile Phe Phe Arg Glu Lys Ser Leu Lys Asn
785 790 795 800
Asp His Pro Thr His Pro Ala Gly Lys Pro Ile Lys Lys Lys Ser Arg
805 810 815
Gln Lys Lys Gly Glu Glu Ser Leu Phe Glu Tyr Asp Leu Val Lys Asp
820 825 830
Arg Arg Tyr Thr Met Asp Lys Phe Gln Phe His Val Pro Ile Thr Met
835 840 845
Asn Phe Lys Cys Ser Ala Gly Ser Lys Val Asn Asp Met Val Asn Ala
850 855 860
His Ile Arg Glu Ala Lys Asp Met His Val Ile Gly Ile Asp Arg Gly
865 870 875 880
Glu Arg Asn Leu Leu Tyr Ile Cys Val Ile Asp Ser Arg Gly Thr Ile
885 890 895
Leu Asp Gln Ile Ser Leu Asn Thr Ile Asn Asp Ile Asp Tyr His Asp
900 905 910
Leu Leu Glu Ser Arg Asp Lys Asp Arg Gln Gln Glu His Arg Asn Trp
915 920 925
Gln Thr Ile Glu Gly Ile Lys Glu Leu Lys Gln Gly Tyr Leu Ser Gln
930 935 940
Ala Val His Arg Ile Ala Glu Leu Met Val Ala Tyr Lys Ala Val Val
945 950 955 960
Ala Leu Glu Asp Leu Asn Met Gly Phe Lys Arg Gly Arg Gln Lys Val
965 970 975
Glu Ser Ser Val Tyr Gln Gln Phe Glu Lys Gln Leu Ile Asp Lys Leu
980 985 990
Asn Tyr Leu Val Asp Lys Lys Lys Arg Pro Glu Asp Ile Gly Gly Leu
995 1000 1005
Leu Arg Ala Tyr Gln Phe Thr Ala Pro Phe Lys Ser Phe Lys Glu
1010 1015 1020
Met Gly Lys Gln Asn Gly Phe Leu Phe Tyr Ile Pro Ala Trp Asn
1025 1030 1035
Thr Ser Asn Ile Asp Pro Thr Thr Gly Phe Val Asn Leu Phe His
1040 1045 1050
Val Gln Tyr Glu Asn Val Asp Lys Ala Lys Ser Phe Phe Gln Lys
1055 1060 1065
Phe Asp Ser Ile Ser Tyr Asn Pro Lys Lys Asp Trp Phe Glu Phe
1070 1075 1080
Ala Phe Asp Tyr Lys Asn Phe Thr Lys Lys Ala Glu Gly Ser Arg
1085 1090 1095
Ser Met Trp Ile Leu Cys Thr His Gly Ser Arg Ile Lys Asn Phe
1100 1105 1110
Arg Asn Ser Gln Lys Asn Gly Gln Trp Asp Ser Glu Glu Phe Ala
1115 1120 1125
Leu Thr Glu Ala Phe Lys Ser Leu Phe Val Arg Tyr Glu Ile Asp
1130 1135 1140
Tyr Thr Ala Asp Leu Lys Thr Ala Ile Val Asp Glu Lys Gln Lys
1145 1150 1155
Asp Phe Phe Val Asp Leu Leu Lys Leu Phe Lys Leu Thr Val Gln
1160 1165 1170
Met Arg Asn Ser Trp Lys Glu Lys Asp Leu Asp Tyr Leu Ile Ser
1175 1180 1185
Pro Val Ala Gly Ala Asp Gly Arg Phe Phe Asp Thr Arg Glu Gly
1190 1195 1200
Asn Lys Ser Leu Pro Lys Asp Ala Asp Ala Asn Gly Ala Tyr Asn
1205 1210 1215
Ile Ala Leu Lys Gly Leu Trp Ala Leu Arg Gln Ile Arg Gln Thr
1220 1225 1230
Ser Glu Gly Gly Lys Leu Lys Leu Ala Ile Ser Asn Lys Glu Trp
1235 1240 1245
Leu Gln Phe Val Gln Glu Arg Ser Tyr Glu Lys Asp
1250 1255 1260
<210> 14
<211> 1324
<212> PRT
<213> 解糖胨普雷沃菌(Prevotella disiens)
<400> 14
Met Glu Asn Tyr Gln Glu Phe Thr Asn Leu Phe Gln Leu Asn Lys Thr
1 5 10 15
Leu Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Cys Glu Leu Leu Glu
20 25 30
Glu Gly Lys Ile Phe Ala Ser Gly Ser Phe Leu Glu Lys Asp Lys Val
35 40 45
Arg Ala Asp Asn Val Ser Tyr Val Lys Lys Glu Ile Asp Lys Lys His
50 55 60
Lys Ile Phe Ile Glu Glu Thr Leu Ser Ser Phe Ser Ile Ser Asn Asp
65 70 75 80
Leu Leu Lys Gln Tyr Phe Asp Cys Tyr Asn Glu Leu Lys Ala Phe Lys
85 90 95
Lys Asp Cys Lys Ser Asp Glu Glu Glu Val Lys Lys Thr Ala Leu Arg
100 105 110
Asn Lys Cys Thr Ser Ile Gln Arg Ala Met Arg Glu Ala Ile Ser Gln
115 120 125
Ala Phe Leu Lys Ser Pro Gln Lys Lys Leu Leu Ala Ile Lys Asn Leu
130 135 140
Ile Glu Asn Val Phe Lys Ala Asp Glu Asn Val Gln His Phe Ser Glu
145 150 155 160
Phe Thr Ser Tyr Phe Ser Gly Phe Glu Thr Asn Arg Glu Asn Phe Tyr
165 170 175
Ser Asp Glu Glu Lys Ser Thr Ser Ile Ala Tyr Arg Leu Val His Asp
180 185 190
Asn Leu Pro Ile Phe Ile Lys Asn Ile Tyr Ile Phe Glu Lys Leu Lys
195 200 205
Glu Gln Phe Asp Ala Lys Thr Leu Ser Glu Ile Phe Glu Asn Tyr Lys
210 215 220
Leu Tyr Val Ala Gly Ser Ser Leu Asp Glu Val Phe Ser Leu Glu Tyr
225 230 235 240
Phe Asn Asn Thr Leu Thr Gln Lys Gly Ile Asp Asn Tyr Asn Ala Val
245 250 255
Ile Gly Lys Ile Val Lys Glu Asp Lys Gln Glu Ile Gln Gly Leu Asn
260 265 270
Glu His Ile Asn Leu Tyr Asn Gln Lys His Lys Asp Arg Arg Leu Pro
275 280 285
Phe Phe Ile Ser Leu Lys Lys Gln Ile Leu Ser Asp Arg Glu Ala Leu
290 295 300
Ser Trp Leu Pro Asp Met Phe Lys Asn Asp Ser Glu Val Ile Asp Ala
305 310 315 320
Leu Lys Gly Phe Tyr Ile Glu Asp Gly Phe Glu Asn Asn Val Leu Thr
325 330 335
Pro Leu Ala Thr Leu Leu Ser Ser Leu Asp Lys Tyr Asn Leu Asn Gly
340 345 350
Ile Phe Ile Arg Asn Asn Glu Ala Leu Ser Ser Leu Ser Gln Asn Val
355 360 365
Tyr Arg Asn Phe Ser Ile Asp Glu Ala Ile Asp Ala Gln Asn Ala Glu
370 375 380
Leu Gln Thr Phe Asn Asn Tyr Glu Leu Ile Ala Asn Ala Leu Arg Ala
385 390 395 400
Lys Ile Lys Lys Glu Thr Lys Gln Gly Arg Lys Ser Phe Glu Lys Tyr
405 410 415
Glu Glu Tyr Ile Asp Lys Lys Val Lys Ala Ile Asp Ser Leu Ser Ile
420 425 430
Gln Glu Ile Asn Glu Leu Val Glu Asn Tyr Val Ser Glu Phe Asn Ser
435 440 445
Asn Ser Gly Asn Met Pro Arg Lys Val Glu Asp Tyr Phe Ser Leu Met
450 455 460
Arg Lys Gly Asp Phe Gly Ser Asn Asp Leu Ile Glu Asn Ile Lys Thr
465 470 475 480
Lys Leu Ser Ala Ala Glu Lys Leu Leu Gly Thr Lys Tyr Gln Glu Thr
485 490 495
Ala Lys Asp Ile Phe Lys Lys Asp Glu Asn Ser Lys Leu Ile Lys Glu
500 505 510
Leu Leu Asp Ala Thr Lys Gln Phe Gln His Phe Ile Lys Pro Leu Leu
515 520 525
Gly Thr Gly Glu Glu Ala Asp Arg Asp Leu Val Phe Tyr Gly Asp Phe
530 535 540
Leu Pro Leu Tyr Glu Lys Phe Glu Glu Leu Thr Leu Leu Tyr Asn Lys
545 550 555 560
Val Arg Asn Arg Leu Thr Gln Lys Pro Tyr Ser Lys Asp Lys Ile Arg
565 570 575
Leu Cys Phe Asn Lys Pro Lys Leu Met Thr Gly Trp Val Asp Ser Lys
580 585 590
Thr Glu Lys Ser Asp Asn Gly Thr Gln Tyr Gly Gly Tyr Leu Phe Arg
595 600 605
Lys Lys Asn Glu Ile Gly Glu Tyr Asp Tyr Phe Leu Gly Ile Ser Ser
610 615 620
Lys Ala Gln Leu Phe Arg Lys Asn Glu Ala Val Ile Gly Asp Tyr Glu
625 630 635 640
Arg Leu Asp Tyr Tyr Gln Pro Lys Ala Asn Thr Ile Tyr Gly Ser Ala
645 650 655
Tyr Glu Gly Glu Asn Ser Tyr Lys Glu Asp Lys Lys Arg Leu Asn Lys
660 665 670
Val Ile Ile Ala Tyr Ile Glu Gln Ile Lys Gln Thr Asn Ile Lys Lys
675 680 685
Ser Ile Ile Glu Ser Ile Ser Lys Tyr Pro Asn Ile Ser Asp Asp Asp
690 695 700
Lys Val Thr Pro Ser Ser Leu Leu Glu Lys Ile Lys Lys Val Ser Ile
705 710 715 720
Asp Ser Tyr Asn Gly Ile Leu Ser Phe Lys Ser Phe Gln Ser Val Asn
725 730 735
Lys Glu Val Ile Asp Asn Leu Leu Lys Thr Ile Ser Pro Leu Lys Asn
740 745 750
Lys Ala Glu Phe Leu Asp Leu Ile Asn Lys Asp Tyr Gln Ile Phe Thr
755 760 765
Glu Val Gln Ala Val Ile Asp Glu Ile Cys Lys Gln Lys Thr Phe Ile
770 775 780
Tyr Phe Pro Ile Ser Asn Val Glu Leu Glu Lys Glu Met Gly Asp Lys
785 790 795 800
Asp Lys Pro Leu Cys Leu Phe Gln Ile Ser Asn Lys Asp Leu Ser Phe
805 810 815
Ala Lys Thr Phe Ser Ala Asn Leu Arg Lys Lys Arg Gly Ala Glu Asn
820 825 830
Leu His Thr Met Leu Phe Lys Ala Leu Met Glu Gly Asn Gln Asp Asn
835 840 845
Leu Asp Leu Gly Ser Gly Ala Ile Phe Tyr Arg Ala Lys Ser Leu Asp
850 855 860
Gly Asn Lys Pro Thr His Pro Ala Asn Glu Ala Ile Lys Cys Arg Asn
865 870 875 880
Val Ala Asn Lys Asp Lys Val Ser Leu Phe Thr Tyr Asp Ile Tyr Lys
885 890 895
Asn Arg Arg Tyr Met Glu Asn Lys Phe Leu Phe His Leu Ser Ile Val
900 905 910
Gln Asn Tyr Lys Ala Ala Asn Asp Ser Ala Gln Leu Asn Ser Ser Ala
915 920 925
Thr Glu Tyr Ile Arg Lys Ala Asp Asp Leu His Ile Ile Gly Ile Asp
930 935 940
Arg Gly Glu Arg Asn Leu Leu Tyr Tyr Ser Val Ile Asp Met Lys Gly
945 950 955 960
Asn Ile Val Glu Gln Asp Ser Leu Asn Ile Ile Arg Asn Asn Asp Leu
965 970 975
Glu Thr Asp Tyr His Asp Leu Leu Asp Lys Arg Glu Lys Glu Arg Lys
980 985 990
Ala Asn Arg Gln Asn Trp Glu Ala Val Glu Gly Ile Lys Asp Leu Lys
995 1000 1005
Lys Gly Tyr Leu Ser Gln Ala Val His Gln Ile Ala Gln Leu Met
1010 1015 1020
Leu Lys Tyr Asn Ala Ile Ile Ala Leu Glu Asp Leu Gly Gln Met
1025 1030 1035
Phe Val Thr Arg Gly Gln Lys Ile Glu Lys Ala Val Tyr Gln Gln
1040 1045 1050
Phe Glu Lys Ser Leu Val Asp Lys Leu Ser Tyr Leu Val Asp Lys
1055 1060 1065
Lys Arg Pro Tyr Asn Glu Leu Gly Gly Ile Leu Lys Ala Tyr Gln
1070 1075 1080
Leu Ala Ser Ser Ile Thr Lys Asn Asn Ser Asp Lys Gln Asn Gly
1085 1090 1095
Phe Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Lys Ile Asp Pro
1100 1105 1110
Val Thr Gly Phe Thr Asp Leu Leu Arg Pro Lys Ala Met Thr Ile
1115 1120 1125
Lys Glu Ala Gln Asp Phe Phe Gly Ala Phe Asp Asn Ile Ser Tyr
1130 1135 1140
Asn Asp Lys Gly Tyr Phe Glu Phe Glu Thr Asn Tyr Asp Lys Phe
1145 1150 1155
Lys Ile Arg Met Lys Ser Ala Gln Thr Arg Trp Thr Ile Cys Thr
1160 1165 1170
Phe Gly Asn Arg Ile Lys Arg Lys Lys Asp Lys Asn Tyr Trp Asn
1175 1180 1185
Tyr Glu Glu Val Glu Leu Thr Glu Glu Phe Lys Lys Leu Phe Lys
1190 1195 1200
Asp Ser Asn Ile Asp Tyr Glu Asn Cys Asn Leu Lys Glu Glu Ile
1205 1210 1215
Gln Asn Lys Asp Asn Arg Lys Phe Phe Asp Asp Leu Ile Lys Leu
1220 1225 1230
Leu Gln Leu Thr Leu Gln Met Arg Asn Ser Asp Asp Lys Gly Asn
1235 1240 1245
Asp Tyr Ile Ile Ser Pro Val Ala Asn Ala Glu Gly Gln Phe Phe
1250 1255 1260
Asp Ser Arg Asn Gly Asp Lys Lys Leu Pro Leu Asp Ala Asp Ala
1265 1270 1275
Asn Gly Ala Tyr Asn Ile Ala Arg Lys Gly Leu Trp Asn Ile Arg
1280 1285 1290
Gln Ile Lys Gln Thr Lys Asn Lys Asp Asp Leu Asn Leu Ser Ile
1295 1300 1305
Ser Ser Thr Glu Trp Leu Asp Phe Val Arg Glu Lys Pro Tyr Leu
1310 1315 1320
Lys
<210> 15
<211> 1484
<212> PRT
<213> 未知
<220>
<223> Peregrinibacteria细菌
<220>
<221> misc_feature
<222> (1073)..(1073)
<223> Xaa可以是任何天然存在的氨基酸
<400> 15
Met Ser Asn Phe Phe Lys Asn Phe Thr Asn Leu Tyr Glu Leu Ser Lys
1 5 10 15
Thr Leu Arg Phe Glu Leu Lys Pro Val Gly Asp Thr Leu Thr Asn Met
20 25 30
Lys Asp His Leu Glu Tyr Asp Glu Lys Leu Gln Thr Phe Leu Lys Asp
35 40 45
Gln Asn Ile Asp Asp Ala Tyr Gln Ala Leu Lys Pro Gln Phe Asp Glu
50 55 60
Ile His Glu Glu Phe Ile Thr Asp Ser Leu Glu Ser Lys Lys Ala Lys
65 70 75 80
Glu Ile Asp Phe Ser Glu Tyr Leu Asp Leu Phe Gln Glu Lys Lys Glu
85 90 95
Leu Asn Asp Ser Glu Lys Lys Leu Arg Asn Lys Ile Gly Glu Thr Phe
100 105 110
Asn Lys Ala Gly Glu Lys Trp Lys Lys Glu Lys Tyr Pro Gln Tyr Glu
115 120 125
Trp Lys Lys Gly Ser Lys Ile Ala Asn Gly Ala Asp Ile Leu Ser Cys
130 135 140
Gln Asp Met Leu Gln Phe Ile Lys Tyr Lys Asn Pro Glu Asp Glu Lys
145 150 155 160
Ile Lys Asn Tyr Ile Asp Asp Thr Leu Lys Gly Phe Phe Thr Tyr Phe
165 170 175
Gly Gly Phe Asn Gln Asn Arg Ala Asn Tyr Tyr Glu Thr Lys Lys Glu
180 185 190
Ala Ser Thr Ala Val Ala Thr Arg Ile Val His Glu Asn Leu Pro Lys
195 200 205
Phe Cys Asp Asn Val Ile Gln Phe Lys His Ile Ile Lys Arg Lys Lys
210 215 220
Asp Gly Thr Val Glu Lys Thr Glu Arg Lys Thr Glu Tyr Leu Asn Ala
225 230 235 240
Tyr Gln Tyr Leu Lys Asn Asn Asn Lys Ile Thr Gln Ile Lys Asp Ala
245 250 255
Glu Thr Glu Lys Met Ile Glu Ser Thr Pro Ile Ala Glu Lys Ile Phe
260 265 270
Asp Val Tyr Tyr Phe Ser Ser Cys Leu Ser Gln Lys Gln Ile Glu Glu
275 280 285
Tyr Asn Arg Ile Ile Gly His Tyr Asn Leu Leu Ile Asn Leu Tyr Asn
290 295 300
Gln Ala Lys Arg Ser Glu Gly Lys His Leu Ser Ala Asn Glu Lys Lys
305 310 315 320
Tyr Lys Asp Leu Pro Lys Phe Lys Thr Leu Tyr Lys Gln Ile Gly Cys
325 330 335
Gly Lys Lys Lys Asp Leu Phe Tyr Thr Ile Lys Cys Asp Thr Glu Glu
340 345 350
Glu Ala Asn Lys Ser Arg Asn Glu Gly Lys Glu Ser His Ser Val Glu
355 360 365
Glu Ile Ile Asn Lys Ala Gln Glu Ala Ile Asn Lys Tyr Phe Lys Ser
370 375 380
Asn Asn Asp Cys Glu Asn Ile Asn Thr Val Pro Asp Phe Ile Asn Tyr
385 390 395 400
Ile Leu Thr Lys Glu Asn Tyr Glu Gly Val Tyr Trp Ser Lys Ala Ala
405 410 415
Met Asn Thr Ile Ser Asp Lys Tyr Phe Ala Asn Tyr His Asp Leu Gln
420 425 430
Asp Arg Leu Lys Glu Ala Lys Val Phe Gln Lys Ala Asp Lys Lys Ser
435 440 445
Glu Asp Asp Ile Lys Ile Pro Glu Ala Ile Glu Leu Ser Gly Leu Phe
450 455 460
Gly Val Leu Asp Ser Leu Ala Asp Trp Gln Thr Thr Leu Phe Lys Ser
465 470 475 480
Ser Ile Leu Ser Asn Glu Lys Leu Lys Ile Ile Thr Asp Ser Gln Thr
485 490 495
Pro Ser Glu Ala Leu Leu Lys Met Ile Phe Asn Asp Ile Glu Lys Asn
500 505 510
Met Glu Ser Phe Leu Lys Glu Thr Asn Asp Ile Ile Thr Leu Lys Lys
515 520 525
Tyr Lys Gly Asn Lys Glu Gly Thr Glu Lys Ile Lys Gln Trp Phe Asp
530 535 540
Tyr Thr Leu Ala Ile Asn Arg Met Leu Lys Tyr Phe Leu Val Lys Glu
545 550 555 560
Asn Lys Ile Lys Gly Asn Ser Leu Asp Thr Asn Ile Ser Glu Ala Leu
565 570 575
Lys Thr Leu Ile Tyr Ser Asp Asp Ala Glu Trp Phe Lys Trp Tyr Asp
580 585 590
Ala Leu Arg Asn Tyr Leu Thr Gln Lys Pro Gln Asp Glu Ala Lys Glu
595 600 605
Asn Lys Leu Lys Leu Asn Phe Asp Asn Pro Ser Leu Ala Gly Gly Trp
610 615 620
Asp Val Asn Lys Glu Cys Ser Asn Phe Cys Val Ile Leu Lys Asp Lys
625 630 635 640
Asn Glu Lys Lys Tyr Leu Ala Met Ile Lys Lys Gly Glu Asn Thr Leu
645 650 655
Phe Gln Lys Glu Trp Thr Glu Gly Arg Gly Lys Asn Leu Thr Lys Lys
660 665 670
Ser Asn Pro Leu Phe Glu Ile Asn Asn Cys Glu Ile Leu Ser Lys Met
675 680 685
Glu Tyr Asp Phe Trp Ala Asp Val Ser Lys Met Ile Pro Lys Cys Ser
690 695 700
Thr Gln Leu Lys Ala Val Val Asn His Phe Lys Gln Ser Asp Asn Glu
705 710 715 720
Phe Ile Phe Pro Ile Gly Tyr Lys Val Thr Ser Gly Glu Lys Phe Arg
725 730 735
Glu Glu Cys Lys Ile Ser Lys Gln Asp Phe Glu Leu Asn Asn Lys Val
740 745 750
Phe Asn Lys Asn Glu Leu Ser Val Thr Ala Met Arg Tyr Asp Leu Ser
755 760 765
Ser Thr Gln Glu Lys Gln Tyr Ile Lys Ala Phe Gln Lys Glu Tyr Trp
770 775 780
Glu Leu Leu Phe Lys Gln Glu Lys Arg Asp Thr Lys Leu Thr Asn Asn
785 790 795 800
Glu Ile Phe Asn Glu Trp Ile Asn Phe Cys Asn Lys Lys Tyr Ser Glu
805 810 815
Leu Leu Ser Trp Glu Arg Lys Tyr Lys Asp Ala Leu Thr Asn Trp Ile
820 825 830
Asn Phe Cys Lys Tyr Phe Leu Ser Lys Tyr Pro Lys Thr Thr Leu Phe
835 840 845
Asn Tyr Ser Phe Lys Glu Ser Glu Asn Tyr Asn Ser Leu Asp Glu Phe
850 855 860
Tyr Arg Asp Val Asp Ile Cys Ser Tyr Lys Leu Asn Ile Asn Thr Thr
865 870 875 880
Ile Asn Lys Ser Ile Leu Asp Arg Leu Val Glu Glu Gly Lys Leu Tyr
885 890 895
Leu Phe Glu Ile Lys Asn Gln Asp Ser Asn Asp Gly Lys Ser Ile Gly
900 905 910
His Lys Asn Asn Leu His Thr Ile Tyr Trp Asn Ala Ile Phe Glu Asn
915 920 925
Phe Asp Asn Arg Pro Lys Leu Asn Gly Glu Ala Glu Ile Phe Tyr Arg
930 935 940
Lys Ala Ile Ser Lys Asp Lys Leu Gly Ile Val Lys Gly Lys Lys Thr
945 950 955 960
Lys Asn Gly Thr Trp Ile Ile Lys Asn Tyr Arg Phe Ser Lys Glu Lys
965 970 975
Phe Ile Leu His Val Pro Ile Thr Leu Asn Phe Cys Ser Asn Asn Glu
980 985 990
Tyr Val Asn Asp Ile Val Asn Thr Lys Phe Tyr Asn Phe Ser Asn Leu
995 1000 1005
His Phe Leu Gly Ile Asp Arg Gly Glu Lys His Leu Ala Tyr Tyr
1010 1015 1020
Ser Leu Val Asn Lys Asn Gly Glu Ile Val Asp Gln Gly Thr Leu
1025 1030 1035
Asn Leu Pro Phe Thr Asp Lys Asp Gly Asn Gln Arg Ser Ile Lys
1040 1045 1050
Lys Glu Lys Tyr Phe Tyr Asn Lys Gln Glu Asp Lys Trp Glu Ala
1055 1060 1065
Lys Glu Val Asp Xaa Trp Asn Tyr Asn Asp Leu Leu Asp Ala Met
1070 1075 1080
Ala Ser Asn Arg Asp Met Ala Arg Lys Asn Trp Gln Arg Ile Gly
1085 1090 1095
Thr Ile Lys Glu Ala Lys Asn Gly Tyr Val Ser Leu Val Ile Arg
1100 1105 1110
Lys Ile Ala Asp Leu Ala Val Asn Asn Glu Arg Pro Ala Phe Ile
1115 1120 1125
Val Leu Glu Asp Leu Asn Thr Gly Phe Lys Arg Ser Arg Gln Lys
1130 1135 1140
Ile Asp Lys Ser Val Tyr Gln Lys Phe Glu Leu Ala Leu Ala Lys
1145 1150 1155
Lys Leu Asn Phe Leu Val Asp Lys Asn Ala Lys Arg Asp Glu Ile
1160 1165 1170
Gly Ser Pro Thr Lys Ala Leu Gln Leu Thr Pro Pro Val Asn Asn
1175 1180 1185
Tyr Gly Asp Ile Glu Asn Lys Lys Gln Ala Gly Ile Met Leu Tyr
1190 1195 1200
Thr Arg Ala Asn Tyr Thr Ser Gln Thr Asp Pro Ala Thr Gly Trp
1205 1210 1215
Arg Lys Thr Ile Tyr Leu Lys Ala Gly Pro Glu Glu Thr Thr Tyr
1220 1225 1230
Lys Lys Asp Gly Lys Ile Lys Asn Lys Ser Val Lys Asp Gln Ile
1235 1240 1245
Ile Glu Thr Phe Thr Asp Ile Gly Phe Asp Gly Lys Asp Tyr Tyr
1250 1255 1260
Phe Glu Tyr Asp Lys Gly Glu Phe Val Asp Glu Lys Thr Gly Glu
1265 1270 1275
Ile Lys Pro Lys Lys Trp Arg Leu Tyr Ser Gly Glu Asn Gly Lys
1280 1285 1290
Ser Leu Asp Arg Phe Arg Gly Glu Arg Glu Lys Asp Lys Tyr Glu
1295 1300 1305
Trp Lys Ile Asp Lys Ile Asp Ile Val Lys Ile Leu Asp Asp Leu
1310 1315 1320
Phe Val Asn Phe Asp Lys Asn Ile Ser Leu Leu Lys Gln Leu Lys
1325 1330 1335
Glu Gly Val Glu Leu Thr Arg Asn Asn Glu His Gly Thr Gly Glu
1340 1345 1350
Ser Leu Arg Phe Ala Ile Asn Leu Ile Gln Gln Ile Arg Asn Thr
1355 1360 1365
Gly Asn Asn Glu Arg Asp Asn Asp Phe Ile Leu Ser Pro Val Arg
1370 1375 1380
Asp Glu Asn Gly Lys His Phe Asp Ser Arg Glu Tyr Trp Asp Lys
1385 1390 1395
Glu Thr Lys Gly Glu Lys Ile Ser Met Pro Ser Ser Gly Asp Ala
1400 1405 1410
Asn Gly Ala Phe Asn Ile Ala Arg Lys Gly Ile Ile Met Asn Ala
1415 1420 1425
His Ile Leu Ala Asn Ser Asp Ser Lys Asp Leu Ser Leu Phe Val
1430 1435 1440
Ser Asp Glu Glu Trp Asp Leu His Leu Asn Asn Lys Thr Glu Trp
1445 1450 1455
Lys Lys Gln Leu Asn Ile Phe Ser Ser Arg Lys Ala Met Ala Lys
1460 1465 1470
Arg Lys Lys Lys Arg Pro Ala Ala Thr Lys Lys
1475 1480
<210> 16
<211> 1245
<212> PRT
<213> Porphyromonas macacae
<400> 16
Met Lys Thr Gln His Phe Phe Glu Asp Phe Thr Ser Leu Tyr Ser Leu
1 5 10 15
Ser Lys Thr Ile Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Leu Glu
20 25 30
Asn Ile Lys Lys Asn Gly Leu Ile Arg Arg Asp Glu Gln Arg Leu Asp
35 40 45
Asp Tyr Glu Lys Leu Lys Lys Val Ile Asp Glu Tyr His Glu Asp Phe
50 55 60
Ile Ala Asn Ile Leu Ser Ser Phe Ser Phe Ser Glu Glu Ile Leu Gln
65 70 75 80
Ser Tyr Ile Gln Asn Leu Ser Ile Ser Glu Ala Arg Ala Lys Ile Glu
85 90 95
Lys Thr Met Arg Asp Thr Leu Ala Lys Ala Phe Ser Glu Asp Glu Arg
100 105 110
Tyr Lys Ser Ile Phe Lys Lys Glu Leu Val Lys Lys Asp Ile Pro Val
115 120 125
Trp Cys Pro Ala Tyr Lys Ser Leu Cys Lys Lys Phe Asp Asn Phe Thr
130 135 140
Thr Ser Leu Val Pro Phe His Glu Asn Arg Lys Asn Leu Tyr Thr Ser
145 150 155 160
Asn Glu Ile Thr Ala Ser Ile Pro Tyr Arg Ile Val His Val Asn Leu
165 170 175
Pro Lys Phe Ile Gln Asn Ile Glu Ala Leu Cys Glu Leu Gln Lys Lys
180 185 190
Met Gly Ala Asp Leu Tyr Leu Glu Met Met Glu Asn Leu Arg Asn Val
195 200 205
Trp Pro Ser Phe Val Lys Thr Pro Asp Asp Leu Cys Asn Leu Lys Thr
210 215 220
Tyr Asn His Leu Met Val Gln Ser Ser Ile Ser Glu Tyr Asn Arg Phe
225 230 235 240
Val Gly Gly Tyr Ser Thr Glu Asp Gly Thr Lys His Gln Gly Ile Asn
245 250 255
Glu Trp Ile Asn Ile Tyr Arg Gln Arg Asn Lys Glu Met Arg Leu Pro
260 265 270
Gly Leu Val Phe Leu His Lys Gln Ile Leu Ala Lys Val Asp Ser Ser
275 280 285
Ser Phe Ile Ser Asp Thr Leu Glu Asn Asp Asp Gln Val Phe Cys Val
290 295 300
Leu Arg Gln Phe Arg Lys Leu Phe Trp Asn Thr Val Ser Ser Lys Glu
305 310 315 320
Asp Asp Ala Ala Ser Leu Lys Asp Leu Phe Cys Gly Leu Ser Gly Tyr
325 330 335
Asp Pro Glu Ala Ile Tyr Val Ser Asp Ala His Leu Ala Thr Ile Ser
340 345 350
Lys Asn Ile Phe Asp Arg Trp Asn Tyr Ile Ser Asp Ala Ile Arg Arg
355 360 365
Lys Thr Glu Val Leu Met Pro Arg Lys Lys Glu Ser Val Glu Arg Tyr
370 375 380
Ala Glu Lys Ile Ser Lys Gln Ile Lys Lys Arg Gln Ser Tyr Ser Leu
385 390 395 400
Ala Glu Leu Asp Asp Leu Leu Ala His Tyr Ser Glu Glu Ser Leu Pro
405 410 415
Ala Gly Phe Ser Leu Leu Ser Tyr Phe Thr Ser Leu Gly Gly Gln Lys
420 425 430
Tyr Leu Val Ser Asp Gly Glu Val Ile Leu Tyr Glu Glu Gly Ser Asn
435 440 445
Ile Trp Asp Glu Val Leu Ile Ala Phe Arg Asp Leu Gln Val Ile Leu
450 455 460
Asp Lys Asp Phe Thr Glu Lys Lys Leu Gly Lys Asp Glu Glu Ala Val
465 470 475 480
Ser Val Ile Lys Lys Ala Leu Asp Ser Ala Leu Arg Leu Arg Lys Phe
485 490 495
Phe Asp Leu Leu Ser Gly Thr Gly Ala Glu Ile Arg Arg Asp Ser Ser
500 505 510
Phe Tyr Ala Leu Tyr Thr Asp Arg Met Asp Lys Leu Lys Gly Leu Leu
515 520 525
Lys Met Tyr Asp Lys Val Arg Asn Tyr Leu Thr Lys Lys Pro Tyr Ser
530 535 540
Ile Glu Lys Phe Lys Leu His Phe Asp Asn Pro Ser Leu Leu Ser Gly
545 550 555 560
Trp Asp Lys Asn Lys Glu Leu Asn Asn Leu Ser Val Ile Phe Arg Gln
565 570 575
Asn Gly Tyr Tyr Tyr Leu Gly Ile Met Thr Pro Lys Gly Lys Asn Leu
580 585 590
Phe Lys Thr Leu Pro Lys Leu Gly Ala Glu Glu Met Phe Tyr Glu Lys
595 600 605
Met Glu Tyr Lys Gln Ile Ala Glu Pro Met Leu Met Leu Pro Lys Val
610 615 620
Phe Phe Pro Lys Lys Thr Lys Pro Ala Phe Ala Pro Asp Gln Ser Val
625 630 635 640
Val Asp Ile Tyr Asn Lys Lys Thr Phe Lys Thr Gly Gln Lys Gly Phe
645 650 655
Asn Lys Lys Asp Leu Tyr Arg Leu Ile Asp Phe Tyr Lys Glu Ala Leu
660 665 670
Thr Val His Glu Trp Lys Leu Phe Asn Phe Ser Phe Ser Pro Thr Glu
675 680 685
Gln Tyr Arg Asn Ile Gly Glu Phe Phe Asp Glu Val Arg Glu Gln Ala
690 695 700
Tyr Lys Val Ser Met Val Asn Val Pro Ala Ser Tyr Ile Asp Glu Ala
705 710 715 720
Val Glu Asn Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe
725 730 735
Ser Pro Tyr Ser Lys Gly Ile Pro Asn Leu His Thr Leu Tyr Trp Lys
740 745 750
Ala Leu Phe Ser Glu Gln Asn Gln Ser Arg Val Tyr Lys Leu Cys Gly
755 760 765
Gly Gly Glu Leu Phe Tyr Arg Lys Ala Ser Leu His Met Gln Asp Thr
770 775 780
Thr Val His Pro Lys Gly Ile Ser Ile His Lys Lys Asn Leu Asn Lys
785 790 795 800
Lys Gly Glu Thr Ser Leu Phe Asn Tyr Asp Leu Val Lys Asp Lys Arg
805 810 815
Phe Thr Glu Asp Lys Phe Phe Phe His Val Pro Ile Ser Ile Asn Tyr
820 825 830
Lys Asn Lys Lys Ile Thr Asn Val Asn Gln Met Val Arg Asp Tyr Ile
835 840 845
Ala Gln Asn Asp Asp Leu Gln His Gly Ile Asp Arg Gly Glu Arg Asn
850 855 860
Leu Leu Tyr Ile Ser Arg Ile Asp Thr Arg Gly Asn Leu Leu Glu Gln
865 870 875 880
Phe Ser Leu Asn Val Ile Glu Ser Asp Lys Gly Asp Leu Arg Thr Asp
885 890 895
Tyr Gln Lys Ile Leu Gly Asp Arg Glu Gln Glu Arg Leu Arg Arg Arg
900 905 910
Gln Glu Trp Lys Ser Ile Glu Ser Ile Lys Asp Leu Lys Asp Gly Tyr
915 920 925
Met Ser Gln Val Val His Lys Ile Cys Asn Met Val Val Glu His Lys
930 935 940
Ala Ile Val Val Leu Glu Asn Leu Asn Leu Ser Phe Met Lys Gly Arg
945 950 955 960
Lys Lys Val Glu Lys Ser Val Tyr Glu Lys Phe Glu Arg Met Leu Val
965 970 975
Asp Lys Leu Asn Tyr Leu Val Val Asp Lys Lys Asn Leu Ser Asn Glu
980 985 990
Pro Gly Gly Leu Tyr Ala Ala Tyr Gln Leu Thr Asn Pro Leu Phe Ser
995 1000 1005
Phe Glu Glu Leu His Arg Tyr Pro Gln Ser Gly Ile Leu Phe Phe
1010 1015 1020
Val Asp Pro Trp Asn Thr Ser Leu Thr Asp Pro Ser Thr Gly Phe
1025 1030 1035
Val Asn Leu Leu Gly Arg Ile Asn Tyr Thr Asn Val Gly Asp Ala
1040 1045 1050
Arg Lys Phe Phe Asp Arg Phe Asn Ala Ile Arg Tyr Asp Gly Lys
1055 1060 1065
Gly Asn Ile Leu Phe Asp Leu Asp Leu Ser Arg Phe Asp Val Arg
1070 1075 1080
Val Glu Thr Gln Arg Lys Leu Trp Thr Leu Thr Thr Phe Gly Ser
1085 1090 1095
Arg Ile Ala Lys Ser Lys Lys Ser Gly Lys Trp Met Val Glu Arg
1100 1105 1110
Ile Glu Asn Leu Ser Leu Cys Phe Leu Glu Leu Phe Glu Gln Phe
1115 1120 1125
Asn Ile Gly Tyr Arg Val Glu Lys Asp Leu Lys Lys Ala Ile Leu
1130 1135 1140
Ser Gln Asp Arg Lys Glu Phe Tyr Val Arg Leu Ile Tyr Leu Phe
1145 1150 1155
Asn Leu Met Met Gln Ile Arg Asn Ser Asp Gly Glu Glu Asp Tyr
1160 1165 1170
Ile Leu Ser Pro Ala Leu Asn Glu Lys Asn Leu Gln Phe Asp Ser
1175 1180 1185
Arg Leu Ile Glu Ala Lys Asp Leu Pro Val Asp Ala Asp Ala Asn
1190 1195 1200
Gly Ala Tyr Asn Val Ala Arg Lys Gly Leu Met Val Val Gln Arg
1205 1210 1215
Ile Lys Arg Gly Asp His Glu Ser Ile His Arg Ile Gly Arg Ala
1220 1225 1230
Gln Trp Leu Arg Tyr Val Gln Glu Gly Ile Val Glu
1235 1240 1245
<210> 17
<211> 1250
<212> PRT
<213> Smithella物种
<400> 17
Met Gln Thr Leu Phe Glu Asn Phe Thr Asn Gln Tyr Pro Val Ser Lys
1 5 10 15
Thr Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Lys Asp Phe Ile
20 25 30
Glu Gln Lys Gly Leu Leu Lys Lys Asp Glu Asp Arg Ala Glu Lys Tyr
35 40 45
Lys Lys Val Lys Asn Ile Ile Asp Glu Tyr His Lys Asp Phe Ile Glu
50 55 60
Lys Ser Leu Asn Gly Leu Lys Leu Asp Gly Leu Glu Lys Tyr Lys Thr
65 70 75 80
Leu Tyr Leu Lys Gln Glu Lys Asp Asp Lys Asp Lys Lys Ala Phe Asp
85 90 95
Lys Glu Lys Glu Asn Leu Arg Lys Gln Ile Ala Asn Ala Phe Arg Asn
100 105 110
Asn Glu Lys Phe Lys Thr Leu Phe Ala Lys Glu Leu Ile Lys Asn Asp
115 120 125
Leu Met Ser Phe Ala Cys Glu Glu Asp Lys Lys Asn Val Lys Glu Phe
130 135 140
Glu Ala Phe Thr Thr Tyr Phe Thr Gly Phe His Gln Asn Arg Ala Asn
145 150 155 160
Met Tyr Val Ala Asp Glu Lys Arg Thr Ala Ile Ala Ser Arg Leu Ile
165 170 175
His Glu Asn Leu Pro Lys Phe Ile Asp Asn Ile Lys Ile Phe Glu Lys
180 185 190
Met Lys Lys Glu Ala Pro Glu Leu Leu Ser Pro Phe Asn Gln Thr Leu
195 200 205
Lys Asp Met Lys Asp Val Ile Lys Gly Thr Thr Leu Glu Glu Ile Phe
210 215 220
Ser Leu Asp Tyr Phe Asn Lys Thr Leu Thr Gln Ser Gly Ile Asp Ile
225 230 235 240
Tyr Asn Ser Val Ile Gly Gly Arg Thr Pro Glu Glu Gly Lys Thr Lys
245 250 255
Ile Lys Gly Leu Asn Glu Tyr Ile Asn Thr Asp Phe Asn Gln Lys Gln
260 265 270
Thr Asp Lys Lys Lys Arg Gln Pro Lys Phe Lys Gln Leu Tyr Lys Gln
275 280 285
Ile Leu Ser Asp Arg Gln Ser Leu Ser Phe Ile Ala Glu Ala Phe Lys
290 295 300
Asn Asp Thr Glu Ile Leu Glu Ala Ile Glu Lys Phe Tyr Val Asn Glu
305 310 315 320
Leu Leu His Phe Ser Asn Glu Gly Lys Ser Thr Asn Val Leu Asp Ala
325 330 335
Ile Lys Asn Ala Val Ser Asn Leu Glu Ser Phe Asn Leu Thr Lys Met
340 345 350
Tyr Phe Arg Ser Gly Ala Ser Leu Thr Asp Val Ser Arg Lys Val Phe
355 360 365
Gly Glu Trp Ser Ile Ile Asn Arg Ala Leu Asp Asn Tyr Tyr Ala Thr
370 375 380
Thr Tyr Pro Ile Lys Pro Arg Glu Lys Ser Glu Lys Tyr Glu Glu Arg
385 390 395 400
Lys Glu Lys Trp Leu Lys Gln Asp Phe Asn Val Ser Leu Ile Gln Thr
405 410 415
Ala Ile Asp Glu Tyr Asp Asn Glu Thr Val Lys Gly Lys Asn Ser Gly
420 425 430
Lys Val Ile Ala Asp Tyr Phe Ala Lys Phe Cys Asp Asp Lys Glu Thr
435 440 445
Asp Leu Ile Gln Lys Val Asn Glu Gly Tyr Ile Ala Val Lys Asp Leu
450 455 460
Leu Asn Thr Pro Cys Pro Glu Asn Glu Lys Leu Gly Ser Asn Lys Asp
465 470 475 480
Gln Val Lys Gln Ile Lys Ala Phe Met Asp Ser Ile Met Asp Ile Met
485 490 495
His Phe Val Arg Pro Leu Ser Leu Lys Asp Thr Asp Lys Glu Lys Asp
500 505 510
Glu Thr Phe Tyr Ser Leu Phe Thr Pro Leu Tyr Asp His Leu Thr Gln
515 520 525
Thr Ile Ala Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr Gln Lys Pro
530 535 540
Tyr Ser Thr Glu Lys Ile Lys Leu Asn Phe Glu Asn Ser Thr Leu Leu
545 550 555 560
Gly Gly Trp Asp Leu Asn Lys Glu Thr Asp Asn Thr Ala Ile Ile Leu
565 570 575
Arg Lys Asp Asn Leu Tyr Tyr Leu Gly Ile Met Asp Lys Arg His Asn
580 585 590
Arg Ile Phe Arg Asn Val Pro Lys Ala Asp Lys Lys Asp Phe Cys Tyr
595 600 605
Glu Lys Met Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro
610 615 620
Lys Val Phe Phe Ser Gln Ser Arg Ile Gln Glu Phe Thr Pro Ser Ala
625 630 635 640
Lys Leu Leu Glu Asn Tyr Ala Asn Glu Thr His Lys Lys Gly Asp Asn
645 650 655
Phe Asn Leu Asn His Cys His Lys Leu Ile Asp Phe Phe Lys Asp Ser
660 665 670
Ile Asn Lys His Glu Asp Trp Lys Asn Phe Asp Phe Arg Phe Ser Ala
675 680 685
Thr Ser Thr Tyr Ala Asp Leu Ser Gly Phe Tyr His Glu Val Glu His
690 695 700
Gln Gly Tyr Lys Ile Ser Phe Gln Ser Val Ala Asp Ser Phe Ile Asp
705 710 715 720
Asp Leu Val Asn Glu Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys
725 730 735
Asp Phe Ser Pro Phe Ser Lys Gly Lys Pro Asn Leu His Thr Leu Tyr
740 745 750
Trp Lys Met Leu Phe Asp Glu Asn Asn Leu Lys Asp Val Val Tyr Lys
755 760 765
Leu Asn Gly Glu Ala Glu Val Phe Tyr Arg Lys Lys Ser Ile Ala Glu
770 775 780
Lys Asn Thr Thr Ile His Lys Ala Asn Glu Ser Ile Ile Asn Lys Asn
785 790 795 800
Pro Asp Asn Pro Lys Ala Thr Ser Thr Phe Asn Tyr Asp Ile Val Lys
805 810 815
Asp Lys Arg Tyr Thr Ile Asp Lys Phe Gln Phe His Ile Pro Ile Thr
820 825 830
Met Asn Phe Lys Ala Glu Gly Ile Phe Asn Met Asn Gln Arg Val Asn
835 840 845
Gln Phe Leu Lys Ala Asn Pro Asp Ile Asn Ile Ile Gly Ile Asp Arg
850 855 860
Gly Glu Arg His Leu Leu Tyr Tyr Ala Leu Ile Asn Gln Lys Gly Lys
865 870 875 880
Ile Leu Lys Gln Asp Thr Leu Asn Val Ile Ala Asn Glu Lys Gln Lys
885 890 895
Val Asp Tyr His Asn Leu Leu Asp Lys Lys Glu Gly Asp Arg Ala Thr
900 905 910
Ala Arg Gln Glu Trp Gly Val Ile Glu Thr Ile Lys Glu Leu Lys Glu
915 920 925
Gly Tyr Leu Ser Gln Val Ile His Lys Leu Thr Asp Leu Met Ile Glu
930 935 940
Asn Asn Ala Ile Ile Val Met Glu Asp Leu Asn Phe Gly Phe Lys Arg
945 950 955 960
Gly Arg Gln Lys Val Glu Lys Gln Val Tyr Gln Lys Phe Glu Lys Met
965 970 975
Leu Ile Asp Lys Leu Asn Tyr Leu Val Asp Lys Asn Lys Lys Ala Asn
980 985 990
Glu Leu Gly Gly Leu Leu Asn Ala Phe Gln Leu Ala Asn Lys Phe Glu
995 1000 1005
Ser Phe Gln Lys Met Gly Lys Gln Asn Gly Phe Ile Phe Tyr Val
1010 1015 1020
Pro Ala Trp Asn Thr Ser Lys Thr Asp Pro Ala Thr Gly Phe Ile
1025 1030 1035
Asp Phe Leu Lys Pro Arg Tyr Glu Asn Leu Asn Gln Ala Lys Asp
1040 1045 1050
Phe Phe Glu Lys Phe Asp Ser Ile Arg Leu Asn Ser Lys Ala Asp
1055 1060 1065
Tyr Phe Glu Phe Ala Phe Asp Phe Lys Asn Phe Thr Glu Lys Ala
1070 1075 1080
Asp Gly Gly Arg Thr Lys Trp Thr Val Cys Thr Thr Asn Glu Asp
1085 1090 1095
Arg Tyr Gln Trp Asn Arg Ala Leu Asn Asn Asn Arg Gly Ser Gln
1100 1105 1110
Glu Lys Tyr Asp Ile Thr Ala Glu Leu Lys Ser Leu Phe Asp Gly
1115 1120 1125
Lys Val Asp Tyr Lys Ser Gly Lys Asp Leu Lys Gln Gln Ile Ala
1130 1135 1140
Ser Gln Glu Ser Ala Asp Phe Phe Lys Ala Leu Met Lys Asn Leu
1145 1150 1155
Ser Ile Thr Leu Ser Leu Arg His Asn Asn Gly Glu Lys Gly Asp
1160 1165 1170
Asn Glu Gln Asp Tyr Ile Leu Ser Pro Val Ala Asp Ser Lys Gly
1175 1180 1185
Arg Phe Phe Asp Ser Arg Lys Ala Asp Asp Asp Met Pro Lys Asn
1190 1195 1200
Ala Asp Ala Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Leu Trp
1205 1210 1215
Cys Leu Glu Gln Ile Ser Lys Thr Asp Asp Leu Lys Lys Val Lys
1220 1225 1230
Leu Ala Ile Ser Asn Lys Glu Trp Leu Glu Phe Val Gln Thr Leu
1235 1240 1245
Lys Gly
1250
<210> 18
<211> 3987
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 18
atggccggga gcaagaagcg ccggataaag caggacacgc agttcgaggg cttcaccaac 60
ctgtaccaag tctccaagac gctccggttc gagcttatcc cgcaagggaa gaccctgaaa 120
cacatccagg aacaaggttt catcgaggag gacaaggccc gcaacgacca ctacaaggag 180
ctcaagccca taatcgatcg gatctacaag acgtacgccg accagtgcct ccaactggtg 240
cagctcgact gggagaacct gagcgccgcc attgacagct accgcaagga aaagacggag 300
gagacgcgca acgcccttat tgaggagcaa gccacctacc gcaacgccat ccacgactac 360
ttcatcgggc gcaccgacaa cctgacggac gcgatcaaca agcgccacgc ggaaatctac 420
aagggccttt tcaaggccga gctcttcaac gggaaggtcc taaaacagct cgggactgtc 480
acgacaaccg agcatgagaa cgccctcctt cgcagcttcg acaagttcac cacatacttc 540
tcgggcttct accggaaccg caagaacgtt ttcagcgccg aggacatctc caccgccatc 600
ccgcacagga tcgtccagga caacttcccc aagttcaagg agaactgcca catcttcacg 660
cgcctgatta cagccgtacc ttcacttcgt gagcacttcg agaacgtcaa aaaggccatc 720
gggatcttcg tctccacgtc catcgaggag gtattctctt tcccgttcta taaccagctc 780
ctgacccaga cgcagatcga cctctacaac cagctactgg gcggcatcag ccgggaggcc 840
gggaccgaga aaataaaggg cctcaacgaa gttctcaacc tggccatcca gaagaacgac 900
gagaccgcgc atatcatcgc atccctgccg catcgcttca ttcctttgtt caagcagata 960
ttgagcgacc ggaacaccct ctcgttcatc ctcgaagaat tcaagagcga cgaggaggtc 1020
attcagtctt tctgcaagta caagacgctc ctacggaatg agaatgtgct ggagaccgcg 1080
gaggcactct tcaatgagct gaactccatt gacctgaccc acatcttcat tagccacaag 1140
aaactggaga cgatctccag cgccctgtgc gaccactggg acactctccg caacgccctc 1200
tacgaacgcc ggatctccga acttaccggc aagataacta agtcggctaa ggagaaggtg 1260
caacggagcc tcaagcacga ggacatcaac cttcaggaaa tcatctcagc cgcgggcaag 1320
gagctgagcg aggcgtttaa gcagaaaaca tcggagatac tgagccacgc gcacgcggcc 1380
ctggatcaac cgctgccgac gactctcaag aagcaagagg agaaggaaat ccttaagtcc 1440
cagctcgact cgctgctcgg cctctatcac ttgctcgact ggttcgcggt tgatgagtcc 1500
aacgaggtgg acccggagtt ctccgcgcgc ctcacgggta ttaagctgga gatggagcca 1560
agcttaagct tctacaacaa ggcccgcaac tacgcgacca aaaaaccgta ctcagtcgag 1620
aaattcaagc tgaatttcca gatgcctaca ttggcgaggg ggtgggacgt gaaccgcgag 1680
aagaacaatg gagccatcct gttcgtcaaa aatgggttgt actacctggg catcatgccc 1740
aagcagaagg gccgttacaa ggccctgtca ttcgagccta ccgagaagac ctcggagggc 1800
ttcgacaaga tgtactacga ctatttcccg gacgccgcca agatgatccc gaagtgctcc 1860
acgcagctca aagccgtcac ggcccacttc cagacgcata ccacgccgat acttctgagc 1920
aacaacttca ttgagccgct agagatcacg aaggagatat acgacctaaa caaccccgaa 1980
aaggagccca agaagttcca gacagcctac gctaagaaga caggtgatca gaagggatat 2040
agggaggcac tctgcaagtg gatcgacttc acgcgcgact tcctgtcgaa atatacaaag 2100
acgaccagca ttgacctaag ttctctccgc ccatcctccc agtacaagga tctgggcgag 2160
tattatgcgg agctgaaccc attgctgtac cacatcagct tccagaggat cgccgagaag 2220
gagattatgg acgcggtgga gacggggaaa ctatacctgt tccaaatata taacaaggac 2280
ttcgctaaag ggcaccacgg gaagcccaac ctgcacacac tctactggac gggcttgttt 2340
tcgccagaaa atttggccaa gacttcgatc aagctcaacg gccaggcgga gttgttttac 2400
cgtcccaagt ctcgcatgaa gcgcatggcg catcgcctcg gagagaaaat gcttaacaag 2460
aagctcaagg atcagaagac gcccatacct gatacgttgt accaggaatt gtacgactac 2520
gtgaaccacc gcctatcgca cgacctctca gacgaggccc gcgccctcct cccaaacgtg 2580
attactaagg aggtttccca tgaaataatc aaggaccgac ggttcaccag cgacaaattt 2640
tttttccacg tgcctatcac gctcaattac caggcggcca actccccatc gaagttcaac 2700
cagcgcgtga acgcctacct taaggagcac ccggagaccc caatcatcgg gatcgaccgt 2760
ggcgagcgga acctgatcta tattacggtg atcgatagca ccgggaagat cctggagcag 2820
cgctccctga acacaatcca gcagtttgac taccagaaga aactcgacaa ccgggagaag 2880
gagcgcgtcg cagcccggca agcatggagt gtggtcggca ccataaagga cctgaaacag 2940
ggttacctaa gtcaagttat ccacgagatc gttgacctga tgatacacta tcaagccgta 3000
gtcgtgctgg agaacctcaa cttcgggttt aagtccaagc gcaccggcat cgcggagaag 3060
gcggtgtacc agcagttcga gaagatgctg atcgacaagc tgaactgcct ggtgctcaag 3120
gactaccctg cggagaaggt cggcggggtc ttgaacccgt accagctaac cgaccagttc 3180
acgagcttcg ccaaaatggg cacgcagtcc ggattcttgt tttatgtccc ggctccatat 3240
acaagtaaga tcgacccgct gacagggttt gttgacccat tcgtgtggaa gaccatcaag 3300
aaccacgaga gcaggaaaca cttcttagag ggcttcgact tcctgcatta cgacgttaag 3360
acaggcgact tcatcctgca cttcaagatg aaccgcaacc tgtcgttcca gaggggcctg 3420
cccggcttca tgcccgcctg ggatatcgtc tttgagaaga atgagacgca gttcgacgcg 3480
aaggggacgc cgttcatcgc tggaaagcgg atcgtgccgg tcatcgagaa ccaccgcttc 3540
acgggtcgct accgagattt ataccccgcc aacgaactaa ttgcgctgct ggaggagaag 3600
gggatcgtgt tccgagatgg cagcaacatt ctcccgaagc tgctggagaa cgacgactcg 3660
cacgctattg acacgatggt cgccctcata cggagcgtgc ttcagatgcg gaacagtaac 3720
gctgccacgg gcgaggacta cattaactcc cccgtccgcg acctcaacgg ggtctgcttc 3780
gatagccgct tccagaaccc ggagtggcct atggatgcgg acgcgaacgg ggcctaccac 3840
atcgccctca agggccaact cctgctcaac cacttgaagg aaagcaaaga cctcaaattg 3900
cagaatggca tcagtaacca ggactggctc gcgtacatcc aggaactgag aaacgggtcc 3960
aagaagcggc gtatcaagca agattga 3987
<210> 19
<211> 3987
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 19
atggcgggaa gcaaaaagcg ccggattaag caagacacgc agttcgaggg cttcacgaac 60
ctctaccaag tcagcaagac cctccggttc gagctgatac cacagggaaa gacgctcaag 120
cacatccagg aacagggctt catcgaggag gacaaggcgc gcaacgacca ctacaaggag 180
ttgaaaccga tcatcgaccg catctacaag acgtacgccg accagtgcct ccagctcgtg 240
cagctcgact gggagaacct ctccgccgcc attgactcgt accggaagga gaagactgag 300
gagacccgca acgccctgat cgaggagcaa gcaacctacc ggaacgccat ccacgactac 360
ttcatcggcc gcaccgacaa cctcaccgac gcgatcaaca agcggcacgc ggagatatac 420
aaagggctgt tcaaggcgga gctgttcaac ggcaaggtgc tcaagcagct agggacggtg 480
accacgaccg agcacgagaa cgcgctcctc cgcagcttcg acaagttcac cacctacttc 540
agcggcttct accggaaccg caagaatgtg ttcagcgcgg aggacatcag cacggccatc 600
ccgcaccgca tcgtccagga caacttcccg aagttcaagg agaactgcca catcttcacc 660
cgcctgataa ccgccgtccc ctccctgcgg gagcacttcg agaacgtcaa aaaggcaatt 720
gggatcttcg tctcgaccag cattgaggag gtgttcagct tccccttcta caaccagctc 780
ctcacccaga cgcagatcga cctgtacaat cagttgctcg gcgggataag ccgcgaggcg 840
ggaaccgaaa aaatcaaggg gctgaacgaa gtgttgaacc tcgccatcca gaagaacgac 900
gagaccgcgc acatcatcgc ctccctgccc caccggttca tcccgctgtt caagcagatc 960
ctctctgacc ggaacaccct gtccttcatt cttgaggagt tcaagtcgga cgaggaggtc 1020
atccagagct tctgcaagta caagacgctg ctacggaacg agaacgtgct ggagacggcg 1080
gaggcactgt tcaacgagct aaacagcatc gacctcacgc acatcttcat cagtcacaag 1140
aaactggaga ccatctcctc cgcgctgtgc gaccactggg acacgctcag gaacgcgctc 1200
tacgagcgcc gaatcagtga gctgacgggc aagatcacga agtccgcgaa ggagaaggtg 1260
cagcggtccc tcaagcacga ggacatcaac ctccaggaga tcatctcagc ggctgggaaa 1320
gagctgtccg aggcgttcaa gcagaaaacg agcgaaatcc tgtcccacgc gcacgcggcc 1380
ctggatcagc ctctgccgac gaccctcaag aaacaagaag aaaaggaaat cctcaagtcg 1440
cagctcgact cgctgctggg cctgtaccat ctcctcgact ggttcgccgt ggacgagagc 1500
aacgaggtgg accccgagtt ctccgcgcgg cttacgggga tcaagctgga gatggagccc 1560
agcctgtcct tctacaacaa ggcgcgcaac tacgccacca agaagcccta cagcgtggag 1620
aagttcaagc tcaacttcca gatgcccact ctcgcacgtg ggtgggacgt caaccgcgaa 1680
aaaaataatg gggcgatcct gttcgtcaag aacggcctgt actacttggg catcatgccg 1740
aaacagaagg gccgctacaa ggccctgagc ttcgaaccga ccgagaaaac gagcgagggg 1800
ttcgacaaga tgtactacga ctacttcccc gacgccgcga agatgattcc aaagtgctcc 1860
acgcagctta aggccgtgac ggcccacttc cagacgcaca cgaccccgat cctcctcagc 1920
aacaacttca tcgagcccct ggagatcacg aaggagatat acgacctgaa caacccggag 1980
aaggagccca agaaattcca gaccgcctac gccaagaaga caggcgacca aaagggttac 2040
agggaggccc tctgcaagtg gatcgacttc actagggact tcctgtccaa gtacaccaag 2100
actacctcta tcgacctgtc cagcctccgc ccgtcgtccc agtacaaaga tttgggcgag 2160
tattacgcgg agctgaaccc actgctctac cacatcagct tccagcgcat cgcggagaag 2220
gagatcatgg acgcagtgga gacgggcaag ctatacctat ttcagatata caacaaagac 2280
ttcgctaagg gacaccacgg caagcctaac ctgcacaccc tctactggac ggggctcttc 2340
agcccggaga acctcgccaa gacctcgatc aagctcaacg gccaggccga gctgttctac 2400
cggcccaagt cccgcatgaa gcggatggcc caccggctcg gggagaaaat gctcaacaag 2460
aaattgaagg accaaaaaac gccgataccc gacaccctat accaggagct gtacgactat 2520
gtgaaccacc gcctgagcca cgacctcagc gacgaggcgc gggccctcct gccgaacgtc 2580
atcacaaagg aggtcagcca cgagatcatc aaggaccggc gcttcacctc cgacaagttt 2640
ttctttcacg tgcccatcac gctcaactac caggccgcca actcgccgtc caagttcaac 2700
cagcgcgtga acgcctacct caaggagcac cccgagaccc cgatcatcgg gattgaccga 2760
ggggagcgga acctcatcta catcaccgtc atcgacagca ccgggaagat ccttgaacag 2820
cggtcgctca acaccatcca gcagttcgac taccagaaga aactcgacaa ccgggagaag 2880
gagagagtgg cggcccgcca ggcttggtcc gtcgtcggga cgattaagga cttgaaacaa 2940
ggttacctgt cgcaagtgat ccacgagatc gttgacctga tgatccacta ccaagccgtc 3000
gtggtcctgg agaacctcaa cttcggcttc aagagcaaac gaaccggcat cgcggagaag 3060
gccgtgtacc agcagttcga aaaaatgctg atcgacaagc tgaactgcct cgtgctcaag 3120
gactaccccg ctgagaaggt cggcggggtg ctgaacccgt accagctcac tgaccagttc 3180
accagcttcg caaagatggg cacccagtcc ggcttcctgt tctacgtgcc tgcgccatac 3240
acctcgaaga tcgacccgct caccgggttc gtggacccct tcgtctggaa gaccatcaag 3300
aaccacgaga gccgcaagca cttcctggag ggcttcgact tcctccacta cgacgtcaag 3360
accggggact tcatcctgca cttcaagatg aaccgcaacc tcagtttcca gcgcggcctg 3420
ccggggttca tgcccgcttg ggatatagtc ttcgagaaga atgagacgca gttcgacgcg 3480
aagggcaccc cgttcatcgc cgggaagcgc atcgtgccgg tcatcgagaa ccaccggttc 3540
accgggcgct accgcgacct atacccggcg aacgagttga tcgccctcct ggaggagaag 3600
ggcatcgtgt tccgcgacgg ctccaacatc ctcccgaagc tgctcgaaaa cgacgactcc 3660
cacgccatcg acacgatggt cgcgctgatc cggtcggtgc tccagatgcg gaactccaac 3720
gccgcgacgg gcgaggacta catcaacagt ccggtccgcg atctgaacgg cgtctgcttc 3780
gactcccggt tccagaaccc cgagtggccg atggacgcgg acgcgaacgg cgcataccac 3840
atcgccctaa aagggcaatt gctgctcaac cacctcaagg aatccaaaga cctaaagctc 3900
cagaacggca tctccaacca ggactggctg gcgtacatcc aggaactgcg gaacgggagc 3960
aaaaaacgtc ggatcaagca agattga 3987
<210> 20
<211> 3987
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 20
atggcgggct ccaagaaacg ccggattaag caagataccc agttcgaggg gttcacgaac 60
ctctaccaag tgagcaagac cctccgattc gaactgattc ctcaggggaa gaccctcaag 120
cacatccagg agcaagggtt catcgaggag gacaaggcgc ggaacgacca ctacaaggaa 180
ctcaaaccca tcatcgaccg catctacaag acctacgccg atcagtgcct ccagctcgtg 240
cagttggact gggagaacct cagcgcggcc attgactcct accggaagga gaaaacggag 300
gagacgcgca acgcgctcat cgaggaacag gcaacctatc gcaacgccat ccacgactac 360
ttcatcggga ggactgacaa cctcactgac gcgattaaca agcgccacgc ggagatatac 420
aagggactct tcaaagcgga gctgtttaac ggcaaggttc tcaagcaact cggcactgtg 480
accacgaccg agcatgagaa cgccctgctc cgctccttcg acaagttcac cacctacttc 540
tccgggttct accgcaaccg caagaatgtc ttcagcgcgg aggacatcag cacggccatt 600
ccacatcgaa tcgtccaaga taacttcccg aagttcaagg agaactgcca catcttcacc 660
cgactcatta ctgctgtacc gtcgttacgc gaacacttcg agaacgtcaa gaaggcaatt 720
ggaatcttcg tctctacgtc aatagaggag gtgttcagct tccctttcta caaccagctc 780
cttacgcaga cccagataga cctgtacaat cagctcctcg gtgggatcag ccgggaggcg 840
gggactgaga agattaaagg gctcaacgag gtcttgaacc tggccatcca aaaaaacgat 900
gagacggcgc acatcatcgc ctcgctgccc caccggttca tcccgctgtt caagcagatc 960
ctcagtgaca ggaacacctt gagctttatc ctagaggagt tcaagagcga cgaggaggtg 1020
atccagagct tctgcaagta caaaaccctg ctgaggaacg agaacgtcct ggagacggcg 1080
gaggcgctgt tcaacgagct gaactctatc gacttaactc acatattcat ctcgcacaag 1140
aagctggaga ctattagctc tgcactctgc gaccactggg acaccctccg caacgcgctc 1200
tacgagcgcc gcatctcgga gctgaccggg aagatcacca aatccgcgaa ggaaaaggtc 1260
cagcgttccc tcaaacacga ggatattaac ttacaggaga ttatctcagc ggctgggaag 1320
gagttgtcag aggcgttcaa gcagaaaact tccgagatcc tgagccacgc gcacgcagcg 1380
ctcgaccagc ctctgcccac caccctcaaa aagcaggaag aaaaagagat cctcaagagc 1440
cagttggact ccctgctggg gctctatcac cttctcgact ggttcgccgt cgatgagtcg 1500
aacgaggtgg accccgagtt ctccgcccgg ctgaccggca tcaagctaga gatggagccg 1560
tccctcagct tctacaataa ggcccgcaac tacgcgacca aaaaacccta cagcgtggag 1620
aagttcaagc tgaacttcca gatgccgacc ttagcacgcg gttgggacgt aaacagggag 1680
aagaacaatg gagccatcct gttcgtcaag aacgggcttt actacctcgg gataatgccc 1740
aagcagaagg gccgctacaa ggccctttcc ttcgagccga cggagaaaac ctccgagggg 1800
ttcgacaaga tgtactacga ctacttcccc gacgccgcca agatgatccc gaagtgctca 1860
acgcagctaa aagccgtgac cgcccacttc cagacccaca cgacgccgat cctgctgagc 1920
aacaacttca tcgagcccct tgagatcact aaggagatat acgacctgaa caaccccgag 1980
aaggagccca agaagtttca aaccgcctac gccaaaaaaa ctggcgacca aaagggctac 2040
agggaggcgc tgtgtaagtg gatcgacttc acacgcgact tcctttcgaa gtatacgaag 2100
acaacctcta ttgacctgag cagcctgcgt cctagctccc agtacaaaga tttgggcgag 2160
tactacgcgg agcttaatcc actactctac cacatctcat tccagcgcat cgctgagaag 2220
gaaatcatgg acgcggtgga gacaggcaaa ctgtacctct tccagatata caacaaagac 2280
ttcgctaagg ggcaccacgg gaagcccaac cttcatacgc tctactggac gggcctattc 2340
agccccgaaa atctggccaa gacctccatc aagctgaacg gccaagcgga gctgttctac 2400
agacccaaga gccggatgaa gcggatggcc cacaggctcg gcgagaaaat gcttaacaaa 2460
aagttgaagg accagaaaac ccctatcccc gacaccctct accaggaact gtacgactac 2520
gtgaaccaca ggctctcgca cgacctttcc gacgaggccc gtgccctact cccgaacgtc 2580
attaccaaag aggtttcgca cgagatcatc aaggaccggc ggttcacgag cgacaagttt 2640
ttctttcacg tccccatcac ccttaactac caggcggcca actccccatc caagttcaac 2700
cagcgtgtga atgcctacct caaggagcac ccagagaccc cgatcattgg gatcgaccgg 2760
ggcgagcgga acctgatcta catcaccgtc atcgactcga cgggcaagat tcttgagcag 2820
agatcgttga ataccataca gcagttcgac taccagaaga aactcgacaa ccgcgagaag 2880
gagcgcgtgg cggcccgcca ggcgtggtcc gtcgttggga cgattaagga cttgaaacaa 2940
ggttatctgt cccaagtcat ccacgagatc gttgatctga tgatccacta tcaggcagtg 3000
gtggtgctgg agaatctcaa cttcggcttc aagagtaagc ggacgggaat cgccgagaag 3060
gccgtgtacc agcagttcga gaagatgctg atcgacaagc tcaactgcct tgtgctgaaa 3120
gactacccgg ccgagaaggt cggcggcgtc ctcaacccgt accaacttac cgaccagttc 3180
acctccttcg ccaagatggg cactcagtcc gggttcttgt tctacgtccc cgcaccttac 3240
acctctaaga tcgaccctct gactggcttc gtagatccat tcgtgtggaa gaccattaag 3300
aaccacgaga gccgcaagca cttcctggag ggcttcgact tcctgcacta cgacgtgaag 3360
accggggact tcatccttca cttcaagatg aaccggaacc tcagcttcca gcggggcctg 3420
ccggggttca tgcccgcctg ggacatcgtg ttcgagaaga acgagaccca gttcgacgcg 3480
aagggcacgc ccttcatcgc cgggaagcgt atcgtgccgg tgatcgagaa ccatcgtttc 3540
acgggtcgct accgtgacct ctacccggcg aacgagctta tcgcactcct ggaggagaag 3600
ggcatcgtct tccgggacgg ctccaacatc ctcccgaaac tgctggaaaa cgacgactct 3660
cacgccatcg acacgatggt ggccctcatc cggtccgtgc tccaaatgcg gaacagcaac 3720
gccgccaccg gtgaggacta catcaacagc ccggtccggg atctgaacgg ggtgtgcttc 3780
gattcgcggt tccagaatcc tgagtggccg atggacgcgg atgcaaacgg ggcgtaccac 3840
atcgcgctca agggccagtt acttctgaac caccttaagg agtctaaaga tttgaaactc 3900
cagaacggga tctcgaacca ggactggctg gcctacatcc aagagttgcg gaacggcagc 3960
aagaagcggc ggattaagca agattag 3987
<210> 21
<211> 1592
<212> DNA
<213> 蒺藜苜蓿(Medicago truncatula)
<400> 21
actgttaata atttttaaac gtcagcgcac taaaaaaacg aaaagacgga cacgtgaaaa 60
taaaaaacac acactagttt atgacgcaat actattttac ttatgatttg ggtacattag 120
acaaaaccgt gaaagagatg tatcagctat gaaacctgta tacttcaata cagagactta 180
ctcatatcgg atacgtacgc acgaagtatc atattaatta ttttaatttt taataaatat 240
tttatcggat acttatgtga tactctacat atacacaagg atatttctaa gatactttat 300
agatacgtat cctagaaaaa catgaagagt aaaaaagtga gacaatgttg taaaaattca 360
ttataaatgt atatgattca attttagata tgcatcagta taattgattc tcgatgaaac 420
acttaaaatt atatttcttg tggaagaacg tagcgagaga ggtgattcag ttagacaaca 480
ttaaataaaa ttaatgttaa gttcttttaa tgatgtttct ctcaatatca catcatatga 540
aaatgtaata tgatttataa gaaaattttt aaaaaattta ttttaataat cacatgtact 600
attttttaaa aattgtatct tttataataa tacaataata aagagtaatc agtgttaatt 660
tttcttcaaa tataagtttt attataaatc attgttaacg tatcataagt cattaccgta 720
tcgtatctta attttttttt aaaaaccgct aattcacgta cccgtattgt attgtacccg 780
cacctgtatc acaatcgatc ttagttagaa gaattgtctc gaggcggtgc aagacagcat 840
ataatagacg tggactctct tataccaaac gttgtcgtat cacaaagggt taggtaacaa 900
gtcacagttt gtccacgtgt cacgttttaa ttggaagagg tgccgttggc gtaatataac 960
agccaatcga tttttgctat aaaagcaaat caggtaaact aaacttcttc attcttttct 1020
tccccatcgc tacaaaaccg gttcctttgg aaaagagatt cattcaaacc tagcacccaa 1080
ttccgtttca aggtataatc tactttctat tcttcgatta ttttattatt attagctact 1140
atcgtttaat cgatcttttc ttttgatccg tcaaatttaa attcaattag ggttttgttc 1200
ttttctttca tctgattgaa atccttctga attgaaccgt ttacttgatt ttactgttta 1260
ttgtatgatt taatcctttg tttttcaaag acagtcttta gattgtgatt aggggttcat 1320
ataaattttt agatttggat ttttgtattg tatgattcaa aaaatacgtc ctttaattag 1380
attagtacat ggatattttt tacccgattt attgattgtc agggagaatt tgatgagcaa 1440
gtttttttga tgtctgttgt aaattgaatt gattataatt gctgatctgc tgcttccagt 1500
tttcataacc catattcttt taaccttgtt gtacacacaa tgaaaaattg gtgattgatt 1560
catttgtttt tctttgtttt ggattataca gg 1592
<210> 22
<211> 2000
<212> DNA
<213> 玉米
<400> 22
gtcgtgcccc tctctagaga taaagagcat tgcatgtcta aagtataaaa aattaccaca 60
tatttttttg tcacacttat ttgaagtgta gtttatctat ctctatacat atatttaaac 120
ttcactctac aaataatata gtctataata ctaaaataat attagtgttt tagaggatca 180
tataaataaa ctgctagaca tggtctaaag gataattgaa tattttgaca atctacagtt 240
ttatcttttt agtgtgcatg tgatctctct gttttttttg caaatagctt gacctatata 300
atacttcatc cattttatta gtacatccat ttaggattta gggttgatgg tttctataga 360
ctaattttta gtacatccat tttattcttt ttagtctcta aattttttaa aactaaaact 420
ctattttagt tttttattta ataatttaga tataaaatga aataaaataa attgactaca 480
aataaaacaa atacccttta agaaataaaa aaactaagca aacatttttc ttgtttcgag 540
tagataatga caggctgttc aacgccgtcg acgagtctaa cggacaccaa ccagcgaacc 600
agcagcgtcg cgtcgggcca agcgaagcag acggcacggc atctctgtag ctgcctctgg 660
acccctctcg agagttccgc tccaccgttg gacttgctcc gctgtcggca tccagaaatt 720
gcgtggcgga gcggcagacg tgaggcggca cggcaggcgg cctcttcctc ctctcacggc 780
accggcagct acgggggatt cctttcccac cgctccttcg ctttcccttc ctcgcccgcc 840
gtaataaata gacaccccct ccacaccctc tttccccaac ctcgtgttcg ttcggagcgc 900
acacacacgc aaccagatct cccccaaatc cagccgtcgg cacctccgct tcaaggtacg 960
ccgctcatcc tccccccccc cctctctcta ccttctctag atcggcgatc cggtccatgg 1020
ttagggcccg gtagttctac ttctgttcat gtttgtgtta gagcaaacat gttcatgttc 1080
atgtttgtga tgatgtggtc tggttgggcg gtcgttctag atcggagtag gatactgttt 1140
caagctacct ggtggattta ttaattttgt atctgtatgt gtgtgccata catcttcata 1200
gttacgagtt taagatgatg gatggaaata tcgatctagg ataggtatac atgttgatgc 1260
gggttttact gatgcatata cagagatgct ttttttctcg cttggttgtg atgatatggt 1320
ctggttgggc ggtcgttcta gatcggagta gaatactgtt tcaaactacc tggtggattt 1380
attaaaggat aaagggtcgt tctagatcgg agtagaatac tgtttcaaac tacctggtgg 1440
atttattaaa ggatctgtat gtatgtgcct acatcttcat agttacgagt ttaagatgat 1500
ggatggaaat atcgatctag gataggtata catgttgatg cgggttttac tgatgcatat 1560
acagagatgc tttttttcgc ttggttgtga tgatgtggtc tggttgggcg gtcgttctag 1620
atcggagtag aatactgttt caaactacct ggtggattta ttaattttgt atctttatgt 1680
gtgtgccata catcttcata gttacgagtt taagatgatg gatggaaata ttgatctagg 1740
ataggtatac atgttgatgt gggttttact gatgcatata catgatggca tatgcggcat 1800
ctattcatat gctctaacct tgagtaccta tctattataa taaacaagta tgttttataa 1860
ttattttgat cttgatatac ttggatgatg gcatatgcag cagctatatg tggatttttt 1920
agccctgcct tcatacgcta tttatttgct tggtactgtt tcttttgtcc gatgctcacc 1980
ctgttgtttg gtgatacttc 2000
<210> 23
<211> 228
<212> PRT
<213> 褐家鼠
<400> 23
Ser Ser Glu Thr Gly Pro Val Ala Val Asp Pro Thr Leu Arg Arg Arg
1 5 10 15
Ile Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro Arg Glu Leu Arg
20 25 30
Lys Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp Gly Gly Arg His Ser
35 40 45
Ile Trp Arg His Thr Ser Gln Asn Thr Asn Lys His Val Glu Val Asn
50 55 60
Phe Ile Glu Lys Phe Thr Thr Glu Arg Tyr Phe Cys Pro Asn Thr Arg
65 70 75 80
Cys Ser Ile Thr Trp Phe Leu Ser Trp Ser Pro Cys Gly Glu Cys Ser
85 90 95
Arg Ala Ile Thr Glu Phe Leu Ser Arg Tyr Pro His Val Thr Leu Phe
100 105 110
Ile Tyr Ile Ala Arg Leu Tyr His His Ala Asp Pro Arg Asn Arg Gln
115 120 125
Gly Leu Arg Asp Leu Ile Ser Ser Gly Val Thr Ile Gln Ile Met Thr
130 135 140
Glu Gln Glu Ser Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr Ser Pro
145 150 155 160
Ser Asn Glu Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu
165 170 175
Tyr Val Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu
180 185 190
Asn Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile Ala
195 200 205
Leu Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp Ala
210 215 220
Thr Gly Leu Lys
225
<210> 24
<211> 199
<212> PRT
<213> 智人
<400> 24
Met Glu Ala Ser Pro Ala Ser Gly Pro Arg His Leu Met Asp Pro His
1 5 10 15
Ile Phe Thr Ser Asn Phe Asn Asn Gly Ile Gly Arg His Lys Thr Tyr
20 25 30
Leu Cys Tyr Glu Val Glu Arg Leu Asp Asn Gly Thr Ser Val Lys Met
35 40 45
Asp Gln His Arg Gly Phe Leu His Asn Gln Ala Lys Asn Leu Leu Cys
50 55 60
Gly Phe Tyr Gly Arg His Ala Glu Leu Arg Phe Leu Asp Leu Val Pro
65 70 75 80
Ser Leu Gln Leu Asp Pro Ala Gln Ile Tyr Arg Val Thr Trp Phe Ile
85 90 95
Ser Trp Ser Pro Cys Phe Ser Trp Gly Cys Ala Gly Glu Val Arg Ala
100 105 110
Phe Leu Gln Glu Asn Thr His Val Arg Leu Arg Ile Phe Ala Ala Arg
115 120 125
Ile Tyr Asp Tyr Asp Pro Leu Tyr Lys Glu Ala Leu Gln Met Leu Arg
130 135 140
Asp Ala Gly Ala Gln Val Ser Ile Met Thr Tyr Asp Glu Phe Lys His
145 150 155 160
Cys Trp Asp Thr Phe Val Asp His Gln Gly Cys Pro Phe Gln Pro Trp
165 170 175
Asp Gly Leu Asp Glu His Ser Gln Ala Leu Ser Gly Arg Leu Arg Ala
180 185 190
Ile Leu Gln Asn Gln Gly Asn
195
<210> 25
<211> 621
<212> DNA
<213> 海七鳃鳗(Petromyzon marinus)
<400> 25
acagatgcag agtatgtgag aattcacgaa aagctggaca tctatacctt caagaagcag 60
ttctttaaca ataagaagtc tgtgagccat aggtgctacg tgctgttcga gctgaagaga 120
aggggtgaaa gaagggcatg tttttggggg tatgctgtga acaagcccca gtctggaact 180
gagagaggca ttcacgccga aattttcagc atcagaaagg tggaggaata cctgagggat 240
aaccctggac agtttacaat taattggtat tctagctggt ctccatgcgc tgactgtgcc 300
gagaagatcc tggaatggta caaccaggag ctgagaggaa atggccatac cctgaagatt 360
tgggcctgca agctgtacta tgaaaagaac gcaagaaatc agatcggact gtggaacctg 420
agggataatg gtgtggggct gaacgtgatg gtgtccgagc actatcagtg ctgtagaaag 480
attttcattc agtcctcaca taatcagctg aacgagaata gatggctgga aaagactctg 540
aagagggctg agaagagaag gtccgaactg tcaattatga tccaggtgaa gatcctgcac 600
accactaagt cacctgccgt g 621
<210> 26
<211> 160
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 26
Phe Glu Arg Asn Tyr Asp Pro Arg Glu Leu Arg Lys Glu Thr Tyr Leu
1 5 10 15
Leu Tyr Glu Ile Lys Trp Gly Lys Ser Gly Lys Leu Trp Arg His Trp
20 25 30
Cys Gln Asn Asn Arg Thr Gln His Ala Glu Val Tyr Phe Leu Glu Asn
35 40 45
Ile Phe Asn Ala Arg Arg Phe Asn Pro Ser Thr His Cys Ser Ile Thr
50 55 60
Trp Tyr Leu Ser Trp Ser Pro Cys Ala Glu Cys Ser Gln Lys Ile Val
65 70 75 80
Asp Phe Leu Lys Glu His Pro Asn Val Leu Glu Ile Tyr Val Ala Arg
85 90 95
Leu Tyr Tyr His Glu Asp Glu Arg Asn Arg Gln Gly Leu Arg Asp Leu
100 105 110
Val Asn Ser Gly Val Thr Ile Arg Ile Met Asp Leu Pro Asp Tyr Asn
115 120 125
Tyr Cys Trp Lys Thr Phe Val Ser Asp Gln Gly Gly Asp Glu Asp Tyr
130 135 140
Trp Pro Gly His Phe Ala Pro Trp Ile Lys Gln Tyr Ser Leu Lys Leu
145 150 155 160
<210> 27
<211> 207
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 27
Thr Asp Ala Glu Tyr Val Arg Ile His Glu Lys Leu Asp Ile Tyr Thr
1 5 10 15
Phe Lys Lys Gln Phe Ser Asn Asn Lys Lys Ser Val Ser His Arg Cys
20 25 30
Tyr Val Leu Phe Glu Leu Lys Arg Arg Gly Glu Arg Arg Ala Cys Phe
35 40 45
Trp Gly Tyr Ala Val Asn Lys Pro Gln Ser Gly Thr Glu Arg Gly Ile
50 55 60
His Ala Glu Ile Phe Ser Ile Arg Lys Val Glu Glu Tyr Leu Arg Asp
65 70 75 80
Asn Pro Gly Gln Phe Thr Ile Asn Trp Tyr Ser Ser Trp Ser Pro Cys
85 90 95
Ala Asp Cys Ala Glu Lys Ile Leu Glu Trp Tyr Asn Gln Glu Leu Arg
100 105 110
Gly Asn Gly His Thr Leu Lys Ile Trp Val Cys Lys Leu Tyr Tyr Glu
115 120 125
Lys Asn Ala Arg Asn Gln Ile Gly Leu Trp Asn Leu Arg Asp Asn Gly
130 135 140
Val Gly Leu Asn Val Met Val Ser Glu His Tyr Gln Cys Cys Arg Lys
145 150 155 160
Ile Phe Ile Gln Ser Ser His Asn Gln Leu Asn Glu Asn Arg Trp Leu
165 170 175
Glu Lys Thr Leu Lys Arg Ala Glu Lys Arg Arg Ser Glu Leu Ser Ile
180 185 190
Met Phe Gln Val Lys Ile Leu His Thr Thr Lys Ser Pro Ala Val
195 200 205
<210> 28
<211> 228
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 28
Ser Ser Lys Thr Gly Pro Val Ala Val Asp Pro Thr Leu Arg Arg Arg
1 5 10 15
Ile Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro Arg Glu Leu Arg
20 25 30
Lys Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp Gly Gly Arg His Ser
35 40 45
Ile Trp Arg His Thr Ser Gln Asn Thr Asn Lys His Val Glu Val Asn
50 55 60
Phe Ile Glu Lys Phe Thr Thr Glu Arg Tyr Phe Cys Pro Asn Thr Arg
65 70 75 80
Cys Ser Ile Thr Trp Phe Leu Ser Trp Ser Pro Cys Gly Glu Cys Ser
85 90 95
Arg Ala Ile Thr Glu Phe Leu Ser Arg Tyr Pro Asn Val Thr Leu Phe
100 105 110
Ile Tyr Ile Ala Arg Leu Tyr His Leu Ala Asn Pro Arg Asn Arg Gln
115 120 125
Gly Leu Arg Asp Leu Ile Ser Ser Gly Val Thr Ile Gln Ile Met Thr
130 135 140
Glu Gln Glu Ser Gly Tyr Cys Trp His Asn Phe Val Asn Tyr Ser Pro
145 150 155 160
Ser Asn Glu Ser His Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu
165 170 175
Tyr Val Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu
180 185 190
Asn Ile Leu Arg Arg Lys Gln Ser Gln Leu Thr Ser Phe Thr Ile Ala
195 200 205
Leu Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp Ala
210 215 220
Thr Gly Leu Lys
225
<210> 29
<211> 162
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 29
Ser Phe Glu Arg Asn Tyr Asp Pro Arg Glu Leu Arg Lys Glu Thr Tyr
1 5 10 15
Leu Leu Tyr Glu Ile Lys Trp Gly Lys Ser Gly Lys Leu Trp Arg His
20 25 30
Trp Cys Gln Asn Asn Arg Thr Gln His Ala Glu Val Tyr Phe Leu Glu
35 40 45
Asn Ile Phe Asn Ala Arg Arg Phe Asn Pro Ser Thr His Cys Ser Ile
50 55 60
Thr Trp Tyr Leu Ser Trp Ser Pro Cys Ala Glu Cys Ser Gln Lys Ile
65 70 75 80
Val Asp Phe Leu Lys Glu His Pro Asn Val Asn Leu Glu Ile Tyr Val
85 90 95
Ala Arg Leu Tyr Tyr Pro Glu Asn Glu Arg Asn Arg Gln Gly Leu Arg
100 105 110
Asp Leu Val Asn Ser Gly Val Thr Ile Arg Ile Met Asp Leu Pro Asp
115 120 125
Tyr Asn Tyr Cys Trp Lys Thr Phe Val Ser Asp Gln Gly Gly Asp Glu
130 135 140
Asp Tyr Trp Pro Gly His Phe Ala Pro Trp Ile Lys Gln Tyr Ser Leu
145 150 155 160
Lys Leu
<210> 30
<211> 166
<212> PRT
<213> 大肠杆菌(Escherichia coli)
<400> 30
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
1 5 10 15
Leu Ala Lys Arg Ala Trp Asp Glu Arg Glu Val Pro Val Gly Ala Val
20 25 30
Leu Val His Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Pro Ile
35 40 45
Gly Arg His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
50 55 60
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
65 70 75 80
Val Thr Leu Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
85 90 95
Arg Ile Gly Arg Val Val Phe Gly Ala Arg Asp Ala Lys Thr Gly Ala
100 105 110
Ala Gly Ser Leu Met Asp Val Leu His His Pro Gly Met Asn His Arg
115 120 125
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
130 135 140
Ser Asp Phe Phe Arg Met Arg Arg Gln Glu Ile Lys Ala Gln Lys Lys
145 150 155 160
Ala Gln Ser Ser Thr Asp
165
<210> 31
<211> 166
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 31
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
1 5 10 15
Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val
20 25 30
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile
35 40 45
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
50 55 60
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
65 70 75 80
Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
85 90 95
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ala Lys Thr Gly Ala
100 105 110
Ala Gly Ser Leu Met Asp Val Leu His Tyr Pro Gly Met Asn His Arg
115 120 125
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
130 135 140
Cys Tyr Phe Phe Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys
145 150 155 160
Ala Gln Ser Ser Thr Asp
165
<210> 32
<211> 166
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 32
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
1 5 10 15
Leu Ala Lys Arg Ala Trp Asp Glu Arg Glu Val Pro Val Gly Ala Val
20 25 30
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ser Ile
35 40 45
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
50 55 60
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
65 70 75 80
Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
85 90 95
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ala Lys Thr Gly Ala
100 105 110
Ala Gly Ser Leu Met Asp Val Leu His Tyr Pro Gly Met Asn His Arg
115 120 125
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
130 135 140
Cys Tyr Phe Phe Arg Met Arg Arg Gln Val Phe Asn Ala Gln Lys Lys
145 150 155 160
Ala Gln Ser Ser Thr Asp
165
<210> 33
<211> 166
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 33
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
1 5 10 15
Leu Ala Lys Arg Ala Leu Asp Glu Arg Glu Val Pro Val Gly Ala Val
20 25 30
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile
35 40 45
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
50 55 60
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
65 70 75 80
Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
85 90 95
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ala Lys Thr Gly Ala
100 105 110
Ala Gly Ser Leu Met Asp Val Leu His Tyr Pro Gly Met Asn His Arg
115 120 125
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Asn Ala Leu Leu
130 135 140
Cys Tyr Phe Phe Arg Met Arg Arg Gln Val Phe Asn Ala Gln Lys Lys
145 150 155 160
Ala Gln Ser Ser Thr Asp
165
<210> 34
<211> 166
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 34
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
1 5 10 15
Leu Ala Lys Arg Ala Leu Asp Glu Arg Glu Val Pro Val Gly Ala Val
20 25 30
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile
35 40 45
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
50 55 60
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
65 70 75 80
Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
85 90 95
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ala Lys Thr Gly Ala
100 105 110
Ala Gly Ser Leu Met Asp Val Leu His Tyr Pro Gly Met Asn His Arg
115 120 125
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Asn Ala Leu Leu
130 135 140
Cys Tyr Phe Phe Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys
145 150 155 160
Ala Gln Ser Ser Thr Asp
165
<210> 35
<211> 1763
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 35
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
1 5 10 15
Leu Ala Lys Arg Ala Trp Asp Glu Arg Glu Val Pro Val Gly Ala Val
20 25 30
Leu Val His Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Pro Ile
35 40 45
Gly Arg His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
50 55 60
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
65 70 75 80
Val Thr Leu Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
85 90 95
Arg Ile Gly Arg Val Val Phe Gly Ala Arg Asp Ala Lys Thr Gly Ala
100 105 110
Ala Gly Ser Leu Met Asp Val Leu His His Pro Gly Met Asn His Arg
115 120 125
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
130 135 140
Ser Asp Phe Phe Arg Met Arg Arg Gln Glu Ile Lys Ala Gln Lys Lys
145 150 155 160
Ala Gln Ser Ser Thr Asp Ser Gly Gly Ser Ser Gly Gly Ser Ser Gly
165 170 175
Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Gly
180 185 190
Gly Ser Ser Gly Gly Ser Ser Glu Val Glu Phe Ser His Glu Tyr Trp
195 200 205
Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu
210 215 220
Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu
225 230 235 240
Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr Ala His Ala Glu
245 250 255
Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu
260 265 270
Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys Val Met Cys Ala
275 280 285
Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg
290 295 300
Asn Ala Lys Thr Gly Ala Ala Gly Ser Leu Met Asp Val Leu His Tyr
305 310 315 320
Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp
325 330 335
Glu Cys Ala Ala Leu Leu Cys Tyr Phe Phe Arg Met Pro Arg Gln Val
340 345 350
Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Thr Asp Ser Gly Gly Ser
355 360 365
Ser Gly Gly Ser Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala
370 375 380
Thr Pro Glu Ser Ser Gly Gly Ser Ser Gly Gly Ser Asp Lys Lys Tyr
385 390 395 400
Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile
405 410 415
Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn
420 425 430
Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe
435 440 445
Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg
450 455 460
Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile
465 470 475 480
Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu
485 490 495
Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro
500 505 510
Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro
515 520 525
Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala
530 535 540
Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg
545 550 555 560
Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val
565 570 575
Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu
580 585 590
Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser
595 600 605
Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu
610 615 620
Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser
625 630 635 640
Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp
645 650 655
Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn
660 665 670
Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala
675 680 685
Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn
690 695 700
Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr
705 710 715 720
Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln
725 730 735
Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn
740 745 750
Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr
755 760 765
Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu
770 775 780
Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe
785 790 795 800
Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala
805 810 815
Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg
820 825 830
Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly
835 840 845
Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser
850 855 860
Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly
865 870 875 880
Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn
885 890 895
Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr
900 905 910
Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly
915 920 925
Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val
930 935 940
Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys
945 950 955 960
Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser
965 970 975
Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu
980 985 990
Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu
995 1000 1005
Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp
1010 1015 1020
Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe
1025 1030 1035
Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly
1040 1045 1050
Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys
1055 1060 1065
Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe
1070 1075 1080
Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr
1085 1090 1095
Phe Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp
1100 1105 1110
Ser Leu His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile
1115 1120 1125
Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val
1130 1135 1140
Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile Glu Met
1145 1150 1155
Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg
1160 1165 1170
Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser
1175 1180 1185
Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn
1190 1195 1200
Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr
1205 1210 1215
Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val
1220 1225 1230
Asp His Ile Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp
1235 1240 1245
Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp
1250 1255 1260
Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp
1265 1270 1275
Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp
1280 1285 1290
Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys
1295 1300 1305
Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr
1310 1315 1320
Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr
1325 1330 1335
Asp Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu
1340 1345 1350
Lys Ser Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr
1355 1360 1365
Lys Val Arg Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr
1370 1375 1380
Leu Asn Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys
1385 1390 1395
Leu Glu Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val
1400 1405 1410
Arg Lys Met Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr
1415 1420 1425
Ala Lys Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr
1430 1435 1440
Glu Ile Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile
1445 1450 1455
Glu Thr Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg
1460 1465 1470
Asp Phe Ala Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn
1475 1480 1485
Ile Val Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu
1490 1495 1500
Ser Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys
1505 1510 1515
Lys Asp Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr
1520 1525 1530
Val Ala Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys
1535 1540 1545
Ser Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile
1550 1555 1560
Met Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu
1565 1570 1575
Ala Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu
1580 1585 1590
Pro Lys Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met
1595 1600 1605
Leu Ala Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu
1610 1615 1620
Pro Ser Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu
1625 1630 1635
Lys Leu Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe
1640 1645 1650
Val Glu Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile
1655 1660 1665
Ser Glu Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp
1670 1675 1680
Lys Val Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg
1685 1690 1695
Glu Gln Ala Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu
1700 1705 1710
Gly Ala Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg
1715 1720 1725
Lys Arg Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile
1730 1735 1740
His Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser
1745 1750 1755
Gln Leu Gly Gly Asp
1760
<210> 36
<211> 1565
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 36
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
1 5 10 15
Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val
20 25 30
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile
35 40 45
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
50 55 60
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
65 70 75 80
Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
85 90 95
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala
100 105 110
Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg
115 120 125
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
130 135 140
Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys
145 150 155 160
Ala Gln Ser Ser Ile Asn Ser Gly Gly Ser Ser Gly Gly Ser Ser Gly
165 170 175
Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Gly
180 185 190
Gly Ser Ser Gly Gly Ser Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile
195 200 205
Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val
210 215 220
Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile
225 230 235 240
Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala
245 250 255
Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg
260 265 270
Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala
275 280 285
Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val
290 295 300
Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Ile Val
305 310 315 320
Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg
325 330 335
Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr
340 345 350
Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu
355 360 365
Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln
370 375 380
Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala
385 390 395 400
Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser
405 410 415
Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn
420 425 430
Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn
435 440 445
Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser
450 455 460
Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly
465 470 475 480
Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala
485 490 495
Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala
500 505 510
Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp
515 520 525
Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr
530 535 540
Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile
545 550 555 560
Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile
565 570 575
Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg
580 585 590
Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro
595 600 605
His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu
610 615 620
Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile
625 630 635 640
Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn
645 650 655
Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro
660 665 670
Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe
675 680 685
Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val
690 695 700
Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu
705 710 715 720
Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe
725 730 735
Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr
740 745 750
Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys
755 760 765
Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe
770 775 780
Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp
785 790 795 800
Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile
805 810 815
Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg
820 825 830
Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln Leu
835 840 845
Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile
850 855 860
Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu
865 870 875 880
Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp
885 890 895
Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly
900 905 910
Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly Ser Pro
915 920 925
Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu
930 935 940
Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile Glu Met
945 950 955 960
Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu
965 970 975
Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile
980 985 990
Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu
995 1000 1005
Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp Gln
1010 1015 1020
Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile
1025 1030 1035
Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val
1040 1045 1050
Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro
1055 1060 1065
Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu
1070 1075 1080
Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr
1085 1090 1095
Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe
1100 1105 1110
Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val
1115 1120 1125
Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn
1130 1135 1140
Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys
1145 1150 1155
Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg
1160 1165 1170
Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala
1175 1180 1185
Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser
1190 1195 1200
Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met
1205 1210 1215
Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr
1220 1225 1230
Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr
1235 1240 1245
Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn
1250 1255 1260
Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala
1265 1270 1275
Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys
1280 1285 1290
Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu
1295 1300 1305
Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp
1310 1315 1320
Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr
1325 1330 1335
Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys
1340 1345 1350
Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg
1355 1360 1365
Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly
1370 1375 1380
Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr
1385 1390 1395
Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser
1400 1405 1410
Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys
1415 1420 1425
Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys
1430 1435 1440
Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln
1445 1450 1455
His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe
1460 1465 1470
Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu
1475 1480 1485
Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala
1490 1495 1500
Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro
1505 1510 1515
Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr
1520 1525 1530
Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser
1535 1540 1545
Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly
1550 1555 1560
Gly Asp
1565
<210> 37
<211> 1565
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 37
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
1 5 10 15
Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val
20 25 30
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile
35 40 45
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
50 55 60
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Tyr Asp Ala Thr Leu Tyr
65 70 75 80
Ser Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
85 90 95
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ala Lys Thr Gly Ala
100 105 110
Ala Gly Ser Leu Met Asp Val Leu His His Pro Gly Met Asn His Arg
115 120 125
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
130 135 140
Cys Arg Phe Phe Arg Met Pro Arg Arg Val Phe Asn Ala Gln Lys Lys
145 150 155 160
Ala Gln Ser Ser Thr Asp Ser Gly Gly Ser Ser Gly Gly Ser Ser Gly
165 170 175
Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Gly
180 185 190
Gly Ser Ser Gly Gly Ser Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile
195 200 205
Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val
210 215 220
Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile
225 230 235 240
Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala
245 250 255
Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg
260 265 270
Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala
275 280 285
Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val
290 295 300
Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Ile Val
305 310 315 320
Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg
325 330 335
Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr
340 345 350
Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu
355 360 365
Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln
370 375 380
Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala
385 390 395 400
Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser
405 410 415
Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn
420 425 430
Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn
435 440 445
Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser
450 455 460
Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly
465 470 475 480
Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala
485 490 495
Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala
500 505 510
Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp
515 520 525
Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr
530 535 540
Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile
545 550 555 560
Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile
565 570 575
Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg
580 585 590
Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro
595 600 605
His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu
610 615 620
Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile
625 630 635 640
Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn
645 650 655
Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro
660 665 670
Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe
675 680 685
Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val
690 695 700
Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu
705 710 715 720
Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe
725 730 735
Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr
740 745 750
Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys
755 760 765
Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe
770 775 780
Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp
785 790 795 800
Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile
805 810 815
Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg
820 825 830
Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln Leu
835 840 845
Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile
850 855 860
Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu
865 870 875 880
Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp
885 890 895
Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly
900 905 910
Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly Ser Pro
915 920 925
Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu
930 935 940
Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile Glu Met
945 950 955 960
Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu
965 970 975
Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile
980 985 990
Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu
995 1000 1005
Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp Gln
1010 1015 1020
Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile
1025 1030 1035
Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val
1040 1045 1050
Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro
1055 1060 1065
Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu
1070 1075 1080
Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr
1085 1090 1095
Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe
1100 1105 1110
Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val
1115 1120 1125
Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn
1130 1135 1140
Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys
1145 1150 1155
Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg
1160 1165 1170
Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala
1175 1180 1185
Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser
1190 1195 1200
Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met
1205 1210 1215
Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr
1220 1225 1230
Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr
1235 1240 1245
Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn
1250 1255 1260
Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala
1265 1270 1275
Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys
1280 1285 1290
Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu
1295 1300 1305
Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp
1310 1315 1320
Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr
1325 1330 1335
Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys
1340 1345 1350
Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg
1355 1360 1365
Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly
1370 1375 1380
Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr
1385 1390 1395
Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser
1400 1405 1410
Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys
1415 1420 1425
Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys
1430 1435 1440
Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln
1445 1450 1455
His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe
1460 1465 1470
Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu
1475 1480 1485
Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala
1490 1495 1500
Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro
1505 1510 1515
Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr
1520 1525 1530
Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser
1535 1540 1545
Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly
1550 1555 1560
Gly Asp
1565
<210> 38
<211> 364
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 38
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
1 5 10 15
Leu Ala Lys Arg Ala Trp Asp Glu Arg Glu Val Pro Val Gly Ala Val
20 25 30
Leu Val His Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Pro Ile
35 40 45
Gly Arg His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
50 55 60
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
65 70 75 80
Val Thr Leu Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
85 90 95
Arg Ile Gly Arg Val Val Phe Gly Ala Arg Asp Ala Lys Thr Gly Ala
100 105 110
Ala Gly Ser Leu Met Asp Val Leu His His Pro Gly Met Asn His Arg
115 120 125
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
130 135 140
Ser Asp Phe Phe Arg Met Arg Arg Gln Glu Ile Lys Ala Gln Lys Lys
145 150 155 160
Ala Gln Ser Ser Thr Asp Ser Gly Gly Ser Ser Gly Gly Ser Ser Gly
165 170 175
Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Gly
180 185 190
Gly Ser Ser Gly Gly Ser Ser Glu Val Glu Phe Ser His Glu Tyr Trp
195 200 205
Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu
210 215 220
Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu
225 230 235 240
Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr Ala His Ala Glu
245 250 255
Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu
260 265 270
Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys Val Met Cys Ala
275 280 285
Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg
290 295 300
Asn Ala Lys Thr Gly Ala Ala Gly Ser Leu Met Asp Val Leu His Tyr
305 310 315 320
Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp
325 330 335
Glu Cys Ala Ala Leu Leu Cys Tyr Phe Phe Arg Met Pro Arg Gln Val
340 345 350
Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Thr Asp
355 360
<210> 39
<211> 167
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 39
Met Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu
1 5 10 15
Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala
20 25 30
Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala
35 40 45
Ile Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg
50 55 60
Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Tyr Asp Ala Thr Leu
65 70 75 80
Tyr Ser Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His
85 90 95
Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ala Lys Thr Gly
100 105 110
Ala Ala Gly Ser Leu Met Asp Val Leu His His Pro Gly Met Asn His
115 120 125
Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu
130 135 140
Leu Cys Arg Phe Phe Arg Met Pro Arg Arg Val Phe Asn Ala Gln Lys
145 150 155 160
Lys Ala Gln Ser Ser Thr Asp
165
<210> 40
<211> 167
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 40
Met Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu
1 5 10 15
Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala
20 25 30
Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala
35 40 45
Ile Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg
50 55 60
Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu
65 70 75 80
Tyr Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His
85 90 95
Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly
100 105 110
Ala Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His
115 120 125
Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu
130 135 140
Leu Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys
145 150 155 160
Lys Ala Gln Ser Ser Ile Asn
165
<210> 41
<211> 83
<212> PRT
<213> 芽孢杆菌噬菌体AR9
<400> 41
Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val
1 5 10 15
Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile
20 25 30
Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu
35 40 45
Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr
50 55 60
Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile
65 70 75 80
Lys Met Leu
<210> 42
<211> 19
<212> RNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<220>
<221> misc_feature
<222> (1)..(19)
<223> n是a, c, g或u
<400> 42
nnnnnnnnnn nnnnnnnnn 19
<210> 43
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<220>
<221> misc_feature
<222> (1)..(3)
<223> n是a, c, g或t
<220>
<221> misc_feature
<222> (4)..(22)
<223> n是a, c, g或t
<400> 43
nnnnnnnnnn nnnnnnnnna aa 22
<210> 44
<211> 22
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<220>
<221> misc_feature
<222> (4)..(22)
<223> n是a, c, g或t
<400> 44
tttnnnnnnn nnnnnnnnnn nn 22
<210> 45
<211> 24
<212> PRT
<213> 人工序列
<220>
<223> 合成肽
<400> 45
Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala Arg
1 5 10 15
Leu Lys Lys Gly Ser Gly Ser Gly
20
<210> 46
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 46
Glu Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala
1 5 10 15
Arg Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu Ser Lys Asn
20 25 30
Tyr His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser
35 40 45
Gly Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala
50 55 60
Arg Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu Ser Lys Asn
65 70 75 80
Tyr His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser
85 90 95
Gly Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala
100 105 110
Arg Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu Ser Lys Asn
115 120 125
Tyr His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser
130 135 140
Gly Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala
145 150 155 160
Arg Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu Ser Lys Asn
165 170 175
Tyr His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser
180 185 190
Gly Glu Glu Leu Leu Ser Lys Asn Tyr His Leu Glu Asn Glu Val Ala
195 200 205
Arg Leu Lys Lys Gly Ser Gly Ser Gly Glu Glu Leu Leu Ser Lys Asn
210 215 220
Tyr His Leu Glu Asn Glu Val Ala Arg Leu Lys Lys Gly Ser Gly Ser
225 230 235 240
Gly
<210> 47
<211> 277
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 47
Met Gly Pro Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala
1 5 10 15
Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ser Ser Thr Gly Ala
20 25 30
Val Thr Thr Ser Asn Tyr Ala Ser Trp Val Gln Glu Lys Pro Gly Lys
35 40 45
Leu Phe Lys Gly Leu Ile Gly Gly Thr Asn Asn Arg Ala Pro Gly Val
50 55 60
Pro Ser Arg Phe Ser Gly Ser Leu Ile Gly Asp Lys Ala Thr Leu Thr
65 70 75 80
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Phe Cys Ala Leu
85 90 95
Trp Tyr Ser Asn His Trp Val Phe Gly Gln Gly Thr Lys Val Glu Leu
100 105 110
Lys Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
115 120 125
Ser Ser Gly Gly Gly Ser Glu Val Lys Leu Leu Glu Ser Gly Gly Gly
130 135 140
Leu Val Gln Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Val Ser Gly
145 150 155 160
Phe Ser Leu Thr Asp Tyr Gly Val Asn Trp Val Arg Gln Ala Pro Gly
165 170 175
Arg Gly Leu Glu Trp Ile Gly Val Ile Trp Gly Asp Gly Ile Thr Asp
180 185 190
Tyr Asn Ser Ala Leu Lys Asp Arg Phe Ile Ile Ser Lys Asp Asn Gly
195 200 205
Lys Asn Thr Val Tyr Leu Gln Met Ser Lys Val Arg Ser Asp Asp Thr
210 215 220
Ala Leu Tyr Tyr Cys Val Thr Gly Leu Phe Asp Tyr Trp Gly Gln Gly
225 230 235 240
Thr Leu Val Thr Val Ser Ser Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
245 250 255
Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
260 265 270
Gly Gly Gly Gly Ser
275
<210> 48
<211> 66
<212> DNA
<213> 贝酵母(Saccharomyces bayanus)
<400> 48
ttcttgtcgt acttatagat cgctacgtta tttcaatttt gaaaatctga gtcctgggag 60
tgcgga 66
<210> 49
<211> 605
<212> PRT
<213> 智人
<400> 49
Met Ser Gly Trp Glu Ser Tyr Tyr Lys Thr Glu Gly Asp Glu Glu Ala
1 5 10 15
Glu Glu Glu Gln Glu Glu Asn Leu Glu Ala Ser Gly Asp Tyr Lys Tyr
20 25 30
Ser Gly Arg Asp Ser Leu Ile Phe Leu Val Asp Ala Ser Lys Ala Met
35 40 45
Phe Glu Ser Gln Ser Glu Asp Glu Leu Thr Pro Phe Asp Met Ser Ile
50 55 60
Gln Cys Ile Gln Ser Val Tyr Ile Ser Lys Ile Ile Ser Ser Asp Arg
65 70 75 80
Asp Leu Leu Ala Trp Phe Tyr Gly Thr Glu Lys Asp Lys Asn Ser Val
85 90 95
Asn Phe Lys Ile Tyr Val Leu Gln Glu Leu Asp Asn Pro Gly Ala Lys
100 105 110
Arg Ile Leu Glu Leu Asp Gln Phe Lys Gly Gln Gln Gly Gln Lys Arg
115 120 125
Phe Gln Asp Met Met Gly His Gly Ser Asp Tyr Ser Leu Ser Glu Val
130 135 140
Leu Trp Val Cys Ala Asn Leu Phe Ser Asp Val Gln Phe Lys Met Ser
145 150 155 160
His Lys Arg Ile Met Leu Phe Thr Asn Glu Asp Asn Pro His Gly Asn
165 170 175
Asp Ser Ala Lys Ala Ser Arg Ala Arg Thr Lys Ala Gly Asp Leu Arg
180 185 190
Asp Thr Gly Ile Phe Leu Asp Leu His Leu Lys Lys Pro Gly Gly Phe
195 200 205
Asp Ile Ser Leu Phe Tyr Arg Asp Ile Ile Ser Ile Ala Glu Asp Glu
210 215 220
Asp Leu Arg Val His Phe Glu Glu Ser Ser Lys Leu Glu Asp Leu Leu
225 230 235 240
Arg Lys Val Arg Ala Lys Glu Thr Arg Lys Arg Ala Leu Ser Arg Leu
245 250 255
Lys Leu Lys Leu Asn Lys Asp Ile Val Ile Ser Val Gly Ile Tyr Asn
260 265 270
Leu Val Gln Lys Ala Leu Lys Pro Pro Pro Ile Lys Leu Tyr Arg Glu
275 280 285
Thr Asn Glu Pro Val Lys Thr Lys Thr Arg Thr Phe Asn Thr Ser Thr
290 295 300
Gly Gly Leu Leu Leu Pro Ser Asp Thr Lys Arg Ser Gln Ile Tyr Gly
305 310 315 320
Ser Arg Gln Ile Ile Leu Glu Lys Glu Glu Thr Glu Glu Leu Lys Arg
325 330 335
Phe Asp Asp Pro Gly Leu Met Leu Met Gly Phe Lys Pro Leu Val Leu
340 345 350
Leu Lys Lys His His Tyr Leu Arg Pro Ser Leu Phe Val Tyr Pro Glu
355 360 365
Glu Ser Leu Val Ile Gly Ser Ser Thr Leu Phe Ser Ala Leu Leu Ile
370 375 380
Lys Cys Leu Glu Lys Glu Val Ala Ala Leu Cys Arg Tyr Thr Pro Arg
385 390 395 400
Arg Asn Ile Pro Pro Tyr Phe Val Ala Leu Val Pro Gln Glu Glu Glu
405 410 415
Leu Asp Asp Gln Lys Ile Gln Val Thr Pro Pro Gly Phe Gln Leu Val
420 425 430
Phe Leu Pro Phe Ala Asp Asp Lys Arg Lys Met Pro Phe Thr Glu Lys
435 440 445
Ile Met Ala Thr Pro Glu Gln Val Gly Lys Met Lys Ala Ile Val Glu
450 455 460
Lys Leu Arg Phe Thr Tyr Arg Ser Asp Ser Phe Glu Asn Pro Val Leu
465 470 475 480
Gln Gln His Phe Arg Asn Leu Glu Ala Leu Ala Leu Asp Leu Met Glu
485 490 495
Pro Glu Gln Ala Val Asp Leu Thr Leu Pro Lys Val Glu Ala Met Asn
500 505 510
Lys Arg Leu Gly Ser Leu Val Asp Glu Phe Lys Glu Leu Val Tyr Pro
515 520 525
Pro Asp Tyr Asn Pro Glu Gly Lys Val Thr Lys Arg Lys His Asp Asn
530 535 540
Glu Gly Ser Gly Ser Lys Arg Pro Lys Val Glu Tyr Ser Glu Glu Glu
545 550 555 560
Leu Lys Thr His Ile Ser Lys Gly Thr Leu Gly Lys Phe Thr Val Pro
565 570 575
Leu Lys Glu Ala Cys Arg Ala Tyr Gly Leu Lys Ser Gly Leu Lys Lys
580 585 590
Gln Glu Leu Leu Glu Ala Leu Thr Lys His Phe Gln Asp
595 600 605
<210> 50
<211> 482
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 50
Met Val Arg Ser Gly Asn Lys Ala Ala Trp Leu Cys Met Asp Val Gly
1 5 10 15
Phe Thr Met Ser Asn Ser Ile Pro Gly Ile Glu Ser Pro Phe Glu Gln
20 25 30
Ala Lys Lys Val Ile Thr Met Phe Val Gln Arg Gln Val Phe Ala Glu
35 40 45
Asn Lys Asp Glu Ile Ala Leu Val Leu Phe Gly Thr Asp Gly Thr Asp
50 55 60
Asn Pro Leu Ser Gly Gly Asp Gln Tyr Gln Asn Ile Thr Val His Arg
65 70 75 80
His Leu Met Leu Pro Asp Phe Asp Leu Leu Glu Asp Ile Glu Ser Lys
85 90 95
Ile Gln Pro Gly Ser Gln Gln Ala Asp Phe Leu Asp Ala Leu Ile Val
100 105 110
Ser Met Asp Val Ile Gln His Glu Thr Ile Gly Lys Lys Phe Glu Lys
115 120 125
Arg His Ile Glu Ile Phe Thr Asp Leu Ser Ser Arg Phe Ser Lys Ser
130 135 140
Gln Leu Asp Ile Ile Ile His Ser Leu Lys Lys Cys Asp Ile Ser Glu
145 150 155 160
Arg His Ser Ile His Trp Pro Cys Arg Leu Thr Ile Gly Ser Asn Leu
165 170 175
Ser Ile Arg Ile Ala Ala Tyr Lys Ser Ile Leu Gln Glu Arg Val Lys
180 185 190
Lys Thr Thr Trp Asp Ala Lys Thr Leu Lys Lys Glu Asp Ile Gln Lys
195 200 205
Glu Thr Val Tyr Cys Leu Asn Asp Asp Asp Glu Thr Glu Val Leu Lys
210 215 220
Glu Asp Ile Ile Gln Gly Phe Arg Tyr Gly Ser Asp Ile Val Pro Phe
225 230 235 240
Ser Lys Val Asp Glu Glu Gln Met Lys Tyr Lys Ser Glu Gly Lys Cys
245 250 255
Phe Ser Val Leu Gly Phe Cys Lys Ser Ser Gln Val Gln Arg Arg Phe
260 265 270
Phe Met Gly Asn Gln Val Leu Lys Val Phe Ala Ala Arg Asp Asp Glu
275 280 285
Ala Ala Ala Val Ala Leu Ser Ser Leu Ile His Ala Leu Asp Asp Leu
290 295 300
Asp Ile Trp Ala Ile Val Arg Tyr Ala Tyr Asp Lys Arg Ala Asn Pro
305 310 315 320
Gln Val Gly Val Ala Phe Pro His Ile Lys His Asn Tyr Glu Cys Leu
325 330 335
Val Tyr Val Gln Leu Pro Phe Met Glu Asp Leu Arg Gln Tyr Met Phe
340 345 350
Ser Ser Leu Lys Asn Ser Lys Lys Tyr Ala Pro Thr Glu Ala Gln Leu
355 360 365
Asn Ala Val Asp Ala Leu Ile Asp Ser Met Ser Leu Ala Lys Lys Asp
370 375 380
Glu Lys Thr Asp Thr Leu Glu Asp Leu Phe Pro Thr Thr Lys Ile Pro
385 390 395 400
Asn Pro Arg Phe Gln Arg Leu Phe Gln Cys Leu Leu His Arg Ala Leu
405 410 415
His Pro Arg Glu Pro Leu Pro Pro Ile Gln Gln His Ile Trp Asn Met
420 425 430
Leu Asn Pro Pro Ala Glu Val Thr Thr Lys Ser Gln Ile Pro Leu Ser
435 440 445
Lys Ile Lys Thr Leu Phe Pro Leu Ile Glu Ala Lys Lys Lys Asp Gln
450 455 460
Val Thr Ala Gln Glu Ile Phe Gln Asp Asn His Glu Asp Gly Pro Thr
465 470 475 480
Ala Lys
<210> 51
<211> 10
<212> DNA
<213> Methanobacterium thermoautotrophicum
<400> 51
aatttttgga 10
<210> 52
<211> 83
<212> PRT
<213> Methanobacterium thermoautotrophicum
<400> 52
Gly Ser Val Ile Asp Val Ser Ser Gln Arg Val Asn Val Gln Arg Pro
1 5 10 15
Leu Asp Ala Leu Gly Asn Ser Leu Asn Ser Pro Val Ile Ile Lys Leu
20 25 30
Lys Gly Asp Arg Glu Phe Arg Gly Val Leu Lys Ser Phe Asp Leu His
35 40 45
Met Asn Leu Val Leu Asn Asp Ala Glu Glu Leu Glu Asp Gly Glu Val
50 55 60
Thr Arg Arg Leu Gly Thr Val Leu Ile Arg Gly Asp Asn Ile Val Tyr
65 70 75 80
Ile Ser Pro
<210> 53
<211> 25
<212> DNA
<213> 噬菌体MS2
<400> 53
gcgcacatga ggatcaccca tgtgc 25
<210> 54
<211> 116
<212> PRT
<213> 噬菌体MS2
<400> 54
Met Ala Ser Asn Phe Thr Gln Phe Val Leu Val Asp Asn Gly Gly Thr
1 5 10 15
Gly Asp Val Thr Val Ala Pro Ser Asn Phe Ala Asn Gly Ile Ala Glu
20 25 30
Ile Ser Ser Asn Ser Arg Ser Gln Ala Tyr Lys Val Thr Cys Ser Val
35 40 45
Arg Gln Ser Ser Ala Gln Asn Arg Lys Tyr Thr Ile Lys Val Glu Val
50 55 60
Pro Lys Gly Ala Trp Arg Ser Tyr Leu Asn Met Glu Leu Thr Ile Pro
65 70 75 80
Ile Phe Ala Thr Asn Ser Asp Cys Glu Leu Ile Val Lys Ala Met Gln
85 90 95
Gly Leu Leu Lys Asp Gly Asn Pro Ile Pro Ser Ala Ile Ala Ala Asn
100 105 110
Ser Gly Ile Tyr
115
<210> 55
<211> 26
<212> DNA
<213> 噬菌体PP7
<400> 55
ataaggagtt tatatggaaa ccctta 26
<210> 56
<211> 127
<212> PRT
<213> 噬菌体PP7
<400> 56
Met Ser Lys Thr Ile Val Leu Ser Val Gly Glu Ala Thr Arg Thr Leu
1 5 10 15
Thr Glu Ile Gln Ser Thr Ala Asp Arg Gln Ile Phe Glu Glu Lys Val
20 25 30
Gly Pro Leu Val Gly Arg Leu Arg Leu Thr Ala Ser Leu Arg Gln Asn
35 40 45
Gly Ala Lys Thr Ala Tyr Arg Val Asn Leu Lys Leu Asp Gln Ala Asp
50 55 60
Trp Asp Cys Ser Thr Ser Val Cys Gly Glu Leu Pro Lys Val Arg Tyr
65 70 75 80
Thr Gln Val Trp Ser His Asp Val Thr Ile Val Ala Asn Ser Thr Glu
85 90 95
Ala Ser Arg Lys Ser Leu Tyr Asp Leu Thr Lys Ser Leu Val Ala Thr
100 105 110
Ser Gln Val Glu Asp Leu Val Val Asn Leu Val Pro Leu Gly Arg
115 120 125
<210> 57
<211> 19
<212> DNA
<213> 志贺氏杆菌噬菌体
<400> 57
ctgaatgcct gcgagcatc 19
<210> 58
<211> 62
<212> PRT
<213> 志贺氏杆菌噬菌体
<400> 58
Met Lys Ser Ile Arg Cys Lys Asn Cys Asn Lys Leu Leu Phe Lys Ala
1 5 10 15
Asp Ser Phe Asp His Ile Glu Ile Arg Cys Pro Arg Cys Lys Arg His
20 25 30
Ile Ile Met Leu Asn Ala Cys Glu His Pro Thr Glu Lys His Cys Gly
35 40 45
Lys Arg Glu Lys Ile Thr His Ser Asp Glu Thr Val Arg Tyr
50 55 60
<210> 59
<211> 1367
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 59
Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly
1 5 10 15
Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys
20 25 30
Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly
35 40 45
Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys
50 55 60
Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr
65 70 75 80
Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe
85 90 95
Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His
100 105 110
Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His
115 120 125
Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser
130 135 140
Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met
145 150 155 160
Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp
165 170 175
Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn
180 185 190
Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys
195 200 205
Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu
210 215 220
Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu
225 230 235 240
Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp
245 250 255
Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp
260 265 270
Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu
275 280 285
Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile
290 295 300
Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met
305 310 315 320
Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala
325 330 335
Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp
340 345 350
Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln
355 360 365
Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly
370 375 380
Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys
385 390 395 400
Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly
405 410 415
Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu
420 425 430
Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro
435 440 445
Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met
450 455 460
Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val
465 470 475 480
Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn
485 490 495
Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu
500 505 510
Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr
515 520 525
Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys
530 535 540
Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val
545 550 555 560
Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser
565 570 575
Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr
580 585 590
Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn
595 600 605
Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu
610 615 620
Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His
625 630 635 640
Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr
645 650 655
Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys
660 665 670
Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala
675 680 685
Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys
690 695 700
Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His
705 710 715 720
Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile
725 730 735
Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg
740 745 750
His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr
755 760 765
Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu
770 775 780
Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val
785 790 795 800
Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln
805 810 815
Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu
820 825 830
Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys Asp
835 840 845
Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly
850 855 860
Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn
865 870 875 880
Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe
885 890 895
Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys
900 905 910
Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys
915 920 925
His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu
930 935 940
Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys
945 950 955 960
Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu
965 970 975
Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val Val
980 985 990
Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val
995 1000 1005
Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys
1010 1015 1020
Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr
1025 1030 1035
Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn
1040 1045 1050
Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr
1055 1060 1065
Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg
1070 1075 1080
Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu
1085 1090 1095
Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg
1100 1105 1110
Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys
1115 1120 1125
Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu
1130 1135 1140
Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser
1145 1150 1155
Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser Phe
1160 1165 1170
Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu
1175 1180 1185
Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe
1190 1195 1200
Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly Glu
1205 1210 1215
Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val Asn
1220 1225 1230
Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro
1235 1240 1245
Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His
1250 1255 1260
Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg
1265 1270 1275
Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr
1280 1285 1290
Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile
1295 1300 1305
Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe
1310 1315 1320
Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr
1325 1330 1335
Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly
1340 1345 1350
Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp
1355 1360 1365
<210> 60
<211> 1367
<212> PRT
<213> 人工序列
<220>
<223> 合成多肽
<400> 60
Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly
1 5 10 15
Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys
20 25 30
Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly
35 40 45
Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys
50 55 60
Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr
65 70 75 80
Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe
85 90 95
Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His
100 105 110
Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His
115 120 125
Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser
130 135 140
Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met
145 150 155 160
Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp
165 170 175
Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn
180 185 190
Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys
195 200 205
Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu
210 215 220
Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu
225 230 235 240
Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp
245 250 255
Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp
260 265 270
Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu
275 280 285
Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile
290 295 300
Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met
305 310 315 320
Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala
325 330 335
Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp
340 345 350
Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln
355 360 365
Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly
370 375 380
Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys
385 390 395 400
Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly
405 410 415
Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu
420 425 430
Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro
435 440 445
Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met
450 455 460
Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val
465 470 475 480
Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn
485 490 495
Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu
500 505 510
Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr
515 520 525
Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys
530 535 540
Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val
545 550 555 560
Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser
565 570 575
Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr
580 585 590
Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn
595 600 605
Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu
610 615 620
Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His
625 630 635 640
Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr
645 650 655
Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys
660 665 670
Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala
675 680 685
Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys
690 695 700
Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His
705 710 715 720
Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile
725 730 735
Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg
740 745 750
His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr
755 760 765
Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu
770 775 780
Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val
785 790 795 800
Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln
805 810 815
Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu
820 825 830
Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Ala Asp
835 840 845
Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly
850 855 860
Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn
865 870 875 880
Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe
885 890 895
Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys
900 905 910
Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys
915 920 925
His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu
930 935 940
Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys
945 950 955 960
Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu
965 970 975
Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val Val
980 985 990
Gly Thr Ala Leu Ile Lys Lys Tyr Pro Ala Leu Glu Ser Glu Phe Val
995 1000 1005
Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys
1010 1015 1020
Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr
1025 1030 1035
Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn
1040 1045 1050
Gly Glu Ile Arg Lys Ala Pro Leu Ile Glu Thr Asn Gly Glu Thr
1055 1060 1065
Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg
1070 1075 1080
Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu
1085 1090 1095
Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg
1100 1105 1110
Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys
1115 1120 1125
Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu
1130 1135 1140
Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser
1145 1150 1155
Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser Phe
1160 1165 1170
Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu
1175 1180 1185
Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe
1190 1195 1200
Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly Glu
1205 1210 1215
Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val Asn
1220 1225 1230
Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro
1235 1240 1245
Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His
1250 1255 1260
Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg
1265 1270 1275
Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr
1280 1285 1290
Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile
1295 1300 1305
Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe
1310 1315 1320
Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr
1325 1330 1335
Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly
1340 1345 1350
Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp
1355 1360 1365
<210> 61
<211> 4101
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 61
gacaagaagt acagcatcgg gctggcgatc gggaccaact ccgtcggctg ggctgtgatt 60
accgacgagt acaaggtgcc atccaagaag ttcaaggtcc tcggcaacac tgaccggcac 120
agcattaaga agaacctgat tggggcgctg ctgttcgatt cgggggagac tgcggaggcg 180
accaggctga agcggactgc gcgccggagg tacaccagga ggaagaatcg gatctgctac 240
ctccaggaga ttttctcgaa tgagatggcc aaggtggacg attccttctt ccatcgcctg 300
gaggagtcgt tcctcgttga ggaggacaag aagcatgaga ggcatcccat tttcgggaat 360
atcgttgacg aggtggctta ccatgagaag tacccgacca tctaccatct gcggaagaag 420
ctcgtcgatt cgaccgataa ggccgacctg cggctgatct acctggccct cgcgcacatg 480
attaagttcc ggggccattt cctcatcgag ggcgacctca acccggacaa ctcggacgtg 540
gataagctct tcattcagct cgtgcagaca tacaaccagc tcttcgagga gaatcccatt 600
aacgcctcgg gggtcgacgc taaggctatt ctctcggctc ggctgtcgaa gtcgcgccgg 660
ctggagaatc tcattgccca gctcccaggc gagaagaaga acggcctctt cggcaacctg 720
attgccctgt cgctggggct cacaccgaat ttcaagtcga acttcgacct cgccgaggac 780
gctaagctcc agctcagcaa ggatacttac gatgatgacc tcgataacct gctcgcccag 840
attggggatc agtacgcgga tctgttcctc gcggccaaga atctcagcga tgctattctc 900
ctgtcggaca ttctccgcgt caacacagag attactaagg ccccactgtc ggcgagcatg 960
attaagaggt acgatgagca tcatcaggac ctgacactgc tcaaggcgct ggtccggcag 1020
cagctccccg agaagtacaa ggagattttc ttcgatcagt caaagaatgg gtacgcgggc 1080
tacattgatg gcggcgcgtc ccaggaggag ttctacaagt tcattaagcc catcctggag 1140
aagatggacg ggaccgagga gctgctggtg aagctcaatc gggaggacct gctccggaag 1200
cagcgcacat tcgacaatgg ctcgattcct caccagattc acctgggcga gctgcacgcc 1260
attctccgca ggcaggagga cttctacccg ttcctcaagg acaaccgcga gaagatcgag 1320
aagatcctga ccttccggat tccatactac gtggggccgc tcgcgcgggg gaactcccgg 1380
ttcgcgtgga tgactcgcaa gtccgaagaa acgattacac cgtggaattt cgaggaggtc 1440
gtcgacaagg gcgctagtgc gcagtcattc attgagagga tgaccaattt cgataagaac 1500
ctgcctaacg agaaggtgct gccgaagcat tcgctgctct acgagtactt caccgtttac 1560
aatgagctga ccaaggtgaa gtatgtgact gagggcatga ggaagccagc gttcctgagc 1620
ggcgagcaga agaaggctat cgtggacctg ctcttcaaga ctaaccggaa ggtgactgtg 1680
aagcagctca aggaggacta cttcaagaag attgagtgct tcgattccgt tgagattagc 1740
ggggtggagg atcggttcaa tgcttcgctc gggacatacc acgatctcct gaagatcatt 1800
aaggataagg acttcctcga caacgaggag aacgaggaca ttctcgaaga tattgtcctg 1860
accctcaccc tcttcgagga tcgggagatg atcgaggaga ggctcaagac atacgctcat 1920
ctgttcgatg ataaggtcat gaagcagctg aagcgcaggc ggtacacagg gtgggggcgg 1980
ctgagccgga agctgatcaa cgggattcgg gataagcagt ccgggaagac aattctcgac 2040
ttcctcaagt ccgacgggtt cgctaaccgg aacttcatgc agctcattca tgatgactcg 2100
ctgacattca aggaggatat tcagaaggcg caggtttcgg ggcagggcga ctcgctccac 2160
gagcatattg cgaatctggc gggctccccc gcgattaaga agggcattct gcaaaccgtc 2220
aaggtggttg atgagctggt caaggtcatg gggcggcata agccagagaa tattgtcatc 2280
gagatggcgc gggagaatca gaccacacag aaggggcaga agaactcacg ggagcggatg 2340
aagcgcatcg aggagggcat caaggagctg gggtcgcaga tcctgaagga gcatcccgtg 2400
gagaacactc agctgcaaaa tgagaagctg tacctctact acctccagaa cgggagggac 2460
atgtatgtgg atcaggagct ggatattaat aggctgagcg attacgatgt cgaccacatt 2520
gtcccacagt cgttcctgaa ggacgacagc attgacaaca aggtgctgac ccgctcggat 2580
aagaacaggg gcaagagcga taatgttcca agcgaggagg ttgtgaagaa gatgaagaac 2640
tactggcggc agctcctgaa cgcgaagctc atcacacagc ggaagttcga caacctcacc 2700
aaggctgagc gcgggggcct gagcgagctg gacaaggcgg ggttcattaa gaggcagctg 2760
gtcgagacac ggcagattac aaagcatgtt gcgcagattc tcgattcccg gatgaacacc 2820
aagtacgatg agaacgataa gctgattcgg gaggtcaagg taattaccct gaagtccaag 2880
ctggtgtccg acttcaggaa ggacttccag ttctacaagg ttcgggagat caacaactac 2940
caccacgcgc atgatgccta cctcaacgcg gtcgtgggga ccgctctcat caagaagtac 3000
ccaaagctgg agtcagagtt cgtctacggg gattacaagg tttacgacgt gcggaagatg 3060
atcgctaaga gcgagcagga gattggcaag gctaccgcta agtacttctt ctactccaac 3120
atcatgaact tcttcaagac agagattacc ctcgcgaatg gcgagatccg gaagaggccc 3180
ctcatcgaga caaatgggga gacaggggag attgtctggg ataaggggcg ggatttcgcg 3240
accgtccgga aggtcctgtc gatgccccag gttaatattg tcaagaagac tgaggtccag 3300
actggcggct tctcaaagga gtcgattctc ccaaagagga actccgataa gctcattgct 3360
cggaagaagg attgggaccc caagaagtac gggggattcg actcccccac tgttgcttac 3420
tctgttctgg ttgttgctaa ggtggagaag gggaagtcga agaagctgaa gagcgtgaag 3480
gagctgctcg ggattacaat tatggagagg tcatccttcg agaagaatcc catcgacttc 3540
ctggaggcca agggctacaa ggaggtgaag aaggacctga ttattaagct gcccaagtac 3600
tcgctcttcg agctggagaa tgggcggaag cggatgctgg cgtccgcggg ggagctgcaa 3660
aaggggaacg agctggcgct cccctccaag tatgtgaact tcctctacct ggcgtcgcac 3720
tacgagaagc tgaaggggtc cccagaggat aatgagcaga agcagctctt cgtcgagcag 3780
cataagcact acctggacga gattatcgag cagattagcg agttctcgaa gcgggtcatc 3840
ctcgcggatg cgaacctgga taaggtgctc agcgcctaca ataagcaccg ggacaagccg 3900
attcgggagc aggcggagaa tattattcac ctcttcacac tcaccaacct cggggcacca 3960
gctgcgttca agtacttcga cactactatc gaccggaagc ggtacacctc gacgaaggag 4020
gtgctcgacg ccaccctcat tcaccagtcg atcacaggcc tgtacgagac acggattgac 4080
ctgtcccagc tcgggggcga c 4101
<210> 62
<211> 4101
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 62
gacaagaagt actccattgg cctggcgatt gggacaaact cggtggggtg ggccgtgatt 60
acggatgagt acaaggttcc aagcaagaag ttcaaggtcc tcgggaacac agatcggcat 120
tcgattaaga agaatctcat tggggcgctc ctcttcgact cgggggagac agcggaggct 180
accaggctca agcggacagc caggcggcgg tacacaaggc ggaagaatcg catctgctac 240
ctccaggaga ttttctcgaa tgagatggcg aaggtggacg acagcttctt ccatcggctg 300
gaggagtcct tcctggtgga ggaggataag aagcacgaga ggcatccaat tttcgggaac 360
atcgtggacg aggttgcgta ccatgagaag taccctacaa tctaccatct gcggaagaag 420
ctggttgact ccacagacaa ggcggacctg aggctgatct acctcgctct ggcccacatg 480
attaagttcc gcgggcattt cctgatcgag ggggacctga atcccgacaa ttcggatgtg 540
gacaagctct tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaatcccatc 600
aatgcgtcgg gcgttgacgc taaggccatt ctgtccgcta ggctgtcgaa gagcaggagg 660
ctggagaacc tgatcgccca gctgccaggc gagaagaaga atgggctctt cgggaatctg 720
attgcgctct ccctggggct gacaccgaac ttcaagagca atttcgatct ggctgaggac 780
gcgaagctcc agctctcgaa ggacacttac gacgatgacc tcgataacct cctcgcgcag 840
atcggggacc agtacgctga tctcttcctc gccgctaaga acctctcgga tgctatcctg 900
ctctccgaca ttctccgggt taataccgag attacaaagg ccccactgtc ggcgtccatg 960
atcaagcggt acgatgagca tcatcaggat ctcaccctgc tcaaggccct cgtgcggcag 1020
cagctgcccg agaagtacaa ggagattttc ttcgaccaga gcaagaatgg gtacgctggc 1080
tacattgacg gcggggcctc acaggaggag ttctacaagt tcatcaagcc aatcctggag 1140
aagatggatg ggacagagga gctgctggtg aagctcaacc gggaggatct gctcaggaag 1200
cagcggacgt tcgacaacgg gtcgattccc catcagatcc acctggggga gctgcacgcg 1260
atcctgcgcc ggcaggagga tttctaccct ttcctgaagg ataatcggga gaagatcgag 1320
aagattctca ccttccggat tccctactac gtcgggccac tcgcgcgggg caatagcagg 1380
ttcgcctgga tgacacggaa gagcgaggag acaatcaccc cctggaactt cgaggaggtt 1440
gtcgacaagg gggcgtccgc ccagtcattc attgagcgga tgaccaattt cgacaagaat 1500
ctgccaaatg agaaggttct cccaaagcat agcctcctct acgagtactt cactgtttac 1560
aacgagctga ccaaggtgaa gtatgtgacc gagggcatgc ggaagcccgc gttcctgtcc 1620
ggcgagcaga agaaggccat tgtggacctc ctgttcaaga ccaatcgcaa ggtcacagtc 1680
aagcagctca aggaggatta cttcaagaag atcgagtgct tcgactcggt tgagattagc 1740
ggggtggagg atcggttcaa cgcgagcctc ggcacttacc acgacctcct gaagatcatc 1800
aaggataagg acttcctcga caacgaggag aacgaggata ttctggagga catcgtgctc 1860
accctgacgc tgttcgagga tcgggagatg atcgaggagc gcctgaagac ctacgctcat 1920
ctcttcgatg ataaggtcat gaagcagctg aagaggaggc ggtacaccgg gtggggccgc 1980
ctgagcagga agctcattaa cgggatcagg gacaagcaga gcggcaagac catcctggac 2040
ttcctcaaga gcgatggctt cgccaaccgg aatttcatgc agctcatcca cgacgactcc 2100
ctcaccttca aggaggacat tcagaaggct caggtcagcg gccagggcga ctcgctgcat 2160
gagcacatcg ctaacctggc gggcagccca gccatcaaga agggcatcct ccagacagtg 2220
aaggtcgtgg atgagctggt gaaggtcatg ggccggcata agcccgagaa tattgtgatt 2280
gagatggcgc gggagaatca gaccactcag aagggccaga agaactcgcg ggagcgcatg 2340
aagaggatcg aggaggggat taaggagctg ggcagccaga ttctcaagga gcaccccgtg 2400
gagaataccc agctccagaa cgagaagctg tacctctact acctccagaa tgggcgggac 2460
atgtatgttg atcaggagct ggacatcaat cgcctctcgg attacgacgt ggaccacatc 2520
gtgccccaga gcttcctgaa ggatgatagc atcgacaata aggtcctgac ccgctccgac 2580
aagaatcgcg gcaagagcga caacgtgccg agcgaggagg tcgtgaagaa gatgaagaac 2640
tactggcggc agctgctgaa cgcgaagctc attacacagc ggaagttcga taacctgacg 2700
aaggcggaga ggggcggcct ctccgagctg gacaaggcgg gcttcattaa gaggcagctc 2760
gtggagactc gccagatcac caagcacgtg gctcagatcc tcgatagccg gatgaatacg 2820
aagtacgatg agaatgacaa gctcatccgg gaggtgaagg taatcaccct gaagtcaaag 2880
ctcgttagcg atttccggaa ggacttccag ttctacaagg tgcgggagat taacaactac 2940
catcatgcgc acgatgcgta cctcaatgcg gtggtgggca cagccctgat taagaagtac 3000
cccaagctgg agagcgagtt cgtctacggg gactacaagg tgtacgatgt tcggaagatg 3060
atcgccaaga gcgagcagga gattgggaag gccaccgcta agtacttctt ctactcgaat 3120
attatgaatt tcttcaagac cgagatcaca ctcgctaatg gggagattcg gaagcggccc 3180
ctcatcgaga ctaacgggga gactggcgag attgtgtggg acaaggggcg cgacttcgct 3240
accgtgcgca aggtcctctc gatgccccag gttaatattg ttaagaagac agaggtgcag 3300
acgggcgggt tctccaagga gtctatcctg ccgaagcgga actcggacaa gctgatcgcc 3360
cgcaagaagg attgggaccc caagaagtac gggggattcg atagcccaac cgtggcttac 3420
agcgtcctgg tggtcgccaa ggttgagaag gggaagtcga agaagctcaa gagcgttaag 3480
gagctgctgg gcatcaccat catggagcgg tccagcttcg agaagaatcc tatcgacttc 3540
ctggaggcta aggggtacaa ggaggtcaag aaggacctga tcattaagct gcccaagtac 3600
tctctgttcg agctggagaa cgggaggaag cggatgctgg cgtctgctgg cgagctacag 3660
aagggcaatg agctggcgct cccctcgaag tatgtcaact tcctctacct ggcttcccat 3720
tacgagaagc tgaagggctc gcccgaggat aatgagcaga agcagctctt cgtggagcag 3780
cacaagcact acctcgacga gatcattgag cagatttcgg agttctcgaa gcgggtcatt 3840
ctcgcggacg cgaacctcga caaggtcctc tcggcgtaca acaagcaccg ggacaagccc 3900
atccgggagc aggccgagaa cattatccac ctcttcacac tgaccaacct cggcgctccc 3960
gccgcgttca agtacttcga caccaccatt gaccgcaaga gatacacatc caccaaggag 4020
gtgctggacg cgaccctcat ccaccagagc atcacaggcc tctacgagac acggatcgac 4080
ctctcgcagc tcgggggcga t 4101
<210> 63
<211> 4092
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 63
gacaagaagt actcgatcgg cctggcgatt ggcacaaaca gcgtggggtg ggctgtgatc 60
actgatgagt acaaggtgcc atcgaagaag ttcaaggtgc tggggaatac agaccggcat 120
tcgatcaaga agaatctcat tggcgctctc ctcttcgatt ccggcgagac tgctgaggcg 180
acccgcctga agcgcaccgc ccggcggcgc tacactcggc ggaagaatag gatttgctac 240
ctccaggaga ttttctcgaa tgagatggcc aaggtggatg acagcttctt ccaccgcctg 300
gaggagtcgt tcctggtcga ggaggacaag aagcatgagc ggcaccctat cttcgggaat 360
atcgttgatg aggtcgccta ccacgagaag taccccacta tctaccatct ccgcaagaag 420
ctcgtggaca gcacagataa ggccgacctc cgcctgatct acctcgccct cgcgcacatg 480
attaagttcc gggggcactt cctcattgag ggggatctga atcccgataa ctccgacgtg 540
gacaagctgt tcatccagct ggtgcagaca tacaaccagc tgttcgagga gaatcccatc 600
aacgcgagcg gcgtggacgc taaggccatt ctgtcggcta ggctctcgaa gtcgaggcgg 660
ctggagaacc tgattgcgca gctccccggc gagaagaaga acgggctgtt cgggaatctc 720
atcgccctct ccctcggcct cacaccaaac ttcaagagca atttcgacct ggctgaggac 780
gctaagctgc aactctcaaa ggatacatac gatgacgacc tggacaatct cctggctcag 840
atcggcgacc agtacgctga cctgttcctc gcggccaaga atctgtcgga cgcgattctc 900
ctcagcgaca tcctgcgcgt caataccgag attacgaagg ctccactgtc tgcgtcaatg 960
attaagcggt acgatgagca tcaccaggat ctgaccctcc tgaaggcgct cgtgcggcag 1020
cagctgcccg agaagtacaa ggagattttc ttcgatcaga gcaagaatgg ctacgccggc 1080
tacatcgacg ggggcgcgag ccaggaggag ttctacaagt tcatcaagcc catcctggag 1140
aagatggacg gcaccgagga gctactcgtg aagctcaatc gggaggatct cctccggaag 1200
cagcggacat tcgataacgg gtctatccca caccagatcc acctcggcga gctgcatgcg 1260
attctgcggc ggcaggagga tttctaccct ttcctgaagg acaaccggga gaagatcgag 1320
aagatcctca cattccggat tccatactac gtcggccccc tggcgagggg caatagccgg 1380
ttcgcgtgga tgacaaggaa gtccgaggag actattaccc cgtggaattt cgaggaggtg 1440
gttgacaagg gcgcttccgc gcagagcttc attgagcgga tgacaaactt cgacaagaat 1500
ctccccaacg agaaggtcct gccgaagcat agcctcctgt acgagtactt caccgtctac 1560
aatgagctaa ctaaggtcaa gtatgtgaca gagggcatga ggaagccagc cttcctctca 1620
ggcgagcaga agaaggccat tgtggacctc ctgttcaaga caaaccgcaa ggtgacagtg 1680
aagcagctga aggaggatta cttcaagaag attgagtgct tcgactcagt ggagatttca 1740
ggcgtggagg atcggttcaa cgcgagcctg gggacttacc acgacctgct gaagattatt 1800
aaggacaagg acttcctgga taacgaggag aatgaggaca tcctggagga tattgtgctc 1860
accctcaccc tgttcgagga cagggagatg attgaggaga ggctcaagac ctacgcgcac 1920
ctgttcgatg acaaggtcat gaagcagctg aagaggcggc gctacactgg gtggggccgc 1980
ctgtcgcgga agctgatcaa cggcattcgg gataagcagt ccgggaagac cattctggat 2040
ttcctgaagt cggacggctt cgccaacagg aatttcatgc agctgatcca cgacgactcc 2100
ctcaccttca aggaggacat tcagaaggcc caggttagcg gccaggggga ctcactccac 2160
gagcatattg ccaatctggc cggctctcca gctatcaaga agggcatcct gcaaacagtt 2220
aaggttgttg acgagctggt taaggtcatg gggcggcata agcccgagaa cattgtcatc 2280
gagatggctc gggagaacca gacaactcag aagggccaga agaactccag ggagcgcatg 2340
aagcggattg aggagggcat taaggagctg gggtcccaga tcctcaagga gcaccctgtc 2400
gagaacactc agctgcaaaa cgagaagctc tacctgtact acctccagaa cgggcgggat 2460
atgtatgtgg atcaggagct ggacatcaac aggctctccg actacgacgt ggatcacatt 2520
gtcccacagt ctttcctcaa ggatgattcc atcgacaaca aggtgctgac gcgcagcgac 2580
aagaataggg ggaagtcgga caacgttccg agcgaggagg tcgtgaagaa gatgaagaat 2640
tactggaggc agctcctgaa tgcgaagctg atcactcaga ggaagttcga caatctgaca 2700
aaggcggaga ggggcgggct ctcggagctg gataaggcgg gcttcatcaa gcggcagctc 2760
gttgaaaccc ggcagatcac caagcatgtc gcccagatcc tcgatagccg catgaacacc 2820
aagtacgatg agaacgacaa gctcattcgg gaggttaagg tcattacgct gaagtccaag 2880
ctcgtcagcg acttcaggaa ggatttccag ttctacaagg ttcgggagat taacaactac 2940
caccacgcgc atgatgcgta cctgaacgct gttgtcggca ctgctctcat caagaagtac 3000
ccaaagctgg agtccgagtt cgtctacggg gactacaagg tctacgatgt ccggaagatg 3060
atcgccaagt cggagcagga gatcgggaag gctactgcga agtacttctt ctacagcaac 3120
attatgaatt tcttcaagac ggagattacg ctggcgaacg gggagattag gaagaggccc 3180
ctcattgaga ctaatgggga gacaggcgag attgtttggg acaagggccg cgacttcgcg 3240
actgtgcgga aggtcctgtc catgccacag gtgaatattg ttaagaagac agaggtgcag 3300
actgggggct tctcgaagga gagcattctc ccaaagcgga acagcgataa gctcatcgcg 3360
cgcaagaagg attgggaccc taagaagtac ggcggcttcg attctcccac tgtggcctac 3420
tccgttctcg tggttgccaa ggttgagaag gggaagtcga agaagctgaa gtcggtcaag 3480
gagctgctcg ggattacaat catggagcgg agcagcttcg agaagaaccc tattgatttc 3540
ctggaggcca agggctacaa ggaggttaag aaggatctca ttatcaagct ccctaagtac 3600
tctctgttcg agctggagaa tggccggaag aggatgctgg cctcggctgg cgagctacag 3660
aaggggaatg agctggccct cccgtcgaag tatgtgaatt tcctgtacct cgcgtcgcac 3720
tacgagaagc tcaagggcag cccggaggat aatgagcaga agcagctctt cgtggagcag 3780
cataagcact acctggacga gatcattgag cagatcagcg agttctcgaa gcgggttatt 3840
ctggctgatg ctaacctgga caaggttctg agcgcctaca ataagcatcg cgacaagccg 3900
attcgcgagc aggcggagaa tattatccac ctgttcaccc tcactaacct cggggctccc 3960
gcggccttca agtacttcga taccacaata gataggaagc ggtacacctc gacgaaggag 4020
gtcctcgacg ccacactcat ccatcagtcg attacaggcc tgtacgagac acggattgac 4080
ctctcgcagc tg 4092
<210> 64
<211> 4101
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 64
gacaagaagt attccatagg cctggctatc ggcaccaaca gcgtgggctg ggccgtcatc 60
accgacgagt acaaagtgcc gagtaaaaag ttcaaagtgc tcggcaacac cgaccgccac 120
tccataaaga aaaacctgat cggggcgctc ctgttcgaca gcggcgagac ggcggaggcc 180
acccgcttga aacgcacggc ccgacggcgc tacacgcggc gcaagaaccg gatctgttac 240
ctacaggaga ttttctctaa cgagatggcg aaggtggacg actcgttctt tcaccgcctc 300
gaagagtcct tcctcgtgga ggaggacaag aaacacgagc gccacccgat cttcggcaac 360
atcgtggacg aggtggccta ccacgagaag tacccgacca tctaccacct ccggaagaaa 420
ctcgtggaca gcacggacaa ggccgacctg aggctcatct acctcgccct ggcgcacatg 480
attaagttcc ggggccactt cctgatcgag ggcgacctga acccggacaa cagcgacgtg 540
gacaagctgt tcatccagct agtccagacc tacaaccagc ttttcgagga aaaccccatc 600
aacgccagcg gggtggacgc gaaggcgatc ctgtccgccc ggctgagcaa gtcccggcgg 660
ctggagaacc tcatcgcgca gttgcccggc gagaagaaga acgggctgtt cgggaacctg 720
atcgccctct ccctggggct caccccgaac ttcaagtcca acttcgacct cgccgaggac 780
gccaaactac agctgagcaa ggacacctac gacgacgacc tcgacaacct gctggcccag 840
atcggggacc agtacgcaga cctgttcctc gccgccaaga acctctccga cgccatcctg 900
ctgtcggaca tcctgcgggt gaacacggag atcacgaagg ccccgctctc ggcctcgatg 960
attaaacgct acgacgagca ccaccaggac ttgaccctcc tcaaggcgct ggtccgccag 1020
cagcttcccg agaagtacaa ggaaatcttt ttcgatcaga gcaagaacgg gtacgccggg 1080
tacatcgacg gcggggcgtc ccaggaggag ttctacaagt tcatcaagcc catcctggag 1140
aaaatggacg ggaccgagga gctgctcgtg aagctcaacc gcgaagattt gctccgcaag 1200
cagcgcacgt tcgacaacgg gtcgatcccg caccagatcc acctgggcga gctgcacgcg 1260
atcctcaggc gtcaggaaga cttctacccc ttcctcaagg acaaccgcga gaagatagag 1320
aagattctga ccttcagaat tccttattac gtgggcccgc tggctcgggg caactcgcgc 1380
ttcgcctgga tgacgcgcaa gtccgaggag accatcaccc cgtggaactt cgaggaggtg 1440
gtggataagg gtgcctcggc ccagtccttc atcgagcgga tgaccaactt cgacaagaac 1500
ctgccgaacg agaaggtgct ccccaagcac agcctgctct acgaatattt cacggtgtac 1560
aacgagctga cgaaggtcaa gtacgtgacc gagggaatga ggaaacctgc attcctctcc 1620
ggggagcaga agaaagccat agtcgacctc ctgttcaaga ccaaccggaa ggtcaccgtc 1680
aagcagctca aggaggacta cttcaagaag atcgagtgct tcgattcagt ggagatcagc 1740
ggcgtcgagg accggttcaa cgccagcctg ggcacctacc acgacctgct caagatcatc 1800
aaggacaagg acttcctcga caacgaggag aacgaggaca tcctggagga catcgtgctg 1860
accctgacgc tcttcgagga ccgcgagatg atcgaggagc gcctcaagac ctacgcccac 1920
ctgttcgacg acaaggtgat gaagcagctc aagcggcgga gatatactgg gtggggccgc 1980
ctctcccgga agctcattaa cggtatcagg gataagcagt ccgggaagac gatcctcgac 2040
ttcctcaagt cggacgggtt cgccaaccgc aacttcatgc agctcatcca cgacgactcc 2100
ctgacgttca aggaggacat ccagaaggcc caagtgtctg gtcaaggtga ctcgctccac 2160
gagcacatcg ccaacctcgc gggcagcccg gccatcaaga agggaatact ccagaccgtc 2220
aaggtggtgg acgagctggt gaaggtcatg ggccgccaca agccggagaa catcgtcatc 2280
gagatggcgc gggagaacca gaccacgcag aaggggcaga aaaatagccg tgagcgcatg 2340
aagcgcatcg aggaggggat taaggagttg ggcagccaga tcctcaagga gcaccctgtg 2400
gagaacacgc agttgcaaaa cgagaagctc tacctgtact acctccagaa cgggagggat 2460
atgtacgtgg accaagaact ggacatcaac cgcctgtccg actacgacgt ggaccacatc 2520
gtgccgcaga gcttcctcaa ggacgacagc atcgacaaca aggtgctcac ccggtccgac 2580
aagaatcggg gcaagtccga caacgtgccc agcgaggagg tcgtcaaaaa gatgaaaaac 2640
tactggcgac aactactgaa cgccaagctc atcacccagc gcaagttcga caacctcaca 2700
aaagccgagc gcggcgggtt gagcgagctg gacaaggccg ggttcatcaa gcgccagctc 2760
gtcgagacgc gccagatcac gaagcacgtc gcgcagatac tcgacagccg gatgaacacc 2820
aagtacgacg agaacgacaa gctcatccgg gaggtgaagg tcatcaccct caagtcgaag 2880
ctcgtgagcg acttccgcaa ggacttccag ttctacaagg tccgggagat caacaactac 2940
caccacgccc acgatgctta tcttaacgcc gtggtgggga cggccctcat taagaaatac 3000
ccgaagctgg agtcggagtt cgtgtacggc gactacaagg tgtacgacgt caggaagatg 3060
atcgccaagt ccgaacagga gatcgggaag gccacggcga aatacttctt ctacagcaac 3120
atcatgaact tcttcaagac cgagatcacc ctcgccaacg gcgagatccg caagcgcccg 3180
ctcatcgaga cgaacgggga gaccggcgag atcgtctggg acaaggggcg cgacttcgcc 3240
actgtgcgga aggtgctgtc gatgccccag gtcaacatcg tcaagaagac ggaggtccag 3300
acgggcgggt tcagcaagga gagcatcctg ccgaagcgca acagcgacaa gctgatcgcc 3360
cgcaaaaagg actgggatcc aaaaaagtac ggcggcttcg acagccccac cgtcgcctac 3420
agcgtcctcg tcgtcgctaa agtcgagaag ggcaagtcca aaaagctcaa gagcgtcaag 3480
gagctgctcg ggatcaccat catggagcgg tccagcttcg agaagaaccc aattgatttc 3540
ctggaggcga agggctacaa ggaggtcaag aaagacctca tcataaagct gccgaagtac 3600
tcactcttcg agctggagaa cgggcgcaag cggatgctgg cgtcggccgg agagctccaa 3660
aagggcaacg agctggcgct gccgagcaag tacgtgaact tcctctacct ggcgtcccac 3720
tacgagaagc tcaagggcag tccagaggat aacgagcaga agcagctatt cgtggagcag 3780
cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gcgcgtcatc 3840
ctggcggacg ccaacctgga caaggtgctg tccgcgtaca acaagcaccg cgacaagccg 3900
atccgcgagc aagccgagaa catcatccac ctgttcaccc tcacgaacct cggggcaccc 3960
gccgccttca aatatttcga cacgaccatc gaccgcaagc gctacaccag cacgaaggag 4020
gtgctcgacg ccaccctgat ccaccagagc atcaccgggc tgtacgagac ccgcatcgac 4080
ctctcgcagc tcggcgggga c 4101
<210> 65
<211> 4101
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 65
gacaagaagt acagtattgg attggccatc gggacgaaca gcgtgggctg ggccgtcatc 60
accgacgagt acaaggtgcc atccaagaag tttaaggttc tggggaatac cgaccgccac 120
tcgatcaaga aaaatctcat cggggcgctg cttttcgaca gcggcgagac ggcggaagcg 180
acgcggctca agcggacggc tcgtcgccgt tacacccggc gtaagaaccg catctgttac 240
ctccaggaga tattcagcaa cgagatggcg aaggtggacg actccttttt ccaccgtctt 300
gaggagtcct tcctggtcga ggaggacaag aagcacgagc gccacccgat cttcgggaac 360
atcgtggacg aggtggccta ccacgagaag taccccacga tctaccacct ccgcaaaaaa 420
ctcgtggact caactgacaa ggccgatttg aggcttatct acctcgccct cgcccacatg 480
attaagttcc gtgggcactt cctaatcgag ggtgacctca accccgacaa ctctgacgtg 540
gacaagctgt tcatccagct tgtgcagacc tacaatcagc tctttgagga gaatccgatc 600
aacgcatctg gtgtggacgc aaaggccatc ctcagcgcgc ggctgagcaa gtctaggcgg 660
ttggagaacc tgatcgccca actgcccggc gagaagaaaa atggcctctt cggcaacctg 720
atcgccctgt cgctggggct cacgccgaac ttcaagagta actttgacct ggcggaggac 780
gctaagctcc agctatctaa ggacacatac gacgacgacc tggacaacct gctggcccag 840
atcggcgacc agtacgccga cctcttccta gccgccaaga acctgtccga cgccatcctc 900
ctcagcgaca tcctgcgcgt gaacacggag atcacgaagg ctccgctcag cgcctccatg 960
attaagcggt acgacgagca ccaccaagac ctaactttac tcaaagccct cgtgcggcag 1020
cagcttcccg agaagtacaa agagatattt tttgatcagt ccaagaacgg ttatgcgggc 1080
tacatcgacg gcggcgcgag ccaggaggag ttctacaagt tcatcaagcc catcctggag 1140
aagatggacg gcacggagga gctgctcgtg aagctcaacc gtgaagacct cctgcgaaag 1200
cagcgaacct tcgacaacgg ttcgatcccg caccagatcc acctcgggga gctgcacgcc 1260
atcctgaggc gacaggagga cttctaccct ttcctaaagg acaaccgcga gaagattgaa 1320
aaaatcctga cgtttcgcat accctactac gtcggcccgc tggcgcgcgg caactcccgg 1380
ttcgcctgga tgacccgtaa gagcgaggag acgatcaccc cgtggaactt cgaggaggtc 1440
gtggacaagg gcgcgagcgc gcagagcttc atcgagcgca tgaccaactt cgacaagaac 1500
ctcccgaacg agaaggtgct cccaaagcac tccctcctgt acgagtattt caccgtgtac 1560
aacgagttga caaaggtgaa gtacgtgacg gagggaatgc ggaagcctgc gttcctctcg 1620
ggcgagcaga agaaggcaat cgtggacctg ctcttcaaga ccaaccggaa ggtgacggtg 1680
aagcagctca aggaggacta cttcaaaaaa atcgagtgct tcgactccgt ggagataagc 1740
ggcgtggagg accgattcaa cgcctccctc ggcacctacc acgacctcct taagatcatc 1800
aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctc 1860
accctgaccc tcttcgagga ccgggagatg atcgaggagc gcctcaagac gtacgcccac 1920
ttgttcgacg acaaggtgat gaagcagctc aagcggcggc gatacaccgg gtggggccgc 1980
ctatcccgca aacttatcaa cggcatccgc gacaagcagt ccggcaagac gatcctggat 2040
ttcctcaagt cggacgggtt cgccaaccgg aacttcatgc agctcatcca cgacgacagc 2100
ctcacgttca aggaggacat ccagaaggcc caagtgagcg gtcaagggga cagcctccac 2160
gagcacattg cgaaccttgc tgggagccct gcgatcaaga aggggatatt gcaaaccgtg 2220
aaggtcgtgg acgagttggt gaaggtcatg gggcgacaca agcccgagaa catcgtgatc 2280
gagatggcca gggaaaatca gaccacgcag aagggccaaa aaaacagccg cgagcggatg 2340
aagcggatcg aggagggcat caaggagctg gggtcgcaga tcctcaagga gcacccggtg 2400
gagaacacgc agctccagaa cgagaagctg tacctctatt acctacagaa cgggcgggat 2460
atgtacgtgg accaggagct agacatcaac cgcctgtccg actacgacgt ggaccatatc 2520
gtcccgcagt cgttcttgaa ggacgacagc atcgacaaca aggtgctcac aagatcggat 2580
aagaatcgag gcaagtccga caacgtgccc tcggaggagg tggtcaagaa aatgaaaaac 2640
tactggcggc agttgctgaa cgccaagctc attacgcagc ggaagttcga caacctgacg 2700
aaggctgaac gtggtgggct cagcgagcta gacaaggcgg ggttcatcaa gcggcagctc 2760
gtcgagaccc ggcagatcac caagcacgtg gcgcagatcc tggactcgcg catgaacacc 2820
aagtacgacg agaacgacaa gctcatccgt gaggtgaagg tcatcaccct taagtctaag 2880
ctggtcagtg acttccgcaa ggacttccag ttctacaagg tccgggagat caacaactac 2940
caccacgcgc acgacgccta cctcaacgcg gtggtgggga cggcgcttat taagaaatat 3000
cccaagctgg aaagcgagtt cgtttacggc gactacaagg tgtacgacgt ccgcaagatg 3060
atcgcaaagt cggaacagga aatcggaaag gcgacggcca aatatttctt ttactccaac 3120
atcatgaatt tttttaagac ggagatcacc ctggcgaacg gggagatccg caagcggccc 3180
ctcatcgaga ccaacgggga gacgggcgag atcgtctggg acaagggccg ggacttcgcc 3240
accgtgcgga aggtgctttc tatgcctcaa gtcaatatcg tcaaaaagac agaggtgcag 3300
accggcgggt tcagcaagga gtctatcctg ccgaagcgca actcggacaa gctcatcgcg 3360
cgcaagaaag actgggaccc caaaaaatat ggcgggttcg actcgccgac cgtcgcctac 3420
agcgtcctcg tggtggctaa ggtcgagaag ggcaagagca aaaagctaaa gtcggtgaag 3480
gagctgctgg gcatcaccat catggagcgc tcgtctttcg agaagaatcc aatcgacttc 3540
ctagaggcga aggggtacaa ggaggtcaaa aaggatctta tcatcaaact gccgaagtac 3600
agtctgttcg agctggagaa cgggcggaag cggatgctgg ctagtgcggg cgagttgcag 3660
aagggcaacg agttggcact gccctccaag tacgtgaact tcctgtacct ggcctcccac 3720
tacgagaagc tcaaggggag ccccgaggac aacgagcaga agcagctatt cgtcgagcag 3780
cacaagcact acctggacga gatcatcgag cagatcagtg agttctccaa gcgggtcatc 3840
ctcgcggacg ccaacctgga caaggtgctg agcgcgtaca acaagcacag ggacaagcca 3900
atcagggaac aggccgagaa catcatccac ctgttcaccc tgaccaacct gggtgcaccg 3960
gctgccttca agtactttga cacgaccatc gaccggaagc gctacacctc cacgaaggag 4020
gtgctggacg ccacgctgat ccaccagagc atcaccgggc tctacgagac acggatcgac 4080
ctgagccagc ttggcgggga c 4101
<210> 66
<211> 4092
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 66
gacaaaaagt attccattgg actcgctatc ggcacgaaca gcgtcgggtg ggcggtcatc 60
actgacgagt acaaggtgcc gagcaagaag tttaaggtgc tgggaaacac cgacaggcac 120
tcgatcaaga aaaatcttat cggggcccta ctcttcgact ccggagaaac cgccgaggcc 180
acccggttga agcgcacggc ccgccgtcgc tacaccaggc gcaagaaccg gatctgctac 240
ctccaggaga tattcagcaa tgagatggcg aaggtggacg actcgttttt tcacaggcta 300
gaggagtctt tcctcgtgga ggaggacaag aaacacgagc gccaccccat cttcggcaac 360
atcgtggatg aggtggcata tcacgagaag tacccaacca tctaccacct ccgcaaaaag 420
ctcgtggact ctaccgacaa ggccgacctc cgtctgatct acctcgcgct ggcccacatg 480
attaagttcc gaggacactt tctgatcgag ggcgacctga acccagacaa cagcgacgtg 540
gacaagctgt tcatccaact tgtccagacc tacaatcagc tcttcgagga gaaccctatc 600
aacgcctcgg gcgtggacgc gaaggccatc ctgtccgccc gcctgagcaa gtcgcggcgg 660
ctggagaacc tgatcgccca gctccccggc gaaaaaaaga acggcctctt cggcaacctc 720
atcgcgttgt cgctggggct caccccgaac ttcaagtcca acttcgacct ggccgaggac 780
gctaaactcc agctctcgaa ggatacctac gacgacgacc tcgacaacct gctggcccag 840
atcggcgacc agtacgcgga ccttttcctg gcggccaaga acctgagcga cgcgatcctc 900
cttagcgaca tactccgtgt gaacaccgag atcacgaagg ccccgctctc cgcgtccatg 960
attaagcgct acgacgagca ccaccaagac cttaccctgc ttaaggcgct ggtcaggcag 1020
cagttaccgg agaagtacaa ggagatcttt tttgatcaat ctaagaacgg ttacgccggg 1080
tacatcgacg gcggcgcgtc ccaggaggag ttctacaagt tcatcaagcc gatcttggag 1140
aaaatggacg ggaccgagga gctgctcgtg aagctcaacc gcgaagacct cctccgcaag 1200
cagcgcacct tcgacaacgg gagcatcccg caccagatcc acctgggaga gctgcacgcg 1260
atcctgcgga gacaagagga cttctacccc ttcctcaagg acaaccggga gaagattgaa 1320
aaaatactta cttttcgtat cccgtactac gtcgggcccc ttgcgagggg caactccaga 1380
ttcgcgtgga tgacccgcaa gtccgaggag accatcaccc cgtggaactt cgaggaggtg 1440
gtggacaagg gcgcgtcggc ccagtcgttc atcgagcgca tgaccaactt cgacaagaac 1500
cttccgaacg agaaggtgct cccgaagcac agcctgctct acgaatattt tactgtgtac 1560
aacgagctga cgaaggtcaa gtacgttacg gaggggatga ggaagcccgc cttcctctcc 1620
ggcgagcaga agaaagccat tgtggatctc ctgttcaaga ccaaccgcaa ggtgacggtg 1680
aaacagctca aagaggacta cttcaagaag atcgagtgct tcgactccgt agagatcagc 1740
ggggtcgagg accgcttcaa cgcctcgctg ggcacgtacc acgacctgct aaagattatc 1800
aaggacaaag acttcctaga caatgaggag aacgaggaca ttctggagga catcgtgctg 1860
actctgacgc tgttcgaaga ccgcgagatg atcgaggagc ggcttaagac gtacgcccac 1920
ctgttcgacg acaaggtgat gaagcagttg aaacggcggc gctacaccgg gtggggccgc 1980
ctctcccgca agctcatcaa cggcatccgc gacaagcagt cggggaagac gatcctggac 2040
ttcctcaaga gcgacggctt cgccaaccga aacttcatgc agctaatcca cgacgacagc 2100
ctgacgttca aggaggacat ccagaaggcc caagtgagcg gccagggaga ctcgctacac 2160
gagcatatcg ccaacctggc tggcagcccg gcgattaaga aaggaatcct ccaaaccgtc 2220
aaagtggtgg acgagctggt gaaggtgatg ggccgccaca agcccgagaa cattgtgatc 2280
gagatggcgc gggagaacca gacgacgcag aagggccaaa aaaatagcag ggaaaggatg 2340
aagcgaatag aggaggggat caaggagctg gggagccaga ttctcaaaga gcacccggtc 2400
gagaacacac agctccagaa cgagaagctg tacctctact acctccaaaa cggccgcgat 2460
atgtacgtgg accaggaact agacatcaac cggctgagcg actatgacgt ggaccacatc 2520
gtgccgcagt ccttcctcaa ggacgactcg attgacaaca aagtgctcac tagatccgac 2580
aagaacagag gcaagagcga taacgtcccg tcggaggagg tcgtcaagaa aatgaaaaac 2640
tactggcggc agctcctaaa cgccaagctc atcacgcagc gtaagttcga caacctgacg 2700
aaggcggagc ggggcgggct gagcgagctg gacaaagcgg ggttcatcaa gcggcagctc 2760
gttgagacgc ggcagatcac aaagcacgtc gcgcaaatcc tcgactcccg catgaacacc 2820
aagtacgacg agaacgacaa gctcatccgg gaggtgaagg tcattaccct taaatcgaag 2880
ctcgtcagcg actttcgtaa ggacttccag ttctacaagg tcagagagat caacaactac 2940
caccacgccc acgacgccta tctgaacgcc gtggtgggca ccgcgcttat taagaagtac 3000
cccaagctgg agtccgagtt cgtgtacggc gactacaagg tttatgacgt caggaagatg 3060
atcgccaagt cggaacagga gatcggaaaa gctaccgcca aatatttctt ctatagcaac 3120
atcatgaact tcttcaaaac cgagatcacc ctcgccaacg gcgagatccg gaagcgcccg 3180
ctcatcgaga ccaacgggga gaccggggag atcgtctggg acaaggggcg ggacttcgct 3240
actgtccgaa aggtgctctc catgccacaa gtgaatatcg tcaagaaaac agaggtgcag 3300
accggagggt tcagtaagga gtccatcctg cccaagcgga actccgacaa gctaattgct 3360
cgcaaaaagg attgggatcc taaaaaatat ggcggcttcg actcgcccac ggtcgcctac 3420
tctgtgctgg tcgtggcgaa ggtggagaag ggcaagtcca agaagctcaa gagcgtcaag 3480
gagctgctgg ggatcacgat catggagcgt agttcgtttg agaagaatcc catcgacttc 3540
ctggaggcta agggctacaa ggaggtcaaa aaggacctca tcattaagct gccgaagtac 3600
agcctcttcg agctggagaa cgggcggaag cgtatgctcg cctccgctgg ggagttacaa 3660
aaggggaacg agctggcgct gccgtctaag tacgtcaact tcctgtacct ggcctcccac 3720
tacgagaagc tcaaggggtc gccggaggac aacgagcaga agcagctctt cgtagagcag 3780
cacaagcact acctggacga gatcatcgag cagatttcag agttctcaaa gcgggtcatc 3840
ctcgccgacg ccaacctgga caaggtgctc tcggcctaca acaagcaccg ggacaagccg 3900
atccgcgaac aggccgaaaa catcatccac ctgttcacgc tcaccaacct cggtgccccg 3960
gcggccttca agtactttga cacgaccatc gaccggaagc gctatacctc gacgaaggag 4020
gtgctggacg ccaccctgat ccaccagtcc atcaccgggc tttacgagac ccggatcgac 4080
ctctcgcagc ta 4092
<210> 67
<211> 4101
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 67
gacaagaagt atagtattgg actcgccatc ggaaccaact ctgtggggtg ggctgttatt 60
acagatgaat ataaggtgcc atccaaaaag tttaaagttc tgggcaatac tgatagacac 120
tcaatcaaga agaatctgat aggtgcactt ctgtttgata gtggagagac tgccgaggca 180
accagactta aaaggactgc aagaagaaga tataccagaa gaaagaatag gatttgctat 240
ttgcaggaaa tcttcagcaa cgaaatggcc aaggttgatg actcattttt ccataggttg 300
gaggagagtt ttcttgtgga ggaagataag aagcacgaaa gacacccaat tttcgggaat 360
atagtggacg aggtggctta tcatgagaag tatcccacta tctaccacct gagaaagaaa 420
cttgtggact caaccgataa ggctgatctt aggcttatat acttggccct tgcacatatg 480
atcaaattca ggggccattt tcttatcgaa ggcgatctta atcccgataa ctcagatgtg 540
gacaagctgt ttatacaact tgtgcaaacc tacaatcaac tcttcgagga gaatcccatt 600
aacgcctccg gcgtggatgc aaaagccata ctgtcagcca gactgagcaa aagtaggaga 660
ctggagaatc ttatagccca actgcccggt gaaaagaaga atgggctctt cggaaatctg 720
atcgctcttt cattggggtt gacacccaac tttaagagta actttgactt ggcagaagat 780
gcaaagttgc agctcagtaa agacacatat gacgatgacc ttgacaatct cttggcacaa 840
ataggggatc aatacgctga ccttttcctc gctgccaaga acctcagcga cgctatactg 900
ttgtccgaca ttcttagggt taataccgaa attacaaagg cccctcttag tgcaagtatg 960
atcaaaaggt atgatgagca tcaccaagac cttacactgc tgaaggctct ggttagacag 1020
caactccctg aaaagtataa ggaaatattc ttcgaccaaa gtaagaacgg gtacgccggt 1080
tatattgatg ggggcgcaag tcaagaagaa ttttacaaat tcatcaagcc aattcttgaa 1140
aagatggacg ggactgagga attgctggtg aaactgaata gagaggacct tcttagaaaa 1200
cagaggacat ttgacaatgg gtccatccca caccagattc atctggggga actccacgca 1260
atattgagga gacaagaaga cttttaccca ttccttaagg ataatagaga gaaaatcgaa 1320
aaaatcctga ctttcaggat tccttactat gttgggccac tggccagggg gaactcaaga 1380
ttcgcttgga tgacaaggaa gtcagaagaa accataaccc cttggaattt tgaagaggtg 1440
gttgataagg gggcatcagc ccagtctttc atagagagga tgaccaactt tgataaaaat 1500
cttccaaatg agaaggtttt gccaaaacat agtcttttgt acgagtactt tactgtttat 1560
aacgaattga ccaaggtgaa gtatgtgacc gagggaatga ggaagccagc atttttgtcc 1620
ggggagcaaa agaaagcaat cgttgatctt ctcttcaaga ccaacagaaa agtgaccgtg 1680
aaacaactga aggaagacta cttcaaaaag atagaatgtt tcgattcagt ggaaattagc 1740
ggtgttgaag acaggttcaa tgcttcattg ggtacttacc acgacctgtt gaagataatc 1800
aaagacaagg actttctcga taatgaggag aacgaagaca tcttggaaga cattgtgctt 1860
acactcactt tgtttgagga cagggaaatg attgaggaaa gactcaaaac ttacgctcat 1920
ttgtttgatg ataaggttat gaaacaacta aaaagaagaa ggtacaccgg ctggggaaga 1980
ttgagtagga aactgatcaa cggtattaga gataaacaat ccggaaagac tatcctcgat 2040
ttccttaaga gtgatggctt tgcaaatagg aattttatgc agctgattca tgacgactca 2100
cttaccttca aagaagacat ccaaaaagct caggtgtctg ggcaaggcga cagtctgcat 2160
gaacatatag ctaacttggc tgggagtccc gccatcaaga aggggatact tcaaacagtt 2220
aaagttgtgg acgaattggt gaaggtaatg ggaaggcaca agcctgaaaa tatagtgata 2280
gaaatggcaa gggaaaatca aacaacccag aagggacaga agaacagtag ggaaaggatg 2340
aaaaggatag aagaggggat caaagagctt ggtagccaga tcctcaagga acatccagtg 2400
gagaataccc aacttcaaaa cgagaaactc tatttgtact acttgcagaa cggaagagat 2460
atgtatgtgg accaagagct tgatattaac aggctgagcg attatgacgt tgaccacata 2520
gtgccccaat cattcctcaa ggatgactct attgataata aggtgctgac aaggagtgac 2580
aagaatagag ggaaatccga caacgttcca tccgaggaag ttgtgaagaa gatgaagaac 2640
tactggaggc agttgctgaa cgctaagctc attacccaga ggaaattcga taacctgacc 2700
aaagcagaga gaggcgggct gagcgaactc gataaagcag gtttcatcaa gagacaactc 2760
gtggagacta ggcaaattac taagcacgtg gctcaaatac tcgacagcag gatgaacaca 2820
aagtacgacg agaacgacaa gctcattaga gaggttaagg ttattactct gaaaagtaaa 2880
ttggttagcg atttcagaaa ggatttccaa ttctataagg ttagagagat caacaattat 2940
catcatgcac atgatgccta tctgaatgct gtggttggta cagcccttat caagaagtac 3000
cctaagctag agagcgagtt tgtgtacgga gattataagg tgtatgatgt gaggaaaatg 3060
atcgctaaaa gtgagcaaga gattggaaag gctaccgcca aatacttctt ttattccaat 3120
attatgaatt tcttcaagac agaaatcacc ctggctaacg gcgagataag gaagaggccg 3180
cttatcgaaa ctaatgggga gacaggcgaa atagtgtggg acaaagggag ggatttcgca 3240
actgtgagga aggttttgag catgcctcag gtgaatatcg ttaagaaaac cgaagttcaa 3300
actggagggt tctctaagga aagcattctc cccaagagga actccgacaa gctgattgct 3360
agaaagaaag actgggaccc caagaagtat ggcggattcg actcacccac tgtggcatat 3420
agcgttctcg tggtggcaaa ggttgaaaag ggtaaatcca aaaaactcaa atccgtgaag 3480
gaactccttg gcataactat tatggaaagg agtagctttg aaaagaatcc catcgacttt 3540
ctcgaagcta agggctataa ggaagttaag aaggacctta taatcaaact tccaaaatac 3600
tccctttttg agttggaaaa cggcagaaag agaatgttgg ccagtgccgg ggagcttcaa 3660
aagggcaacg aactggctct gcctagcaaa tatgtgaact ttttgtatct ggcatcacac 3720
tacgagaaac ttaaaggctc tcctgaggac aacgagcaaa aacagctctt tgttgaacag 3780
cataagcact acctcgacga gattattgag cagatcagcg agttctcaaa gagagttatt 3840
ctggctgacg ctaatcttga caaggttttg tccgcttaca acaaacacag ggataagcca 3900
atcagggagc aggcagaaaa cataatccat ctctttaccc tgacaaacct cggtgccccc 3960
gctgctttca agtattttga tactaccatt gacaggaaga gatatacttc cactaaggaa 4020
gtgctcgacg caaccctcat acaccaaagt atcacaggcc tctatgaaac taggatagat 4080
ttgtctcaac ttgggggcga t 4101
<210> 68
<211> 4101
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 68
gacaaaaagt attccatcgg gcttgctatc ggaaccaact ctgtggggtg ggcagttatt 60
accgacgaat acaaggtgcc cagcaagaag tttaaggttc tggggaacac agatagacat 120
agcataaaga aaaacctgat aggcgcactg ttgttcgact ccggggaaac agccgaagct 180
accaggctga agagaactgc aagaagaagg tacaccagaa gaaaaaacag aatatgttat 240
ctccaagaga ttttctctaa cgagatggcc aaggtggacg actcattctt tcacagactg 300
gaagaatctt tccttgtgga agaagataag aaacacgaga ggcaccctat ttttggcaat 360
atcgtggatg aggtggctta ccacgaaaaa taccctacaa tataccacct caggaaaaaa 420
ttggttgata gtacagacaa ggccgacctc aggctcatct atttggccct ggcccatatg 480
attaaattca gggggcactt tctcatcgag ggagatttga accccgacaa cagtgatgtt 540
gataagctct ttattcagct cgtgcagact tacaatcagt tgtttgagga aaaccccatt 600
aatgcttccg gggtggacgc caaggcaatc ctttctgcaa gactctcaaa gtcaaggaga 660
ctcgaaaatc tgatagcaca gcttccagga gagaagaaga acgggctctt tggaaacctg 720
atcgctctgt cactcggact cacacccaat ttcaaaagca attttgattt ggcagaggac 780
gctaagctgc aactcagtaa ggatacctac gacgatgact tggataatct gctcgcacaa 840
attggggacc agtatgcaga cctgtttctc gcagctaaga acttgagtga cgccatattg 900
ctcagtgaca tcctcagggt taataccgag attacaaaag ctccactctc tgcaagcatg 960
atcaagaggt atgacgagca ccatcaagac ctgacactcc ttaaggcgtt ggttaggcag 1020
caacttcctg aaaagtataa ggaaatcttc ttcgatcaaa gcaaaaacgg ctacgccggc 1080
tatatagacg ggggagcatc ccaagaagaa ttttataagt tcataaaacc tatattggag 1140
aagatggacg ggacagagga attgctcgtg aaactgaaca gggaggatct cctcaggaag 1200
caaaggacct tcgacaatgg ctccatccca catcagattc acctcggcga actgcacgca 1260
atactgagaa gacaagagga cttttatcct ttcctgaagg acaacaggga gaaaatcgag 1320
aaaatcttga cattcagaat cccatactac gttgggcctc tggccagagg taacagtagg 1380
ttcgcctgga tgactaggaa atcagaggag actattacac cctggaactt tgaagaagtt 1440
gttgataagg gagcttcagc acaatcattc atcgaaagaa tgacaaactt tgacaaaaat 1500
ctgcctaatg agaaagtgct cccaaaacat tccctgctgt atgagtattt taccgtttat 1560
aacgagctta ccaaggtgaa atacgttact gaaggtatga gaaagccagc ttttctttca 1620
ggggagcaaa agaaggctat cgtggatctt ctctttaaga ccaacagaaa ggttaccgtg 1680
aagcagctta aggaagacta ctttaaaaag atcgagtgtt ttgactcagt ggaaataagc 1740
ggtgttgaag atagattcaa cgcatccttg ggaacttatc atgatcttct taagataatc 1800
aaggataaag actttctcga caacgaggaa aacgaagata tactggagga catagttctg 1860
acacttactt tgttcgagga tagggagatg atcgaggaaa gactgaaaac atatgctcac 1920
cttttcgacg acaaagttat gaaacaactc aagagaagga gatatacagg gtgggggaga 1980
ttgagcagga aactgattaa tggtatcaga gacaaacagt caggaaaaac aatactcgac 2040
tttttgaaat cagacgggtt cgcaaatagg aatttcatgc agcttataca cgacgattca 2100
cttactttta aagaggacat tcaaaaggct caagttagtg gacaaggtga ctccctccac 2160
gaacacatcg caaatctcgc tggcagccct gcaattaaga agggtatact ccagacagtt 2220
aaggttgttg acgagctggt taaagtgatg ggaagacaca aacccgagaa catagtgata 2280
gagatggcca gggaaaacca aaccactcaa aaagggcaga aaaattccag agagaggatg 2340
aaaaggattg aagaaggtat caaggagctg ggtagccaaa ttctgaaaga acatcctgtg 2400
gaaaacactc aactccagaa tgagaaactc tatctgtact atctgcaaaa tgggagagat 2460
atgtatgtgg accaggaact ggacataaac aggctctcag attacgatgt ggatcatatc 2520
gtgccacagt cctttcttaa ggatgatagc atcgacaata aggtgcttac caggtccgac 2580
aagaacaggg gaaagtcaga taacgtgcct tctgaagaag ttgttaaaaa gatgaagaac 2640
tactggagac agctgcttaa cgctaagctc ataacacaga ggaagtttga caacttgacc 2700
aaggccgaga gaggcggact ctcagaattg gataaggcag ggttcataaa aaggcagctg 2760
gtggaaacaa ggcagataac taaacatgtg gctcagatcc tcgatagtag gatgaataca 2820
aaatacgatg agaacgacaa gctcataagg gaggttaaag tgataactct gaaatccaaa 2880
ctggttagcg attttaggaa ggatttccag ttttacaaag ttagggagat caacaattat 2940
catcacgccc acgatgccta cttgaacgca gttgtgggta ctgcacttat caaaaagtac 3000
cctaagctgg aatccgagtt tgtttatgga gactataagg tgtacgacgt tagaaaaatg 3060
attgcaaagt cagagcagga gatagggaaa gccactgcaa aatatttctt ttatagcaat 3120
atcatgaatt tctttaagac agaaatcaca ctggccaatg gggaaataag gaagaggccc 3180
ctgatcgaaa ctaatggcga gacaggggag attgtgtggg ataaaggtag ggactttgca 3240
acagtgagga aagtgctgag catgccccaa gttaatatcg ttaaaaagac cgaggttcaa 3300
acagggggct ttagtaagga aagcattttg cccaagagga atagtgacaa attgattgct 3360
aggaaaaaag attgggaccc caaaaagtat ggcggatttg atagccccac tgttgcttac 3420
tccgtgctcg tggttgcaaa ggtggagaag ggaaagagca agaaactgaa gtcagttaag 3480
gaactccttg gtatcactat catggaaaga agctcctttg agaagaaccc tattgacttc 3540
ctggaggcta aagggtacaa agaggttaag aaagacctta tcattaaatt gcccaaatat 3600
agtcttttcg agcttgaaaa cggaagaaag aggatgcttg catccgctgg cgaattgcaa 3660
aagggcaatg agcttgctct cccttccaag tatgtgaact tcctttatct tgcctcacac 3720
tatgaaaaac tcaaaggttc acccgaagac aacgaacaaa agcaactatt tgtggaacaa 3780
cacaagcact acctggacga aatcattgag caaatttctg agttttcaaa aagggtaatc 3840
ttggctgacg caaatctcga caaagttttg tcagcttaca acaaacatag agataagcca 3900
attagagagc aagctgagaa tatcatccat ctgtttaccc tgactaacct tggagcgcct 3960
gctgctttta aatatttcga caccacaatc gacaggaaga ggtacactag cactaaggaa 4020
gttctcgacg ccaccctcat ccaccagagt attacaggcc tgtacgagac aagaattgat 4080
ctttctcaac ttggtggtga c 4101
<210> 69
<211> 4101
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 69
gataagaagt actcaatcgg tctggcaatc ggaaccaact ctgtgggttg ggcagtgatt 60
acagatgagt ataaggtgcc aagcaaaaaa ttcaaggtgc tgggtaatac cgacagacac 120
agcattaaga agaatttgat tggagcactc ctctttgact caggggaaac agcagaggca 180
acaaggctga agaggacagc aaggcggagg tacacaaggc ggaaaaacag gatatgctac 240
ctccaggaaa tctttagcaa cgagatggct aaagtggatg atagcttttt ccatagactc 300
gaagaatcct ttcttgttga agaggacaaa aagcatgaaa ggcatcccat cttcggcaat 360
atagttgatg aggttgcata ccatgagaag taccccacaa tctaccacct cagaaagaaa 420
cttgtggact ccacagataa agcagacctg aggctcatat acctcgcact cgcacacatg 480
atcaagttca gagggcactt tctcatcgaa ggtgacctga atccagataa ttcagatgtg 540
gataaactgt ttatacagct ggtgcaaaca tacaaccaac ttttcgagga aaacccaatc 600
aatgcctccg gtgttgatgc aaaggccatc ctgtcagcaa gactcagcaa aagcaggcgg 660
ctcgaaaacc tcatcgccca gcttcccggt gaaaagaaga acgggctctt tggtaatctc 720
atcgcattga gccttggtct tactccaaac ttcaagagca attttgatct ggcagaggat 780
gctaaactgc aactctcaaa ggacacatat gacgatgacc ttgacaatct gttggcccag 840
atcggggacc aatatgcaga cctcttcctg gccgcaaaga atctgtcaga tgcaatcctc 900
ttgtccgaca tactgagagt taacactgag atcacaaagg cacctctgtc cgcctccatg 960
attaagagat acgatgagca tcaccaggat ctgactttgc tcaaagccct cgttagacag 1020
cagttgccag aaaagtacaa agaaatattc tttgatcaat caaaaaacgg atatgcaggg 1080
tacatcgacg gtggggcaag ccaggaagag ttctacaaat tcatcaaacc tatcctggaa 1140
aagatggatg ggacagaaga gctgctggtt aagctgaata gggaagacct cctcagaaag 1200
cagaggacat ttgataacgg gagcatccct catcaaatcc acctcggtga actccatgct 1260
atcctgagaa ggcaggaaga cttttatcca tttttgaagg acaataggga gaaaatcgaa 1320
aaaatcctga cattcagaat cccatactac gttggtcctc tggcaagagg taacagtagg 1380
ttcgcatgga tgacaaggaa aagcgaggag acaatcacac cctggaattt tgaggaagtt 1440
gttgacaagg gtgccagcgc acaatccttt atcgaaagaa tgacaaattt cgacaagaat 1500
ctgcctaacg aaaaggttct cccaaagcat tcactcctgt acgaatattt tacagtttat 1560
aacgaactga ctaaagttaa atacgttacc gagggtatga ggaagccagc attcctttcc 1620
ggggaacaga agaaagctat tgtggacctc ctgttcaaga caaatagaaa agtgacagtt 1680
aagcaactca aagaggatta cttcaaaaag atcgaatgtt ttgactctgt ggagatcagc 1740
ggggtggagg atagattcaa cgccagcctg ggtacatatc atgatctcct gaaaatcatt 1800
aaagacaagg acttccttga caacgaggag aacgaggaca ttctggaaga cattgttctg 1860
accctcacac tctttgagga tagggagatg attgaggaaa gactgaagac ctacgcccac 1920
ctctttgacg ataaagtgat gaaacagctc aagagaagaa ggtatacagg ttgggggaga 1980
ctgagcagga agttgatcaa tgggattagg gacaaacagt ccgggaaaac aatcctcgat 2040
tttctgaagt cagacggttt cgcaaacaga aattttatgc agctcattca cgatgacagc 2100
ttgacattca aggaagacat ccaaaaggct caagtgagcg gccaagggga tagcctccac 2160
gagcatattg caaatctggc aggttcacca gccatcaaaa agggcatact tcagacagtt 2220
aaggttgtgg acgaattggt taaagttatg ggcaggcata agccagagaa tatcgttatc 2280
gaaatggcaa gggagaacca aacaactcaa aaagggcaga aaaatagcag agagaggatg 2340
aaaagaatcg aggaagggat caaggaactt gggtcccaaa tcctcaagga gcacccagtt 2400
gaaaatactc aactgcaaaa cgagaagctc tatctctact atctccaaaa cgggagggat 2460
atgtatgttg accaggagct ggatattaac agactgtcag attatgatgt tgatcatatc 2520
gtgccccagt cattcctgaa ggacgattcc atcgacaaca aagttctcac aaggtccgat 2580
aaaaacaggg gcaagtccga taacgttcca agcgaagaag tggtgaaaaa gatgaaaaac 2640
tattggagac aacttctgaa tgcaaagttg attactcaga gaaagtttga caacctcaca 2700
aaagcagaaa gaggcgggct tagcgaactc gataaggcag ggtttatcaa aagacagctg 2760
gttgagacaa ggcagatcac aaaacatgtg gcacagatcc ttgactcaag gatgaatacc 2820
aagtatgatg agaatgataa gttgatcagg gaggttaaag ttatcacact caaatccaaa 2880
ctggtgtcag acttcaggaa agactttcaa ttttataagg tgagggagat caataactac 2940
caccatgcac atgacgccta cctgaacgca gtggtgggta cagcattgat taaaaaatac 3000
cctaagctgg agtctgagtt tgtgtacggg gactacaagg tgtacgacgt gaggaaaatg 3060
atagccaagt ccgagcagga gatcgggaaa gcaacagcta agtatttctt ttacagtaat 3120
atcatgaatt tctttaaaac tgagattact ctggcaaacg gggagatcag gaaaagaccc 3180
ctcatcgaga ctaatggtga aacaggtgag atcgtttggg acaaggggag ggattttgct 3240
actgttagaa aagttctgag tatgccacaa gtgaatattg tgaaaaagac agaagttcag 3300
acaggtgggt tctccaaaga atccatcctg cccaagagaa attcagacaa gctcatcgca 3360
agaaagaagg actgggaccc taagaagtac ggaggatttg acagccccac cgtggcctat 3420
tccgtgcttg ttgtggcaaa ggtggagaaa gggaagagca aaaaactgaa atccgtgaaa 3480
gaactgctgg gaattaccat catggaaaga agctcctttg agaagaaccc aatcgacttc 3540
ctggaagcaa aaggatataa ggaagtgaaa aaggacctca ttatcaagct cccaaaatac 3600
tcacttttcg agttggagaa cggtagaaag aggatgctgg caagcgcagg ggaacttcag 3660
aaaggcaatg agctggcatt gccatcaaag tatgtgaact tcctctactt ggccagccat 3720
tacgagaaac ttaaaggtag cccagaagat aacgagcaaa aacagctctt tgtggaacag 3780
cataagcatt atctggatga gatcatagaa caaatctcag agttttccaa gagagttatc 3840
ctcgcagatg caaacctgga taaggttctc tcagcctata ataagcatag agacaagcca 3900
attagagagc aagcagagaa cattatccac ttgttcactc ttacaaacct gggggcacca 3960
gccgccttca aatatttcga tacaacaata gacagaaaga ggtataccag caccaaagaa 4020
gttctcgacg ccacactgat ccatcaatca atcacaggcc tttacgaaac taggatcgac 4080
ttgtcacaac tgggtgggga t 4101
<210> 70
<211> 3307
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 70
gagcaaggac acctacgacg acgacttgga caacctattg gcccagatag gtgaccagta 60
tgcagacctc ttccttgcgg ccaagaactt gagtgacgct atactgctca gtgacatcct 120
gagggtgaac actgagatca ctaaggcccc tctctctgcc tcaatgatta agcgttacga 180
cgagcatcac caggatctca ccctgcttaa ggcccttgtt cggcagcagc tccctgagaa 240
gtacaaggag atattttttg accagtctaa gaacggctac gccggttaca ttgacggtgg 300
ggcaagccag gaggagttct acaagttcat caagccgatc cttgagaaga tggacggcac 360
cgaggagcta cttgtcaagt tgaaccggga agacctgctc cggaaacagc gtacattcga 420
caacggcagc atccctcacc agatccacct gggcgaacta cacgccatcc tccgacgtca 480
ggaggacttc tatccattct tgaaagataa cagggaaaaa atcgaaaaaa tacttacgtt 540
tcgaatacct tactacgtgg ggccccttgc tcggggaaac tccagattcg catggatgac 600
caggaagtca gaggagacca tcacaccctg gaactttgag gaggtggttg acaaaggtgc 660
ttctgcccag tccttcattg agcggatgac taacttcgac aagaacctgc ccaacgagaa 720
ggtgctgcca aagcacagcc tgctctacga atactttact gtgtacaatg agctgacgaa 780
ggtgaagtac gtgacagagg ggatgcggaa gcccgctttc ctgagcggcg agcaaaaaaa 840
agcaatcgtg gacctactgt tcaagaccaa ccgaaaggtg acagtgaagc agctcaagga 900
ggactacttc aaaaaaatcg agtgcttcga ctctgttgag ataagcggcg tggaggaccg 960
attcaacgcc tcattgggaa cctatcacga cctgctcaag atcattaagg acaaggactt 1020
cctggataat gaggagaatg aggacatcct ggaggatatt gtgctgaccc ttactctatt 1080
cgaggacagg gagatgatcg aggagcgact caagacctac gctcacctgt tcgacgacaa 1140
ggttatgaag caattgaagc gtaggcgata cacggggtgg ggaagactct cccgaaaact 1200
gataaacggc atcagggaca agcagtcagg gaagacgatc ttggacttcc tgaaatccga 1260
cgggttcgcc aaccgcaact tcatgcagct cattcacgac gactcactaa cgttcaaaga 1320
ggacattcag aaggctcaag tcagtggaca aggcgactcc ctgcacgagc acattgcaaa 1380
ccttgcgggc tccccggcga ttaaaaaggg cattctccaa acggttaagg tggtggacga 1440
gctggtgaag gtgatgggcc gacacaagcc tgagaacatc gtgatcgaga tggccaggga 1500
gaaccagact acccagaagg gtcagaagaa ctctcgggaa cgtatgaagc gtattgagga 1560
ggggattaag gagttgggct ctcaaatcct caaggagcac cctgtggaga acactcagct 1620
ccaaaacgag aagctgtacc tgtactacct gcaaaacggg cgcgatatgt acgtggatca 1680
ggagttggac atcaacaggc ttagcgatta cgacgtggac cacatcgtgc cacagtcatt 1740
cttaaaggac gacagcatcg acaacaaggt tctgacgagg agcgacaaga atcgagggaa 1800
aagtgacaat gttccatccg aggaggtggt caagaaaatg aagaactatt ggcgtcagct 1860
tctgaacgcc aagctcatca cccagcggaa attcgacaac ctgactaagg ctgagcgagg 1920
cggactctcc gagcttgaca aggctggctt catcaagcgg cagttggtcg aaacccgaca 1980
gataacgaag cacgttgccc agatacttga ctcccgtatg aacaccaagt acgacgagaa 2040
cgacaagctc atcagggagg tgaaggtcat tacccttaag tccaaactcg tcagcgactt 2100
tcgtaaggac ttccagttct acaaggtgcg cgagatcaat aactaccacc acgcacacga 2160
cgcctacctg aacgcagtgg ttggaaccgc gttgattaaa aagtacccca agttggagtc 2220
ggagttcgtt tacggggact acaaggtgta cgacgttcgg aagatgatcg ccaagtctga 2280
acaggagatc gggaaagcaa ccgccaagta tttcttctat agcaacatca tgaacttctt 2340
taaaaccgag atcacacttg ccaatggcga gatccgtaag aggccgctga tcgagacaaa 2400
tggggagact ggcgagatcg tgtgggacaa gggccgcgac ttcgcaaccg ttcggaaagt 2460
cttgtccatg cctcaagtca acatcgtcaa gaagactgag gtgcaaacag gcgggttctc 2520
gaaggagtcc atactgccca agaggaactc agacaagctc atagcacgca aaaaagactg 2580
ggatccaaag aaatacggcg ggttcgactc gccgacagtc gcatactccg tgttagtggt 2640
ggctaaagtg gaaaagggga agtccaagaa gctcaagtcc gtcaaggagt tgctcgggat 2700
caccattatg gaacggtcct cattcgagaa gaatcccatt gacttcctag aggcgaaggg 2760
ctacaaagag gtcaaaaagg acctaattat taagctcccc aagtattcac tcttcgaact 2820
tgaaaatggt cgtaagcgga tgttggcaag cgctggagag cttcagaagg ggaacgagct 2880
tgcactgcct tccaagtacg tgaacttcct gtacctcgcc tctcattacg agaagttgaa 2940
gggctcaccg gaggacaacg agcagaagca gttgttcgtg gagcagcaca agcactacct 3000
cgacgagatc attgagcaga taagtgagtt cagcaaacgg gtgatccttg ccgacgctaa 3060
cctggacaag gtgctgagcg cctacaacaa gcacagagac aagccgatcc gagagcaagc 3120
ggagaacatc atacacctgt tcaccctcac gaacctcggg gctcccgcag ccttcaaata 3180
ttttgacacg accatcgacc gtaaacgcta cactagcacg aaggaggtgc tggacgctac 3240
ccttatccac cagtccatca ccggcctgta cgagacgaga atcgacttgt cgcagctcgg 3300
tggtgac 3307
<210> 71
<211> 4101
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 71
gacaaaaaat actcaattgg tctggcaatt gggaccaaca gtgtcggatg ggccgtgatt 60
accgacgagt acaaggtgcc gtccaaaaaa ttcaaggtgc ttgggaacac cgaccgccac 120
tcgatcaaga aaaacctaat cggtgcgttg cttttcgaca gtggggagac cgccgaggca 180
acacgcttaa aacgcacagc taggaggaga tatacacggc gcaagaaccg aatatgctac 240
ttacaggaga tattctccaa tgagatggcg aaggtggacg actctttctt ccatcggctt 300
gaggaatcct tcctggtcga ggaggacaag aagcacgagc gacacccgat attcgggaac 360
atcgttgatg aggtggcgta ccacgagaag tacccaacga tataccactt acgcaagaag 420
ctcgtggact ctacggacaa ggccgacttg cgccttatct acttggcact ggcccacatg 480
attaagttcc gaggccactt ccttatcgag ggtgacctga accccgataa ctccgacgtg 540
gacaagctct tcatccaact cgtccagaca tacaaccagc tattcgagga gaatcctatc 600
aacgcctctg gggtggacgc taaagctatc ctctcagccc gcctgtcaaa gtcgaggagg 660
ttggagaacc taatcgccca gcttccaggc gagaagaaaa atgggctgtt cggaaacctt 720
atcgcactct cactgggcct aaccccgaac ttcaagtcca acttcgacct ggcagaggac 780
gcgaaattgc agttgtcgaa agacacctat gacgatgacc tggacaacct gttggcccag 840
ataggggacc agtacgccga cctgttccta gcggccaaga acctgtccga cgccatcttg 900
ctgtcggata tactgcgggt gaacaccgag atcactaaag cacctctctc cgccagcatg 960
attaagcgtt acgacgagca ccaccaagat ttgaccctgc taaaggcact tgtacggcag 1020
cagcttcccg agaagtacaa ggagatcttt ttcgaccaaa gcaagaacgg ctacgccggg 1080
tacatcgacg gaggtgccag ccaggaggag ttctacaagt tcattaagcc catcctggag 1140
aagatggacg ggactgagga actacttgtg aagctgaacc gggaagactt actacggaag 1200
cagcgtacct tcgacaacgg ttctatccca catcagatcc atcttgggga gttgcacgcg 1260
atcctgcgac gccaggagga cttttacccc ttcctgaaag acaaccgcga gaaaatcgag 1320
aagatactga ccttcagaat accttactac gtcggacccc ttgcgcgagg caactcaaga 1380
ttcgcgtgga tgaccaggaa atcagaggag accatcacac cctggaattt cgaggaggtg 1440
gttgacaagg gtgcctccgc ccagtccttt atcgaacgaa tgaccaactt cgacaagaac 1500
ttgcccaacg agaaggtgct ccccaaacac agcctcctct acgaatattt cacagtgtac 1560
aacgagctta ctaaagttaa gtatgttact gagggcatga ggaaacccgc cttcctgtca 1620
ggcgagcaga agaaagctat tgtggacctc cttttcaaga ccaaccggaa ggtgacagtg 1680
aagcagctca aggaggacta cttcaagaag atagagtgct tcgacagcgt ggagatcagc 1740
ggggtggagg acagattcaa tgcctctctc ggaacatacc acgacttgct taagatcatc 1800
aaggacaagg acttcctcga caacgaggaa aacgaggata ttctggagga tattgttctg 1860
actcttaccc tgttcgagga ccgggagatg atcgaggagc gtctcaagac ctacgcccac 1920
ctgttcgacg acaaagttat gaagcagctc aagcgtcgga gatataccgg atggggccgt 1980
ctgtctcgga agctcatcaa cgggatcagg gacaagcagt cagggaagac gatcttagac 2040
ttccttaagt ctgacggctt cgccaacagg aacttcatgc agttgatcca cgacgacagc 2100
cttaccttca aggaggacat ccagaaggcc caagtgagtg gccagggtga cagcctccac 2160
gagcatattg ctaatcttgc gggttcccca gcgattaaaa agggcatact tcaaaccgtt 2220
aaggtggtgg acgagcttgt caaggtgatg gggcgacaca agcccgagaa catcgtgatc 2280
gagatggcca gggagaacca gaccacccag aaggggcaga agaatagccg agaacgcatg 2340
aagcgcatcg aggaggggat taaggagcta gggagccaga tcctcaagga acatcccgtc 2400
gagaacaccc agctccagaa cgagaagcta tacctctact acttgcaaaa cgggagggat 2460
atgtacgtgg atcaggagtt ggacattaac cgcctaagcg actacgacgt agatcacatc 2520
gtgcctcagt cattcctcaa agacgacagc attgacaaca aagtcttgac ccgatccgac 2580
aagaaccgag gaaaatccga caatgtgccc tcagaggagg tcgtcaagaa aatgaagaac 2640
tattggaggc agctacttaa cgccaaactc ataacccagc ggaagttcga caacctgaca 2700
aaggctgagc ggggtgggct cagcgagctt gacaaggctg gcttcatcaa gcggcagttg 2760
gtggagacaa gacagataac gaagcacgtg gctcagatcc tggactctcg catgaacacg 2820
aagtacgacg agaacgacaa attgatccgc gaggtcaagg ttattacgct caagagcaaa 2880
cttgtcagcg atttccgcaa ggacttccag ttctacaagg tgagggagat taacaactac 2940
caccatgcac atgatgccta cttgaacgca gtggtgggga ccgcgcttat taaaaagtac 3000
cctaagttgg agtcagagtt cgtttatggg gactacaagg tgtacgacgt ccggaagatg 3060
attgcaaagt ctgaacagga aatcgggaag gccaccgcca aatatttctt ctacagtaac 3120
attatgaatt tttttaagac tgaaattact ctcgcaaacg gcgagatcag gaagcgtccc 3180
ctcatcgaga caaacgggga gaccggggag atagtctggg acaaggggcg ggacttcgct 3240
acggtgagga aggtgctctc gatgccacaa gtgaacatcg tcaaaaagac agaggtgcag 3300
accggtggct tctcaaagga gtcaatcctg ccaaaacgta acagcgacaa gctcatcgcc 3360
cgcaagaaag actgggaccc taagaagtat ggtgggttcg actcaccgac ggtcgcatac 3420
tccgttctgg tcgtggcaaa ggtggaaaag ggcaagtcca aaaaactgaa atccgtgaag 3480
gagttgcttg gcattaccat catggaacgc agcagcttcg agaagaaccc cattgacttc 3540
ctggaggcta aagggtacaa ggaggtcaag aaagatttaa ttattaagct acctaagtac 3600
agcttgttcg agctggagaa cggccgaaaa cgaatgctcg catccgccgg ggaacttcaa 3660
aagggcaacg agcttgcgct gccctccaag tacgtgaact tcctgtactt ggcatcccac 3720
tacgagaaac tcaagggtag cccagaggac aacgagcaga agcagctatt cgtggagcag 3780
cacaagcact acctcgacga gataatcgag cagatcagtg agttcagtaa gcgggtgata 3840
ctcgcggacg ccaacttgga caaggtgctt agtgcctaca acaagcaccg tgacaagccc 3900
atccgagaac aggctgagaa catcatccac cttttcactc tgacaaacct cggtgctccc 3960
gccgccttca aatacttcga cactaccatc gacaggaagc gctacacatc tacgaaggaa 4020
gttcttgacg ctacgcttat tcatcagtct atcacagggc tgtacgagac aaggatcgac 4080
cttagccaac tcggcgggga t 4101
<210> 72
<211> 13170
<212> DNA
<213> 大豆
<400> 72
gcttaataat agaagagtaa tgggacagtg atgcttgtgt gtagatatga gtaaaataag 60
agtacatgtg atgacaatat aaataaggtt gaatgctaca aaggtaaata gaaaataatc 120
cattttattt tatttattat acataaatga taattaaaat ataattcaaa tgttattttc 180
ttttcctttt ctaattaaac ccaattaaaa tatttcttct tcttttattt actcacaaaa 240
aatatatttc cataactacc ctcacacttc ttcttttata tatatatata tatatataaa 300
gataaaaatt aagtatcgaa acaaaactga ttcacagtat ttattttaag taattataga 360
aaatatagaa gaaataatga tttgatttat cattggcaat acatttttct atttttttaa 420
aaattatcaa aaatattttt taatgactca atctaattat ctaatccaac ccaatcaatt 480
gaattggatt gctaaaaaaa taaatgcaac taaacctaat tcaattatgt ttaattgagt 540
tatattgtaa actagaagtg tttccaatgt attttgtaca tgcagttaca taatataaag 600
aaagagagca aatgcaatac agtacaaaac ataaaattac ttatattgca taacaaagta 660
acacgtgtaa tgtagttgga tagattttat atatatgttg agatatatat gttgagatta 720
gattaaattt aactcaatcc acttatatct gtaatttttg tcttatattt tatctaaaat 780
ttgtattata aataatttaa tatcttatat ttgagtcaat tataatttaa ttttaatgtg 840
cctccaatat tagttatgct ggtgtctatt gaaacgccga ggccgaggga tcgaagtgta 900
cctacacgcc acactgcaag agtgtcgcca agttgggttt gatgaacgaa aatgaccaat 960
ccatccaaca aaattttaat taattttcaa taataataat aaaaaaaggc aaagaaagtt 1020
acgcacaaaa gcctcgtttt gttctgtccc tttattcctt ccttcctttc ttcttcttcc 1080
ttacctcttt ccctggctga tttagggttt tctggcaata atctcctttc ggtacttttt 1140
acctctgaat cctcatcatc gctattgctg attacactcc tcacaatgct cactcaacgc 1200
ctcgttcgaa aaacgtgaaa tcttctcttc tgcaatttct agggtttcga attgcggaac 1260
cgattcattc aactcaacac cacaaagact cgatcgatcc atggtagcgt tgtgattggc 1320
gctcggtggc gagcgagttg atagattgcg ggttggctgg attttgtatg gaaactcgga 1380
gccggaagcg ggcggaggct tcctcagctg ccccttcatc cccttcctct ggtcccacca 1440
ctcgttcaag caagcgcgca cgtctctctt cttcttcttc cgcttccgcc gccgtgaaca 1500
ctcggtcccg tgcatccaat actaaggaac ccttgccccc taaaaatccc cctcctcctc 1560
ttcctcccat ggactctgcc aacgaatcct ctggctcccg ccgtgatcgc cgcaacaaca 1620
aagaaaactc ctccgacaag gggaaggaga aagagcatga tgtcaggatt agggacaggg 1680
atgcggcatt gaacatggat ggcagtggtg gagacgagga cgatgacaat gacaatgaca 1740
gtgaaggcgg tgtggggatt ttgcatcaaa acttgacctc tgcgagtagt gcacttcagg 1800
ggcttcttcg gaaactcggt gctggtttgg atgatctgct tccatcgtcg gctatgggct 1860
ctgcatcttc ctctcaccag agtgggaggc tgaaaaagat cctctttgga ttgcgggctg 1920
acggcgaaga agggcggcag gtcgaagcat tgacacagct ctgtgagatg ctttcgattg 1980
ggactgaaga atcgcttagt acattctctg ttgattcgtt tgttcctgtg ctcgtggggt 2040
tgttgaatca cgagagcaac cctgacatca tgcttcttgc tgccagagca ttgacccatc 2100
tctgcgatgt gctcccttca tcttgtgctg cggttgtgca ttatggtgca gtctcaatct 2160
tttgtgccag gttgctcaca atagaataca tggacttggc tgagcaggtt gttctttaat 2220
gggaatggtg ttagaagaaa cttatgaaat ataaatattt gtcttgaact ctggtgctgc 2280
ctaattgttt tttaaaacta tgtatattgc aatgtttctg ggttcagttg tagaagaatc 2340
atgcattttg aaaatgaaat tcttatagtt tgattactgt gtttgatggc atgaacagtc 2400
ccttcaagct ttgaagaaga tctctctgga gcatcccact gcctgcctac gagcaggtgc 2460
tcttatggcc gtgctttctt atttggactt cttttcaaca ggagttcagg taaattgtca 2520
gtgaagtacc taaatttaga ggatgattat attatatggt ccttttggat agtcataaat 2580
catttggcca cagatgcctt tagtgagcaa gtgtgtgtat gttgacatta ttctttgttt 2640
cagcgggttg cattgtctac tgctgcaaat atgtgcaaga agcttccttc agatgcagct 2700
gattttgtga tggaagctgt tcctcttctg acaaaccttc ttcagtacca tgattccaag 2760
gtaaggtcat gttttttgtt ttaagtcttg tcacataatg gaaactgtat ttttcctttt 2820
gcatctataa cttttcccgt taaggaagtt ttttttagct tgagaggagg gagattttga 2880
tccctgtatc ccatagtata aaatattttt ctatagcttc accttatgca ttatcaattt 2940
ttgtaggttc tggagcacgc ctctgtttgt ttgactcgaa tagctgaagc ttttgcatca 3000
tctccagaca aattagatga gttgtgcaac catggacttg taacacaagc tacctccctc 3060
atttctaaca gcagttctgg aggtggtcag gcttctctca gcacaccgac atataccgtg 3120
agtgcaattt ttactttaag tcggatgcat tttgcatgta gttttggtcc ttgacttggg 3180
catatgcagg gtttaatccg acttctttca acttgtgcga gtggatccct cttggagcta 3240
aaaccttatt acttcttgga attagtggta ttcttaaaga tattctatct ggttctggag 3300
tttcttctaa tgcctctgtt tctctgcatt aagtaggcca ccagaacagg tatagtataa 3360
caccagaact tttcttttag tcattcatgt gtagttttat ctcgtaatgt tcattaagca 3420
gcctttgacc ttaaatacac cattatttag atccttgata tttccagttc tcttacattt 3480
gtgaaggaat gaaattattc taattctctg tttcaaatat atagatattt gagattgtaa 3540
acctggcgaa tgagcttctg cccccattgc cacatggaac aatttctctc cctatcatct 3600
ccaacatgtt tttgaaaggg cccattgtaa agaagtctcc ttctggtagc tctgggaaac 3660
aagaagacac taatggaaat gttcctgaga tatcggctcg tgagaaacta ttaaatgatc 3720
agcctgaact acttaagcaa tttgcgatgg atctcctccc agttttaata caggttgaca 3780
tttgtgcatt gatcatttgc ttaaattttg cttgataaat ttgttaaatt aaaaaagaag 3840
ttctgatatt gctcctttgc ttccttattt atcttctttg gttaattgat ggtattggct 3900
tgcttttgat acagatatat ggttctagtg tcaatggtcc tgttcggcac aaatgccttt 3960
ctgtcattgg gaaattgatg tatttcagca cggcagagat gatccagtct ttgttgagtg 4020
tgacaaatat atcaaggtat gtagaagttt aattgggttg atattgcttg atatcctact 4080
gattttatgg gtttagaaaa tttatgcatt gttgctttga tcatatagtt tcttagctgg 4140
cgtcctagca tggaaagatc cacatgtttt gcttcctgcc ttgaaaattg ctgaaattct 4200
tatggaaaag cttcccggga cattctccaa gatgttcatt agagaaggtg ttgtgcatgc 4260
agtggaccaa cttattttag cctcaaattc aaccaatatc tctacacagg catctcctgc 4320
tgagaaggat aatgattcta tatctggagc atcatctcgc tctaggcgtt atcggcgacg 4380
gagtgggaat tccaatcccg atggaaatcc tttggacgat ttgaaaactc cagtttcagt 4440
aaatgttggt tcacctccaa gttctgtgga tatgccaaca ttaaattcca gtattcggtt 4500
atctgttagt acagctgcca aagcttttaa agataagtac tttccttcag atcctggggc 4560
tgctgaagtg ggtattactg atgatctttt gcatctgaaa aatctttgca tgaagttaaa 4620
tgctggtgat gatgaacaaa ggactaatgg aaagggggaa tctaaaactt ctggatttgg 4680
tccggaagag tatttaattg ggatcatagc taacatgcta aaggaacttg gcaaaggaga 4740
tggtgtatct acttttgaat tcattggtag tggtgttgtt gcagctctgt tgaattattt 4800
ttcttgtggg tatttctcta aagatcgacc attagaaacc caccttccca agcttcgcca 4860
acaagcactt acaaggttta agttatttat agcagttgca ctaccttcta ctattgaagt 4920
tgggactgtg gctcctatga ctgtcttggt ccagaagctt caaaatgcct tgtcctcctt 4980
ggagcgtttc cctgttgtgc tgagtcattc atctaggtca tctagtggga gtgcacgcct 5040
ctcctctgga ctaagtgctt tatctcagcc cttcaagttg cggctttgtc gagcccaggg 5100
tgaaaagtca cttagggatt attcatccaa cgttgtactg gttgatccat tagcaagttt 5160
agcagcgatt gaggaatttg tttggcctcg tatccaacgt agtgaatctg gtcagaagtc 5220
cactgtagcc actgggaatt ctgaatctgg gacaactcct gcaggagctg gtgtatcctc 5280
tcctactacc cgtcgccatt ctactagatc cagatcatct gttaatatag gtgatacatc 5340
tagaaaggaa ataactcaag ataaaagtac aagttcttcc aagggtaagg gaaaagttgt 5400
attaaagcct gcacaagagg aggcaagagg acctcagacc agaaatgcta ctcgcagaag 5460
agcagctctt gataaagatg ctcaaatgaa gcctgtaaac gctgactcta cttctgaggt 5520
ataccgatgt aaactgagtt agggactgtt atgacagtta tctgtagaga ataactgctt 5580
tagaaatcaa tgttagaaac tcaaatatct gtcttagaaa acagtactgt attagaaact 5640
tcaggatagt agttatgtat ttggagtata tgtctgttag tatgtggttt taattaggca 5700
gtttaaagtt atttttaagt tcgtggacag agaatactct gtaggagact tctgtctctg 5760
ggacagtaag tgctgttttt gtatgtgata ggagactcta gtctttagta cagcaaatgt 5820
attgctgtat ttgtatgtga taggaaactt ttgtctctgg gacagcaagt gtattgctgt 5880
attgtgtgtg atttaagtgt tctatatatt ttaggcaaat gctagggcgc tgtttgagga 5940
agtaaaaagg gaaagttttt tattgggatg cgtaaaatgc actcccccta tattttattt 6000
ctttacttta tcatttggtg ttctatcata tactttgaaa tcctagcgat gagtttacaa 6060
tgctacaact tttttcttct gttaatttat aactttcagt tatttagttt ttgttggatg 6120
gttttcttct tttgctctcc acttccttct ggctatgtat gatctgacta tgaatgttgc 6180
tgttttcatg ttggtttata cagtgaaata tgaggagaat acttaataat gccaatataa 6240
ggtgtaaaga tgcattatta gattttgcat cgcaagcttg aaaagtcatc tggaatttct 6300
ctgcaacttt ggacacactg gattttgttg tctgtagatc cttgaaatgt tttacctctg 6360
ccatttagtt ttataaatgg cttttggtta caattatcta tttagatata tctcttatgt 6420
gagaaacact acctaagtgt tcttttgatt attatgttga acagttgctt ttttgcacca 6480
ttgatttctt tacatacatt aaaattaagc tgttataaga tttccctgca aaagtgttga 6540
gttactacaa aagtgaggaa cattgccaag cctcatcctt ctagcattaa gtgtatgtgt 6600
caggattgtt ttagattatt tgtgggtggg ttgcatattt tttctattta ggttatactt 6660
gttttgtttt tcattgcttt tgattatggc agtgtttaca tgcatgttgt ttagctaggt 6720
tcttgtggtt gttaaactgg tggaaggatc tttagagtat aaaatacaac acacattcta 6780
tatgcagttg atagggctca acttggctgc taaactcatc tttttactgc tatgctttgg 6840
actaaatcat gaacctaatc caagtatcct gaagtatcta aatattagtc tcttgttttt 6900
tcttttcttt ttttaccctt ctattgacat ttttcattgt ctacattgta ggatgaagat 6960
ttggatatat cccctgttga gattgatgag gcattggtga ttgaagatga cgatatttct 7020
gatgatgaag atgatgacca cgaagatgta tgttatctac tgttttgtct acttttctta 7080
cttggtgatg agtatcacat ttaagtaatt actgtgttac tgtaattttt ttttttgctt 7140
ttctgacaac ccccagcttt ttctacgttt aacacggctt ttaatatatt aaaatctatt 7200
tttggctacc aacgctggtg tccttcaatc ttgtttacat ttttttcaaa atgatatgct 7260
gctgtctttg tatcatttct ttgctttcta ttttttcttc tgggtataat gatgcaatga 7320
agttttagtt tatgatctta tgcgaactat ccaagcctct gaccaatagc tcaatctagg 7380
tgttttattt ccaaaatttt aattttagct accatcatta ttgttaacca agattgaatg 7440
ttatagttaa attcagttgt atctattctt aaaatattgg tgcttgtttt atttttattg 7500
atgcaggtac tgagggatga ttctcttcct gtttgttcac ctgacaaagt acacgatgtg 7560
aaattaggcg acttagcgga ggagagtaat gttgctcctg caacaagtga tggccaggct 7620
aatgctgcct caggttctag tagcaaagct ggtacagtca ggggatcaga ctctactgat 7680
tttaggagtg gctataactc aagctcaaga ggtgcaatgt catttgctgc tgctgctatg 7740
gctggacttg gatctgccaa tagcagaggt atcaggggtg gaagagatcg actagggcgt 7800
ccattgtttg gtagttctaa tgatcctcca aagttgatat ttactgctgg tgggaagcag 7860
cttaataggc atttgactat ttatcaggca attcaaaggc agcttgtgct agatgatgat 7920
gaaagatttg ctggcagtag tgactatgta tccagtgacg gaagcaggtt gtggggtgat 7980
atttatacta taacttatca tagggctgag aaccagacag ataggactcc ccctggaggt 8040
tcaacctcta atgcttcaaa atcttgcaaa tctgggtctg tatcgaattc tagttccgaa 8100
gccaagctgc atcagacatc tgtactagat agtatcttgc agggagaatt gccctgtgaa 8160
ctggagaaat ctaatcctac atacaatata ttggcattat tgcgggtgct tgagggtttg 8220
aaccaacttg catctcgttt gagggcccaa gtggttactg atagctttgc tgagggaaaa 8280
attttggatt tagatgagct aagtgttacc agcggtgcta gggttcctac agaggaattt 8340
ataagcagca aacttactcc aaaattagct aggcaaatac aagatgccct tgccttatgc 8400
agtgggagtc ttccctcgtg gtgttaccag ttatctaaag cgtgcccttt tttgtttcct 8460
tttgagaccc ggcgacagta tttttattca actgcctttg ggttatctcg tgcattgtat 8520
cgccttcagc agcagcaggg tgctgatggt catggatcaa caaatgaaag agaggtcagg 8580
gttgggagat tgcagcgtca aaaggttcgt gtctctcgaa atcgcatttt ggattctgct 8640
gctaaagtga tggagttgta ttctagtcaa aaggctgtac ttgaagtaga atattttggt 8700
gaagttggca ccggtctggg tcccactctg gagttctaca cacttctcag tcatgactta 8760
caaaaaatta tacttgaaat gtggagatca ggttcttcag agaaatatca aatgaaaatt 8820
gatggagatg aaaagaaaat gaaacgtagt gaaggctctt ttgttggaga tggagaactt 8880
gttcaagctc ctcttgggct gtttcctcga ccttggcctg caaatgctga tgcatcagag 8940
ggtacccaat tttttaaagt gattgaatat ttccgactat taggccgtgt aatggctaag 9000
gctcttcaag atggacgcct attggattta ccaatgtcag tggcatttta taagcttgtt 9060
cttggtcaag taagttatga aatgttgatg tcttatctga tttgatgtgc atcttaaggt 9120
tgatttttag tatgtatata tttaaccctt gatatattgc aggagcttga tttgcatgac 9180
attcttttca ttgatgctga gcttgggaaa actttgcaag agttaaatgc ccttgtttgc 9240
cgaaaacatt atatacaatc tactggtggt agctacaccg atacctttgc taatttgcat 9300
tttcgtgggg ccccaataga agatctctgc ttggacttca cacttcctgg ttatccagag 9360
tacatcttga aacctggaga tgaaattgta tgtattcagt ctgtttctat tacctggttt 9420
ttgttttggt tctgattctg tctgttatga aaattgcttt gaacttactg tcaaacttcc 9480
aggttgacat caataatcta gaggagtaca tatccatggt ggttgaggca actgttaaga 9540
ctggaatcat gcgtcaaatg gaagcattta gagcagggtt caatcaggtt atatgctgtc 9600
tcaattaatt catgtaactt ttgtctttga ctgtgcacct tgtttggtga tgctgtgtaa 9660
gctcttatct aaaatgtcat gtatttttta actgtgatca gaaagttctg taatatcatt 9720
gttgtctgtc ttttacttgc acattgtcaa ttcttattta aaacatcatg tattttttaa 9780
ttgtgatcag aaagttctgt aatatcattg tctgtctttt acttgcacat tgtccattct 9840
ttttggtgtt ccttctctca aacaatttta ttgaaaagta aacttgactg gggtttaatt 9900
tgataatatt gagcatggat tcttcagctt ttagattctt aaggggcatt gttttttcta 9960
ataaaaatta ttacttttgg ttatatcttg agcagtgaac tgcatatata gcctgaatat 10020
cggatgtatt aattaagaaa ttacaaaatg tttgtttttg acttttactt atgtttgcta 10080
ccaaccttgt gtcccccttc ccctggcccc tgcacatcaa gaatggaaga tagaaaatac 10140
acattttttt ttatattgat gtcaaatatt cagttattta atgtcaaaat tttacaagtg 10200
aaataagcta ctggctgagg aagagatggg ctgggctggg caatagttgg aacttggaaa 10260
aactagtttg atgatttgtt catttgcttg agtccaaaaa ctaaacattt gtcacttgcc 10320
cacttgttct tttcaattca ggtttttgac atctcatctt tacaaatttt ttctccccaa 10380
gaactggatt acttgctttg tggccggaga gaattgtgga aggtattctt ttttgtgcaa 10440
aagtattact gctgcttaca acaatatttt aaatgttacc atgaataatg tagttataaa 10500
tttatttttc tcttttctgc agactgagac actagccgat catataaaat ttgaccatgg 10560
ttatactgcc aagagccctg ccatagttaa tgtatggttc ttttccccat agaaggacat 10620
ttgtgttttt ggaaatttag gcttgctata tttgatgctg aacctctatg atgctatttt 10680
cagttactcg gaattatggg agaattcaca ccagagcaac agcgtgcctt ctgtcaattt 10740
gttactggtg cacctaggtt gcctcctggt ggactggcag ttctaaatcc aaaattaacg 10800
attgtgagga aggtattgaa aaaatatttt tgatcacttg caacctgtgc tatttattca 10860
tgcggccttc acgcaatttt gtacttgaat gttaaagtgg ctttcctcta cttttccttt 10920
ttacctctta aaagggggct tacaaaatgc aagtttgtaa gtgattacat ttacatgtct 10980
gggtgcagct ttcatcaagt gcagctaatg cttcatctaa tgggaacggt ccttcagaat 11040
tagcggatga tgacttgcca agtgtgatga cgtgtgcaaa ttaccttaag cttcctcctt 11100
attctaccaa ggtagaacac tgcaaagcat tgttggtaat atgatcatgc atgtcaaagt 11160
gtcttttggt gttgaactgt tgatatttga tttccatttt aaaacaggaa attatgtaca 11220
agaagctact ctatgcaatc agtgaaggcc agggatcctt tgatttatcg tgagtttctg 11280
gaactaacca acccggccct gtatgttaat atgatgatga atcagggttt actcttgaga 11340
cttgattatt ttttggttat atgtactttc ctccctgctt ctcaaataat ggcctctccc 11400
gggtcacaca tcaaaggtgg tgacatggtt caggtgagtt taccattgtt gtgaaacctc 11460
actatgattc ttgacactac tagaccattg ttgtgagctt accaagagcc tcttattatt 11520
ttgttcattt tcatttgttg gtaaactaat taatatcatt tctttgatta ctggcagttg 11580
aatcggatcg cctcgttcta aaggtgtaaa ggaatttgcg gagctcgttt tagctgttac 11640
aacatgtaaa tagaatagga atgttgagta attccctccc cctctaaatt ttccatcatt 11700
ctgttttcaa ctcaaattat tggcagttag ttattataca tgcatggaag atggtaaaag 11760
gaatagaaat aatatcaaat actatgcttt attatttcat gttagatctg ttacgagctt 11820
ttgttgaggt aaaatgtgga tatcactcta tgctgatgct atttgctatg attgccccaa 11880
aatgaggcgc tctatagcat aaataagtgc tgtttgggtt gccgattggt tagtcgctaa 11940
tgtacttttg ctaaagcatc tcttttgtta cttctccgtt gaaatgcatt gaaattcccg 12000
caaccaaggt ttgaatggga attcaaacat tccctaaatt tcatgtacta gctagggcta 12060
tctaaactgt gatagcgcca attctaatgc actttaaatt ttccccaagt tattagaata 12120
ttatgttcta atgtgttgaa acggttcaaa tagacttttt atattataag attaaattac 12180
tcatttgatt tcttgattta tattttttta gttcctataa tttatatttt aatttgtttt 12240
tagttcttat agtttgaaag tgatttttta gtctttataa tttatatttt aattatcttt 12300
tatcacaaaa aatatataaa aataattaat tataaattat taattttttt tataaattat 12360
cttgtgataa attagttata aattatttgt tattattttt ataattaatc ttaattgata 12420
atattagtta tattttgatg gtaaggatta aaaaaaatta aaatataaaa tatactgatt 12480
aaaaatataa cttaaatcat aaaaattaaa aaaaataaaa tataaattat aagaattaaa 12540
aaaattacat caattaaaag aaaatggtct agtatgacaa taagagaaat gcaaattcct 12600
agctatagta cgacacaata aatttttggt agctccaatc agagtgaagg acgagtgcca 12660
ttggatatct attcctccag tcgaggatgg cgtgcatgtt gatctgaaaa agacgtcccc 12720
agtctatgac acggctcagg gatcaaccaa agcacccatt ccatgttttg gtgtaaatac 12780
tctatctcca caatcattaa ttctatgcta ctttgttgag tacccaacta cctgtcgtag 12840
tcgtggttgt gaaaaacaac tcccataccc ataactgtca tactgtgagt tgctgcaatg 12900
caatatggat gctcttattg agcaacgaaa caagtggtta taggaatatg tcagagtcaa 12960
aggataaaat aattaaagtt cattaataat ttttttatta gttttataaa ataaaaagac 13020
ttccattttt gacaaatgtc atatatgctg tcaaataagt ttggtatata actcttattt 13080
aaagtaaatt ttgttttaaa attttcttta ggtgtcattc attgttggac tggcggccct 13140
gcccgttgtc aaagagcatc gttatcgact 13170
<210> 73
<211> 16141
<212> DNA
<213> 大豆
<400> 73
tttttagaaa atattaaata attgataaga tatatgataa ttataataac aaataattta 60
aaatatcaaa tattttatct gtatatcatt atataattat atttatgtat cttaactcaa 120
ataattagta aaaataatct ttaattaaat attttttata aataggtata aattatgtga 180
taattttata aatatataaa aacataagca tgactgatat tattaaatta tgaatattat 240
atatatatat atatgcgtgt gtgtgcaaaa aaaaaatcaa tttgaattgg attagatcta 300
ttttaaaaat taaatatgat ccaatccaat aaaaaaattg attttttaaa tttttaatct 360
atttggattt tgatttttta aagttttatt ggattagatc gatttatgaa caccgtctaa 420
tatactaggt tcaaacctta gtgtgattat tgcaaaacaa aattatatga gagatttact 480
tggtaagata tatatctttc aattcaactt tagtttagat taattcaaat ttaacctcac 540
ttttagttct ctctcttcaa tgaaaaagaa actcactttt aattattgat gatctcataa 600
aggtaaaaga cttttcaata tggaccaatt tatttaaact aaaaaagagt tatttaaaaa 660
actaaaatat gtttttaaaa aaaatagttt agacagattc agattttttt gcttgtttta 720
ttaaaaaaaa ttaagtcaag ctttaataaa caggtcatat gtcatctttg ttattaaatt 780
aaaatcactt tgaaatttga ttgatttcat tggatgattg actgacttaa aaactttgtt 840
attaattttt aaaaatgatt ttataagaca aaatttaaaa tcaaattttt aaagagtaag 900
agtgagaaaa aaatatgaaa aagtatgact caagctcttg tctgttttta cataaagttt 960
taagagctag attattttta tatacacttt tttatgatat attttatctc aagatcttaa 1020
gcaacaaata ataatttcaa ggcaaatata tataatagga cgcacaaaac aaaaaataga 1080
aatcaacata caccaaattt tagttagttt gtctgaatgg aataatattt taaagcttga 1140
atgttggatc gattagtgtt atttccaaaa gcaaaactat gtccctggtt tatgtttgaa 1200
tattaactta tgtttgtgtt ttatttccaa aaaaaaaaaa gaagtacagt cacaatttat 1260
atttgaagtt gctaaactat actaataagg gaggaaaact aggttatttg aacatgaccg 1320
gccccttgtc gacaacaact gaaagctcga taaatgatca tcactaaatc actatagtag 1380
cccgataact tctaaaaaac atttagaaag tcacctttca gtttttaaat gtgtaaatat 1440
taatgattat gatgattttt tttttaaaaa ttctaaacaa gtcaagtgca aagattattt 1500
atttcattca ggttaagtta aaatgggatt tgtgattgtg atttcctaaa gttagtgtga 1560
tccgtaatac atgtccagta ttccgaaaag tacaaacaat tactgatcta ataaacaata 1620
tccttaaaac tttcaagact ttatggagac aaatagcaat caaatgtaca tgcaaaaaaa 1680
aaaaaaaaca gaaatatatc caggatttaa gcattttcaa taggaagact atgcattgta 1740
tttgcacata aaagtaggcc gtaacaaaaa gtcctatccc cgctcctctt tccccgatcg 1800
ataagaatag aacatccaat aatccattaa gggatttcta tcaacgttaa ttctagctag 1860
aaacctctgc atgagaacac tatctagtac tctactcgtg tctttcatta catgttcaag 1920
tgagcatgca aagtaggtgg gggaagtaat aaaccagaca atcaccaacc ttttccatga 1980
gagagacata tatattagca aagcaaggaa ccaaagactt caaatttgca ctctcagtgg 2040
gaaggtctgg tatataggca actctggggt acattggggc gtaatttgtt ttaaaaatat 2100
aactattatt gaaaattaaa ataaaatgtt gtagctattt agagtatata gttaaaggta 2160
aaaatattta gaagaaaaat gacaattatt gcttttataa taattataga tattgatagt 2220
gatatataat agatttatat agtttttatt ttttatttgt ttttgaatat tttataaata 2280
aaaaattaca agtgtttaaa atatacttct atcaaaaaaa tcaaattaaa atgtgtatat 2340
aaacattaca tgattattgt ttcaattttt tagtatttta agaattattg gtttgtgtgt 2400
tataagtatt aattgctact gacttataag aattaagaac taaaattatt gaaacaaagc 2460
tgatttggca gtatttattt aaagcagtta tagaaaaatt gaagaaataa tgtgttgatt 2520
tatcattggc aatacatttt tttaattttt tttaattact caatccaatt atctaatcca 2580
acggccaatt aataccagcc ttatgtttaa ttggattata ttgtaaatta gattaaacta 2640
aactcaatcc aaccagttac atcttatatt tttatctaaa atttgtatta taatacattt 2700
tctattaaat acaatttaat aatttaactt tattgtgcaa gtgcaacgta tattaataca 2760
atgcaggtgt ctattgaaac gacgacgttg agggatcgaa gtgtacgcgc cacattgcga 2820
cagtgccacc gaggagcgta gctagttcag ttccgtttga tgaacgaaaa tgaccaatcc 2880
accgagcgaa attgtaattt tcaacaataa aaaaggcaaa gaaagttaca tatgaaagcc 2940
tgttttgttc tgtccctttt tattcttccc tttctttctt acttacccct ttccctggct 3000
tagggttttc tgcccccctg aatcctcatc gctattcatg attacactcc ttacaattct 3060
cactcagcga ctcgttcgaa atacgtgaaa tccccttatc tccaatttct agggtttcga 3120
ttcattcaac tcaccacaaa gattggatcc atggtagcgt gattggcgct cggtggcgag 3180
cgagttgata attatcgggt tgggtggatt ttgtatggaa actcggagcc ggaagcgggc 3240
ggaggcttcc tcagctgccc cttcatcccc ttcctctggt cccaccactc gttccagcaa 3300
gcgtgcacgc ctctcttctt cttcctccgc ttccgccgcc gctgccgccg catccgtttc 3360
ttccgtgaac actcgctccc gttcatcccg gactaaggaa cccttgcccc ctaagaatcc 3420
tcctcccatg gactctgcca atgaatcctc tggctcccgc cgcgatcgcc gcaacaaaga 3480
caactccgac aaggggaagg agaaagagca tgatgttagg attagggaca gggatgctga 3540
cagagggctg gcattgaaca tggatggtgg cggagacgat gatgacaatg acagcgaagg 3600
cggtgtgggg attttgcatc aaaacttgac ctctgcgagt agtgcacttc aggggcttct 3660
tcggaaactt ggtgctggtt tggatgatct gcttccatca tcggctatgg gttctgcgtc 3720
ttcctctcac cagagtggga ggctgaaaaa gatcctcttc ggattgcggg cggatggcga 3780
agaagggcgg caggttgaag cattgacaca gctctgtgag atgctttcga ttgggactga 3840
agagtcactt agtacattct ctgttgattc gtttgttccc gtgcttgtgg ggttgttgaa 3900
tcacgagagc aaccctgaca tcatgcttct tgctgccaga gcattgaccc atctctgtga 3960
tgtgctccct tcatcttgtg ctgctgttgt gcattatggt gccgtgtcaa tcttttgtgc 4020
gaggttgctc accatagaat acatggacct ggctgagcag gttgttcctt ggttcatgac 4080
tattgtggaa ttaatgaaaa tggtgctaga aagttataaa ataaatattt gtgtttcagt 4140
gttgaaccct ggtgctgcct aattgttttt aaaactatat atttcgatgt ttctgggtca 4200
aattttcagt tgtagaagaa ttatgccttt tttttttaat ctaagactta gcattttgaa 4260
aatgaaattc atatagtttg attactgtat ttgatggcat gaacagtccc ttcaagctct 4320
gaagaagata tctcaggagc atcccactgc ctgcctacga gcaggtgctc ttatggccgt 4380
gctttcctat ttggacttct tttcaacagg agttcaggta aattgtcagt gaagtaccta 4440
catttagagg atgattgcat cccactggtc cttttggaca gtcataaatc atttggccac 4500
aggtgccatt agtgagcaag tgtatgttga cattattctt tgtttcagcg ggttgcattg 4560
tctactgctg caaatatgtg caaaaagctt ccttcagatg cagctgattt tgtgatggaa 4620
gctgttcctc ttctgacaaa ccttcttcag taccatgatt ccaaggtaag gtcatgtttt 4680
gttgcaagtc ttgtcacata atggaaactg tatttttcct tttgcatcca taaacttgcc 4740
ttttaaggaa gtttagcttg agaagaggga aattttgatc cctatatccc atggtataaa 4800
atatttatct atagcttcac cttatgcatt ttcaattttt gtaggttctg gagcacgcct 4860
ctgtttgttt gactcgaata gctgaagctt ttgcgtcatc tccagacaaa ttagatgaat 4920
tgtgcaacca tggacttgta acacaagctg cctccctcat ttctaacagc agttctgggg 4980
gtggtcaggc ttctctcagc acgccgacat atactgtaag tgcaattttt acttttagtt 5040
agatgcattt tgcctatagt ttggtccttg acctgggtat atgcagggtt taatccgact 5100
tctttcaact tgtgcgagtg gatctcctct tggagctaaa accttactac ttcttggaat 5160
tagtggtatt cttaaagata ttctatctgg ttctggagtt tcttctaagg cctctgtttc 5220
tcctgcatta agtaggccgc cagaacaggt atagtataac atcagaactt ttcttttggt 5280
cattcatgtg tagttttatc tcgtaatgtt cattaaacag acactgacct taaatccatc 5340
attattctaa ttcttggttt caaatatata gatatttgag attgtaaacc tcacgaatga 5400
gcttctgcct ccattgccac atggaacaat ttctctccct atcatctcca acatgttttt 5460
gaaagggccc attgtaaaga agtctcctgc tggtagctct ggaaaacaag aagacacaaa 5520
tggaaatgtt cctgagatat cggctcgtga gaaactatta aatgatcagc ctgaactact 5580
taagcaattt gcgatggatc tccttccagt tttaatacag gttgatattt gtgcatcaat 5640
tgcttaaact ttgcttgata aatttgttaa attgaaaaaa atgttctgat aaattgctcc 5700
tttgcttcct tatttgtctt ctttggttaa ttgatgatat tggcttgctg ttgatataga 5760
tatatggttc tagtgtcaat ggtcctgttc ggcacaaatg cctttctgtc attggaaaat 5820
tgatgtattt cagcacagca gagatgatcc agtctttgtt gagtgtgaca aatatatcaa 5880
ggtatgttga aatttaattg agttgatatt gcttgatacc ctcactgatt ttatgggttt 5940
agaaaattta tgcattgttg ctttgatcat atagtttctt agctggcgtc ttagcatgga 6000
aagatccaca tgttttgctt cctgccttga aaattgctga aattcttatg gaaaagcttc 6060
ctgggacatt ctccaagatg ttcattagag aaggtgttgt gcatgcagtg gaccaactta 6120
ttttaactgg aaattcgacc aatatctcta cacaggcatc ttctgccgag aaggataatg 6180
attctatatc tggagcatca tctcgctcta ggcgttatcg gcgacgcagt gggaattcca 6240
atcctgatgg aaatcctttg gacgatttga aaactccagt ttcagtaaat gttggttcac 6300
ctcctaattc tgtggatatg ccaacagtaa attccagtat tcggttatct gttagtacag 6360
ctgccaaagc ttttaaagat aagtattttc cttcagatcc tggggctgct gaagtgggta 6420
ttactgatga tcttttgcat ctgaaaaatc tttgcatgaa gttaaatgct ggtgctgatg 6480
aacaaaggac caatggaaag gggaaatcta aaacttctgg atttggtctg gaagagtatt 6540
taattgggat catagctgac atgctaaagg aacttggcaa aggagatggg gtatctactt 6600
ttgaattcat tggtagtggt gttgttgcag ctctgttgaa ttatttttct tgtgggtatt 6660
tctctaaaga tcgaccatta gaaacccacc ttcccaagct tcgccaacaa gcacttacaa 6720
ggtttaagtt atttatagct gttgcactac cttctactac tgaagatggg actgtggctc 6780
ctatgactgt cttggtccag aagcttcaaa atgccttgtc ctccttggag cgtttccctg 6840
ttgtgctgag tcattcatct aggtcatcta gtgggagtgc acgcctctcc tctggactaa 6900
gtgcattatc tcagcccttc aagttgcggc tttgtcgagc ccagggtgaa aggtcactta 6960
gggattattc atccaatgtt gtactggttg atccattagc aagtttagca gcaattgagg 7020
aatttgtttg gcctcgtatc caacgaagtg aattgggtca gaagtccact gtacctgctg 7080
ggaattctga atctgggaca actcctacag gagctggtgt atcctctcca actacccatc 7140
gccattctac tagatccaga tcatctgtta atataggtga tacatctaga aaggaaataa 7200
gtcaagataa aagcacaagc tcttccaagg gtaagggaaa agctgtatta aagcctgcac 7260
aagaggaggc aagaggacct cagaccagaa atgctactcg cagaagagaa gctcttgata 7320
aagatgctca aataaagcct gtaaacggtg actctacttc tgaggtatac tgatggaaac 7380
cgagttaggg gcagatatga cagttatctg tagaaaataa ctgctttaga aatcaattct 7440
gtataagaaa ctcaaataac tgtcttagaa aacagtactg tattagaaac tgcaggatgg 7500
tagttatgtg tagaaaataa ctgctggtag ttatgtatta ggagtatatg tctgttagta 7560
tgtggtgtaa ttgggcagtg taaagtcatt tttaagtttg tggatagaga atactctgta 7620
ggagacttct gtctctggga cagctagtgc tgttttttgt atatgatagg agactttagt 7680
ctctaggacg gcaggtgtat tgctgtattt gtatgtgata ggagactttt gtctctggga 7740
cagcaagtgt attgctgtat tgtgtgattt cagtgttcta tatattttag gtaaatgcta 7800
gggctctggt tgaggaagta aaaagaggga tgcgtaaaaa tgtactcctc ctatgatttc 7860
caaaaaaaaa cttttctttt ttaattcctt aaccagtgcc tggcactggt tagcaagacc 7920
ctatatttta ttactttatc atttggtgtt ctatcatata ctgtgaaatc ctagcgatga 7980
ctttacaatg cttcaacttt tttcttctgt taatttataa cttccttctg gctatgtatg 8040
acctgactat gaatgctctg ttttcatgtt ggcttataaa gtgaaatacg aagagaatac 8100
ttgataatgc caatataaga tgtaaagatg cattattaca tttttcattg caagcttgaa 8160
aagacatctt acatttctct gtatctgtaa ctttggacat gctggatttt gttgtctgta 8220
gatccttaaa atgttacctc tgccatttag ttttataaat ggtttttgat tataattatc 8280
tatttaagat atcatctcta atatgagaaa cactgcctag gttttctttg gattatgttg 8340
aacagttgct tttttgcacc attgatttct ttacatacat taaaattaag ctgttgtaag 8400
gtttccctgc aaaagtgttg atttactaaa aaattgaggc agagctagca ctgagaggat 8460
aacttatcta ttttgcgttg aagtaacatt gctgtaagta atagtgtatt tggatgtaat 8520
cagatggata ctttgtatat gctgatcctc atccttcttt cattaagtgc atgtgtcaga 8580
atttttagta tggtaccact caaaaagtca aaatcgattc ctattttcag tttagcttag 8640
tttttaatta ttttcttata acattaaact tgtctgtcat gaagttgtag tttgtatgac 8700
tctcattggg gaatatttta gattatttgt gggcgggtta catatttttt ccgtttaggt 8760
gaagctttct tgttttgttt tttgttgctt ttgattatgg cagtgtttac atgcatgttg 8820
tttagctagg ttcttgtggt tgttaaactg gtggaagcat ctttgagtat aaattttttt 8880
tttggaaggc gtctttgagt ataaatatta gtcatttgtt tttcctgcta tgctttggac 8940
taaatcatga acctaatcca agtatcttga agtagtcatt tgtttttcct tttctttttt 9000
tacccttcta gtgacatttt ccaatgtcta cattgtagga tgaagatttg gatatatccc 9060
ctgttgagat tgatgaggca ttggtgattg aagatgatga tatttctgat gatgaagatg 9120
atgaccatga tgatgtatgt tatctactgt ttcttcttct ttggctagga ttttcttact 9180
tccttggtga tgagtatctc atttaagtaa ttactgtgtt tctgtctttt tttttttttt 9240
gcttttctga catcatttcc ttttttaaca tggcttttaa atatgttaaa atctgtgtgc 9300
atctttattt ttttatttcc agtgctggtg tccttcaatc ttgtttacaa ttttttcaaa 9360
atgagttgct gctgtctttc tatcatttct ttgttttcta tttttctttt ctgggtataa 9420
tgatgcaatg aagttttggt ttatgacctt atgcaaacta tccataatcc aagtctctga 9480
ccaatagctc aatcctggtg ttttatttcc aaatttttaa ttttagctac catcattatt 9540
gttaaccaag attgaacatt ataattaaat tcagttgtat ctattaaaat ttttgtgctt 9600
gttttgtttt ttctgatgca caggtactga gggatgattc tcttcctgtt tgttcacctg 9660
acaaagtaca tgatgtgaaa ttgggcgaca tagtggagga gagtaatgtt gctcctgcaa 9720
ctagtgatgg tggccagact aatgctgcct caggttctag tagcaaagct ggtacagtca 9780
ggggatcaga ctctgctgat tttaggagtg gctatacctc aagctcaaga ggtgcaatgt 9840
catttgctgc tgctgctatg gctggacttg gatctgccaa tagcagaggt atcaggggtg 9900
gaagagatcg actagggcgt ccattgtttg gtagttctaa tgatcctcca aagttgatat 9960
ttactgctgg tgggaagcag cttaataggc atttgactat ttatcaggca attcaaaggc 10020
agcttgtgct agatgaagat gatgaggaga gatttgctgg cagtagtgac tatgtatcca 10080
gtgatggaag caggttgtgg ggtgatattt atactataac ttatcagagg gcagagaacc 10140
agacagatag gactccccct ggaggttcaa cctctaatgc ttcaaaatct ggcaaatctg 10200
ggtctgtatt gaattctagt tctgaagaca agctaaatca gacatctgta ttagatagta 10260
ttttgcaggg agaattgccc tgtgaactgg agaaatctaa tcctacatac aatatattgg 10320
cattattgcg ggtgcttgag ggtttgaacc aacttgcatc tcgtttgagg gcccaagtgg 10380
ttactgatag ctttgcagag ggaaaaattt tggatttagt tgagctaagt tttaccagtg 10440
gtgctagggt tcctacagag gaatttataa gcagcaaact tactccaaaa ttagctaggc 10500
aaatacaaga tgcccttgcc ttatgtagtg ggagtcttcc ctcatggtgt taccagttat 10560
ctaaagcgtg cccttttttg tttccttttg agacccggcg acagtatttt tattcaactg 10620
cctttgggtt atctcgtgca ttgtatcgcc ttcagcagca gcagggtgct gatggtcatg 10680
gatcaacaaa tgaaagagag gtcagggttg ggagattgca gcgtcaaaag gttcgtgtct 10740
ctcgaaatcg cattttggat tctgctgcta aggtgatgga gttgtattct agtcaaaagg 10800
ctgtacttga agtagaatat tttggtgaag ttggcaccgg tctgggtccc actctggagt 10860
tctacacact tctcagtcat gacttacaaa aagttgtact tcaaatgtgg agatcaggtt 10920
cttcagagaa atatcaaatg gaaattgatg gagatgaaaa gaaaatgaaa aatagtgaag 10980
gctcttttgt tggagatgga gaactcgttc aagctcctct tgggctgttt cctcgacctt 11040
ggcctgcaaa tgctgatgca tcagagggta cccaaatttt caaagtgatt gaatatttcc 11100
gattattagg ccgtgtaatg gctaaagctc ttcaagatgg acgcttattg gatttaccat 11160
tgtcagtggc attttataag cttgttcttg gtcaagtaag ttatgaaatg ttgatgtctt 11220
gtctgatttc atgtgtatct taaggttgat ttttagtctc tatatattta gcctttgata 11280
tattgcagga gcttgatttg catgacattc ttttcattga tgctgaactt gggaaaactt 11340
tgcaagagtt aaatgccctt gtttgccgga aatgttttat agaatctatt ggtggtagct 11400
acactgatac ctttgctaat ttgcattttc gtggggcccc aatagaagat ctctgcttgg 11460
acttcacact tcctggttat ccagagtaca tcttgaaacc tggagatgaa attgtatgta 11520
ttcagtctgt tttttttacc tggtttttgt tttggttctg attctgtctg taataaaaat 11580
tgctttgaac ttactgtcaa actttcaggt tgacatcaat aatctagagg agtacatatc 11640
catggtggtt gaggcaacgg ttaagactgg aatcatgcgt caaatggaag cttttagagc 11700
agggtttaat caggttatat gttgtctcaa taaattcatg taactttgtc tttgactgtg 11760
catcttgttt ggtgatgctg agtataaaaa atatcatgta ttttttaact gattaatggt 11820
tcattctttt tggtattcct tttctagttt ctctcaaaca attttattga aaactaaact 11880
tgactggggt ttaatttgaa aatattgagt atggattttt cagcttttag attcttaagg 11940
ggcattgttt tctactaaaa attgttactt ttggttatgt cttgagcagt gaactgtata 12000
tatatcctga atctcggatg tatcaattaa gaaattacta aatgtttgtt tctgactttt 12060
acttatgttt gctaccaacc ttgtatcccc cttcccctgc agatgaagaa tggaagttag 12120
aaaatacaca ttttttttgt attgatgtca aatattcagt tatttaatgt caaaatttta 12180
caagtgaaat gagctactag cctaacttat atggaagaga tgggctgggc aatagtttga 12240
acttggaaca actagttgga taatttgttc atttgcttga gtccaaaaac taaacatttg 12300
tcactttccc acttgttctt gtcaattcag gtttttgaca tctcatcttt acaaattttt 12360
tctccccaag aactggatta cttgctttgc ggccggagag aattgtggaa ggtattcttt 12420
tttatacaaa agtattactg ctgcttacaa caatctttta gatgttacca tggataatgt 12480
agttataatt ttttttctcc tatctgcaga ctgagacact agctgatcat ataaaatttg 12540
accatggtta tactgccaag agccctgcca tagttaatgt atgttttttt attcctgtag 12600
aaggacaatt gtgtttttgg aaatttaggc ttgttatatt tggtgctgaa cctgtatgat 12660
gctattttca gttactcgaa attatgggag aattcacacc agagcagcag cgtgccttct 12720
gtcaatttgt tactggtgca cctaggctgc ctcctggtgg actggcagtt ctaaatccaa 12780
aattaacgat tgtgaggaag gtattgaaaa atatttttga tcacttgcaa cctgtgttat 12840
tcattcatgc cttcatgcaa ttttgtactt gatatcttga atgttaaagt tttttggggg 12900
gcgaggatct atttgaactt cggtagcaag atgtgtctgg atttactgcc tgacatatgt 12960
tgctccacta ttcctttacc tcttgaaggg gggttttcaa aatgcaatgt tagtaagtga 13020
ttacatttac atgtctgggt gcagctttcg tcaagtgcag ctaatgcttc atctaacggg 13080
aatgggcctt cagaattagc agatgatgac ttgccaagtg tgatgacgtg tgcaaattac 13140
ctgaagcttc ctccttattc taccaaggta gaacactgca aagcattgtt ggttatatga 13200
tcatgcatgt caaagtgtct tttgatcttt gatttccatt ttaaaacagg aaattatgta 13260
caagaagcta ctctatgcaa tcagtgaagg ccagggatcc tttgatttat catgagttct 13320
ggaactaccc aacccggccc tgtatgttaa tatgatgatg aatcagggtt tactcgagac 13380
ttaattattt tttggttgca tgtactttcc tccctgcttc tcaaaataat ggcctctctc 13440
gggtcacgca catcaaaggt ggtgacatgg ttcaggtgag tttaccattg ttgtgaaacc 13500
tcactatgat tcttgacatt actagaccat tgttgtgagc ttaccaagag cctcttatta 13560
ttttgttcat tttcatttgt tggtaaacta aattaatatt aattctttga ttattggcag 13620
ttgaatcaga tcgcctcgtt tttaaaggtg taaaggaatt tgcggagctc gttttagctg 13680
ttacatgtaa atagaatagg aatgttgagt aattccctcc caaatttttc atcattctgt 13740
tttcaactca aagtattggc agttagttat catacctgta tggaagatgg taaaaggaat 13800
agaaataata tcgtatacta tgctttatga tttcatgtta gatctgttac gagcttttgt 13860
tgaggtaaaa tgcgaattat tctatgcttg tttgctatga ttgtcccaaa atgaggcgtt 13920
ctagcataaa taagtactgt ttgagttacc gattaattaa tttgttggtc gctaatgagt 13980
cgaggatgcg ttgtatgttt tgatctgaaa aagacgtccc aactccccag tctaagacac 14040
agaccaggaa ccaacctacc acccattctt tggtgtaaat ggcagtctat ctccaaaatc 14100
aataattctg ctactttgag ttgagtaccc atagcacttg tgattttgaa aacagtctca 14160
tacccatacc tcttatttcc aaaaggtgta attattgtgc acactttttt ttttcattct 14220
gaatgccaca aaaaagtttg atatataaag aaggttttag aaattatata agataaaatg 14280
tgttacaatc tcattaaatc aattcacatt tttgatccaa ttaatttttc atcctgattt 14340
attgttttat aatatgtagt ttgtgcttca gattatattt ttagtatgat tttaaatttt 14400
acaattttga ataaaaatat aatatgttaa gattatatat tgaatcaata tgaccgagtc 14460
ataataataa aataattata tatcactgga aattaaaaat gctgtacgta taataaatga 14520
tctctaaaaa gttattgctg taattctaag aataatgaag ttgaagaaat ttaatgtttg 14580
aattaaaatt tgaaaagtat ttttatggat tctaatgcat gattatgaac ttcaatgcat 14640
caaatgaaga agagaagttt ccgaataaaa gagagaataa aaaaataata gatataaaaa 14700
gttattaatg gtgttttata tatatgggtg ttcatatatt atctctatac acaattaaca 14760
ttaactcaca aatcattagg actataacag caggtgaaat gtttgataat ttggaggaac 14820
ttaaagattt aaacttcatt atcatatatt attatatttc aaaagggaaa aacaacctat 14880
aacttttttc ttattaatta tctagttcaa ataaaactaa cttagttctt attaattatc 14940
tagttctaat ttaccaaaaa gaatggacca tttgtttaat ttgctaaatt caatatgaac 15000
aagtctattt gaaccgtttc agctcactac aacctaacaa caacaacaac aacaacttgg 15060
aattagttat ctttgtgttg catttaaatt gtgttctaat tatttcctgg attgatacat 15120
gtttgatacg tagtagaaaa tttaaaattg atagtgtttt tatatgatcc aaacatattg 15180
gcacatcgtt aaataaaagt taaactttta agtgattgta gaatttccac tttgggttct 15240
aaaagtggta atttatgtta attttaaaac ttgctagtga attaatactt ttaaagatta 15300
aaataacaca agttgtaaat ttgaaggatt aaaatggtca ttgagtagtt gtcggaaaaa 15360
atggtcaatg agcttaaata tatcttgacg atggatcaag attgttttgt gctttttata 15420
taataaacta cactatatat atatatatat atatatattt attaaagaga ctatgttcta 15480
tctctttccc attaagaaac aaacaaaaca aagcaccgct acttggtaca aaacacagta 15540
ttctgtacaa atagtgagta ataagtttgg aaaaaagaag gaacaataat catacatgga 15600
gtgaaggaac aatgatagaa aggaatatga aacaattatt ccaagaaatt gacaagttgc 15660
cggggaacat taaacaatta tttggggtta agcattttcg acatcttatc tgataaaata 15720
tttctattgc atgaacgcgt aagatctcca atacctcaca aaatgaaaat gtacgaatat 15780
tggacaagta atcttatcga aagttatttg taagtgaaac atcttgaaat cttataccaa 15840
acatgaaaat tcaattcatc attttcatat gtaaggattc ttattgagca acgtaacaag 15900
tacctaagga aacccaacaa aattaagtgg aaattagtgg acttctaaca agaaacccaa 15960
gggtaataat acaaactacc aaaagttatt caatcgtgtg cattgttaac taataatgag 16020
gagtgtgagg ttagtacttt ttgtcatatg ttagcgagtt ttaaagagtt tataactttt 16080
tttgttttgg tgataaacgg aataattaat gaactctttt gaattcaaat gaaccaagaa 16140
a 16141
<210> 74
<211> 12233
<212> DNA
<213> 大豆
<400> 74
tactatgcga tttagttaat ctttgacttt cttgtttgat atccaatttt aaaagtcagt 60
ttacatttca taatgaataa aatcaaaata agatatttaa aaaataaaac atgttactgt 120
attattttaa catattttta aaataatttt aagttaaaaa atgatttaat attttaagaa 180
aacaaaaata ctattaaaca tgtaaatcaa ttcacataaa attatttgag tttaattaat 240
attctttaat tcgagattta tacatatgaa cacatgatta aacatctact atcataaata 300
ctatcttaat acccgttata tttgtagaaa attatttact tgtttagatg aacgaaatat 360
tttttttttc agagacgaat ggaagaatga gttaaaataa acaaattata taatttttat 420
aaagaaaaaa atatatttaa aaattaaaca tgttactaca tcatttctaa caattttttt 480
tatattttta tgttaaaaaa gatttaatat ttcagaaaaa aaacactgaa tcatgtaaaa 540
aaattgacat gaaattgttt aagtttattc atcttttttc cttttcatca aaccaaataa 600
taagtgagag aaggagtgga gagttagcaa tttggattat ttatattttg ttttttaaat 660
aaaaaatcat atttaatatt tttgtgtcta ttttaattta tttatcttta atcaaacaat 720
cttcattttc atcatatttt ttacctatta acttttgttt tctccccttt atttctcaca 780
actaaaacaa acaaggagag attaaattaa attgataaat aaaagataca gtgttgctgc 840
ttttgttgag gttgagaggg caagtattta atatataaaa aatgggtgat atgaaatgta 900
ataattttca atgagctgaa aacgacgcgg agggagcaag cgagttgtga cttctgactc 960
agtgttttag agtgacgagt ggatgtatgt gcaaacaacg tatataaggc gtatttgata 1020
tatctctgtc tttcttgttc cttttttatt ccctctccct ctcaccttag ggttttcatg 1080
caacaatttc ctcctctctc tgtacactat attgggaatt ttcctttgat ttagggtttt 1140
cgccgatttt aatttaatac atttaatctg agatgcaagg gattaatcca atcgatcgat 1200
ctaggatagg tgagacaaat tgaagggtga ttctgggttt tgtatggaaa ctcgtagtcg 1260
taagcgggcg gaggcttcct cagctgcccc ttcatcatcc tcgtccacca ccaccaccac 1320
cgcctctcgt tccgccaagt gctctcgtct ttcttcttct tcttcttcca tcccaaacac 1380
aactactgct aatacacgtt ctcgttccgc caggaacaac aacaacaact ccgtttctcc 1440
catggacccc accaatgaat cctccggttc cagacgtgat cgccgcggca agaatttcga 1500
tagggacaat tcggacaaag ggaaggagaa agaacatgat gttaggatta gggatgcgga 1560
gcgggagcga gccttggcgt tgaacttgga ggctgaagat gttggggatg acgacgatga 1620
taatgatagt gagggcggtg tcggaattct gcaccagaat ttgatttctg ccagtacctt 1680
tcgagggctt cttcggaaac ttggtgctgg tttggatgat ttgcttcctg ctacggctat 1740
gggcggttct gtgccctctt ctcaccagac tggcggactc aagcagatat tgtctggttt 1800
gcgtgccgat ggggaagaag gtcgtcaggt tgaggcattg acgcatcttt gtgacatgct 1860
ctccattggc actgaagatt cattaagtac attttcggtt gattcatttg ttcctgtgct 1920
agtgggcttg cttaatcatg agagcaatcc cgatgtcatg cttcttgcgg ccagggcgct 1980
aacccattta tgtgatgtgc ttccttcatc ctgtgctgct gttgtgcatt atggtgcagt 2040
ctctatcttc tgtgcgaggc tgcttaccat agagtatatg gacttggctg agcaggttat 2100
tttctcactc aatctctttt atgcctttag ttattatgta tctggaattc tttttagaat 2160
tttgtcttac tttgaattct gatagatgtg attgttgatg tgagcagtct cttcaagcac 2220
taaagaagat ttctcaggag cacccaactg cctgtcttcg agctggagct ctgatggctg 2280
ttctttctta cttggacttc ttctcgacag gagttcaggt aactcatcaa tgaaatccta 2340
atatcaaagg gtgatgattc attttgcatt tgccaccctt gattggcaag cctatattga 2400
cacaatcatg tttcagcggg tggcattgtc tactgctgca aatatgtgca agaagcttcc 2460
tcctgatgca gctgactttg tgatggaagc tgttccactt ttgacaaacc tccttcagta 2520
ccacgactcc aaggtaaggc caaggccatg tttgcttgtc ataaaaatga aaactgcact 2580
ttccttgtca ttccttccct ttaaggaaat tttgtttatg taaatattga ttcctacaat 2640
gttgtcaatt atctatttga gtcttcataa tattcaatgt taatttttgt aggtcctgga 2700
acatgcctct gtttgtttga cacgaatagc taaagcgttt gcatcatctc cagacaaatt 2760
agatgaattg tgcaatcatg gactggtaac acaagctgcc tctctcattt ctaccagcgg 2820
ttctggaggt gggcaggctt ctctcagcac cccaacatat actgtaagtg caatctttgc 2880
cactaaatct gatattttta tcctttggtg ggttgttcat ttgacttcat tggtgcttgc 2940
agggtttgat ccgccttctt tccacatgtg caagtgggtc tcctcttgga gctaaaacgt 3000
tgcttctcca tggagctagt ggcatactta aagatatact atccggttcc ggtgtttctt 3060
ctaacacctc tgtttcgcct gcattgagta ggccagcgga tcaggtatga tgtgcttttg 3120
agttcttttg gtctattaga tgtagttggt atctctatag ttcatttgat agactgtgac 3180
attgatctca tcatttaaat ctcagaaaac ttttcagttc tctttttgtg aaagatgagt 3240
ttatcctaat tctcttttct aatatttaga tatttgagat tgtgaacctg gcaaatgaac 3300
ttctgcctcc attgcctcaa ggaaccattt ctcttcctgt cagctccaac ttgtttgtga 3360
aagagtctgt tgtgaaaaaa tctcctcctt ctgggaatcc cgggatacaa gaagacacaa 3420
atggaaatgt tcatgaaata tcagctcgtg caaaattatt aaatgataag cctgagttac 3480
ttaagcaatt tgggatggat ctcctcccag ttttaatgca ggttcaatgt ttaaatttac 3540
tcatcgttaa aatgctcaaa ttgtattttg tgatttgttt ataaattttt aattttttaa 3600
ataaataacc agctctattt tttatatcct tgacttcttc attccttcac tcctgtgact 3660
tattaatagt tttgtcttac ttttgaagta gatatatggt gctagcgtca atggtccagt 3720
tcggcacaaa tgtctttctg tcattggaaa attgatgtat ttcagcacag ctgagatgat 3780
ccagtctttg ttgagtgtaa caaatatatc aaggtattta gaacttcaat tgggttgctg 3840
ttgctctatg ttctctgtag aaaacttatg cattgtcact ttgatgatat agtttcttag 3900
ctggtgtgtt agcatggaaa gatccacatg ttttggttcc tgccttgcaa atttcagaaa 3960
ttcttatgga aaagcttcct ggaattttct ctaaaatgtt tgtcagagaa ggtgtggttc 4020
atgcagttga ccaacttatt ttggctggaa atgcaactaa tatatctaca caaacatcat 4080
ctgctgagaa ggatactgat tctgtatctg gaacttcatc tcgctctaga cgctatcgcc 4140
tgcgcagtgg taattcaaat cccgatgcga accgttcgga tgatttgaag agtccagttc 4200
cagtaaatgt tggtttgcca ccaagttctg tggaaactcc aacaactaat tctagtatcc 4260
gtgcatccat tagctcagtt gctaatgctt ttaaagacaa gtactttcct tctgatcctg 4320
ggtctgttga agtgggtgtt agtgatgatc ttttgcatct gaaaaatcta tgctcgaagt 4380
tgaacactgg tgttgatgac caaagaagta aggccaaggg aaaagttgaa gcttctggat 4440
ttgatctgga tgatgattct actaacacag aagagtattt gattggggtg atatctgaca 4500
tgctaaagga acttggcaaa ggagatagtg tatctacttt tgaatttatc ggtagtggtg 4560
ttgttgaagc cttgctaaat tatttttctt gtgggtattt ctctaaagat cgaatatcag 4620
aaaccaatct ccccaagctt cgccaacagg cacttacaag gttcaagtca tttgttgctg 4680
ttgcattacc tttgagcatt gacaatgggg ctgttgctcc tatgactgtc ttggttcaga 4740
agcttcaaaa tgtgttgtcc tccttggagc gtttccctgt aatgctgagt aattcatctc 4800
ggtcatctag tggaagtgga cgtctctcct ctgggctaag tgcattatct cagcccataa 4860
aattacgttt ctgtcgagcc cagggtgaaa agtcacttaa ggattattca tccagtgtgg 4920
tactgattga tccgttagca agtctagcag ccatcgagga atttctatgg gctcgtgtcc 4980
agcgtggtga atctggtcta aagtctactg taggcactga aaattctgaa tctggaacaa 5040
ctcctgcagg ggctggtgtt tcatctcctt cctcttatat tccctccact gcctttcgtt 5100
attcaaccgg atccagatcc agatcatctg ttaatatagg agatacacct agaaaagaaa 5160
tatttcaaga caatggcacg agctcatcta agagcaaggg taaagctgta ttaaaacctg 5220
cgcaggagga agcacgggga ccccaaacaa ggaatgcagt gcgcagaaga gcagctctag 5280
ataaagacgc tcaaatgaaa cctgcaaatg gcgactcaac ttctgaggta tgttgctaaa 5340
ttttgggaat gagtcataat agttaaataa ttttaatcta aattggttta taatactgaa 5400
taatttaatc cttgttgtat tttgtttata cttatactta ttgtccttcc ctctttttgt 5460
tagataaaaa accattggtg tgacttcttc atctaatagt tagttgatga tggcttttat 5520
tttactgctt tagattaaaa tgatgctttt cagttgatgt gtttcctgta gcatcttaaa 5580
cttgagtcgt ggcttttatt gtttaagccg ttttttattg tttgacacta ttttccactg 5640
cctttatggg ataggatgaa gaattggata tatctcctgt tgaaattgat gaggctttgg 5700
tgattgaaga tgatgatatt tctgatgatg aggatgaaga ccgtgaagat gtatgtttct 5760
tcttcttctt cttcttcttc ttctaagatt tcatcttatg gttgttatta cttttattca 5820
catctcattt tgtgatctaa atggtaagat ctcacaatct agctgacaca ttcctttctg 5880
aatcacatgt aaaatcttaa atgagtttgt tgtgagattt aaagatcact gaactagtga 5940
gattgtgcaa tccaaaaatc tgtctaattt cgagttatgt ttgaagtatt ttcttttgtg 6000
gtcttctggt tgtgttatgt atgtgactaa ttatgacttg ttttggatgc tacaagtatg 6060
aaatttcaat tttaagtttg ctataatgtt atctactttt ttctttcata ttttctatgc 6120
atacataaac ctatataatt tgtttgctag gattttatga tctacatttt acgattttgc 6180
aaccttcttt caaactaggt agaatctcga tcttgacaaa attggttgct accaggattg 6240
aaaactatca tttaggtgct tccgttcttg gagttgagtt ttacctaata gaaatgttat 6300
tgttgtttcc aagaatggat gctaccgtta attgttgttt agattggata ctatgatttg 6360
gtttagctgt tttgttctag ctattgaatt ttatcaaata gaaatggtat ttattgtttc 6420
ttatattttt aataatcatt aattgttgat tccaagattg gatgatatcg tttgatttgg 6480
ctgcatccat gctattaatt gaatgttatc aaacagaaat gatattattt cactaaccat 6540
catctaatgc atcatcagct tgctttattt atttatttat tttatttttg tgcaggtacg 6600
gagggattat tatcttcctg tctacttgcc tgacgaagtg catgatgtga aattgggtga 6660
ctcagctgag gagagtactg ttgctcctgc aacaagtgat agccagacta atgcagcttc 6720
aggttctagc agcaaagcgg gtacagccag gggttgtgac tctgctgatt ttaggagtgg 6780
gtattcatct agctcaaggg gtgcaatgtc atttgctgct gctgctatgg ctggacttgg 6840
atatgctaat agcagaggtt tcaggggtgg cagagatagg catgggcgcc tgttgtttgg 6900
tagttctaat gatcctccaa agttgatttt tactgctggt gggaagcatc ttaataggaa 6960
tttgactata tatcaggcaa ttcaaagaca gctcatgcta gatgaagatg atgatgagag 7020
acttgctggc agtgaccgtg tatctagtga tggaagcagc ctgtggggtg atatttacac 7080
catcacttat caaagggcag aaaaccagcc agataaggca tccaatggtg gatcaagttc 7140
aaatacttca aaatctgcca aatctgggtc tgcattaaat tccagctcag aagctaaatt 7200
gcatcagaca tctgttctag acagtatatt gcagggagac ttgccatgtg atctagagaa 7260
atctaatcct acctacaata ttttggcact cctgcgtgtg ctggagggtt tgaaccagct 7320
tgcgcctcat ttgaggaccc aaatggtttc tgatagcttc gccaagggaa aaatcttgga 7380
tttagatgag ctaggtgtta caactggtgc tagggtgctt ccagaggaat ttgtgagtgg 7440
taagcttact ccaaaattgg ctaggcaaat acaagatgcc cttgcactat gcagtggtag 7500
tcttccctta tggtgttgcc agttgactaa agcatgccct ttcttgtttc cttttgacac 7560
ccgacgacag tacttttatt ctaccgcatt tgggttatct cgtgcattgt atcgacttca 7620
gcagcagcaa ggcgctgatg gtcatggatc aacaactgag agggaggtga gagttgggag 7680
attgcagcgc caaaaggttc gtgtctctcg aaatcgtgtc ttggattctg ctgcaaaagt 7740
tatggggatg tattctagcc aaaaagctgt acttgaagta gaatattttg gtgaagttgg 7800
gactggcctg ggtcccaccc ttgagtttta tacaattcta agtcatgatt tgcaacaagt 7860
tggattgcaa atgtggagat cttattcttc agaaaaacat caaatggaaa ttgatagaga 7920
tgaaaagaaa aagaaaagtg atggctctgg gcctaatttg gctggagatg gagaacttgt 7980
tgaagctcct ctggggttgt ttcctcggcc ttggcctaca aattctgatg catcagaggg 8040
tagccggttt tcaaaagtcg ttgagtattt ccggctgcta ggtcgtgtta tggctaaagc 8100
tcttcaagac ggacgacttt tggacctgcc attgtcagtg gcattttata agcttgttct 8160
cggccaagta tgttgtgcaa tattgatgtt ttaaccattt tcatttgtat tgttaatctg 8220
atggtcaatt tcttctagtc tatgtaccta accttcactt tgtggcagga tcttgatttg 8280
catgacattc tgtccattga tgctgagctt gggaagactt tgcaagagtt caatgccctt 8340
gtttgtcgga aacattatat agaatctatt ggtggtagct atacagatac aattgttaac 8400
ttgcattttc atggggtgcc aatcgaagat ctttgcttag attttacact ccctggttat 8460
cctgaataca ccttgaagcc aggagatgaa attgtatgga gttaaactct gagctagtct 8520
ttagatattg gttcttgtgt tacagaaatt tcctttaact tactctcaaa ttctcaggtt 8580
gatatcaaca atttggagga gtatatatcc ttggtggcag atgcaactgt caagactgga 8640
atcatgcggc aaatagaagc atttagagca gggtttaacc aggttctatg ctgttcttaa 8700
taattagtaa tttaacttta aacgtatgca ttgttggcgt ggagcatgtt attattctaa 8760
aacaaggtgt ttccctctcc cctttccatt ggaaaatagc ttttattgac attatcatcc 8820
ttgtctatgc atgctcctat acttaagtcc caccacatcc ctgagaattt gtttttaact 8880
gcacacaatg tagatccatg tagctgacac cacctagtgg gataagatta ttgtagttgt 8940
agattggaat tagtttacct gatgcatgac attaatttgt taacatgtca ttgaaatgct 9000
taagtgacat tctgtctcac taaaaattat ttggtgtttt actgagatct taaatatctt 9060
ctgtttatct ctaatgcttg aatattccac ttgggagctt atatatgctc ctgttctttt 9120
acttacattg gctcccaacc accaaccata ctttccctgc acaaaatgga aattgcaaga 9180
ttcagttatt ggatgttaaa ttttttacat tctgaaatct gaatatattg tttgtgaaat 9240
gggtgggtag ttatggttct ggtagactct cttgtggatg agaagaggat attgatgggg 9300
aaattagtta cttttggctc atttaattta gttgaaaact aaacatcact tgtatgcttg 9360
tgctgtttaa tacaggtttt tgacatctcg tctttacaaa tttttactcc tcaagaacta 9420
gataatttgc tttgcggctg cagggagttg tgggaggtat ttttgcccaa ttttattatt 9480
gcttttaata ttagttaaaa tattagaaca aggcattgaa attacaagtt tttttgttat 9540
ttatcttact gcagtctgag acacttgctg atcatataaa attcgaccat gggtacaatg 9600
caaagagccc tgccattatt aatgtatgct tttttcccct aattataagg aatattgtgt 9660
atgacggaaa attagagcta gttatatatg atgctgttct gttggatgct atttcagtta 9720
cttgaaatta tgggagggtt cacaccagag cagcaacgtg ccttctgtca atttgttact 9780
ggtgcaccta ggctgccacc tggagggctg gcagttctaa atccaaaact aacgattgtg 9840
aggaaggtat tcagaaatgg atttttgata acgtgatatt aggtttattc atcttgtact 9900
ttaataattc ttttccatat taatttcatg gttatatatg catgtttatt ttttgttatt 9960
tccattatat tcatgcctta ttttgggtct tggtgtagct ttcgtcaacc gcagttaata 10020
cttcatctaa tgggaatgga ccttcagaat cagcagatga tgacttgcct agtgtgatga 10080
catgtgctaa ttacctgaaa cttcctcctt actctaccaa ggtactgtga tatgacatac 10140
gagaaatgat ttttacgtca caatgtcttc ctacctttac ttacaccttg ttgggggaaa 10200
atgtgttttt ctaagtttct attgttttta ttctccagaa gggaaatggt gcaatgaaaa 10260
tggtgtccaa ataactgttg catcataaat ttattgatta tattatctct catgtccatg 10320
tttggttgga attaattcta gaaaaaattg cttgttttta taatatctca ttttgaaact 10380
agacaaaact aaacaattat ttctccaaaa ttcatacgta tgttcattac tctgatattg 10440
catcttttgg tattaatcta tgttcatttt ttaacaggaa attatgtaca agaagctact 10500
ctatgcaatc aacgagggcc ggggatcctt tgatttatca tgagtttctg aaactaacca 10560
accttaccct gcatgttaat agatggttca gggtttatag actttattct cgaggctcag 10620
ttatttattt attttggttg gatctgcttt gctcctcgtt tctcgcttct cgaataatag 10680
tctcattttc cgtgctcaat gcaaattgca aggttgtgga attgttcagg tgagtaactt 10740
gttacttttt cccccgaact ggctgggagg ttcttctcat attgttctct tttatttctg 10800
acatacttac aagaatttct cccataaatt gcagttgaat cgggtagcca cattttcgaa 10860
gtgtagggga ctttacggag ctcttaattt aattttacct gttagaactt gtaaatagaa 10920
tagcaatgat gaaattttga ttttcttttt tctttttaaa aaatttcatt attctgtttg 10980
taattccaat tattggcaat tagcatgtac gaaaaaatgg taaagggaaa ataaataatt 11040
taaaatattt tgctttataa tctcgttata ttacttagtt acttgtcatt tggagtttta 11100
ttgtaaaatt ttgtcgacct cattcgctta tgttaagtga atgacttcaa gcattatatt 11160
attgattttc ctacgtttgc agtttttatt tttatgcatt aaactaaagt atgttgttag 11220
aaacgtgact tagatccgtt tgttccaatc ttattatttt gaataaaaaa atggtctagc 11280
gttttaaaaa ataagatttt gtttattgtt tttctaaaag ataaatctta cgagtaacat 11340
tttgaataaa aaataaagtt gtgtaatacc tttttttcaa aataatgatt aaaaaggtgg 11400
atttatattt atatatttag gttttgtttg aattcaaatt gtgtgatgcg tgatatactt 11460
ggtaaatgat taaaaagtga ttgttacgac tcaaccagtt taatttatct agctggtttt 11520
aaaagtagaa tgcagatcaa ataaaaagtt aaaaggctga cttaattttt ttattgtaat 11580
ttagtttggt aattattctt attttaaaaa ttgaattgag cattacactt tcttttatat 11640
caattgaaca ttatactatg ttgtaatgta gttattattt tattaagagt ttgtaacgta 11700
gttattatca gtgtttttat aactacttta tatgaactaa ttgatttttt tattcatcta 11760
tgatagtgat tgtgattttt atactgtgtg aacttatttc attagttatc aaattaaaat 11820
aattttaaaa tttataaaac tttttttaaa aaaaggtaaa aaaaaaaact gttgaattca 11880
tcaatcaaat agtaagagat tttgttttat gaaattatat atatttttgt tagagacaat 11940
tcttcttaac tcagatatta tctcttgcat tagttctttt taaatgttta aagatgtata 12000
ttgatgattt aaatttaaga tatttaatag agataaaata atctcatatc aattgattca 12060
ttcatatggg taataagaat ttgaattagt ttgatttaaa ttttacttta cattgactta 12120
aattacacat tatttttagg gcaaaatgtg tttatccttt taaaaaattg aaattctttt 12180
ttgaaatgtg tcacctttga atcttaaatt aaatgctaac ttgaagatag aat 12233
<210> 75
<211> 12516
<212> DNA
<213> 大豆
<400> 75
tatggttatt actaaggatt taacacaaat gattacgtta ttcttattaa ttaaagttac 60
taccactatt tttttggagt gacactacca cataatttgt ttattcatgt tattatatat 120
ttgaggaaaa gtcataaata aattatataa tgaataaaaa taaaataaaa tatttaaaaa 180
ttaaacatgt tactacatta tctttaacat attttctcta tatttttatg ttacaaaaaa 240
ttgtataata tttaagaaaa caaaaacact attaaacaca tagatcaatt gatttaaaac 300
tattaaagtt taagtaatat tttcgaattc gaattctata tatgttacgg catgaaatac 360
ttgtaggaag aattagatgc ttgtggtctc taaaaaaaac ttccctccct caaaattttc 420
cctatggaat gtgaattttt ttgttgggtg atacaaaaat gattatgttt tttctttacc 480
tctttattta ataaatagta aaattttgat gttctttcct tttatttatc attttttcct 540
ttttattaaa ccaaacaata actgagagga ggaatgggga agtagcaatt tggattattc 600
ttttcttgtt ttttttttaa aataaaaaac ataacataaa ataaaaaatt aattttgatg 660
cattgttagt tgaaagatga ttattattat atattaaaag ttaaataata ataataaata 720
atttatgtta atttttaatc taccaaatta ttttaattta aatattattt tctgttttac 780
tttatttatg ttgaatctac caatatttat ttctccccca ccaccccatt ttttacatca 840
acatagatta aaacggatag aatttgtagt ggaagattaa ataaaattga ttaataaaat 900
atatgtatgt ttctggggga caagtattta aatttaaatt tgataagata tgggtgatat 960
gaaatgtagt aattttcaat gagccgaaaa cgacgcggag gcagcaagcg agttctcact 1020
tagagtgttg ttgttgtgca aacaaagtta agttacgtat aaaaggggta tttgatatct 1080
ctgtcttttt attctctccc tctcgctcct caccttaggg ttttcatgca acaatttcct 1140
cctcctcctc ctctctctct ctctctctac actagattgg ggattttcct tggatttagg 1200
gtttccgccg attttgattt taatttaata catttgtctg agatgcaacg gattaatcca 1260
atcgatcgat cggggatagg tgagacaaat tgaagggtga gtgtgggttt tgtatggaaa 1320
ctcggagtcg gaagcgggcg gaggcttcct cagctgcccc ttcatcctcg tccaccaccc 1380
cttctcgttc agccaagcgc tctcgtcttt cttcttcttc ttcttccatc ctacctgtta 1440
atacacgttc tcgttccgcc aggaataaca acaacaacaa caactccggt tccatttctt 1500
tcatggaccc caccaatgaa tcctccgggt cccgacgtga tcgccgtggc aagaatttcg 1560
atagggaaaa ttcggacaaa gggaaggaga aggaacagga tgttaggatt agggatgcgg 1620
agcgggagcg agagcgagcc ttggcgttaa acatggagag tgaagatgtt ggggatgacg 1680
atgataatga tagtgacggc ggtgtgggaa ttctgcacca gaatttgacg tctgccagta 1740
gtgcccttca agggcttctt cggaaacttg gtgctggttt ggatgatttg cttcctgcta 1800
ctgctatggg cggttctgcg tcatctcctc atcagagtgg cagactcaag aagattttgt 1860
ctggcttgcg tgccgatggg gaagaaggtc gtcaggttga ggcattgacg cagctttgtg 1920
acatgctctc cattggcact gaagattccc tcagcacatt ttcggttgac tcatttgttc 1980
ctgtgctagt gggcttgctt aatcacgaga gcaatcccga tgtcatgctt cttgcggcca 2040
gggcgctaac ccatttatgt gatgtgcttc cttcatcctg tgctgctgtt gtgcattatg 2100
gtgctgtctc tatcttctgt gcgaggctgc tcaccataga gtatatggac ttggctgagc 2160
aggttatttt ctcagtcaat ctcttttgtg cctttactta ttatgtatct gtgtcttacg 2220
atgtatgagc ttcctaagtc cttaaccctt agttgttgaa aatgggcttc cgtcttaaac 2280
gaggttctta tcatttgaca caccttacgg ttttctactt tgaattctga tagttgtgat 2340
tgttgatgtg aacagtctct tcaagcacta aagaagattt ctcaggagca cccaactgcc 2400
tgtcttagag ctggagctct gatggctgtt ctttcttact tggacttctt ctcaacagga 2460
gttcaggtaa ctaatcaatg aaatcctaac atcaaagggt gacaattcat tttgcgttta 2520
ccacccttga ttagcaagcc tatattgaca taatcatgtg tttcagcggg tggcattgtc 2580
tactgctgca aatatgtgca agaagcttcc tcctgatgca gctgactttg tgatggaagc 2640
tgttccactt ttgacaaacc tccttcagta ccacgactcc aaggtaaggc atgtttgctt 2700
gtcatatttg tcataaaatg aaaactgcac tttccttgtc attccttcct tttaaggaaa 2760
ttttgtttgt gtaaatattg atccctacaa ctgatgttgt caattatcta tttgcgtttt 2820
cataataaaa aacttcgtac cccattgtcc agaggctctt cactatgcga aggtatgggg 2880
gagggatgtt gtacgcagcc ttacccttgc attttcataa tattcaatgt taatttttgt 2940
aggtcctgga acatgcctct gtttgtttga cacgaatagc tgaagcattt gcatcatctc 3000
cagacaaatt agatgaattg tgcaatcatg gactggtaac acaagctgcc tctctcattt 3060
ctaccagcag ttctggaggt gggcaggctt ctctcagcac tccaacatat actgtaagtg 3120
caatctttgc cactaaatct gatactttta tcctttggtg ggttgttcct ttgacttcat 3180
tggtgcatgc agggtttgat ccgccttctt tccacatgtg caagtgggtc tcctcttgga 3240
gctaaaacgt tgcttctcct tggaactagt ggcattctta aagatatact atccggttcc 3300
ggtgtttctt ctaacacctc tgtttcgcct gcattgagta ggccagcgga tcaggtatgt 3360
gtacttttga gttctttatg tctgttatat gtagttggta tctctatagt tcatttgata 3420
gattgtgaca tcgatctcat tatttaaatc cttgaaaact tttcagttcc ctttttgtga 3480
aagatgagtt tttcctaatt ctctttccta atatttagat atttgagatt gtgaacctgg 3540
caaatgaact tctgcctcca ttgcctcaag gaaccatttc ccttcctgtc agctccaact 3600
tgtttgtgaa agggtctgtt gtgaaaaaat cctcttctgg caattctggg atacaagaag 3660
acacaaatgg aaatgttcat gagatattgg ctcgtgagaa attattaaat gatcagcctg 3720
agttacttca gcaatttggg atggatctcc tcccagtttt aatgcaggtt caatgcttaa 3780
atttacttaa ttgttaaaat gctcaaatta tattttgtga tttgtttatt aatttttaac 3840
ttcttaaata aataactggc tctatttttt atatcctcga cttcttcatt ccttcactcc 3900
cgtcacttat tactagtttt gtcttgcttt tgaaacagat atatggtgct agcgtcaatg 3960
gtccagttcg gcacaaatgt ctttctgtca ttggaaaatt gatgtatttc agcacagctg 4020
agatgatcca gtctttactg agtgtaacaa atatatcaag gtatctagaa cttcaattgg 4080
gttgctgttg ctctatgttc tctgtagaat acttatgcat tgtcactttg atgatatagt 4140
ttcttagctg gtgtgttagc atggaaagat ccacatgttt tggttcctgc cttgcaaatt 4200
tcggaaattc ttatggaaaa gcttcctgga accttctcta agatgtttgt cagagaaggt 4260
gtggttcatg cagttgacca acttattttg gctggaaatt caaccaatat atccacacaa 4320
acatcatctg ctgagaagga taatgattct gtatctggaa cttcatctcg ctctagacgc 4380
tatcgcctgc gcagtggtaa ttcaaatccc gatgcgaacc cttcagatga tttaaagagt 4440
ccagttccag taaatgttgg tttgccacca agttctgtag aaactccaac aactaattct 4500
agtatccgtg catctgttag ctcagttgct agagctttta aagacaagta ctttccttct 4560
gatcctgggt ctgttgaagt gggtgttagt gatgatcttt tgcatctgaa aaatctatgc 4620
acgaagttga tcactggtgt tgatgaccaa agaagcaagg caaagggaaa agttaaagct 4680
tctggatttg gtctggatga taattctagt aacacagaag agtatttgat tggggtgata 4740
tctgacatgc taaaggaact tggcaaagga gatagtgtat ctacttttga atttatcggt 4800
agtggtgttg ttgaagcctt gctaaattat ttttcttgtg ggtatttctc taaagatcga 4860
atatcagaaa ccaatctccc caagcttcgc caacaggcac tttcaaggtt caagtcattt 4920
gtagctgttg cactaccttt gagcattgac aatggggctg ttgctcctat gactgtctta 4980
gttcagaagc ttcaaaatgc gttggcctcc ttggagcgtt tccctgttat gctgagtaat 5040
tcatctcggt catctagtgg aagtgcacgt ctctcctctg ggctaagtgc attatctcag 5100
cccataaaat tacgtctctg ccgagcccag ggtgaaaagt cacttaggga ttattcatcc 5160
aatgtggtac tgattgatcc attagcaagt ctagcagcca tcgaggaatt tctatgggct 5220
cgtgtccagc gtggtgaatc tggtcagaag tctactgtag gcactgaaaa ttctgaatct 5280
ggaacaactc ctgctggggc aggtgtttca tctccttcct cttatactcc ctccactgcc 5340
catcgtcatt ctactagaac cagatcatct gttaatatag gagatacacc tagaaaagaa 5400
acatctcaag acaaaggaac gagctcatca aagagcaagg gtaaagctgt attaaaacct 5460
gcgcaggagg aagcgcaagg accccaaaca aggaatacag tgcgcagaag agcagctctt 5520
gataaagtcg ctcaaatgaa acctgcaaat ggcgactcaa cttctgaggt atgctgctaa 5580
attctgggaa cgagtcataa taattaaata atattaatct aaattggttt ataacattga 5640
ataatttaat ctttgttgga ttttgtttat acttatgctt attctccttc cctctttttg 5700
ttagataaaa aaccattggt gagatttctt catctaatag ttagttgatg ttggctttta 5760
ttttactgct ttggattaaa atgatgcttt cagttgacgt gtctcctgta gcatcttaaa 5820
cttgagtggt ggcttttatt atttaagtca tttttattgt ttgacattat tttcccctgc 5880
ttttatggga taggatgaag aattggatat atctcctgtt gaaattgctg aggctttggt 5940
gattgaagat gatgatattt ctgatgatga ggatgaagac catgaagatg tatgtttctt 6000
cttctgctaa gatttcatct tattgttgtg attactttta ttcatatctc attttgtgat 6060
ctaaattgta agatctcaca atcttgctgc cactttcctt tctgaatcac ttgtaaaatc 6120
ttaaatgagt tcgttgtgag atttcaagat tgctgaactg gtgagattgt gtaatcatgt 6180
gcagtccaaa aatctgtcaa tttcgagtta tgtttcaagt attttctttt gtgacttctg 6240
gttgtgttat gtatgtgact aattatgact tgttttggat gttatggtta tgaaatttca 6300
aggctaagtt tgctataatg ttatctatat ttttctttca tattttctat gcatacataa 6360
acctatataa ttttttggta ggattttatg atctacattt tacaattttg catccccctt 6420
ccaatcttag gtagaatctc aatcttgaca acattggttg ttaccaatat tgaaaactat 6480
catttaggcg catccattct tgcaattgag ttttaccaaa tagaaatgtt attgttgttt 6540
ccaagaatgg atactaccat taattgttgt ttagattgga tattatgatt tggtttagct 6600
gttctgttct agctattgaa ttttatcaaa tagaaatggt actattgttt cttatatttt 6660
taataatcat taattgatga ttccaagatt ggatgctatc atttgattca gttgcatcca 6720
tgctagtaat tgaatttatg atattattcc actaaccatc atctaatgca tcatcagctt 6780
gttttattta ttttatttta ttttatttta tttttgagca ggtgctgagg gatgattctc 6840
ttcctgtctg cttgcctgac aaagtgcatg atgtgaaatt gggtgactca gctgaggaga 6900
gtactgttgc tccagcaaca agtgatagcc agactaatgc agcctcaggt tcaagcagca 6960
aagctggtac agccaggggt tctgactccg ctgattttag gagtgggttt tcatctagct 7020
caaggggtgc aatgtcattt gctgctgctg ctatggctgg acttggatat gctaatagca 7080
gaggtttcag gggcggcaga gataggcatg ggtgcctgtt gtttggtagt tctaatgatc 7140
ctccgaagtt gatttttact actggtggga agcagcttaa taggaatctg agtatatatc 7200
aggcaattca aagacagctt gtgctagatg aagatgatga tgagagattt gctggcagtg 7260
actatgtatc tggtgatgga agcagtctgt ggggtgatat ttacaccatc acttatcaaa 7320
gggcagaaaa ccagccagat aaggcgtcta ctggaggatc aagttcaaat acttcaaaat 7380
ctgccaaatc tgggtctgcc ttaaattcaa gctcagaagc taaattgcat cagacatctg 7440
ttctagacag tatattgcag ggagaattgc catgtgatct agagaaatct aatcccacct 7500
acaatatttt ggcactcctg cgtgtgctgg agggtttcaa ccaacttgcg cctcgtttga 7560
gggtcctaat ggtttctgat agctttgcca agggaaaaat cttggattta gatgagctat 7620
gtgttacaac tggtgctagg gtgcttctag aggaatttgt aagtggtaag cttactccaa 7680
aattggctag gcaaatacaa gatgcccttg cactatgcag tggtaatctt cccttatggt 7740
gttaccagtt gactaaagcg tgccctttct tgtttccttt tgagacccga cgacagtact 7800
tttattctac cgcatttggg ttatctcgtg cactgtatcg acttcaacag cagcaaggtg 7860
ctgatggcca tggttcaaca actgagaggg aggtgagagt tgggagattg cagcgccaaa 7920
aggttcgtgt ctctcgaaat cgtgtcctgg attctgctgc aaaagttatg gagatgtatt 7980
ctagccaaaa agctgtactt gaagtagaat attttggtga agttggcact ggtctgggtc 8040
ccacccttga gttttataca attctaagtc atgatttgca aaaagttgga ctgcaaatgt 8100
ggagatctta ttcttcagac aaacatcaaa tggaaataga tggagatgaa aagaaaaaga 8160
aaagtgaagg ctctgggcct aatttggctg gagatggaga acttgttcaa gctcctctgg 8220
ggttgtttcc tcggccatgg cctacaaatt ctgatgcttc agagagtagc cagttttcaa 8280
aagtcattga gtatttccgg ctactaggtc gtgttatggc taaagctctt caagacggac 8340
gactactgga cctgccattg tcagtggcat tttataagct tgttctctgc caagtatgtt 8400
gtgcaatatt gatgttttaa ccattttcat tttttattgt taatctgatg gtcaatttat 8460
tctagtcaat gaacctaacc ttcactttgt ggcaggatct tgatttgcat gacattctgt 8520
tcattgatgc tgagcttggg aagactttac aagagttcaa tgcccttgtt tgtcggaaac 8580
attatataga atctattggt ggtagctata cagatacaat tgttaacttg tattttcatg 8640
gtgcaccaat cgaagatctt tgcttagatt ttactctccc tggttatcct gaatacacct 8700
tgaagccagg agatgaaatt gtatggagtt aaaccctgag ctagtctata gatcttggtt 8760
cttctgttat gaaaatttcc tttaacttac tctcaaattc tcaggttgat atcaacaatt 8820
tggaggagta tatatccttg gtgatcgatg caactgtcaa gactggaatc atgcggcaaa 8880
tagaagcatt tagagcaggg tttaaccagg ttttatgctg ttcttaataa ttagtaattt 8940
aactttaaat gtacacattg ttggtgtgga gcatgttatt tttctaaaac aaggtgtttc 9000
cctctcccct ttctagtgga aagtatcttt tattgacatt atcctccttg tctatatatg 9060
ctcccatact tatctcccac tgcatccctg agaatttgtt ttttaaccgt gcttaatggt 9120
gttgttagat ccacgtagct gacatctcct tgtgggataa gattattgta gttgtagatt 9180
ggaattagtt tacctgatgc atgacattaa tttgttaaca tgccattgaa attcttaagt 9240
gacattctgt ctcactaaaa gttatttggt gttttaccga gatcttaaat atctttcctt 9300
tatctctcat gcttgaatat tccacttggg atcttataaa atatatgatc ctgttctttt 9360
acttacattg gctcccaatc accaactata ctttccctgt acaaaatgga aattgcaaga 9420
ttcagttatt ggatgttaaa ctttttacat tctgaatata ttgtttgtga aatgggtggg 9480
gtagttatgg ctctgctaga ctctcttgtg gaggatgaga agaggatatt gatggggaaa 9540
ttagttactt ttggctcatt ttatttagtt caaaactaaa catcacttgt atgcttgtgc 9600
tgtttaatgc aggtttttga tatctcatct ttacaaattt ttactcctca agaactagat 9660
aatttgcttt gcggccgcag ggagttgtgg gaggtgattt ttgcccaatt atattattgc 9720
ttttaatatt agttaaaata ttagatttag aacaaggcat tgaaattaaa agttttttgt 9780
ttttaatctt actgcaggct gagacacttg ctgatcatat aaaattcgac catgggtaca 9840
atgcaaagag ccctgccatt gttaatgtat gtttttttcc tcctaattat aagtaatatt 9900
gtgtattata gaaaattaga gctagttata atttatgatg cttgtcctgt tggatgctat 9960
tttagttact tgaaattatg ggagagttca caccagagca gcaacgtgcc ttctgtcaat 10020
ttgttactgg tgcacctagg ctgccacctg gagggctggc agttctaaat ccaaaactaa 10080
cgattgtgag gaaggtattc agaaatgaat ttttgataac gtgatattta ggcttattca 10140
tctatatttt ttttcatatt aatttcatgg ttatatatgc atgtcaatat ttgttattac 10200
cgttgtattc atgccttatt ttttggcctt ggtgcagctt tcgtcaactg cagttaataa 10260
ttcatctaat gggaatggac cttcagaatc agcagatgat gacttgccta gtgtgatgac 10320
atgtgctaat tacctgaaac ttcctcctta ctctaccaag gtactgtgat atggcatatg 10380
agaaatgatt tttacgtcac aatgtcttcc aacccatcct tacaccttgt ttggggaaaa 10440
tgtgtttttt tctaagtttc tattgttttt attttccaga agggaaaatg gtgcaatgaa 10500
aatggtctcc aaataactgt ggcatcataa aattcttgat tatattatct ctcatgtcca 10560
tgtttggctg ggattaattc tcgggaaaaa attgcttatt tttataataa aagctcattt 10620
tgaaacttgg caaaactaag gaattatttc tcaaaaatac atacgtatgt tcattactct 10680
gatattgcat cttttggtat taatctatga tgttcaattt tcaacaggaa attatgtaca 10740
agaagttgct ctatgcaatc agtgagggcc agggatcctt tgatttatca tgagtttctg 10800
aaactaacca accttaccct gcatgttaat agatggatca gggtttattg actttattct 10860
cgagggtcag ttgttttttt tggttggatg tgctttgctc ctcgcttctc gaataatagt 10920
gtctcatttt ccgtgctcaa tgcaaattgc aaggttgtgg aattattcag gtgagttgac 10980
cttttctgcc agtaaggttg tattgatggg attaatcata tctttctgct gcagcacgtg 11040
aataatttgt tacttttttc cctcaaactg gctgggagct tcttctcatg ttgttctctt 11100
ttatttctga catacttata aggatttctc ccataaattg cagttgaatc gggtcgccac 11160
attttcaaag tgtaggggaa tttacggagc tcttatttga ttttacctgt tagaacttgt 11220
aaatagaata gcaatgatga aattttgatt ttcttttttc tttttaaaaa agttcattat 11280
tctgtttgta attccaatta ttggcaatta gcatgtacga agaaaaatgg taaagggaaa 11340
ggaaataatt taaaatatta tgctttataa tctcattcta ttacttagtc actagtcatt 11400
tgttattgta aaattttgtc aatatcgtca catggggaaa tatcgctaat gttaaagcga 11460
atgacatgaa gcattatatt attgattttc ctacgtttgc agatttatct ttatgcatta 11520
aactaaagta tgttgttaga aacatgactt agatccgttt gtttcaatct tattttggag 11580
gaaaaatgtt ttaattagtt ttttaaaatg agatgttaag gccttgtttg gggtggtttt 11640
tagttttgaa tttttaaaaa taacaattag ttttagtttg aaaaaattag tttttaaaat 11700
atgattagtt tcaaataatt tttttatttt aaagcgtaat attatactaa aattagttta 11760
aaagtgattt tgtgtggtga tgattatagt gcttacgagt ttaatttatt ttttctttaa 11820
agataaaata tttttatttt ttaattttag taaagttgga tttgtattta taaaagatga 11880
tcccgtgatt ataaaaggaa ggagggagga aaaagtgatt gggagtttga attcttccaa 11940
tgacattcaa aatgtgtaat gtatgatatc cttgctaaat gattaaaaag tgattgttag 12000
gactagatat tttaaattgt gaagctagtt ttaaaagtag aaagcatatc aaataaaaag 12060
tttaaaaagc tgtttttttt tttttgcaat ttagtttgat aatttattct tattttgaaa 12120
attgaattga acatatgctt tttttaatat caattcaaca ttatatatgt tgatttttct 12180
tttatatata aattacttta ttttaatcta actcacgact cagttttgac aatgttaata 12240
tgtgattcta tatgcgttag acatttttag ttatggaaaa ttattagtat ataatattga 12300
tttcttgttt ttaaatataa atattaatta ttaatttgtc agtgaaaatg atttaaatat 12360
ataaactctt tttttcctcc ttcttaatca ttaatcatta agttaatctt ataatttctt 12420
aatatttaac attcggtaac atataaacta actgattgtg attggtttgt tatattttat 12480
aaaaagagtt tataatgtag ttataatcag tgtttt 12516
<210> 76
<211> 20852
<212> DNA
<213> 玉米
<220>
<221> misc_feature
<222> (1870)..(1969)
<223> n是a, c, g或t
<400> 76
atgagccgag gccaggggta gcggtgtcag cacggacaga ggcagagttg gctcgaaaag 60
aagactgggt cggccggagc ctggccgggt cgtccgttag cgggaccgac gccagagtta 120
atctgccgag gcctcgggct gggctgatgt cttcggggga cagctggtcg aggcctcggg 180
gtgaccagct gagccgcctg ctcgagccgg attcttggag aagaccctgg cggcgatggc 240
ccgggcgtga tgatgatgtc gtccttcaga gtggagatcc tcggaccgcg tcgccgtccg 300
aggctaggtc ggacctctcc gaaggtgtag ttgacgccga gggtgctgct gctccctcca 360
accgtcaaga tccgagcctg caggatcgga ttatcttgtt gtgtgtgttt tctgcggccg 420
ccgaggccca aacacaccct cgccgtgttg taaagctgcg cttcttttcc tcttgtttcg 480
agtatctgga ctttttcgtc ggtaacagaa ttgtctgtgc gagcgagagt tgcttttcat 540
ggaaggtgat gagtgaggta tccgtatccc ggaggcgtag gaatccttcg gctcggtcgg 600
ccttgccgct tacgcgcact cttgcccgtc catagggttc tgtcaccgac gcagtcaaga 660
aggcccgaag aatcgcttca gcagaggagc tttcgagcgt gaagacttgt tcggtccgcg 720
gaatcactta tccgaacgtg agttacttat cgcagaaggt gatgagtgag gtatctgtat 780
cccggaggcg taggagtccc tcggctcggt cagccttggc tgcttacgtg tactccgtcg 840
ttttcgggat ccactttcga agtagtcgaa aaggacgaaa gacattctag cggaaaagat 900
cttttttcga ggaaaaattt cgacgcagag ggggttcccc ccttttagcc cccgagggag 960
ggtcgggctt tgccgaggca aggctgaccc ttccttgatg actaaacttt gcgtgggaat 1020
gaggtatgcg aacaaattga aagcatctta agggtagaag cgacgtagct gttggatgtt 1080
ccaagcgttg ctgtagacct cgccttgact gttggccagc ttgtacgttc cgagcttcag 1140
aactttggcg atgacgaacg tcacttccca gggaggcgtg agcttgtgcc gccctcgggc 1200
gtcctgtcgc agccgaagca ctaggtcgcc cacctggagg tctcgggacc ggacccctcg 1260
ggcgtggtag catcgcaggg actgctggta ccgcgccgag tgtagtaagg ccatgtcccg 1320
agcctcttcc agctggtcca gcgagtcttc tcgactagct tggttgcttt ggtcggcata 1380
ggccctcgtc ctcgaggaac cgtattctaa gtctgtgggc aagatggcct cggccccata 1440
gactagaaag aacgacgtga agcccgtggc tcggctcggc gtcgtcctca ggctccagac 1500
caccgagggg agttccttca tccatcgctt gccgaacttg ttgaggtcgt tgtagatccg 1560
aggcttgagt ccttgtagaa tcatgtcgtt ggcacgctct acttgcccat tcgtcatggg 1620
gtgagccatg gcggcccagt ccacccggat gtggtgatcc tcgcagaagt ctaggaactt 1680
tctgccgatg aactgggtgc cgttgtcggt gatgatggag ttcgggaccc caaaacgatg 1740
gatgatgttg gtgaagaacg ccacctcctg ttcggacctg atgcgtttag gggtcggacc 1800
tcgatccact tggagaattt gtcgatggcg accaacaggt gccgtgtagc cccgggtgcc 1860
ttctgcaagn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1920
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnna agacaaggag 1980
ttaaggaaca caactttgtt catggtacac cttgttctgg agaaatgcaa caattacctt 2040
tttattttct gtttacacat taccctgtct tgaatgctac agagtttcta ctgattagtt 2100
cagtttatga ctatatggtc ttacttattc tctgatacat ttggcaaagg taaaacttca 2160
gggccaacat aggccatgct ctcccctgat cattttagca actctgcaga atatctgaca 2220
gtattttgag ctatattgct aatgaaacat gtaaatatgc ttttggaaac actgaattta 2280
tcttcaaatc acaaccctct ttgcctccta acatcggctg caagctctgc cactggtttt 2340
tgatagaaca tttttctgct ttctattgtt gtttggatct ctagaaataa tatcttcacg 2400
tttgaaatac tatgcataca gataatagac gcatacgtgt accatctctt tgtatttgta 2460
tgcatgctct tgatggtggc ctaacatata tgctacgttt ttttatttag tcattgcaag 2520
ctcttaaaaa aatatccctg gagcatccaa ctgcgtgctt gcgagctggt gcactaatgg 2580
cagtcctatc ataccttgac tttttctcca caggtgttca ggtaatcact ctctgcctat 2640
tatagacttt tgcatcacaa aatactttgt ttgtttcagc tcatttatct catagcttat 2700
tcttatgcat tacttgtcat tatattgtga cagatgaact gtattattta ttcttgttat 2760
ttatttttat tactaactgt tgtttttgtt gatggaatgt tttcagagag ttgcattatc 2820
tacagctgct aatatgtgta ggaaacttcc ttctgatgca tcagattttg taatggaagc 2880
ggttccactt cttactaatc tactgaacta tcatgattca aaagtacaat tgccattcca 2940
aatgctgata tcctctatga taccatttat gtttcagatg ctgatattcc gttttgctta 3000
ttccaggtac tggagcatgc ttctgtctgc cttacccgta ttgcggaatc tttttcacca 3060
tttccagaaa aattggatga gttgtgcagt catggattgg ttgcacaagc tgctagctta 3120
gtgtctgtta gcaactcagc aggacaggca tccttgagta catcaacata tacagtacgc 3180
tccattttgg ctttttgggt acaattattt gttttacgtt tgtgaagttt atgccatgtt 3240
tttatagggt gtgattcgtc ttctctcaat atgcgcaagt ggatcacctc tggcagctaa 3300
aacacttctc cttcttggaa ttagtggcat acttaaagat atcctttcag gctctgggtt 3360
ggttgctggc acgactgtgt cccctgcttt aacaagacca gctgatcagg tgattttgtc 3420
ttttcatagg cacttcaagg ctctttgggt atcatgtgtg cttgatgtct acatttgttg 3480
tttatcttaa caagctaagg ccccgtttgg cactgctcca cttcacaaaa acaagcgcaa 3540
cttcatgaac ttcaaaatat agcagcccag ctttgaagtt gcagagtttt tgaagagttt 3600
ggcttgtaga ccagctttag tttcataaaa aaatataatt aaaaaaaact tgtttcttct 3660
tcctttggcc gtggcggcca gaggcgcacg tgcgcgactt gggaagacag acacaacgct 3720
gcgatggcct ggcgtggctc tggcggccca gcgcgctgca cgcagcctgg cggcacgttg 3780
tcggccgatg cggccctgct cgctagcgca ggcggtggca cgcgcgctgc cagagtttgg 3840
ttcaggcgtg gctctggcag cggcacatgg cctggcggtg cggctgccgg tcggcacggc 3900
cctgctcgct ggcgtaactg tgtgcggcct gagttccctt gagaacagaa cgaaagaaaa 3960
catttcgtga gggattgcgt aactgatggc aaaggtgggt aatttcaccc caacttaatg 4020
agggagttaa aaatacccat tcgtgaagta tacttttggt tgtttcatgg atttggtgaa 4080
gttagggtca acttcacctt ttttggtgaa gctgggagtg tttggctaga gttttataaa 4140
gctaagctag attttgtgaa gtgaagcagt cccaaacatt ccctaagtgt aaattaatgg 4200
agctaggctt actctatgct tcagatagta cttcaaccat tgattccaat ttccaagcct 4260
caggatatca tctctattaa ggcccccttt ggcacggctc tggctctagc ttcttgcagc 4320
cccttggaag gagccctgcc aaacagccat ctaaaacaca gctcctggca aggagccagg 4380
gccagaaaaa tggtttctcc cagctcctta caaacttgat acaaattatt acaaaactgc 4440
cactggacct gttttgacca aaccttttcc aaaacggctt caactggccc aaattataaa 4500
gccagcacaa ttaatcccat acacttccca accttacaag ccccttaata cagcccatct 4560
cttccttcat tctccataca tatctgaaag tcttccatgt tctctctctt ctaccacttc 4620
gtggcgcagt gaggggggct tacttctagg agcgcctaac atgtgctata taacatcttt 4680
acgcacaatg acataggaac aagtgtcgtc aaagaaaaca aataatattt gtgtatatta 4740
aaacattttt tgatgaattt gatgaaatga aaccaaatcg tcccataaca tgtttttctc 4800
ataatgcatt ttgtttttta atcgccacca ataatatact ttgagcttgt agtcaatgtt 4860
cgtttatggt atgacattgt ttttctttaa tgtgagcata ctcatactac ttcccatgtc 4920
gagcatttgt gaacatgatg ttatcctatg ttctattcat gatagcacaa tttatatttg 4980
tgatgaattg tcatctatgt taaattttaa tgggtactgt aacttgtaaa gctactgact 5040
ttttggtagt aatagattat caccacacaa ttccatatgc aaatgatgcc attatgtgtt 5100
ttttggtatt acacagatga atgagattgt gaagcttgca gatgagttac ttccttctct 5160
gcctgttggg accatttctt taccagtgta ttctggtgtt cacatgaaag gttgttctgt 5220
aaagaaatcc acttctagta agcagggtga acatggttca acagcaaatg aactatcagg 5280
tcgggagaag ttattgcgtg atcagcctga acttctgcag caatttggca tggacctgtt 5340
acctaccatg acacaggtca gtcttgtctt attaatgttc tctttgatgt tgttaatggt 5400
aatgctgatg tggtattacg ggacattgtt gctatcatgt tggtgtgata cttaaggact 5460
tcaatacctt gaacaggtgt atggctctag tgtaagtgga ccaatacggc acaggtgctt 5520
atctgtcatt gggaaattaa tgtactatag ctcagctgag atgatccagt ctctccttag 5580
cacaacaaac atttccaggt gcttaacaca atatatttca tcgagttatc tcatgttatt 5640
agaaactatc ttatcacatg aattccaaat atttgtttca agctattaac ttctatatca 5700
accattgcag ttttttggct ggcattttag cttggaaaga tccacaagtg ttgatccctg 5760
ctcttcagat agcagaagtt ttaatggaaa agcttccaga gatttttttg aagatgtttg 5820
tgagggaagg cgttgttcat gctgttgagt cacttatatg cccagaatta tctggtcagg 5880
tgactcctca tgtggattcc attacatctt cacataatag gcgcaaccgc cgtagaaata 5940
atgctgtgaa cactggaaat aacttgcctg atggaccaaa aggttccaat tctatgattg 6000
ccaattcacc cccaagcatg gctgaagttc caaataatag tcttcgtgct ttagttagta 6060
atcatgcaaa gtcatttaag gataaatatt tcccttccga gcctggctca agtgacattg 6120
cagttaccga tgaccttctt aaactgaggg cactttgtgc aaaactgaat acaacagctg 6180
acactatcaa aacaaaagct aaaggaaaat caaaggtagt gtctgacaat agttttgatg 6240
tcttgtgcaa tattgaggag caattggatg atataatagc tgaaatgttg tctgagctta 6300
gcaagggtga tggtgtttcg acatttgagt tcatcggaag tggagttgtt acggctttgc 6360
taacctatct gtcatgtgga acatttggga gggaaaaggt gtctgaggca aacataccaa 6420
atttgcgtca tcaggcagtc agacgataca aagcatttat atcttttgca cttccaaatg 6480
ataaagatgg gaataaaact cctatggcat tcctagtcca taaactgcaa agtgccctat 6540
cttcgttgga acgtttccca gttgtgctca gccattctgg aagggcatcg actttgggag 6600
gatcccgcct gacaacaggt ttaggatctc tgtcgcagcc catcaaattg cgtctttgtc 6660
gagcgcctgg tgagaagtca cttaaggatt tttcgtccaa tgttgttctt attgactcat 6720
tagctagtct ggcagctgtt gaagattttc tttggcctag agtccagcgc actgaaccag 6780
tattgaagcc tccaatgtca tctgcaaata attctggatc tggagcagca agctccacag 6840
cttgtgcgcc ttcaattcca tcagaaactc aatctgtccg tcgcacatcg ttaagatcaa 6900
agtcatcagc tgccaccagt ggtgcaatta agaaggatta tcaggaggga agcataaaca 6960
cctcaaaggg aaaaggaaaa gctgttctta aattgagttt ggatgaacct aaaggcccac 7020
atactaggaa tgctgcgcgc agaaaagcta cttcagagaa agatgttgaa ctgaagccat 7080
cacatggtca catcacttca gaggttagat actcgtgcat tcaatcttat ttgttatttg 7140
cataatatat gcgaaatggc ctctagccta gtggttaaag gcttccgaat agcacctcca 7200
agtctcgggt tcgattcccc tcaggggcga attttccggc ttcctgtgcg ccaccctccg 7260
gttggaccgc tgtagagggg acggttgacg tcagctcgtt agtgatgggg aggggggggg 7320
gcagagtttg gggattttct tggccgagac cattgtttta gtctatcttg atataatacc 7380
gggatggcgg tcattccttc cctggctgag ttttttttat ttgcataata tatcaattga 7440
gccctcctaa gtgagctccg ttgggaagtt tagctttaat ttctggctct aggacacatt 7500
gattgcaaat gtttttttaa caatgccttt tgtttactca gctgacaaat gttatcatag 7560
taatgataca acagtaatgc agctgattta ttttgcctta agtgtatcta gtttgatcag 7620
tactttgttt tagtatgtaa gcaaaggatt tcaaagggat gttcttcata gacatgatgt 7680
acagattgta taactataaa aggagggggg gcagtgctgc aggctccgac atgaggagtc 7740
tggggaaggg ataaaccgag gcaacccttc tcccgtaaat acggagagac actgctttga 7800
acccacaacc tggtgactcc gggagacaat tcacaccact gacgtacaga ttgtataact 7860
ataagtctat aatatcataa atgtataaca atgaaaatgc actgaactgt ggcatttctg 7920
tgtatcatgc atttataata ttatatctga gataggatat aaaaactgaa gcatttttta 7980
taagagcaac aaagattaaa gtagaaatat gcctgtttca ctaatatttt ataatttagt 8040
catattatgt aatgacctga ctacgctttc gtaacttagt atactggact atattgtgga 8100
tactcacaaa tatgaaatat cagtgtaatc ttttactcta attctctaaa taaaagatat 8160
tgttgtattt tgattttaaa tatctaagtg aagttttcag gggaaacaac tttatgaatt 8220
attatttgga atagtacgtg catgttcttt tataagtttg atatctgaat tttagaaatt 8280
aaaaagtttg aaagccctgc aacatatatt tatattatta tacctgcagg atgaagatct 8340
tgatgcttct cctgttgaga ttgatgatgc tttaattctt gatgatgatg acgaggatgt 8400
cccagatgat gaagatgatg atcatgaggc ggtaattatt ttttaaatct tgcttattac 8460
tacaaaggtt tgtattgagg ggacatgatt tggaactcag tagattttgt tgaaatgatc 8520
aggttctccg aggttctctt ccttcctgtg ttcctgagag agtgcatgat gtgaaattag 8580
gagatgctga tgattctagt gttgcctcat tggcaaatga taaccaggca cagccctcat 8640
ctggttctag tacaaaaaat acttctagca ggggattgga cactgctgaa tttagaagtc 8700
cagccacatt tggttcacga ggtgcaatgt cgtttgctgc agctgcaatg gctggattaa 8760
ctccagtagg tggtcgtgga attagaggta gccgagatcg gaatggcctt ccattgggtg 8820
ctcgtgcaac tgagcattac aacaaattga tatttacagc tgctgggaag cagctgaaca 8880
aacatttgac tgtatatcaa gctgttcaaa gacaagtagt tcatgccgag gatgatgaag 8940
atcggttcgg tggatctgat ttacctgatg atggtaacca cttctgggat gatataaggg 9000
gtgatgtgtt cactataacg tatcagaagg ctgataacac agcggagaag gggtctgttg 9060
gaggttcagc ttcagtgcca aaatcttcca aatcagattc ttgcagaact ttgtctgaaa 9120
aacagcgtac ttctcttctt gatagtattt tgcaaggaga gcttccctgt gatttagaga 9180
aatcaaacca aacctacaat atcttatcac tattacatgt gttggagggt ttgaatcagt 9240
tatcacctcg tctgagactg cagtcagcct gtgatgattt tgctgaagga aaagttgcta 9300
cattaaatgg gctatacgat gttggagcta aggtaccctc aaaggagttt atcaacagta 9360
agatgacccc aaaacttgct cggcaaattc aggatgttct tgcactgtgt agtggcagtt 9420
taccatcttg gtgttatcag ctgacgaaag cttgtccttt tctgtttcct ttcgaaacac 9480
gaaggcaata cttctattcc acagcttttg ggttgtctcg tgcacttcat cgtcttcagc 9540
aacaaccggg caatgataat aacactgctt ttgaaagaga agtcaggatt ggtagattgc 9600
aacgccagaa agtccgtgtt tctcgtaacc gtatcttgga ttctgcagct aaagttatgg 9660
agatgttctc taatcaaaag gctgtcctag aagttgaata ctttggtgaa gttggaactg 9720
gtcttggtcc aactttggag ttttatactc tcttaagccg tgagctgcaa agggttgact 9780
taggattgtg gagatctcat tcttcagata attctgggat gcaaattgat gcgaatactg 9840
atgatttaat aagaagtaaa aatcatgaat cagaatcact tactgagagc aggaacatag 9900
tacaatcacc tcttggatta tttcctcagc cttggccacc tactgctgct gcatcagaag 9960
gtagcaaatt cttcaaagtt gtcgagtatt tccgcttagt tggtcgagtg atggcaaaag 10020
cattgcaaga tggaaggctt ctcgatttgc ctttgtcgac agcattttac aaacttttgc 10080
ttggacaagt aagcatgagg gcctccttga gctagtttta tcttatgttg tcttctaaaa 10140
ccttcctctt ttcggttggc acttaacttt ctgtgccttt tccttattaa tttaccaaat 10200
ttgctttaat aggaacttga tttatatgat atattatctt ttgataccga gttcggaaag 10260
acattgcaag aactgcaaat tctcgttgca cgtaaacaat ttttggactc ctgctctagt 10320
gagagccaaa agatagatct atgtttccgt ggtgctcccg ttgaagattt atatttggac 10380
ttcactcttc cgggctatcc tgaatatgtt ctcaaggaag gtggagagaa tgcagaggta 10440
agttatatga taccttagtt tgtttttttc ttctagtgtt gctgcagatg actcattgtt 10500
atcctgtctt ataggtcaac atttgtaact tagaagagta tatttctttg gttgtggatg 10560
ctactgttaa gaccggcata atgcggcaag tagaggcatt taaggcagga ttcaatcagg 10620
tcttctttct tattttgtga tttgtgggca atttctatta ataattgatc ctagaactaa 10680
ctagcacatt tatttattta ttcagaaaaa aatactcatt gtatatttct cacttcaggt 10740
gtttgacata tcatcactgc aaatattttc tcctcaagaa cttgattatc tgatttgtgg 10800
tcgctgtgaa ctttgggagg tattgctttt gtggtcattc ttgatgttgt acttcctctg 10860
tcccagaatg ataagcatag tttgtatagg aaaaagtcaa atttagaatt ttatttccca 10920
gaaattgctt gtattatact agtttatgtg gctttgtgtc ccgctgccat aggtcaatgt 10980
tagctattga aagctagttg ctctagtcta ctgcttccct ttttttttcg ggcggggggg 11040
gggggggggg gggtattctc cctcctttgc agttcacctt ttttctctct taatgaaatg 11100
atacacaatt ctcttttgtg ttcgagaatt ctttttacaa actttggcca acaattaatc 11160
aaattatata catgttttgg acaaacttgc atcaatagat tcatattcag agtgcatttg 11220
agatgtgatt aattgtttag caaacataaa catattgtac gagaaattaa tggtcaattt 11280
ttttaataga ttatatgagt tattgtactc agtctaagtg ttatggaccc tggtcccgcc 11340
catgtaactc cttttccaat atatgcaacg tagcccaccc tgattggagt taacatggta 11400
gcaagcttgg tcttttctct ccctcccttc cctagccgcc acctcctggc tgccatcact 11460
accgtcgggt cgctattgtc gtctgcgcct ccatctcctc cctagcgcct ttcttgctca 11520
tcggccaccc atatttatta aggcagtgtt aataaccatg gggccaccac cttcacttta 11580
gctttctagg cgcgactggt ggccttgttg gccctacggt tggcgccttg cccaccggtg 11640
ctttagtccc ggtcacctcg attgctgccc tagtcgctcc cgtcgccctt gcctttgctg 11700
tggccatcta tggcatgggt gccgccagcc agggcccaga cctagctgac tctggcctag 11760
cttatctttc ggtcgcgtta cttccccctg atgtcgcttt cgctcactcc accttggcca 11820
tagcccccat tgctgccaag acggtgtttg ctgctgctcg ggatcgggaa catgttgctg 11880
cccttgctta ggggcacaaa cgcaccacga cggatgcact cgctcgatag cagaccaatg 11940
ctgagggtca cctgctcggc tcctcctgca tcgagaccct cccagcgctc tagtggtcgc 12000
ccctttgtcg gagtacgagg tcgagatcat caccaacctc tatgccgagg tggcgtgtgt 12060
ccagaacatt tgctctatga ttcccgtcat ccttgacaca acttcctcca actacgcctt 12120
ctggcgggcg atcttgtctt cactctccat cgctacgcct tggactacca cgtcctcacc 12180
aacactgtct ccctcactaa cctatcctgg cggcagatgg atagtgtggt cctctcgtgg 12240
atccccagga ccgccatcgt tgcgctgcat gacgtggttt gcgagcgtgg tggcactatc 12300
cgtcttggac gtcgtcgagt agttccttgg caatcgtgag gcttgcactc tctaccttga 12360
tgtcgccttt cagacttttg tctaggggaa cctctttgtc actgagtact tttgtcagag 12420
gaagggcatg caagcctccc tctatgacct tggggagccc gtctcccata gcatgctcat 12480
cctcaacctc ctgcatggcc ttcacctcca cacttcgatc acttcctcag ttccgttcca 12540
caaggtcaag gtcaacaacg acctcatttc cgaggagctc accaaggggc tgctaccacc 12600
ctctacaact ccaccactgg ggggccagcg catgcaccct cttctactac tatcttggga 12660
ggatcgtcgt tgcgctcgtc tgttcatcaa atctcaagca ttgcacatcc tccccacttg 12720
ccttggtctg aggggagggg tctagggtgg cggtcgacga aaaggcggtc gtggtggtgg 12780
ccatggggac actccttggc cttccatcta caacctgtgg actgatcgca tctccatgtg 12840
gcctggtcct tctccgagag tcccttctca gcgtatcact ccaccgtagc cagctcactt 12900
gacattgctc ggtctggggt atcaccttgg ccatcctacg ccattagcac ctgagtcacc 12960
acttctgtca ctgccacccc acttcgcttt gtcctggaac ccgtggccag gtgggtggga 13020
tcaacagtct cttgctagct ccttcaacac gatgaccttg actcctccca ctgtcaccta 13080
ctaggtggct gattcctggt gcctcctatc acaccacttt ggacgcgagt atgctatatt 13140
cttatccacc tcttcgtcta ctcctaccat tgtgggtaac gagaacattc tctcggtcac 13200
ctctgtcagt gattcggttc ttcctaggcc ctttcacctt cacaatgttc ttgttgcccc 13260
acatatcctt taatatttta tccgttcacc tattgagcat cggtaattct tgttccatag 13320
attttgatcc ttttggcttg tctgtgaagg atcttgctac ctagagtctt cttgcttgtt 13380
ttgatagatc taggcccctg tacacccatg caagcctgcg tctacctccc cacatactgg 13440
tttcgctaca tcatcacatt cgtcctcgct acagtcgcct actttgacta cttccactat 13500
ttgtagtact tggcatcgtc gacttgtcca tcccaacctt gtagcgtagt ccaagctatg 13560
tagtacttta gtaatttctt gtagtagggg cacccttgag catctatgcc agttgtgtaa 13620
gttaggtcgt cattttcgac ttcctttctc tagcttctca caacatacgc tgtgatttgt 13680
ggacctctcc agctctcctg ttaccaatgt tttgggatat aaatattatt tggtgattct 13740
tgatgattgc tctcattgtt tgtggatttt tccattgagt ttaaagtctg acacctttac 13800
gactctgctc ggttacttcg cttgggttta cacttagttt ggttgcacta tgaagatcgt 13860
ccagtgtgac aatggtcgtg agtttgataa cttctctcga tcctttacct cacttggagt 13920
ctaccttcga atgtcttgcc cctacacttc ccatcagagt ggtaaggctg agtgtatgat 13980
ttgcaccacg aataatgtaa tgtgttcctt gatatttcaa tcatctgttt ccgctcgcta 14040
gtgggatggg agcctccaca ctgctaccta cctccttaat tgtcttccta ctacgacaac 14100
tcttatgtca tgccccccac cccactttgc tcgaatggca aggcttagcg catgaattaa 14160
tgtcatgttt ttcttgctat tttactattt taggcatccg ttccttctcc ttactggtct 14220
gagagcctcc acaccaccac ctacctcttt aattgttttc ctattacagc gactcgtgcc 14280
cccacttccc actttgctct ttttgtcaac actccaatga ccatcttcgt gttttgggta 14340
tgtgtgccac cctaaccttt ccaccactac tctccatctt gttcttcgtt ctgctcgctg 14400
tgtctttctc gggtactcat ctgatcacaa agggtactgg tgtcttgacc tcaccactca 14460
tcgccatttg atctctcatc acgtcatatt cgttgggatg gatgtcccct tctcaactaa 14520
cacctagccc accaccgcct cccctttcga gttgtatttt cttcatgatc ctgactccgt 14580
ggtgcccttt gtccaaccac tgtttttcta gttgctgttt tcctctccac tgtttcagcc 14640
ggtggcccct gtgtgcccat ggtggatcat acacccttga attcctttcg ggttgcaccg 14700
ttgcccctcg tcgacctgtg tgtcacctcg tcaacccatg ttgggtccct aggccccggt 14760
ggccccactt ttgacattgc cttcgttgct agggccctct ttattgccct cgcctgtgcc 14820
ctcgccttgt tgctatgccc aacctgtgca tttctatcag cgacatgccc gagtgggtac 14880
accgcctcgc tcttgcgacg agccgacaat gtatcacccc ttcatcgtga ctctcaccac 14940
atccacttga tggcgaattg ccaggccact ggggtcctta gccctgttga ttgcctcatc 15000
ctctccgtga cctcctcatt gtcggtttct catgtgtcgt cttgcgttga ttgcaccctt 15060
tctaatactc attggcgcca cgctatgaag aaagagtatc tgactatcct tgccaatcac 15120
acctgggttc tggtattctg gtgccccatc tttcgtgcga caatgtggtg attgacaagt 15180
gggtctggac acataagtgg caggcggctg gctcgctggg ctactcagtg tcgtgctgtt 15240
gactacgatg agacgttaat tctcgttgtg aagcctgcta ccgaccgtgc catcctcact 15300
ttggccctct cccattttcg gccaattcat cacttggatg tcaaaaatgc ttttctgcat 15360
gggactctca cttagacttg tctactgctg tcagcctact agctgtgttg atcatgcaca 15420
ccccggtatg gtatgcaagc tcaacaaagt atgcaaactc aacaagtccc tctatggttc 15480
gaggcaagcc ccacacgttt ggtacaaccg cttcaccata tatctgctct ctttgttttg 15540
ttgaggccaa gttagacaca tttctgttca tctatcggtg tggtagtgac actgtatacc 15600
tactcctata tgttgatgac atcgtgctca ctacctcctt tctcatgcac tagatcacca 15660
ccgctcttca gcatgagttc accatgaagg atttaggccc ccttcaccac tttatgggga 15720
ttgctgttga gcatcgcttt gatggcctct tttttcagca gcgacgatac accctggaca 15780
tcctcgagcg tgctggcatt ctggattgca agccttgtgt ggcgtcagtg gacatgtagg 15840
ccaagctctc tggcgtcagt gctccagtca gtggccccat cacctaccat agcctcatcg 15900
gtgctcttca ccatctcacc ttcaccagac ctgacatcgt gtatgccatc caacaagtat 15960
gtctttatat gcatgatccc tgtgacctgt acttgggtgt ggtcaagtgg atccttattg 16020
ttgactatgg tctcctcctt tggcgctcct tcatccatga gctaatcatc tatactgacg 16080
ccgatgggcc gagtgtcctg gacacctgtc ggtatacctc ggcgaacctc atctcctggt 16140
cctccaagcg gtagccagtg gtctcccgtt ccagtgctga cgttgagtac tggctgttgc 16200
taatggggtg gctgaggcga gcaagctgca ccaacttctc cagtagcttc acaacccact 16260
caccatgagt accctggtct actacgacaa cgtcagcatc gtctacctct ctgtcaaccc 16320
ttttcaacac caccgtattg accgttttac tattggggac gtccgcgtat gtttcaacga 16380
gctcgtagtt tgatgacacc ttcactaagg gttttccctt gttcttggag tttcggttgt 16440
tggggcggga gacagaaccg gtcctcaact accaattgta ctcaccactc accaagattc 16500
ctaatgcaga gtaaccgact aggaggtgcg aaggcctatg gcggaagggg caagccgcgg 16560
aggccgaccc caacttcggt ccttcctaac aatcgcgaag gctacttgac tacgcaggga 16620
ccttggccaa acctcgacga gggacaaaca cgttcagcga gggcaggcgg gggctaggac 16680
gcgtcgaaga cccccgaagc accgaacgaa gacccaactg ctaccagccg atgttgaccg 16740
aggcgagatc actcctcttt ggtaggccgt cgtgaggcag tttttctcta aggcccccac 16800
gcagaggcca taagacgagg aatgttggat tccgccaagc ggcggctcag ttgtgggcgt 16860
agactcgcct actccacacg tcagcctttg atgtgggtga gaaagatggt gtaatatgga 16920
atgtagccag gggacctgta attaccccgt tgcacacctg ttgcacggta tatttgtggc 16980
atgtagtagg taaccaaggg cattacgata ttttaggcct tgggcccttg gccgccctat 17040
agatagcccc atcctgtagc tggatgggac acacttgaca agacatttgt gctcccagcc 17100
aattgttttg tcgtgccacc tgtaacaccg ctctcgagtg cttgggcaca ctgttctaag 17160
tcccaacatc ggtccagtct caacatcttc cactactaga gtttcatctg cgggggtggg 17220
ggtaggggtg ttagattctt gtgcgttatt gtactcaatt taagtgttat gggccttggc 17280
ccagcccatg taacccatgc aacacagttt accctaattt gggttaggtt tccaacaatt 17340
ttgtttgact tttccattat caaaacacac ttaggccttg ttcgtttacg tcggattgca 17400
cccggaaacg ttccagctaa tcaaagttta tataaattag agaagcaatc cggctaggaa 17460
tcgttccgac ccaccaatcc gacacaaacg aacaagacct taatatttca cgatgaatac 17520
tcaatttaag tgttatgggc cttggcccag cccatgtaac ccatgcaaca cagttcaccc 17580
taatttgggt taggtttcca acaattttgt ttgacttttc cattatcaaa acacacttaa 17640
taagttaata tttcacgatg aatgtagttg ttttggctta ttctgaacaa tttcacatca 17700
tgcagccgga aacactgcct gaacatataa aattcgacca tggttatacc tctaagagtc 17760
ctgcaattgt taatgtaagt ttccctgctt ctttgtaatt catttgtagg tttcgtaagt 17820
ttatttggat gtttctaatg ttgtgttttg cagtttcttg agatcatggc agaatttact 17880
cctgagcagc aacatgcttt ctgccagttt gtgactggtg ctcctcggct tccacctggg 17940
ggtttggctg ctctaaatcc taagttgacc attgttagga aggtaaattg gctatctttg 18000
ttcttatttg tactttatgg ttgacatgcc tgcccacatt gttgaagttt tgaattcttc 18060
ttaaattcca gcactcttct gtggcaaata atagttcaaa tgcaactgga gcgacagagt 18120
ctgcagatga tgatttgcct agtgtcatga cttgcgccaa ctatcttaaa ctaccaccat 18180
actccaccaa ggtatgcttc tttctgcttt ttggctaact gtggttatat ctcctgtatt 18240
gtcttataaa ttgaggattc agaaacccag tcaccaaaga attacttcat atagccttat 18300
cgtaacaggt aactggacaa attttcaact aaggacgtgg aaactaaaat ttaatgtggg 18360
cagcaccttc cagccactca ttagttagaa ttattatttc gttagtttaa atcaaatagc 18420
atattccaca ttgcttgaac cctttatatt gacgcatgct ggtttttttt ctggaaggaa 18480
atatttacag tgctactcgt gctagtgtgt ggcttgttat ctgtgtctga attttgttga 18540
aaacttcttg caggctatca tgctaaagaa actgctttat gcgatcaacg aaggccaagg 18600
gtcatttgat ctttcgtgaa tctcaacact aacataggta ttggtccacc tagaaatctg 18660
cgtcattgtt acccagagtt agtttctacc tcattcatgt atgacatagg ttaaactcag 18720
ctctccggag tcccaccgaa ggtttggagc ccgtaccttt gggtgtggat gtctatactc 18780
tcttttcttg gttgtatatt cttgcggatc tttatagtga ataatagtaa taaattgttt 18840
tgcgcttctt actgtgctaa tcatcagtgt cccacccgaa gcgtcagccg tcagtcgtac 18900
aaattttgct cgatggtttc gctgcccaca aatcggatgg atgggggccc atgaacaagg 18960
gcacgggatt tccggggcta tctgaaatag tgacgggcat gcaaacacac ccaaggtcca 19020
cggcctgctt ttggtcgaca cagtgccacg cgaccgtgct ggatcttatc actggcctgt 19080
cagaggcatc tgaggatgtc aagttgtcaa ccgaggcgcc tatgtgggca cgggactgat 19140
cactttcact cgggtcacag cgttcggtct tggcgcttca gccacaaccc caaggatgac 19200
gatagctggg cacaaggcca cgacagttgc acagatcccc tcgtctcgtt tgcaggtacc 19260
agcgaaattg ctaacgtgcg atgcgacccg ctgcgaaaac tacggatcac gtatcagcgg 19320
tcgttgtcat atatgatcag tcggccgtgc cttggcactg cacaagccaa taaaactccg 19380
ccagaactga ggaaagatgg aaccgtccag gagttgctaa gccatcgatt gcacggatag 19440
tatggcaagc ggtacatcca gtaggtggtt tgacctaaca ttagtgtctg atcccgttag 19500
aacatctcta actccgtctt aaattaatgc actatcataa aatataggac ttataaaaaa 19560
cagttccacc agtgccttaa tctaaaattt ttcatcaaaa aatataggcc aaactatgaa 19620
atgatctaaa tatactacac ccccggctag tgtcctatct ttatgttgta cattattttc 19680
aatattttct tttctaataa tgtaatttac tttccaaatt atatgattta aggcttaact 19740
gttggagtat aatttatttt tagatcccta aacattttaa acgatgtagt attttaattt 19800
ttagggcact attttgagcc aacttgttgg agatgctctt aggttaccta cggattccgg 19860
ataaaaaaat cttgaaattt ctgccataag attctagagg aatctaacac acaggcacaa 19920
aattcaatga atactagaga ttatttttat ccagaatcca gttgatgttg gctattattt 19980
tattcttcta ttttggatcc aaagaaaaga agaagagtaa atccatagaa attcctaact 20040
aaaaatgtga tttctaaaga tttcattgta attcttcgaa ttatcgaaca aatccaatcc 20100
aacataatac aatttttttg aaattagtaa attaagtaat aaagcataga gaaataatta 20160
aagaatacaa tcgttatcca tcttttcttt cttttttgct tcatatacta aaaagaattc 20220
aaaaaggcaa aattttcgtc atgtcatgaa gaattctatc tcttgtatac gaataactag 20280
gatttaaaga aggaaatgac aaagcatatg tgaataaacg attggtttct atcataaacc 20340
tgctaaagcc tcaaaccact gagtacttag cacgggtgct acaaagagat atgtttttac 20400
atatataatc agatactctt gtccaagatc atttgtattt cttttgttta ggctaatttc 20460
ctaactaaaa aagaaaaata aatatcgtat atatagaatg aaccatatag atgagatttt 20520
tctatcaagt gagatgtgaa aaaatgttgt tggctgtgag ctgtaaaaag ctaaaaatag 20580
tttagtggaa gctgctagaa tccactaaaa tttcttctat tatattttta cacggttcca 20640
tctgaaagtt gctaaaaaca agttcatagt gcttccaatt ttgtaaaccg aggcaatcgt 20700
ttttttaaaa agctgattct tgaattgagt ccttcggttt ggcttttggc tcttgagagg 20760
caaaagccaa agccaaaagc caaaccaaac ataccatttg ttattctaca cttaaaacaa 20820
actaatatat aactcgttaa tatcagttcc aa 20852
<210> 77
<211> 14187
<212> DNA
<213> 玉米
<400> 77
tgagaggagt tgagtgcaac tctcgtccgt tacgccacaa cgtggagtag gcaagttttg 60
tacttggccg aaccacggga taaccaccgt gtcatctctg tgattgattt cttgtggtta 120
ttgtgttttg actcctctct agccacttgg acatatttgt gctaacactt aacaagtttt 180
tgtggcttaa gttttgaagt tttacaggat cacctattca cccccccctc taggtgctct 240
cagtatttgc tgtgatcctt gtgtcttgtg tgagttgcta atccctccct tgctctgtgc 300
ttctttgtga acatctttgt aagggcgaga gactccaagt tgtggagatt cctcgcgaac 360
gggatataga aaagaaaagc aaacaccgtg gtattcaagt gggtctttgg accgcttgag 420
aagggttgat tgcaaccctc gtccattggg acgccacaac gtggaagtag gcaagtgttg 480
tacttgaccg aaccacggga taaaccactg tgtctacctg tgttgattct cttgtggtta 540
ttgtgtttcg ctaagactct tatctagcca cttggcaata ctgtgctaac gcttaaccat 600
gtttttgtgg cattaagttc aagttttaca ggatcaccta ttcaccccct ctaggtgctc 660
tcaagagggc gagtggatcc gtcatcagta agacggaata tgcgggaccg ggctcaagtt 720
ggcggctggt gcggacggct ccgtttgagg ccgaggggcg gacgactcca tagatgacgg 780
cgtgcgggcg cgagaacgga tgaaaccggg aggacggtag cgcgcggacc agaagcacat 840
ggcacactag gtcaggggta gtctgacgat agggtgggca aagtgaggca gggtagggcg 900
ggggcatggt tgtgccgtgg tctacattat tgacttaata gatagtagag aacaatagct 960
tatttgagat tatttttggc aaaagaccca ctaccccatg gttatgtaaa tagaaattac 1020
aatatatgcc atcattcttt cctcacttga aataaacaaa caagggtact gctgtcttta 1080
tgaataatct acatttgtat aatctagact accaaacaac tatatgtaga ttataatatg 1140
tatagattat aatctatatt atataattta gattataatc cagattataa aatctataag 1200
atgaaacaaa cagaccctta gagcatctcc agtagttatg taaacaagtc cctaaatatg 1260
aatttaacaa gttttcgata aaaactattc tccaacagtt atgtaaatca gttccctaaa 1320
tgtacaaact ccctacaaaa cacaatttca tccctacatt taccaacctt gttgggactc 1380
actaaatcaa cgttggtact aaatccatca tgaaaataat acttttgcca ctgttttgcg 1440
ctgtcccact gtcctgttga ccacaaatcc atctctaacg cctagatgtc ttgtgccgtc 1500
actgttagac aacctcttat gcggaggcgt cgttagagca catcctgtga tgtcttcgcg 1560
tcatcgccac gtccgtatgg tcttccatcg ataacttcga catcagggaa ggttgttgat 1620
attgatttct accgccgact caattaaaat tagaaagcaa gctcacttat tttttttgtt 1680
gacgtagaca actttaaaat tagcaaccaa taaataggaa actgttggat cacatacatt 1740
tttctacttc ttaaagtgtt ttagcaactt cttaaatcta taatttagga tgctaaattt 1800
acataagtat tgaagatgct cttagtaagc taaaaaacaa caccaaacac tagcacaaga 1860
aagagaggca gataggcatc gagagtggca accgggagat ctgatgaaag gccgtccagg 1920
cggtccggtc caacagtccg tagatccgca tagtgcagct tgattgatgg gatgagagcc 1980
catgaaggga tggccggccc aggcctggag ttgccgacca cgagagatcc tgaccagcag 2040
ttgctgggct gtgtgtgtgt ctgcgtctgt cactctcctc tctcgcgtct tggtcctcct 2100
cctccccttc atcgaacgaa tccatccaag gcaaaaggga acgagggaag caagcaaaaa 2160
atccccaatt cctcctagat cgatccaccg ccgacgggac gaatccaatc caggagctat 2220
cggctggaat tggattcacg cgcacccgcc cgctgccgct ggccgccgcc tagtctcggc 2280
cggccggccg accggccgcc ctccccgcgc gcgcgccttg ccttgtatgg aaacacgcag 2340
cagcagtcgc aagcgcgcag cggccgccgc ctcctcgtcc gcctccaagc gcccccgccg 2400
caactcctct tcctcctcct ctgcgccgcc gcgccagcag ccaccgatgg aaccctcgcc 2460
ctcctcccgc cgccgctccc gcgccgccgc cgccgacaag ggcaaggacc ccgaccccgc 2520
ctcctccgac cacccttccc cacccgcccc tgacgatgac gccgacgtca ccttccccca 2580
gcccttcacc tccgccagca ccgccctcca gggcctcctc cgcaggctcg gcgccggcct 2640
cgacgacctc ctcccatcct cctcctcagc gccctcctcc gccacgtccg gtcacctcaa 2700
gaggatcctc gccggcctgc aggcgcacgg cgacgagtcg cgccagctcc agtccctcat 2760
gcagctctgc gagatgctct ccatcggcac cgaggactcg ctcgccgcct tcccggtcga 2820
tgcttttgtc ccaatcctcg tcgggatgct cggccgcgag gacgagcctg ccaccgctgg 2880
tgccagcccc gacgtcatgc tgctcgcggc gcgggcactc tccaacctcg ttgatgtgct 2940
cccctcctcc tgctccgcgg tcgtgcacta cggggccata cagtgcttct gtgctcgact 3000
gctcaccatc gagtacatgg acctcgctga gcaggtggtg ccttcccttt gtcttcttct 3060
tcctcgtttt atcctgtatc atcgctcatc ctcactatca tctaaaaatg aaatatgatc 3120
agtcatcctc tgcatgcatg cccttagccc cttaccaagg tgctgctaac acatttactt 3180
tactcgcctg ctcagtctct acaagccctt aagaagatct ctctggagca cccaactgcc 3240
tgtttgcgag ctggtgccct aatggctgtt ctatcgtatc tcgacttctt ctccactggg 3300
gttcaggtcg ggtcctctga tctccttaca accgttctta tcttgtgcac tgtttgcatc 3360
tacttacacc tcttcccatg ggggacgtac ttgttgtaga gggtagcctt gtctaccgct 3420
gcaaatattt gcaggaagct gccgtccgat gcttccgagt ttgtcatgga agctgtcccc 3480
ttgctcacga atcttctaaa tcaccatgat tccaaggtac tttgcaagct cagacaacca 3540
tacctctgga ttatgatata ttgaggtata tcatgtattt cccaaccctt catctatgtg 3600
ccttacaggt actagagcat gcttctgttt gtttgacacg cattgctgaa gcttttgcct 3660
attaccccga caaacttgat gaactctgca atcatggatt agttgctcaa gctgctagct 3720
tgatagctgt tggcaactca tctggccaga catcactgag cacatccaca tatacagtat 3780
gggccatttt attttgatac ccttttattg tttgctgtgg cccctctttc tcacacttat 3840
gctaaaatta ggttgcacgc ttttcagggc ttaatccgtc ttctctcaat atgtgcaagt 3900
ggatccctgc tggcagttaa aacactactt cttcttggga tcagcggcac tcttaaagat 3960
atcctttctg gctctgcttt gatttctgga acatcagtct cccctgctgt aacaaggcct 4020
gcagatcagg tgaatatttc tatcactgac ttaggacttg actattctgc tagtgctggc 4080
cttttaaaaa aaaatcaatt tagccacaat ttctctctgt gattggattc aataaaattc 4140
cgatttatca gtgaaaaatt aggctttggc atctcacact ttctgaaatt acatctacca 4200
cacgtatcat gcagactttt agaactctaa aggcgacatc acatgttcat aattatatgt 4260
accattgagc aattgtttgg gtgatttttt tgagcattga acatagatgc taaaatgtcg 4320
ggaattgtgc tcgtcttctt aaaccatgca aataatcatt caaaagagca gtgaactagt 4380
attttcaaag gcctgcatac tttcgttgtg cttttatatt gcctgagtaa gtcttttgaa 4440
atgttttaat gaagcagaaa atgttgattt atggcacaca ggggaacaaa agtctcactt 4500
gtaatatgat agagcaaaat ctcatttgga ttaagtagct gctgtgattc ttgtcgtctt 4560
attttcttcc tagtgtggtg caagtaaatt gtgcttttgg tcaatcgcat ttggtatggt 4620
acagatcgaa gagttttttt tccatgctat acaattgagt atgaatatat aatgccatct 4680
cagcttctgc ttgttgctgt cttgcttaat tgctaaaatc atttgtggta catacacaca 4740
cgagtctggc attttttgag tatgatcgtc tgcaacagct gctgtatctt ctcacctagc 4800
tgctgaaaca ttaattcttg aaacccacac tgtttgactc atcagtttca tgcacctgta 4860
tgttttttgt tgtacaaatg tttttcatat gtttacccca aatattgcag atgtttgaga 4920
ttgtgagcct tgccgatgat ttgcttccgc atatccctgc gagaatcatt aagttaccaa 4980
catattacca tacatataaa ggctcttcaa caaagaaatc tgcttccatc aaacaagagg 5040
ggggtggttc aacaggaaat gaaaggtctg gccgtgagag gctattgcgt gagcaacctg 5100
agcttttaca acagtttggt atggatttgt tacccacaat gacacaggtt agtgctggaa 5160
ttcttttcct gagtcctgac agccatttca attgctattt gtgttttgtc tttgtgtact 5220
ttaatagcct gtgtgcttgt tcaggtgtat ggttctagtg taaatgcacc aatacgtcac 5280
aagtgtttat ctatcattgg aaaattaatg tactacagct ccgccgaaac aatccagtct 5340
ctccttggca caacaaacat atccaggttt tctctctctc tcacctggaa cacacaaaag 5400
agatgcatgt ttcattaata gaaccgaaaa aattctcagt tcatttctag agaatgattt 5460
cagaacaatg gatgttaaaa gacaaaaaga aagtctctca agtctcaatg tacatacttc 5520
ttttccaatc ccttctaagt tatttcattt ctatctgcag cttccttgcg ggcattcttg 5580
catggaaaga tcctcaagtg ttgattcctg ctcttcagat agcagaaata atgatggaaa 5640
agcttccaga gacattttcc aagttatttg tgagggaagg tgtcgttcat gctgtggagt 5700
cactcatatg ttcagaatct tccaacaaaa tgccttctca ggtgccacct caggataaag 5760
ataaagattc tgccatgcca tcacgttcta gacgtcaacg ccggcgtgcg ggtgctgttg 5820
cagcagaaaa tagttcattg gatgaatcaa atagttccaa ccttggtgtt atgtgctcaa 5880
cagcaaccgc ttcagaagct ccaaacacca gccttcgttt cacagttagt gaccatgcaa 5940
agtctttcaa agataaatac tttccggcgg atactgattc aagtgatatc gggtttactg 6000
atgaccttct taaactgagg gcactctgtg caaagttgaa tactgtctct gagaacgtca 6060
aaacaaaagc aaaagggaaa tcaaaagcta taagcactaa ttttttggac atctcaattg 6120
atgttgaaga acaattagac aaaataatat ctgaaatgct ctctgagctt agcaaagtta 6180
atggtgtttc cacatttgaa ttcattagaa gtggtgttgt catagcattg cttgactatt 6240
tgtcctgtgg gacatttggg aaggaaaagg tatctgaagg aaacctatct cagcttcgtc 6300
agcaggccct tagacgatac aaaaccttca tatctgttgc cctttctatt gatcatggac 6360
gagatgaaac tcccatggca ctgttggttc agaaactgca aagtgcattg tcttcgctgg 6420
aacgtttccc tgttgtgctc agccagtcta gtaggattgg tattggaggg tctcgtctga 6480
cttcaggtct tagtgctctg gctcaaccct tcaagcttcg tctttcccga gctcaaggcg 6540
aaaaatcact tcgggattat tcatcaaata ttgtgcttat cgacccattt gctagtcttg 6600
catctgttga agaattcctt tggcctagag ttcaacgcag tgaggttgca tcgaagccta 6660
taattccatc tggtaataat tctgaatctg gggttcctgg caccacggct ggtgcatcat 6720
taacagctgc aatggctcaa tctggtcggc gcccaacaac aagatcaaag tcatctgctg 6780
caggtggtct tacatctaag aaggattctc atgatgaaag cactagtact gccaaaggaa 6840
aaggaaaagc tattgtaaag ccaaactcag atgaatcaaa aggacctaat acaaggaatg 6900
ctgcccacca aaaatctgct tcagagaaag atttggaaat gaagcaggca catggccaca 6960
gtagctctga ggtatttttt ttgtttacat gcctcattcc tggttagttt ggcttgcaaa 7020
gaaaagttac tgtttctatt caagttgggt agcagctctt ctcaaaagtc gcattgtcag 7080
ggcctaagga ggcccacgga ggggctgcgg cagcagcagc catgggccct ccaaggggga 7140
ttagataatg atagttagag ataagattag aagattagtt gagattattt aggagattag 7200
ttgagattat ctaggaaggt tatcagttag tttgttgaga gtttttagga gagttttaag 7260
gagataaagg tctcaagcta gacaagggcg gccacccggt ctatttatgt aatcaatgga 7320
taagaaataa attaggcaag aattagaagg gagaaaccct cctcttgctc ggccgtgggc 7380
agaggccctc ggccgacgtt cctgcccgtc gatacccctc tctaatccct caccgattta 7440
gcgtccataa cacacatcca cttctctggt gtcatatttg gtgccaccta taaagctcat 7500
ggcttgggcg ccaaatgaac atgataattt tagccgcgga tctagttctt aatttatatt 7560
accctcataa tatgtaaaca tgattttgtg agttcaaaac tgtgccaatc taaaaaatat 7620
taggataatg atcaagaagt aggctcattt tctagttgag agtgtaagca tggtaactga 7680
taagtaaata cattctatgt tggtatatgt aggatgaaga gctcgacaca tctcctgttg 7740
agatcgatga tgctttgatg attgatgatg atgacatttc agatgatgat gatgatgacg 7800
acgatgatca tgaggtaact gtttgaaaat tgaatgccca ctcttttttt tgcttcgttg 7860
caaccatccg tagttgctta ggttcatatt caaggtacta aatagcatgg gttcatagca 7920
gtttgatgtt gcggatgtca ggtgctatta cggaccgcta agttccaata gcagtatata 7980
ataatcaaac ataaatttat gtatatatat atcattaata ttaatttaat agacgtttaa 8040
ttaattatac ttagataagt gatgtctcac atcattatta tcaacaaagc cttttagccc 8100
caagcaagtt ggggtaggct agagttgaaa cccatcggaa gccacaaatt aaggttgagg 8160
catgtggaaa tcgatgtctc actctcacat actttaaatt aaataaagga acatgatatt 8220
caacatatta catataagtc atcacacatt ggactggagt ctacaaagaa atatattttg 8280
atagccgtaa aatagtcata aaatcatgtt tcattgtcat gtcagaagta gtaacctcaa 8340
tagtttcatg atagttaaaa tgggataggg ttatttgttg tatatcatac aacactattg 8400
tggacactat gaagaagtgc ggagcactat aacagcctgc tacctcaaat agcagccgct 8460
aagttccaac ctcaaatagt ggtagcggta agagaccgct accgtttttt gcggacgcca 8520
tagcggttgc tactaacgct atttagtacc atgcttatat ttatttactt tatgacaaaa 8580
ataacctttt agttggtcca agatgatcat ttgtgccctt acttgactaa tttaataacc 8640
tctccaaatt cttaggttct ccaagaaggg tctcttccaa tctgttctca agatggtgtg 8700
catgacgtga aactgggtga tgccgatgaa tctaacattg gatcagctac ttttagccat 8760
gcacagccat catctggttc tggggctaga aacacttcta gcaggggacc ggattctact 8820
gaatttcgaa gtatgggcac attcggttca cgaggtgcaa tgtcatttgt tgctgcgacg 8880
atggctggtc tggcttctgt tggtggtcgc agtgttagag gtgtccgaga tcggcgtggt 8940
ctgtcacttg gaggtagcat gaatgatcac aataaactta tattcactgc tggtggcaag 9000
cagcttaaca aacatttgac tgtatatcaa gctatccaac gtcaacttat gctcgacgag 9060
gatgatgaag aaaggtttaa tggatctgat ataccaaatg atggaaaccg cttctggggt 9120
gatgtgttca caataacgta ccagaaggct gataaccaag ttgagaaggg ttgccaaggg 9180
acttccacct cattaaacat aaaatcagat tcttacaggt ccatttctga agcacaaggg 9240
gtttctcttc ttgatagcat cttacaagga gaacttcctt gtgatcttga gaggacaaac 9300
tcaacgtaca acattttagc attattgcgt atacttgagg ggcttaatca gttatcccct 9360
cgtttaagag tacagggagc atctgatgat tttgctgagg gaaaaatcaa taccctggat 9420
gagttatata gaactggagc caaggtacct tctgaagtgt ttgtcaatag caagttaaca 9480
ccaaagcttg ctcggcaaat gcaggacgtt ctcgcactat gtagtggcag tttaccttct 9540
tggtgttatc aaatgaccaa ggcctgccca ttcctgtttc cttttgagac aaggaggcag 9600
tacttctatt ccacagcctt tgggttgtca cgagcattga atcgactcca gcaacagcag 9660
agtgataatc acagctctgg tggtgaaaga gaggtcaggt ttggcaggtt acaacgccag 9720
aaagttcgtg tttcccgtaa ccgtattcta gattctgctg ctaaagttat ggagatgttc 9780
tccagtcaga gggctgttct tgaggtggaa tactttggcg aagttggaac tgggctgggg 9840
cctactctag agttttatac tctcttaagc catgaactac aatgtgctca actgggatta 9900
tggagagcta cttcttcata tgattcagga ttgcacactg atatgaagga tgtgatccgt 9960
ttagatcctg aagatgggtc gtcaggaaaa gaactcaaca cagatctacc aggtgatggc 10020
atgtatttga tacaagctcc tcttggattg ttccctcgac cttggccacc taaagttgat 10080
acttcagaag gtagcagatt cttcaaagtt ttggagtatt tccgcttaat aggtcaagtg 10140
atcgcaaaag tcctgcaaga tggcagactt ttagatttgc ctttatcaac agcgttttat 10200
aagcttatac ttggacaggt atgtacagag aagctctatt tccaagtgct ttgctttcat 10260
tttgttttgc tttgttttat gtattaatta gttgtttgta ttcacttttc acgacttaat 10320
gtgttgcaat tggccaggaa cttgatttgt ttgacatagt gtcatttgat tctgagtttg 10380
ggaagacatt gcaagaactg caagttcttg ttgaacgaaa gcagttcctt gaatccactt 10440
ctggcaagaa ccagctacaa gttgcagatt tgtgtttcca tggcgcttct attgaagatt 10500
tgtgtttaga ttttactctt ccaggttatc ctgattatgt gcttaaagag ggtgaaggag 10560
gcacaattgt aagtgatgaa gttccatatt agcgtgtatt tatcttggtg atttcacttt 10620
gtatgtttta atctgttagt atcatctgca ggttaatatt tataatttag aagagtatat 10680
tactttggtg gtggatgcta cagttaagtc agggataaag cgacaggttg aggcatttag 10740
atcagggttt aatcaggtac agcctcattg gcattgctct aacacaatct tataataatt 10800
ctacttgttc tgattttgtg tagttcagtt tactctaata gttcttgttt attttaatgt 10860
ttcgtaggtg tttgacatgt catccctcca catattttca cctcaagaac ttgactatct 10920
aatatgtggt cgacaagaaa tttgggaggt aatgttctgt cctttttatc caagttaatg 10980
tatgtaaaag tgtggcacta aataaacctt ggaatctgca gtcggaatcc ctggtggata 11040
acataaaatt tgatcatgga tatactgcta aaagtcctgc gatcgtgaat gtaagctcat 11100
tcgtcctcac atgctacatc tcttacatac atccagttgt ggtttttgag aagtgtggtt 11160
ccttgattct tgctcttgtt tgtagctact cgagattttg tcagaattca cccctgagca 11220
gcagcatgcg ttctgccagt ttgtaaccgg tgctcctcgg cttccacatg gtggattggc 11280
tgcccttagc cccaagctta ctatagttag gaaggtacct tttcatggtc tatctatcgt 11340
ggcatttgct ataatgacat gtgcacacct attctgcttg atgttatttt tggggtaact 11400
cggtgttttg attattgcca gcatccttca agtgctgtga atacttcaaa ttcaactggg 11460
gcagtagagt ccgctgatga tgatttgcct agtgtgatga cgtgcgccaa ttatctgaag 11520
ctgcccccct actccacaaa agtacagtgt ccttgctgtt ttatcactaa aaagtcagct 11580
tgacattctg atggttcctg ctgacgggct gttgctattg tttggacagg aaatcatgcg 11640
caagaagttg ctttatgcga tcctggaagg ccgaggatca tttgatcttt cataagaaat 11700
atacaatatc cgaactaaca tacaggggtc agttttattc tggagattta gtttcttggt 11760
ccatgtataa cactgagttg aggttaccag ctcactcagc aaacggcttc aagggcgaga 11820
gccccagtcc tcactcggat gcaggtgctg ctgtcgattt ctcccttctt ttacatttgt 11880
tcagctactg tattctcttc gagggttgcc cttgtcacct gcaattatga atctacagct 11940
tttgtcatcg aggaagttta acagattttg gtttctctac catgatctgt ttccattggg 12000
caggctaagc tatagtaatt gaggatgtgc gccttgagtg aggatgtgcg ccaacaagaa 12060
cccccgtttg tttcagtaca tgtaaataaa agggattttt tgttccagaa ttgttatctg 12120
taacaagaac atctacatct gtgatgactg accattaata aatgtgtccc ggtgtaccat 12180
ttgttttttt tctttgagca ctggcagtag caggatacac aaaagggagt gtatgctata 12240
tactactagt cgcggatagc tcatctggtc atggccacag ttttttctta tggagagaat 12300
aacaaaattt ggccctataa acagaaaaaa ataaagtgca agaatcggca gcatgagaat 12360
tgtttgtacc caagactatc tttagtagct tacccatcct acttatattt aactctacaa 12420
tagtgtagtt tatagtgcag aaaagtatat aaatagtgtt ttaagtaaac tattaaactc 12480
tactctacat ttaaactcta ctattaaact ctacgaatag tgtaatctat aaataatgtt 12540
ttaattaaac tatgagtaaa ctactgactg tctaagtgtt attactgagg attcctacac 12600
cacgataaaa tatacttagt ccgttctagc ttttattttt atgtccatat ttaaatggat 12660
aacaatgaat ctagaatgca tatactaata tcctaaaata aatattattc taaaaaacaa 12720
attataatga ataataataa ctatatatat aatcataggt gcaggataat ttggttatta 12780
taattattat tatataagat aattaatata atactcacac catcttaaaa tataatttat 12840
tttagatttg gagtgtttga atgtactaga actaatagtt agttgactaa aaattgttag 12900
tagaattaga tagctaacaa ataactatct aactattaat taatttatca aaaatagcta 12960
atagctaaac tattagctag agtgtttgga tgtcttaact aattttagcg actaactatt 13020
agctctagtg cattcaaaca cctctaaatc tgttcattgc cggctacctt tcataaataa 13080
cttatgaggc caaactaaag agggcagggc aggcgaatcg aaggaaaagt agtcaggaga 13140
ggcgtttccc tcgcaatcga ccatcacaga tcaaatcgct gcaactagct agggtttcgt 13200
cgcctcatct ctcgtcgatc catcgatggc gtccgagggc gaaaacaaga ggatgatcac 13260
gctcaggagc tccgacttag aggagttcga ggtggaggaa gcggtggcca tggagtcgca 13320
gactctccgc cacatgatcg aggacgactg cgccgacaac ggcatcccgc tccccaacgt 13380
caactccagg atcctctcta aggtcatcga gtactgcaac agtcacgtcc acgccgccgc 13440
caaacccgct gactccgctg cctccgaggg cggcgaggac ctcaagagct gggacgcgaa 13500
gttcgtcaag gtggaccagg ctacgctctt cgacctcatc ctggtcggta acctcccctc 13560
cccttgcttc ttttgcgatt cctgtctatg tataggttag attagatctg ctactgtctt 13620
tctagggttc ggattagatc agtatatatt gattgattgg ttgtgcgtcg tggtggaggt 13680
ccgtaaccaa tcaatttttt ttatctcata agagatgttg tactagcccc tgctgctctg 13740
gaatcaatca accacaaaaa ctgtgatgtg gctaggaaca ctgttgataa atcatgctgt 13800
tgctattatt cgtaacaaca atgcacccgt atcaagctag ctacaggcgg ggatgatatt 13860
tggtttgcag tgagataatt taactagcta gtcagacaga atttggcttg tacttgcgac 13920
ttttgcacac tttggggatt gttcgtttga ccgccacgtg gattgggtgg tattgagtcg 13980
gtttaaatcc atccaaattc attccaatac acttcaatcc acatgtaatt ggacgtgatt 14040
tgtgttattt tctgccattg ctggttgaca taatggaact gtattttcag gctgccaact 14100
atctgaacat caagggattg ctggacctga cctgccagac ggttgctgac atgatcaagg 14160
gcaagactcc ggaggagatc cgcaaga 14187
<210> 78
<211> 1872
<212> PRT
<213> 大豆
<400> 78
Met Glu Thr Arg Ser Arg Lys Arg Ala Glu Ala Ser Ser Ala Ala Pro
1 5 10 15
Ser Ser Pro Ser Ser Gly Pro Thr Thr Arg Ser Ser Lys Arg Ala Arg
20 25 30
Leu Ser Ser Ser Ser Ser Ala Ser Ala Ala Val Asn Thr Arg Ser Arg
35 40 45
Ala Ser Asn Thr Lys Glu Pro Leu Pro Pro Lys Asn Pro Pro Pro Pro
50 55 60
Leu Pro Pro Met Asp Ser Ala Asn Glu Ser Ser Gly Ser Arg Arg Asp
65 70 75 80
Arg Arg Asn Asn Lys Glu Asn Ser Ser Asp Lys Gly Lys Glu Lys Glu
85 90 95
His Asp Val Arg Ile Arg Asp Arg Asp Ala Ala Leu Asn Met Asp Gly
100 105 110
Ser Gly Gly Asp Glu Asp Asp Asp Asn Asp Asn Asp Ser Glu Gly Gly
115 120 125
Val Gly Ile Leu His Gln Asn Leu Thr Ser Ala Ser Ser Ala Leu Gln
130 135 140
Gly Leu Leu Arg Lys Leu Gly Ala Gly Leu Asp Asp Leu Leu Pro Ser
145 150 155 160
Ser Ala Met Gly Ser Ala Ser Ser Ser His Gln Ser Gly Arg Leu Lys
165 170 175
Lys Ile Leu Phe Gly Leu Arg Ala Asp Gly Glu Glu Gly Arg Gln Val
180 185 190
Glu Ala Leu Thr Gln Leu Cys Glu Met Leu Ser Ile Gly Thr Glu Glu
195 200 205
Ser Leu Ser Thr Phe Ser Val Asp Ser Phe Val Pro Val Leu Val Gly
210 215 220
Leu Leu Asn His Glu Ser Asn Pro Asp Ile Met Leu Leu Ala Ala Arg
225 230 235 240
Ala Leu Thr His Leu Cys Asp Val Leu Pro Ser Ser Cys Ala Ala Val
245 250 255
Val His Tyr Gly Ala Val Ser Ile Phe Cys Ala Arg Leu Leu Thr Ile
260 265 270
Glu Tyr Met Asp Leu Ala Glu Gln Ser Leu Gln Ala Leu Lys Lys Ile
275 280 285
Ser Leu Glu His Pro Thr Ala Cys Leu Arg Ala Gly Ala Leu Met Ala
290 295 300
Val Leu Ser Tyr Leu Asp Phe Phe Ser Thr Gly Val Gln Arg Val Ala
305 310 315 320
Leu Ser Thr Ala Ala Asn Met Cys Lys Lys Leu Pro Ser Asp Ala Ala
325 330 335
Asp Phe Val Met Glu Ala Val Pro Leu Leu Thr Asn Leu Leu Gln Tyr
340 345 350
His Asp Ser Lys Val Leu Glu His Ala Ser Val Cys Leu Thr Arg Ile
355 360 365
Ala Glu Ala Phe Ala Ser Ser Pro Asp Lys Leu Asp Glu Leu Cys Asn
370 375 380
His Gly Leu Val Thr Gln Ala Thr Ser Leu Ile Ser Asn Ser Ser Ser
385 390 395 400
Gly Gly Gly Gln Ala Ser Leu Ser Thr Pro Thr Tyr Thr Gly Leu Ile
405 410 415
Arg Leu Leu Ser Thr Cys Ala Ser Gly Ser Pro Leu Gly Ala Lys Thr
420 425 430
Leu Leu Leu Leu Gly Ile Ser Gly Ile Leu Lys Asp Ile Leu Ser Gly
435 440 445
Ser Gly Val Ser Ser Asn Ala Ser Val Ser Pro Ala Leu Ser Arg Pro
450 455 460
Pro Glu Gln Ile Phe Glu Ile Val Asn Leu Ala Asn Glu Leu Leu Pro
465 470 475 480
Pro Leu Pro His Gly Thr Ile Ser Leu Pro Ile Ile Ser Asn Met Phe
485 490 495
Leu Lys Gly Pro Ile Val Lys Lys Ser Pro Ser Gly Ser Ser Gly Lys
500 505 510
Gln Glu Asp Thr Asn Gly Asn Val Pro Glu Ile Ser Ala Arg Glu Lys
515 520 525
Leu Leu Asn Asp Gln Pro Glu Leu Leu Lys Gln Phe Ala Met Asp Leu
530 535 540
Leu Pro Val Leu Ile Gln Ile Tyr Gly Ser Ser Val Asn Gly Pro Val
545 550 555 560
Arg His Lys Cys Leu Ser Val Ile Gly Lys Leu Met Tyr Phe Ser Thr
565 570 575
Ala Glu Met Ile Gln Ser Leu Leu Ser Val Thr Asn Ile Ser Ser Phe
580 585 590
Leu Ala Gly Val Leu Ala Trp Lys Asp Pro His Val Leu Leu Pro Ala
595 600 605
Leu Lys Ile Ala Glu Ile Leu Met Glu Lys Leu Pro Gly Thr Phe Ser
610 615 620
Lys Met Phe Ile Arg Glu Gly Val Val His Ala Val Asp Gln Leu Ile
625 630 635 640
Leu Ala Ser Asn Ser Thr Asn Ile Ser Thr Gln Ala Ser Pro Ala Glu
645 650 655
Lys Asp Asn Asp Ser Ile Ser Gly Ala Ser Ser Arg Ser Arg Arg Tyr
660 665 670
Arg Arg Arg Ser Gly Asn Ser Asn Pro Asp Gly Asn Pro Leu Asp Asp
675 680 685
Leu Lys Thr Pro Val Ser Val Asn Val Gly Ser Pro Pro Ser Ser Val
690 695 700
Asp Met Pro Thr Leu Asn Ser Ser Ile Arg Leu Ser Val Ser Thr Ala
705 710 715 720
Ala Lys Ala Phe Lys Asp Lys Tyr Phe Pro Ser Asp Pro Gly Ala Ala
725 730 735
Glu Val Gly Ile Thr Asp Asp Leu Leu His Leu Lys Asn Leu Cys Met
740 745 750
Lys Leu Asn Ala Gly Asp Asp Glu Gln Arg Thr Asn Gly Lys Gly Glu
755 760 765
Ser Lys Thr Ser Gly Phe Gly Pro Glu Glu Tyr Leu Ile Gly Ile Ile
770 775 780
Ala Asn Met Leu Lys Glu Leu Gly Lys Gly Asp Gly Val Ser Thr Phe
785 790 795 800
Glu Phe Ile Gly Ser Gly Val Val Ala Ala Leu Leu Asn Tyr Phe Ser
805 810 815
Cys Gly Tyr Phe Ser Lys Asp Arg Pro Leu Glu Ala His Leu Pro Lys
820 825 830
Leu Arg Gln Gln Ala Leu Thr Arg Phe Lys Leu Phe Ile Ala Val Ala
835 840 845
Leu Pro Ser Thr Ile Glu Val Gly Thr Val Ala Pro Met Thr Val Leu
850 855 860
Val Gln Lys Leu Gln Asn Ala Leu Ser Ser Leu Glu Arg Phe Pro Val
865 870 875 880
Val Leu Ser His Ser Ser Arg Ser Ser Ser Gly Ser Ala Arg Leu Ser
885 890 895
Ser Gly Leu Ser Ala Leu Ser Gln Pro Phe Lys Leu Arg Leu Cys Arg
900 905 910
Ala Gln Gly Glu Lys Ser Leu Arg Asp Tyr Ser Ser Asn Val Val Leu
915 920 925
Val Asp Pro Leu Ala Ser Leu Ala Ala Ile Glu Glu Phe Val Trp Pro
930 935 940
Arg Ile Gln Arg Ser Glu Ser Gly Gln Lys Ser Thr Val Ala Thr Gly
945 950 955 960
Asn Ser Glu Ser Gly Thr Thr Pro Ala Gly Ala Gly Val Ser Ser Pro
965 970 975
Thr Thr Arg Arg His Ser Thr Arg Ser Arg Ser Ser Val Asn Ile Gly
980 985 990
Asp Thr Ser Arg Lys Glu Ile Thr Gln Asp Lys Ser Thr Ser Ser Ser
995 1000 1005
Lys Gly Lys Gly Lys Val Val Leu Lys Pro Ala Gln Glu Glu Ala
1010 1015 1020
Arg Gly Pro Gln Thr Arg Asn Ala Thr Arg Arg Arg Ala Ala Leu
1025 1030 1035
Asp Lys Asp Ala Gln Met Lys Pro Val Asn Ala Asp Ser Thr Ser
1040 1045 1050
Glu Asp Glu Asp Leu Asp Ile Ser Pro Val Glu Ile Asp Glu Ala
1055 1060 1065
Leu Val Ile Glu Asp Asp Asp Ile Ser Asp Asp Glu Asp Asp Asp
1070 1075 1080
His Glu Asp Val Leu Arg Asp Asp Ser Leu Pro Val Cys Ser Pro
1085 1090 1095
Asp Lys Val His Asp Val Lys Leu Gly Asp Leu Ala Glu Glu Ser
1100 1105 1110
Asn Val Ala Pro Ala Thr Ser Asp Gly Gln Ala Asn Ala Ala Ser
1115 1120 1125
Gly Ser Ser Ser Lys Ala Gly Thr Val Arg Gly Ser Asp Ser Thr
1130 1135 1140
Asp Phe Arg Ser Gly Tyr Asn Ser Ser Ser Arg Gly Ala Met Ser
1145 1150 1155
Phe Ala Ala Ala Ala Met Ala Gly Leu Gly Ser Ala Asn Ser Arg
1160 1165 1170
Gly Ile Arg Gly Gly Arg Asp Arg Leu Gly Arg Pro Leu Phe Gly
1175 1180 1185
Ser Ser Asn Asp Pro Pro Lys Leu Ile Phe Thr Ala Gly Gly Lys
1190 1195 1200
Gln Leu Asn Arg His Leu Thr Ile Tyr Gln Ala Ile Gln Arg Gln
1205 1210 1215
Leu Val Leu Asp Asp Asp Glu Arg Phe Ala Gly Ser Ser Asp Tyr
1220 1225 1230
Val Ser Ser Asp Gly Ser Arg Leu Trp Gly Asp Ile Tyr Thr Ile
1235 1240 1245
Thr Tyr His Arg Ala Glu Asn Gln Thr Asp Arg Thr Pro Pro Gly
1250 1255 1260
Gly Ser Thr Ser Asn Ala Ser Lys Ser Cys Lys Ser Gly Ser Val
1265 1270 1275
Ser Asn Ser Ser Ser Glu Ala Lys Leu His Gln Thr Ser Val Leu
1280 1285 1290
Asp Ser Ile Leu Gln Gly Glu Leu Pro Cys Glu Leu Glu Lys Ser
1295 1300 1305
Asn Pro Thr Tyr Asn Ile Leu Ala Leu Leu Arg Val Leu Glu Gly
1310 1315 1320
Leu Asn Gln Leu Ala Ser Arg Leu Arg Ala Gln Val Val Thr Asp
1325 1330 1335
Ser Phe Ala Glu Gly Lys Ile Leu Asp Leu Asp Glu Leu Ser Val
1340 1345 1350
Thr Ser Gly Ala Arg Val Pro Thr Glu Glu Phe Ile Ser Ser Lys
1355 1360 1365
Leu Thr Pro Lys Leu Ala Arg Gln Ile Gln Asp Ala Leu Ala Leu
1370 1375 1380
Cys Ser Gly Ser Leu Pro Ser Trp Cys Tyr Gln Leu Ser Lys Ala
1385 1390 1395
Cys Pro Phe Leu Phe Pro Phe Glu Thr Arg Arg Gln Tyr Phe Tyr
1400 1405 1410
Ser Thr Ala Phe Gly Leu Ser Arg Ala Leu Tyr Arg Leu Gln Gln
1415 1420 1425
Gln Gln Gly Ala Asp Gly His Gly Ser Thr Asn Glu Arg Glu Val
1430 1435 1440
Arg Val Gly Arg Leu Gln Arg Gln Lys Val Arg Val Ser Arg Asn
1445 1450 1455
Arg Ile Leu Asp Ser Ala Ala Lys Val Met Glu Leu Tyr Ser Ser
1460 1465 1470
Gln Lys Ala Val Leu Glu Val Glu Tyr Phe Gly Glu Val Gly Thr
1475 1480 1485
Gly Leu Gly Pro Thr Leu Glu Phe Tyr Thr Leu Leu Ser His Asp
1490 1495 1500
Leu Gln Lys Ile Ile Leu Glu Met Trp Arg Ser Gly Ser Ser Glu
1505 1510 1515
Lys Tyr Gln Met Lys Ile Asp Gly Asp Glu Lys Lys Met Lys Arg
1520 1525 1530
Ser Glu Gly Ser Phe Val Gly Asp Gly Glu Leu Val Gln Ala Pro
1535 1540 1545
Leu Gly Leu Phe Pro Arg Pro Trp Ser Ala Asn Ala Asp Ala Ser
1550 1555 1560
Glu Gly Thr Gln Phe Phe Lys Val Ile Glu Tyr Phe Arg Leu Leu
1565 1570 1575
Gly Arg Val Met Ala Lys Ala Leu Gln Asp Gly Arg Leu Leu Asp
1580 1585 1590
Leu Pro Met Ser Val Ala Phe Tyr Lys Leu Val Leu Gly Gln Glu
1595 1600 1605
Leu Asp Leu His Asp Ile Leu Phe Ile Asp Ala Glu Leu Gly Lys
1610 1615 1620
Thr Leu Gln Glu Leu Asn Ala Leu Val Cys Arg Lys His Tyr Ile
1625 1630 1635
Gln Ser Thr Gly Gly Ser Tyr Thr Asp Thr Phe Ala Asn Leu His
1640 1645 1650
Phe Arg Gly Ala Pro Ile Glu Asp Leu Cys Leu Asp Phe Thr Leu
1655 1660 1665
Pro Gly Tyr Pro Glu Tyr Ile Leu Lys Pro Gly Asp Glu Ile Val
1670 1675 1680
Asp Ile Asn Asn Leu Glu Glu Tyr Ile Ser Met Val Val Glu Ala
1685 1690 1695
Thr Val Lys Thr Gly Ile Met Arg Gln Met Glu Ala Phe Arg Ala
1700 1705 1710
Gly Phe Asn Gln Val Phe Asp Ile Ser Ser Leu Gln Ile Phe Ser
1715 1720 1725
Pro Gln Glu Leu Asp Tyr Leu Leu Cys Gly Arg Arg Glu Leu Trp
1730 1735 1740
Lys Thr Glu Thr Leu Ala Asp His Ile Lys Phe Asp His Gly Tyr
1745 1750 1755
Thr Ala Lys Ser Pro Ala Ile Val Asn Leu Leu Gly Ile Met Gly
1760 1765 1770
Glu Phe Thr Pro Glu Gln Gln Arg Ala Phe Cys Gln Phe Val Thr
1775 1780 1785
Gly Ala Pro Arg Leu Pro Pro Gly Gly Leu Ala Val Leu Asn Pro
1790 1795 1800
Lys Leu Thr Ile Val Arg Lys Leu Ser Ser Ser Ala Ala Asn Ala
1805 1810 1815
Ser Ser Asn Gly Asn Gly Pro Ser Glu Leu Ala Asp Asp Asp Leu
1820 1825 1830
Pro Ser Val Met Thr Cys Ala Asn Tyr Leu Lys Leu Pro Pro Tyr
1835 1840 1845
Ser Thr Lys Glu Ile Met Tyr Lys Lys Leu Leu Tyr Ala Ile Ser
1850 1855 1860
Glu Gly Gln Gly Ser Phe Asp Leu Ser
1865 1870
<210> 79
<211> 1877
<212> PRT
<213> 大豆
<400> 79
Met Glu Thr Arg Ser Arg Lys Arg Ala Glu Ala Ser Ser Ala Ala Pro
1 5 10 15
Ser Ser Pro Ser Ser Gly Pro Thr Thr Arg Ser Ser Lys Arg Ala Arg
20 25 30
Leu Ser Ser Ser Ser Ser Ala Ser Ala Ala Ala Ala Ala Ala Ser Val
35 40 45
Ser Ser Val Asn Thr Arg Ser Arg Ser Ser Arg Thr Lys Glu Pro Leu
50 55 60
Pro Pro Lys Asn Pro Pro Pro Met Asp Ser Ala Asn Glu Ser Ser Gly
65 70 75 80
Ser Arg Arg Asp Arg Arg Asn Lys Asp Asn Ser Asp Lys Gly Lys Glu
85 90 95
Lys Glu His Asp Val Arg Ile Arg Asp Arg Asp Ala Asp Arg Gly Leu
100 105 110
Ala Leu Asn Met Asp Gly Gly Gly Asp Asp Asp Asp Asn Asp Ser Glu
115 120 125
Gly Gly Val Gly Ile Leu His Gln Asn Leu Thr Ser Ala Ser Ser Ala
130 135 140
Leu Gln Gly Leu Leu Arg Lys Leu Gly Ala Gly Leu Asp Asp Leu Leu
145 150 155 160
Pro Ser Ser Ala Met Gly Ser Ala Ser Ser Ser His Gln Ser Gly Arg
165 170 175
Leu Lys Lys Ile Leu Phe Gly Leu Arg Ala Asp Gly Glu Glu Gly Arg
180 185 190
Gln Val Glu Ala Leu Thr Gln Leu Cys Glu Met Leu Ser Ile Gly Thr
195 200 205
Glu Glu Ser Leu Ser Thr Phe Ser Val Asp Ser Phe Val Pro Val Leu
210 215 220
Val Gly Leu Leu Asn His Glu Ser Asn Pro Asp Ile Met Leu Leu Ala
225 230 235 240
Ala Arg Ala Leu Thr His Leu Cys Asp Val Leu Pro Ser Ser Cys Ala
245 250 255
Ala Val Val His Tyr Gly Ala Val Ser Ile Phe Cys Ala Arg Leu Leu
260 265 270
Thr Ile Glu Tyr Met Asp Leu Ala Glu Gln Ser Leu Gln Ala Leu Lys
275 280 285
Lys Ile Ser Gln Glu His Pro Thr Ala Cys Leu Arg Ala Gly Ala Leu
290 295 300
Met Ala Val Leu Ser Tyr Leu Asp Phe Phe Ser Thr Gly Val Gln Arg
305 310 315 320
Val Ala Leu Ser Thr Ala Ala Asn Met Cys Lys Lys Leu Pro Ser Asp
325 330 335
Ala Ala Asp Phe Val Met Glu Ala Val Pro Leu Leu Thr Asn Leu Leu
340 345 350
Gln Tyr His Asp Ser Lys Val Leu Glu His Ala Ser Val Cys Leu Thr
355 360 365
Arg Ile Ala Glu Ala Phe Ala Ser Ser Pro Asp Lys Leu Asp Glu Leu
370 375 380
Cys Asn His Gly Leu Val Thr Gln Ala Ala Ser Leu Ile Ser Asn Ser
385 390 395 400
Ser Ser Gly Gly Gly Gln Ala Ser Leu Ser Thr Pro Thr Tyr Thr Gly
405 410 415
Leu Ile Arg Leu Leu Ser Thr Cys Ala Ser Gly Ser Pro Leu Gly Ala
420 425 430
Lys Thr Leu Leu Leu Leu Gly Ile Ser Gly Ile Leu Lys Asp Ile Leu
435 440 445
Ser Gly Ser Gly Val Ser Ser Lys Ala Ser Val Ser Pro Ala Leu Ser
450 455 460
Arg Pro Pro Glu Gln Ile Phe Glu Ile Val Asn Leu Thr Asn Glu Leu
465 470 475 480
Leu Pro Pro Leu Pro His Gly Thr Ile Ser Leu Pro Ile Ile Ser Asn
485 490 495
Met Phe Leu Lys Gly Pro Ile Val Lys Lys Ser Pro Ala Gly Ser Ser
500 505 510
Gly Lys Gln Glu Asp Thr Asn Gly Asn Val Pro Glu Ile Ser Ala Arg
515 520 525
Glu Lys Leu Leu Asn Asp Gln Pro Glu Leu Leu Lys Gln Phe Ala Met
530 535 540
Asp Leu Leu Pro Val Leu Ile Gln Ile Tyr Gly Ser Ser Val Asn Gly
545 550 555 560
Pro Val Arg His Lys Cys Leu Ser Val Ile Gly Lys Leu Met Tyr Phe
565 570 575
Ser Thr Ala Glu Met Ile Gln Ser Leu Leu Ser Val Thr Asn Ile Ser
580 585 590
Ser Phe Leu Ala Gly Val Leu Ala Trp Lys Asp Pro His Val Leu Leu
595 600 605
Pro Ala Leu Lys Ile Ala Glu Ile Leu Met Glu Lys Leu Pro Gly Thr
610 615 620
Phe Ser Lys Met Phe Ile Arg Glu Gly Val Val His Ala Val Asp Gln
625 630 635 640
Leu Ile Leu Thr Gly Asn Ser Thr Asn Ile Ser Thr Gln Ala Ser Ser
645 650 655
Ala Glu Lys Asp Asn Asp Ser Ile Ser Gly Ala Ser Ser Arg Ser Arg
660 665 670
Arg Tyr Arg Arg Arg Ser Gly Asn Ser Asn Pro Asp Gly Asn Pro Leu
675 680 685
Asp Asp Leu Lys Thr Pro Val Ser Val Asn Val Gly Ser Pro Pro Asn
690 695 700
Ser Val Asp Met Pro Thr Val Asn Ser Ser Ile Arg Leu Ser Val Ser
705 710 715 720
Thr Ala Ala Lys Ala Phe Lys Asp Lys Tyr Phe Pro Ser Asp Pro Gly
725 730 735
Ala Ala Glu Val Gly Ile Thr Asp Asp Leu Leu His Leu Lys Asn Leu
740 745 750
Cys Met Lys Leu Asn Ala Gly Ala Asp Glu Gln Arg Thr Asn Gly Lys
755 760 765
Gly Lys Ser Lys Thr Ser Gly Phe Gly Leu Glu Glu Tyr Leu Ile Gly
770 775 780
Ile Ile Ala Asp Met Leu Lys Glu Leu Gly Lys Gly Asp Gly Val Ser
785 790 795 800
Thr Phe Glu Phe Ile Gly Ser Gly Val Val Ala Ala Leu Leu Asn Tyr
805 810 815
Phe Ser Cys Gly Tyr Phe Ser Lys Asp Arg Pro Leu Glu Thr His Leu
820 825 830
Pro Lys Leu Arg Gln Gln Ala Leu Thr Arg Phe Lys Leu Phe Ile Ala
835 840 845
Val Ala Leu Pro Ser Thr Thr Glu Asp Gly Thr Val Ala Pro Met Thr
850 855 860
Val Leu Val Gln Lys Leu Gln Asn Ala Leu Ser Ser Leu Glu Arg Phe
865 870 875 880
Pro Val Val Leu Ser His Ser Ser Arg Ser Ser Ser Gly Ser Ala Arg
885 890 895
Leu Ser Ser Gly Leu Ser Ala Leu Ser Gln Pro Phe Lys Leu Arg Leu
900 905 910
Cys Arg Ala Gln Gly Glu Arg Ser Leu Arg Asp Tyr Ser Ser Asn Val
915 920 925
Val Leu Val Asp Pro Leu Ala Ser Leu Ala Ala Ile Glu Glu Phe Val
930 935 940
Trp Pro Arg Ile Gln Arg Ser Glu Leu Gly Gln Lys Ser Thr Val Pro
945 950 955 960
Ala Gly Asn Ser Glu Ser Gly Thr Thr Pro Thr Gly Ala Gly Val Ser
965 970 975
Ser Pro Thr Thr His Arg His Ser Thr Arg Ser Arg Ser Ser Val Asn
980 985 990
Ile Gly Asp Thr Ser Arg Lys Glu Ile Ser Gln Asp Lys Ser Thr Ser
995 1000 1005
Ser Ser Lys Gly Lys Gly Lys Ala Val Leu Lys Pro Ala Gln Glu
1010 1015 1020
Glu Ala Arg Gly Pro Gln Thr Arg Asn Ala Thr Arg Arg Arg Glu
1025 1030 1035
Ala Leu Asp Lys Asp Ala Gln Ile Lys Pro Val Asn Gly Asp Ser
1040 1045 1050
Thr Ser Glu Asp Glu Asp Leu Asp Ile Ser Pro Val Glu Ile Asp
1055 1060 1065
Glu Ala Leu Val Ile Glu Asp Asp Asp Ile Ser Asp Asp Glu Asp
1070 1075 1080
Asp Asp His Asp Asp Val Leu Arg Asp Asp Ser Leu Pro Val Cys
1085 1090 1095
Ser Pro Asp Lys Val His Asp Val Lys Leu Gly Asp Ile Val Glu
1100 1105 1110
Glu Ser Asn Val Ala Pro Ala Thr Ser Asp Gly Gly Gln Thr Asn
1115 1120 1125
Ala Ala Ser Gly Ser Ser Ser Lys Ala Gly Thr Val Arg Gly Ser
1130 1135 1140
Asp Ser Ala Asp Phe Arg Ser Gly Tyr Thr Ser Ser Ser Arg Gly
1145 1150 1155
Ala Met Ser Phe Ala Ala Ala Ala Met Ala Gly Leu Gly Ser Ala
1160 1165 1170
Asn Ser Arg Gly Ile Arg Gly Gly Arg Asp Arg Leu Gly Arg Pro
1175 1180 1185
Leu Phe Gly Ser Ser Asn Asp Pro Pro Lys Leu Ile Phe Thr Ala
1190 1195 1200
Gly Gly Lys Gln Leu Asn Arg His Leu Thr Ile Tyr Gln Ala Ile
1205 1210 1215
Gln Arg Gln Leu Val Leu Asp Glu Asp Asp Glu Glu Arg Phe Ala
1220 1225 1230
Gly Ser Ser Asp Tyr Val Ser Ser Asp Gly Ser Arg Leu Trp Gly
1235 1240 1245
Asp Ile Tyr Thr Ile Thr Tyr Gln Arg Ala Glu Asn Gln Thr Asp
1250 1255 1260
Arg Thr Pro Pro Gly Gly Ser Thr Ser Asn Ala Ser Lys Ser Gly
1265 1270 1275
Lys Ser Gly Ser Val Leu Asn Ser Ser Ser Glu Asp Lys Leu Asn
1280 1285 1290
Gln Thr Ser Val Leu Asp Ser Ile Leu Gln Gly Glu Leu Pro Cys
1295 1300 1305
Glu Leu Glu Lys Ser Asn Pro Thr Tyr Asn Ile Leu Ala Leu Leu
1310 1315 1320
Arg Val Leu Glu Gly Leu Asn Gln Leu Ala Ser Arg Leu Arg Ala
1325 1330 1335
Gln Val Val Thr Asp Ser Phe Ala Glu Gly Lys Ile Leu Asp Leu
1340 1345 1350
Val Glu Leu Ser Phe Thr Ser Gly Ala Arg Val Pro Thr Glu Glu
1355 1360 1365
Phe Ile Ser Ser Lys Leu Thr Pro Lys Leu Ala Arg Gln Ile Gln
1370 1375 1380
Asp Ala Leu Ala Leu Cys Ser Gly Ser Leu Pro Ser Trp Cys Tyr
1385 1390 1395
Gln Leu Ser Lys Ala Cys Pro Phe Leu Phe Pro Phe Glu Thr Arg
1400 1405 1410
Arg Gln Tyr Phe Tyr Ser Thr Ala Phe Gly Leu Ser Arg Ala Leu
1415 1420 1425
Tyr Arg Leu Gln Gln Gln Gln Gly Ala Asp Gly His Gly Ser Thr
1430 1435 1440
Asn Glu Arg Glu Val Arg Val Gly Arg Leu Gln Arg Gln Lys Val
1445 1450 1455
Arg Val Ser Arg Asn Arg Ile Leu Asp Ser Ala Ala Lys Val Met
1460 1465 1470
Glu Leu Tyr Ser Ser Gln Lys Ala Val Leu Glu Val Glu Tyr Phe
1475 1480 1485
Gly Glu Val Gly Thr Gly Leu Gly Pro Thr Leu Glu Phe Tyr Thr
1490 1495 1500
Leu Leu Ser His Asp Leu Gln Lys Val Val Leu Gln Met Trp Arg
1505 1510 1515
Ser Gly Ser Ser Glu Lys Tyr Gln Met Glu Ile Asp Gly Asp Glu
1520 1525 1530
Lys Lys Met Lys Asn Ser Glu Gly Ser Phe Val Gly Asp Gly Glu
1535 1540 1545
Leu Val Gln Ala Pro Leu Gly Leu Phe Pro Arg Pro Trp Pro Ala
1550 1555 1560
Asn Ala Asp Ala Ser Glu Gly Thr Gln Ile Phe Lys Val Ile Glu
1565 1570 1575
Tyr Phe Arg Leu Leu Gly Arg Val Met Ala Lys Ala Leu Gln Asp
1580 1585 1590
Gly Arg Leu Leu Asp Leu Pro Leu Ser Val Ala Phe Tyr Lys Leu
1595 1600 1605
Val Leu Gly Gln Glu Leu Asp Leu His Asp Ile Leu Phe Ile Asp
1610 1615 1620
Ala Glu Leu Gly Lys Thr Leu Gln Glu Leu Asn Ala Leu Val Cys
1625 1630 1635
Arg Lys Cys Phe Ile Glu Ser Ile Gly Gly Ser Tyr Thr Asp Thr
1640 1645 1650
Phe Ala Asn Leu His Phe Arg Gly Ala Pro Ile Glu Asp Leu Cys
1655 1660 1665
Leu Asp Phe Thr Leu Pro Gly Tyr Pro Glu Tyr Ile Leu Lys Pro
1670 1675 1680
Gly Asp Glu Ile Val Asp Ile Asn Asn Leu Glu Glu Tyr Ile Ser
1685 1690 1695
Met Val Val Glu Ala Thr Val Lys Thr Gly Ile Met Arg Gln Met
1700 1705 1710
Glu Ala Phe Arg Ala Gly Phe Asn Gln Val Phe Asp Ile Ser Ser
1715 1720 1725
Leu Gln Ile Phe Ser Pro Gln Glu Leu Asp Tyr Leu Leu Cys Gly
1730 1735 1740
Arg Arg Glu Leu Trp Lys Thr Glu Thr Leu Ala Asp His Ile Lys
1745 1750 1755
Phe Asp His Gly Tyr Thr Ala Lys Ser Pro Ala Ile Val Asn Leu
1760 1765 1770
Leu Glu Ile Met Gly Glu Phe Thr Pro Glu Gln Gln Arg Ala Phe
1775 1780 1785
Cys Gln Phe Val Thr Gly Ala Pro Arg Leu Pro Pro Gly Gly Leu
1790 1795 1800
Ala Val Leu Asn Pro Lys Leu Thr Ile Val Arg Lys Leu Ser Ser
1805 1810 1815
Ser Ala Ala Asn Ala Ser Ser Asn Gly Asn Gly Pro Ser Glu Leu
1820 1825 1830
Ala Asp Asp Asp Leu Pro Ser Val Met Thr Cys Ala Asn Tyr Leu
1835 1840 1845
Lys Leu Pro Pro Tyr Ser Thr Lys Glu Ile Met Tyr Lys Lys Leu
1850 1855 1860
Leu Tyr Ala Ile Ser Glu Gly Gln Gly Ser Phe Asp Leu Ser
1865 1870 1875
<210> 80
<211> 1895
<212> PRT
<213> 大豆
<400> 80
Met Glu Thr Arg Ser Arg Lys Arg Ala Glu Ala Ser Ser Ala Ala Pro
1 5 10 15
Ser Ser Ser Ser Ser Thr Thr Thr Thr Thr Ala Ser Arg Ser Ala Lys
20 25 30
Cys Ser Arg Leu Ser Ser Ser Ser Ser Ser Ile Pro Asn Thr Thr Thr
35 40 45
Ala Asn Thr Arg Ser Arg Ser Ala Arg Asn Asn Asn Asn Asn Ser Val
50 55 60
Ser Pro Met Asp Pro Thr Asn Glu Ser Ser Gly Ser Arg Arg Asp Arg
65 70 75 80
Arg Gly Lys Asn Phe Asp Arg Asp Asn Ser Asp Lys Gly Lys Glu Lys
85 90 95
Glu His Asp Val Arg Ile Arg Asp Ala Glu Arg Glu Arg Ala Leu Ala
100 105 110
Leu Asn Leu Glu Ala Glu Asp Val Gly Asp Asp Asp Asp Asp Asn Asp
115 120 125
Ser Glu Gly Gly Val Gly Ile Leu His Gln Asn Leu Ile Ser Ala Ser
130 135 140
Thr Phe Arg Gly Leu Leu Arg Lys Leu Gly Ala Gly Leu Asp Asp Leu
145 150 155 160
Leu Pro Ala Thr Ala Met Gly Gly Ser Val Pro Ser Ser His Gln Thr
165 170 175
Gly Gly Leu Lys Gln Ile Leu Ser Gly Leu Arg Ala Asp Gly Glu Glu
180 185 190
Gly Arg Gln Val Glu Ala Leu Thr His Leu Cys Asp Met Leu Ser Ile
195 200 205
Gly Thr Glu Asp Ser Leu Ser Thr Phe Ser Val Asp Ser Phe Val Pro
210 215 220
Val Leu Val Gly Leu Leu Asn His Glu Ser Asn Pro Asp Val Met Leu
225 230 235 240
Leu Ala Ala Arg Ala Leu Thr His Leu Cys Asp Val Leu Pro Ser Ser
245 250 255
Cys Ala Ala Val Val His Tyr Gly Ala Val Ser Ile Phe Cys Ala Arg
260 265 270
Leu Leu Thr Ile Glu Tyr Met Asp Leu Ala Glu Gln Ser Leu Gln Ala
275 280 285
Leu Lys Lys Ile Ser Gln Glu His Pro Thr Ala Cys Leu Arg Ala Gly
290 295 300
Ala Leu Met Ala Val Leu Ser Tyr Leu Asp Phe Phe Ser Thr Gly Val
305 310 315 320
Gln Arg Val Ala Leu Ser Thr Ala Ala Asn Met Cys Lys Lys Leu Pro
325 330 335
Pro Asp Ala Ala Asp Phe Val Met Glu Ala Val Pro Leu Leu Thr Asn
340 345 350
Leu Leu Gln Tyr His Asp Ser Lys Val Leu Glu His Ala Ser Val Cys
355 360 365
Leu Thr Arg Ile Ala Lys Ala Phe Ala Ser Ser Pro Asp Lys Leu Asp
370 375 380
Glu Leu Cys Asn His Gly Leu Val Thr Gln Ala Ala Ser Leu Ile Ser
385 390 395 400
Thr Ser Gly Ser Gly Gly Gly Gln Ala Ser Leu Ser Thr Pro Thr Tyr
405 410 415
Thr Gly Leu Ile Arg Leu Leu Ser Thr Cys Ala Ser Gly Ser Pro Leu
420 425 430
Gly Ala Lys Thr Leu Leu Leu His Gly Ala Ser Gly Ile Leu Lys Asp
435 440 445
Ile Leu Ser Gly Ser Gly Val Ser Ser Asn Thr Ser Val Ser Pro Ala
450 455 460
Leu Ser Arg Pro Ala Asp Gln Ile Phe Glu Ile Val Asn Leu Ala Asn
465 470 475 480
Glu Leu Leu Pro Pro Leu Pro Gln Gly Thr Ile Ser Leu Pro Val Ser
485 490 495
Ser Asn Leu Phe Val Lys Glu Ser Val Val Lys Lys Ser Pro Pro Ser
500 505 510
Gly Asn Pro Gly Ile Gln Glu Asp Thr Asn Gly Asn Val His Glu Ile
515 520 525
Ser Ala Arg Ala Lys Leu Leu Asn Asp Lys Pro Glu Leu Leu Lys Gln
530 535 540
Phe Gly Met Asp Leu Leu Pro Val Leu Met Gln Ile Tyr Gly Ala Ser
545 550 555 560
Val Asn Gly Pro Val Arg His Lys Cys Leu Ser Val Ile Gly Lys Leu
565 570 575
Met Tyr Phe Ser Thr Ala Glu Met Ile Gln Ser Leu Leu Ser Val Thr
580 585 590
Asn Ile Ser Ser Phe Leu Ala Gly Val Leu Ala Trp Lys Asp Pro His
595 600 605
Val Leu Val Pro Ala Leu Gln Ile Ser Glu Ile Leu Met Glu Lys Leu
610 615 620
Pro Gly Ile Phe Ser Lys Met Phe Val Arg Glu Gly Val Val His Ala
625 630 635 640
Val Asp Gln Leu Ile Leu Ala Gly Asn Ala Thr Asn Ile Ser Thr Gln
645 650 655
Thr Ser Ser Ala Glu Lys Asp Thr Asp Ser Val Ser Gly Thr Ser Ser
660 665 670
Arg Ser Arg Arg Tyr Arg Leu Arg Ser Gly Asn Ser Asn Pro Asp Ala
675 680 685
Asn Arg Ser Asp Asp Leu Lys Ser Pro Val Pro Val Asn Val Gly Leu
690 695 700
Pro Pro Ser Ser Val Glu Thr Pro Thr Thr Asn Ser Ser Ile Arg Ala
705 710 715 720
Ser Ile Ser Ser Val Ala Asn Ala Phe Lys Asp Lys Tyr Phe Pro Ser
725 730 735
Asp Pro Gly Ser Val Glu Val Gly Val Ser Asp Asp Leu Leu His Leu
740 745 750
Lys Asn Leu Cys Ser Lys Leu Asn Thr Gly Val Asp Asp Gln Arg Ser
755 760 765
Lys Ala Lys Gly Lys Val Glu Ala Ser Gly Phe Asp Leu Asp Asp Asp
770 775 780
Ser Thr Asn Thr Glu Glu Tyr Leu Ile Gly Val Ile Ser Asp Met Leu
785 790 795 800
Lys Glu Leu Gly Lys Gly Asp Ser Val Ser Thr Phe Glu Phe Ile Gly
805 810 815
Ser Gly Val Val Glu Ala Leu Leu Asn Tyr Phe Ser Cys Gly Tyr Phe
820 825 830
Ser Lys Asp Arg Ile Ser Glu Thr Asn Leu Pro Lys Leu Arg Gln Gln
835 840 845
Ala Leu Thr Arg Phe Lys Ser Phe Val Ala Val Ala Leu Pro Leu Ser
850 855 860
Ile Asp Asn Gly Ala Val Ala Pro Met Thr Val Leu Val Gln Lys Leu
865 870 875 880
Gln Asn Val Leu Ser Ser Leu Glu Arg Phe Pro Val Met Leu Ser Asn
885 890 895
Ser Ser Arg Ser Ser Ser Gly Ser Gly Arg Leu Ser Ser Gly Leu Ser
900 905 910
Ala Leu Ser Gln Pro Ile Lys Leu Arg Phe Cys Arg Ala Gln Gly Glu
915 920 925
Lys Ser Leu Lys Asp Tyr Ser Ser Ser Val Val Leu Ile Asp Pro Leu
930 935 940
Ala Ser Leu Ala Ala Ile Glu Glu Phe Leu Trp Ala Arg Val Gln Arg
945 950 955 960
Gly Glu Ser Gly Leu Lys Ser Thr Val Gly Thr Glu Asn Ser Glu Ser
965 970 975
Gly Thr Thr Pro Ala Gly Ala Gly Val Ser Ser Pro Ser Ser Tyr Ile
980 985 990
Pro Ser Thr Ala Phe Arg Tyr Ser Thr Gly Ser Arg Ser Arg Ser Ser
995 1000 1005
Val Asn Ile Gly Asp Thr Pro Arg Lys Glu Ile Phe Gln Asp Asn
1010 1015 1020
Gly Thr Ser Ser Ser Lys Ser Lys Gly Lys Ala Val Leu Lys Pro
1025 1030 1035
Ala Gln Glu Glu Ala Arg Gly Pro Gln Thr Arg Asn Ala Val Arg
1040 1045 1050
Arg Arg Ala Ala Leu Asp Lys Asp Ala Gln Met Lys Pro Ala Asn
1055 1060 1065
Gly Asp Ser Thr Ser Glu Asp Glu Glu Leu Asp Ile Ser Pro Val
1070 1075 1080
Glu Ile Asp Glu Ala Leu Val Ile Glu Asp Asp Asp Ile Ser Asp
1085 1090 1095
Asp Glu Asp Glu Asp Arg Glu Asp Val Arg Arg Asp Tyr Tyr Leu
1100 1105 1110
Pro Val Tyr Leu Pro Asp Glu Val His Asp Val Lys Leu Gly Asp
1115 1120 1125
Ser Ala Glu Glu Ser Thr Val Ala Pro Ala Thr Ser Asp Ser Gln
1130 1135 1140
Thr Asn Ala Ala Ser Gly Ser Ser Ser Lys Ala Gly Thr Ala Arg
1145 1150 1155
Gly Cys Asp Ser Ala Asp Phe Arg Ser Gly Tyr Ser Ser Ser Ser
1160 1165 1170
Arg Gly Ala Met Ser Phe Ala Ala Ala Ala Met Ala Gly Leu Gly
1175 1180 1185
Tyr Ala Asn Ser Arg Gly Phe Arg Gly Gly Arg Asp Arg His Gly
1190 1195 1200
Arg Leu Leu Phe Gly Ser Ser Asn Asp Pro Pro Lys Leu Ile Phe
1205 1210 1215
Thr Ala Gly Gly Lys His Leu Asn Arg Asn Leu Thr Ile Tyr Gln
1220 1225 1230
Ala Ile Gln Arg Gln Leu Met Leu Asp Glu Asp Asp Asp Glu Arg
1235 1240 1245
Leu Ala Gly Ser Asp Arg Val Ser Ser Asp Gly Ser Ser Leu Trp
1250 1255 1260
Gly Asp Ile Tyr Thr Ile Thr Tyr Gln Arg Ala Glu Asn Gln Pro
1265 1270 1275
Asp Lys Ala Ser Asn Gly Gly Ser Ser Ser Asn Thr Ser Lys Ser
1280 1285 1290
Ala Lys Ser Gly Ser Ala Leu Asn Ser Ser Ser Glu Ala Lys Leu
1295 1300 1305
His Gln Thr Ser Val Leu Asp Ser Ile Leu Gln Gly Asp Leu Pro
1310 1315 1320
Cys Asp Leu Glu Lys Ser Asn Pro Thr Tyr Asn Ile Leu Ala Leu
1325 1330 1335
Leu Arg Val Leu Glu Gly Leu Asn Gln Leu Ala Pro His Leu Arg
1340 1345 1350
Thr Gln Met Val Ser Asp Ser Phe Ala Lys Gly Lys Ile Leu Asp
1355 1360 1365
Leu Asp Glu Leu Gly Val Thr Thr Gly Ala Arg Val Leu Pro Glu
1370 1375 1380
Glu Phe Val Ser Gly Lys Leu Thr Pro Lys Leu Ala Arg Gln Ile
1385 1390 1395
Gln Asp Ala Leu Ala Leu Cys Ser Gly Ser Leu Pro Leu Trp Cys
1400 1405 1410
Cys Gln Leu Thr Lys Ala Cys Pro Phe Leu Phe Pro Phe Asp Thr
1415 1420 1425
Arg Arg Gln Tyr Phe Tyr Ser Thr Ala Phe Gly Leu Ser Arg Ala
1430 1435 1440
Leu Tyr Arg Leu Gln Gln Gln Gln Gly Ala Asp Gly His Gly Ser
1445 1450 1455
Thr Thr Glu Arg Glu Val Arg Val Gly Arg Leu Gln Arg Gln Lys
1460 1465 1470
Val Arg Val Ser Arg Asn Arg Val Leu Asp Ser Ala Ala Lys Val
1475 1480 1485
Met Gly Met Tyr Ser Ser Gln Lys Ala Val Leu Glu Val Glu Tyr
1490 1495 1500
Phe Gly Glu Val Gly Thr Gly Leu Gly Pro Thr Leu Glu Phe Tyr
1505 1510 1515
Thr Ile Leu Ser His Asp Leu Gln Gln Val Gly Leu Gln Met Trp
1520 1525 1530
Arg Ser Tyr Ser Ser Glu Lys His Gln Met Glu Ile Asp Arg Asp
1535 1540 1545
Glu Lys Lys Lys Lys Ser Asp Gly Ser Gly Pro Asn Leu Ala Gly
1550 1555 1560
Asp Gly Glu Leu Val Glu Ala Pro Leu Gly Leu Phe Pro Arg Pro
1565 1570 1575
Trp Pro Thr Asn Ser Asp Ala Ser Glu Gly Ser Arg Phe Ser Lys
1580 1585 1590
Val Val Glu Tyr Phe Arg Leu Leu Gly Arg Val Met Ala Lys Ala
1595 1600 1605
Leu Gln Asp Gly Arg Leu Leu Asp Leu Pro Leu Ser Val Ala Phe
1610 1615 1620
Tyr Lys Leu Val Leu Gly Gln Asp Leu Asp Leu His Asp Ile Leu
1625 1630 1635
Ser Ile Asp Ala Glu Leu Gly Lys Thr Leu Gln Glu Phe Asn Ala
1640 1645 1650
Leu Val Cys Arg Lys His Tyr Ile Glu Ser Ile Gly Gly Ser Tyr
1655 1660 1665
Thr Asp Thr Ile Val Asn Leu His Phe His Gly Val Pro Ile Glu
1670 1675 1680
Asp Leu Cys Leu Asp Phe Thr Leu Pro Gly Tyr Pro Glu Tyr Thr
1685 1690 1695
Leu Lys Pro Gly Asp Glu Ile Val Asp Ile Asn Asn Leu Glu Glu
1700 1705 1710
Tyr Ile Ser Leu Val Ala Asp Ala Thr Val Lys Thr Gly Ile Met
1715 1720 1725
Arg Gln Ile Glu Ala Phe Arg Ala Gly Phe Asn Gln Val Phe Asp
1730 1735 1740
Ile Ser Ser Leu Gln Ile Phe Thr Pro Gln Glu Leu Asp Asn Leu
1745 1750 1755
Leu Cys Gly Cys Arg Glu Leu Trp Glu Ser Glu Thr Leu Ala Asp
1760 1765 1770
His Ile Lys Phe Asp His Gly Tyr Asn Ala Lys Ser Pro Ala Ile
1775 1780 1785
Ile Asn Leu Leu Glu Ile Met Gly Gly Phe Thr Pro Glu Gln Gln
1790 1795 1800
Arg Ala Phe Cys Gln Phe Val Thr Gly Ala Pro Arg Leu Pro Pro
1805 1810 1815
Gly Gly Leu Ala Val Leu Asn Pro Lys Leu Thr Ile Val Arg Lys
1820 1825 1830
Leu Ser Ser Thr Ala Val Asn Thr Ser Ser Asn Gly Asn Gly Pro
1835 1840 1845
Ser Glu Ser Ala Asp Asp Asp Leu Pro Ser Val Met Thr Cys Ala
1850 1855 1860
Asn Tyr Leu Lys Leu Pro Pro Tyr Ser Thr Lys Glu Ile Met Tyr
1865 1870 1875
Lys Lys Leu Leu Tyr Ala Ile Asn Glu Gly Arg Gly Ser Phe Asp
1880 1885 1890
Leu Ser
1895
<210> 81
<211> 1891
<212> PRT
<213> 大豆
<400> 81
Met Glu Thr Arg Ser Arg Lys Arg Ala Glu Ala Ser Ser Ala Ala Pro
1 5 10 15
Ser Ser Ser Ser Thr Thr Pro Ser Arg Ser Ala Lys Arg Ser Arg Leu
20 25 30
Ser Ser Ser Ser Ser Ser Ile Leu Pro Val Asn Thr Arg Ser Arg Ser
35 40 45
Ala Arg Asn Asn Asn Asn Asn Asn Asn Ser Gly Ser Ile Ser Phe Met
50 55 60
Asp Pro Thr Asn Glu Ser Ser Gly Ser Arg Arg Asp Arg Arg Gly Lys
65 70 75 80
Asn Phe Asp Arg Glu Asn Ser Asp Lys Gly Lys Glu Lys Glu Gln Asp
85 90 95
Val Arg Ile Arg Asp Ala Glu Arg Glu Arg Glu Arg Ala Leu Ala Leu
100 105 110
Asn Met Glu Ser Glu Asp Val Gly Asp Asp Asp Asp Asn Asp Ser Asp
115 120 125
Gly Gly Val Gly Ile Leu His Gln Asn Leu Thr Ser Ala Ser Ser Ala
130 135 140
Leu Gln Gly Leu Leu Arg Lys Leu Gly Ala Gly Leu Asp Asp Leu Leu
145 150 155 160
Pro Ala Thr Ala Met Gly Gly Ser Ala Ser Ser Pro His Gln Ser Gly
165 170 175
Arg Leu Lys Lys Ile Leu Ser Gly Leu Arg Ala Asp Gly Glu Glu Gly
180 185 190
Arg Gln Val Glu Ala Leu Thr Gln Leu Cys Asp Met Leu Ser Ile Gly
195 200 205
Thr Glu Asp Ser Leu Ser Thr Phe Ser Val Asp Ser Phe Val Pro Val
210 215 220
Leu Val Gly Leu Leu Asn His Glu Ser Asn Pro Asp Val Met Leu Leu
225 230 235 240
Ala Ala Arg Ala Leu Thr His Leu Cys Asp Val Leu Pro Ser Ser Cys
245 250 255
Ala Ala Val Val His Tyr Gly Ala Val Ser Ile Phe Cys Ala Arg Leu
260 265 270
Leu Thr Ile Glu Tyr Met Asp Leu Ala Glu Gln Ser Leu Gln Ala Leu
275 280 285
Lys Lys Ile Ser Gln Glu His Pro Thr Ala Cys Leu Arg Ala Gly Ala
290 295 300
Leu Met Ala Val Leu Ser Tyr Leu Asp Phe Phe Ser Thr Gly Val Gln
305 310 315 320
Arg Val Ala Leu Ser Thr Ala Ala Asn Met Cys Lys Lys Leu Pro Pro
325 330 335
Asp Ala Ala Asp Phe Val Met Glu Ala Val Pro Leu Leu Thr Asn Leu
340 345 350
Leu Gln Tyr His Asp Ser Lys Val Leu Glu His Ala Ser Val Cys Leu
355 360 365
Thr Arg Ile Ala Glu Ala Phe Ala Ser Ser Pro Asp Lys Leu Asp Glu
370 375 380
Leu Cys Asn His Gly Leu Val Thr Gln Ala Ala Ser Leu Ile Ser Thr
385 390 395 400
Ser Ser Ser Gly Gly Gly Gln Ala Ser Leu Ser Thr Pro Thr Tyr Thr
405 410 415
Gly Leu Ile Arg Leu Leu Ser Thr Cys Ala Ser Gly Ser Pro Leu Gly
420 425 430
Ala Lys Thr Leu Leu Leu Leu Gly Thr Ser Gly Ile Leu Lys Asp Ile
435 440 445
Leu Ser Gly Ser Gly Val Ser Ser Asn Thr Ser Val Ser Pro Ala Leu
450 455 460
Ser Arg Pro Ala Asp Gln Ile Phe Glu Ile Val Asn Leu Ala Asn Glu
465 470 475 480
Leu Leu Pro Pro Leu Pro Gln Gly Thr Ile Ser Leu Pro Val Ser Ser
485 490 495
Asn Leu Phe Val Lys Gly Ser Val Val Lys Lys Ser Ser Ser Gly Asn
500 505 510
Ser Gly Ile Gln Glu Asp Thr Asn Gly Asn Val His Glu Ile Leu Ala
515 520 525
Arg Glu Lys Leu Leu Asn Asp Gln Pro Glu Leu Leu Gln Gln Phe Gly
530 535 540
Met Asp Leu Leu Pro Val Leu Met Gln Ile Tyr Gly Ala Ser Val Asn
545 550 555 560
Gly Pro Val Arg His Lys Cys Leu Ser Val Ile Gly Lys Leu Met Tyr
565 570 575
Phe Ser Thr Ala Glu Met Ile Gln Ser Leu Leu Ser Val Thr Asn Ile
580 585 590
Ser Ser Phe Leu Ala Gly Val Leu Ala Trp Lys Asp Pro His Val Leu
595 600 605
Val Pro Ala Leu Gln Ile Ser Glu Ile Leu Met Glu Lys Leu Pro Gly
610 615 620
Thr Phe Ser Lys Met Phe Val Arg Glu Gly Val Val His Ala Val Asp
625 630 635 640
Gln Leu Ile Leu Ala Gly Asn Ser Thr Asn Ile Ser Thr Gln Thr Ser
645 650 655
Ser Ala Glu Lys Asp Asn Asp Ser Val Ser Gly Thr Ser Ser Arg Ser
660 665 670
Arg Arg Tyr Arg Leu Arg Ser Gly Asn Ser Asn Pro Asp Ala Asn Pro
675 680 685
Ser Asp Asp Leu Lys Ser Pro Val Pro Val Asn Val Gly Leu Pro Pro
690 695 700
Ser Ser Val Glu Thr Pro Thr Thr Asn Ser Ser Ile Arg Ala Ser Val
705 710 715 720
Ser Ser Val Ala Arg Ala Phe Lys Asp Lys Tyr Phe Pro Ser Asp Pro
725 730 735
Gly Ser Val Glu Val Gly Val Ser Asp Asp Leu Leu His Leu Lys Asn
740 745 750
Leu Cys Thr Lys Leu Ile Thr Gly Val Asp Asp Gln Arg Ser Lys Ala
755 760 765
Lys Gly Lys Val Lys Ala Ser Gly Phe Gly Leu Asp Asp Asn Ser Ser
770 775 780
Asn Thr Glu Glu Tyr Leu Ile Gly Val Ile Ser Asp Met Leu Lys Glu
785 790 795 800
Leu Gly Lys Gly Asp Ser Val Ser Thr Phe Glu Phe Ile Gly Ser Gly
805 810 815
Val Val Glu Ala Leu Leu Asn Tyr Phe Ser Cys Gly Tyr Phe Ser Lys
820 825 830
Asp Arg Ile Ser Glu Thr Asn Leu Pro Lys Leu Arg Gln Gln Ala Leu
835 840 845
Ser Arg Phe Lys Ser Phe Val Ala Val Ala Leu Pro Leu Ser Ile Asp
850 855 860
Asn Gly Ala Val Ala Pro Met Thr Val Leu Val Gln Lys Leu Gln Asn
865 870 875 880
Ala Leu Ala Ser Leu Glu Arg Phe Pro Val Met Leu Ser Asn Ser Ser
885 890 895
Arg Ser Ser Ser Gly Ser Ala Arg Leu Ser Ser Gly Leu Ser Ala Leu
900 905 910
Ser Gln Pro Ile Lys Leu Arg Leu Cys Arg Ala Gln Gly Glu Lys Ser
915 920 925
Leu Arg Asp Tyr Ser Ser Asn Val Val Leu Ile Asp Pro Leu Ala Ser
930 935 940
Leu Ala Ala Ile Glu Glu Phe Leu Trp Ala Arg Val Gln Arg Gly Glu
945 950 955 960
Ser Gly Gln Lys Ser Thr Val Gly Thr Glu Asn Ser Glu Ser Gly Thr
965 970 975
Thr Pro Ala Gly Ala Gly Val Ser Ser Pro Ser Ser Tyr Thr Pro Ser
980 985 990
Thr Ala His Arg His Ser Thr Arg Thr Arg Ser Ser Val Asn Ile Gly
995 1000 1005
Asp Thr Pro Arg Lys Glu Thr Ser Gln Asp Lys Gly Thr Ser Ser
1010 1015 1020
Ser Lys Ser Lys Gly Lys Ala Val Leu Lys Pro Ala Gln Glu Glu
1025 1030 1035
Ala Gln Gly Pro Gln Thr Arg Asn Thr Val Arg Arg Arg Ala Ala
1040 1045 1050
Leu Asp Lys Val Ala Gln Met Lys Pro Ala Asn Gly Asp Ser Thr
1055 1060 1065
Ser Glu Asp Glu Glu Leu Asp Ile Ser Pro Val Glu Ile Ala Glu
1070 1075 1080
Ala Leu Val Ile Glu Asp Asp Asp Ile Ser Asp Asp Glu Asp Glu
1085 1090 1095
Asp His Glu Asp Val Leu Arg Asp Asp Ser Leu Pro Val Cys Leu
1100 1105 1110
Pro Asp Lys Val His Asp Val Lys Leu Gly Asp Ser Ala Glu Glu
1115 1120 1125
Ser Thr Val Ala Pro Ala Thr Ser Asp Ser Gln Thr Asn Ala Ala
1130 1135 1140
Ser Gly Ser Ser Ser Lys Ala Gly Thr Ala Arg Gly Ser Asp Ser
1145 1150 1155
Ala Asp Phe Arg Ser Gly Phe Ser Ser Ser Ser Arg Gly Ala Met
1160 1165 1170
Ser Phe Ala Ala Ala Ala Met Ala Gly Leu Gly Tyr Ala Asn Ser
1175 1180 1185
Arg Gly Phe Arg Gly Gly Arg Asp Arg His Gly Cys Leu Leu Phe
1190 1195 1200
Gly Ser Ser Asn Asp Pro Pro Lys Leu Ile Phe Thr Thr Gly Gly
1205 1210 1215
Lys Gln Leu Asn Arg Asn Leu Ser Ile Tyr Gln Ala Ile Gln Arg
1220 1225 1230
Gln Leu Val Leu Asp Glu Asp Asp Asp Glu Arg Phe Ala Gly Ser
1235 1240 1245
Asp Tyr Val Ser Gly Asp Gly Ser Ser Leu Trp Gly Asp Ile Tyr
1250 1255 1260
Thr Ile Thr Tyr Gln Arg Ala Glu Asn Gln Pro Asp Lys Ala Ser
1265 1270 1275
Thr Gly Gly Ser Ser Ser Asn Thr Ser Lys Ser Ala Lys Ser Gly
1280 1285 1290
Ser Ala Leu Asn Ser Ser Ser Glu Ala Lys Leu His Gln Thr Ser
1295 1300 1305
Val Leu Asp Ser Ile Leu Gln Gly Glu Leu Pro Cys Asp Leu Glu
1310 1315 1320
Lys Ser Asn Pro Thr Tyr Asn Ile Leu Ala Leu Leu Arg Val Leu
1325 1330 1335
Glu Gly Phe Asn Gln Leu Ala Pro Arg Leu Arg Val Leu Met Val
1340 1345 1350
Ser Asp Ser Phe Ala Lys Gly Lys Ile Leu Asp Leu Asp Glu Leu
1355 1360 1365
Cys Val Thr Thr Gly Ala Arg Val Leu Leu Glu Glu Phe Val Ser
1370 1375 1380
Gly Lys Leu Thr Pro Lys Leu Ala Arg Gln Ile Gln Asp Ala Leu
1385 1390 1395
Ala Leu Cys Ser Gly Asn Leu Pro Leu Trp Cys Tyr Gln Leu Thr
1400 1405 1410
Lys Ala Cys Pro Phe Leu Phe Pro Phe Glu Thr Arg Arg Gln Tyr
1415 1420 1425
Phe Tyr Ser Thr Ala Phe Gly Leu Ser Arg Ala Leu Tyr Arg Leu
1430 1435 1440
Gln Gln Gln Gln Gly Ala Asp Gly His Gly Ser Thr Thr Glu Arg
1445 1450 1455
Glu Val Arg Val Gly Arg Leu Gln Arg Gln Lys Val Arg Val Ser
1460 1465 1470
Arg Asn Arg Val Leu Asp Ser Ala Ala Lys Val Met Glu Met Tyr
1475 1480 1485
Ser Ser Gln Lys Ala Val Leu Glu Val Glu Tyr Phe Gly Glu Val
1490 1495 1500
Gly Thr Gly Leu Gly Pro Thr Leu Glu Phe Tyr Thr Ile Leu Ser
1505 1510 1515
His Asp Leu Gln Lys Val Gly Leu Gln Met Trp Arg Ser Tyr Ser
1520 1525 1530
Ser Asp Lys His Gln Met Glu Ile Asp Gly Asp Glu Lys Lys Lys
1535 1540 1545
Lys Ser Glu Gly Ser Gly Pro Asn Leu Ala Gly Asp Gly Glu Leu
1550 1555 1560
Val Gln Ala Pro Leu Gly Leu Phe Pro Arg Pro Trp Pro Thr Asn
1565 1570 1575
Ser Asp Ala Ser Glu Ser Ser Gln Phe Ser Lys Val Ile Glu Tyr
1580 1585 1590
Phe Arg Leu Leu Gly Arg Val Met Ala Lys Ala Leu Gln Asp Gly
1595 1600 1605
Arg Leu Leu Asp Leu Pro Leu Ser Val Ala Phe Tyr Lys Leu Val
1610 1615 1620
Leu Cys Gln Asp Leu Asp Leu His Asp Ile Leu Phe Ile Asp Ala
1625 1630 1635
Glu Leu Gly Lys Thr Leu Gln Glu Phe Asn Ala Leu Val Cys Arg
1640 1645 1650
Lys His Tyr Ile Glu Ser Ile Gly Gly Ser Tyr Thr Asp Thr Ile
1655 1660 1665
Val Asn Leu Tyr Phe His Gly Ala Pro Ile Glu Asp Leu Cys Leu
1670 1675 1680
Asp Phe Thr Leu Pro Gly Tyr Pro Glu Tyr Thr Leu Lys Pro Gly
1685 1690 1695
Asp Glu Ile Val Asp Ile Asn Asn Leu Glu Glu Tyr Ile Ser Leu
1700 1705 1710
Val Ile Asp Ala Thr Val Lys Thr Gly Ile Met Arg Gln Ile Glu
1715 1720 1725
Ala Phe Arg Ala Gly Phe Asn Gln Val Phe Asp Ile Ser Ser Leu
1730 1735 1740
Gln Ile Phe Thr Pro Gln Glu Leu Asp Asn Leu Leu Cys Gly Arg
1745 1750 1755
Arg Glu Leu Trp Glu Ala Glu Thr Leu Ala Asp His Ile Lys Phe
1760 1765 1770
Asp His Gly Tyr Asn Ala Lys Ser Pro Ala Ile Val Asn Leu Leu
1775 1780 1785
Glu Ile Met Gly Glu Phe Thr Pro Glu Gln Gln Arg Ala Phe Cys
1790 1795 1800
Gln Phe Val Thr Gly Ala Pro Arg Leu Pro Pro Gly Gly Leu Ala
1805 1810 1815
Val Leu Asn Pro Lys Leu Thr Ile Val Arg Lys Leu Ser Ser Thr
1820 1825 1830
Ala Val Asn Asn Ser Ser Asn Gly Asn Gly Pro Ser Glu Ser Ala
1835 1840 1845
Asp Asp Asp Leu Pro Ser Val Met Thr Cys Ala Asn Tyr Leu Lys
1850 1855 1860
Leu Pro Pro Tyr Ser Thr Lys Glu Ile Met Tyr Lys Lys Leu Leu
1865 1870 1875
Tyr Ala Ile Ser Glu Gly Gln Gly Ser Phe Asp Leu Ser
1880 1885 1890
<210> 82
<211> 1840
<212> PRT
<213> 玉米
<400> 82
Met Glu Thr Arg Ser Arg Lys Arg Ala Glu Ala Ser Ser Ser Ser Ala
1 5 10 15
Thr Ser Ser Ser Arg Ser Ser Lys Arg Ser Arg His Asn Pro Asn Pro
20 25 30
Asn Pro Pro Ala Gly Pro Ser Pro Ala Pro Lys Leu Val Pro Leu Pro
35 40 45
Pro Arg Thr Arg Arg Ser Thr Ala Val Asn Pro Leu Pro Pro Met Asp
50 55 60
Ser Ser Gly Asp Asn Asn Ser Asn Pro Val Pro Pro Pro Arg Arg Arg
65 70 75 80
Gly Arg Pro Ser Asn Thr Asp Lys Gly Lys Glu Gln Gln Gln Pro Glu
85 90 95
Pro Ser His Ser Ser Arg Val Arg Glu Ala Glu Arg Leu Leu Gly Leu
100 105 110
Gly Phe Glu Gly Ile Asp Asp Asp Glu Asp Ser Gly Phe Gly Ala Gly
115 120 125
Ala Ile Pro His Ser Leu Thr Ser Ala Ser Thr Ala Leu Gln Gly Leu
130 135 140
Leu Arg Lys Leu Gly Ala Gly Leu Asp Asp Ile Leu Pro Ser Ser Ala
145 150 155 160
Leu Ser Ala Ala Ala Ala Ala Ala Ser Ser Ser Ser Ala Ser Gly Gln
165 170 175
Leu Ser Gly Arg Leu Lys Asn Ile Leu Ala Gly Leu Arg Ala Asp Gly
180 185 190
Glu Asp Gly Arg Gln Val Glu Ala Leu Thr Gln Leu Cys Glu Met Leu
195 200 205
Ser Ile Gly Thr Glu Glu Ser Leu Gly Ala Phe Ser Val Asp Ser Phe
210 215 220
Val Pro Val Leu Val Gly Leu Leu Asn His Glu Ser Asn Pro Asp Ile
225 230 235 240
Met Leu Leu Ala Ala Arg Ala Leu Thr His Leu Cys Asp Val Leu Pro
245 250 255
Ser Ser Cys Ser Ala Val Val His Tyr Gly Ala Val Pro Cys Phe Cys
260 265 270
Ala Arg Leu Leu Thr Ile Glu Tyr Met Asp Leu Ala Glu Gln Arg Val
275 280 285
Ala Leu Ser Thr Ala Ala Asn Met Cys Arg Lys Leu Pro Ser Asp Ala
290 295 300
Ser Asp Phe Val Met Glu Ala Val Pro Leu Leu Thr Asn Leu Leu Asn
305 310 315 320
Tyr His Asp Ser Lys Val Leu Glu His Ala Ser Val Cys Leu Thr Arg
325 330 335
Ile Ala Glu Ser Phe Ser Pro Phe Pro Glu Lys Leu Asp Glu Leu Cys
340 345 350
Ser His Gly Leu Val Ala Gln Ala Ala Ser Leu Val Ser Val Ser Asn
355 360 365
Ser Ala Gly Gln Ala Ser Leu Ser Thr Ser Thr Tyr Thr Gly Val Ile
370 375 380
Arg Leu Leu Ser Ile Cys Ala Ser Gly Ser Pro Leu Ala Ala Lys Thr
385 390 395 400
Leu Leu Leu Leu Gly Ile Ser Gly Ile Leu Lys Asp Ile Leu Ser Gly
405 410 415
Ser Gly Leu Val Ala Gly Thr Thr Val Ser Pro Ala Leu Thr Arg Pro
420 425 430
Ala Asp Gln Met Asn Glu Ile Val Lys Leu Ala Asp Glu Leu Leu Pro
435 440 445
Ser Leu Pro Val Gly Thr Ile Ser Leu Pro Val Tyr Ser Gly Val His
450 455 460
Met Lys Gly Cys Ser Val Lys Lys Ser Thr Ser Ser Lys Gln Gly Glu
465 470 475 480
His Gly Ser Thr Ala Asn Glu Leu Ser Gly Arg Glu Lys Leu Leu Arg
485 490 495
Asp Gln Pro Glu Leu Leu Gln Gln Phe Gly Met Asp Leu Leu Pro Thr
500 505 510
Met Thr Gln Val Tyr Gly Ser Ser Val Ser Gly Pro Ile Arg His Arg
515 520 525
Cys Leu Ser Val Ile Gly Lys Leu Met Tyr Tyr Ser Ser Ala Glu Met
530 535 540
Ile Gln Ser Leu Leu Ser Thr Thr Asn Ile Ser Ser Phe Leu Ala Gly
545 550 555 560
Ile Leu Ala Trp Lys Asp Pro Gln Val Leu Ile Pro Ala Leu Gln Ile
565 570 575
Ala Glu Val Leu Met Glu Lys Leu Pro Glu Ile Phe Leu Lys Met Phe
580 585 590
Val Arg Glu Gly Val Val His Ala Val Glu Ser Leu Ile Cys Pro Glu
595 600 605
Leu Ser Gly Gln Val Thr Pro His Val Asp Ser Ile Thr Ser Ser His
610 615 620
Asn Arg Arg Asn Arg Arg Arg Asn Asn Ala Val Asn Thr Gly Asn Asn
625 630 635 640
Leu Pro Asp Gly Pro Lys Gly Ser Asn Ser Met Ile Ala Asn Ser Pro
645 650 655
Pro Ser Met Ala Glu Val Pro Asn Asn Ser Leu Arg Ala Leu Val Ser
660 665 670
Asn His Ala Lys Ser Phe Lys Asp Lys Tyr Phe Pro Ser Glu Pro Gly
675 680 685
Ser Ser Asp Ile Ala Val Thr Asp Asp Leu Leu Lys Leu Arg Ala Leu
690 695 700
Cys Ala Lys Leu Asn Thr Thr Ala Asp Thr Ile Lys Thr Lys Ala Lys
705 710 715 720
Gly Lys Ser Lys Val Val Ser Asp Asn Ser Phe Asp Val Leu Cys Asn
725 730 735
Ile Glu Glu Gln Leu Asp Asp Ile Ile Ala Glu Met Leu Ser Glu Leu
740 745 750
Ser Lys Gly Asp Gly Val Ser Thr Phe Glu Phe Ile Gly Ser Gly Val
755 760 765
Val Thr Ala Leu Leu Thr Tyr Leu Ser Cys Gly Thr Phe Gly Arg Glu
770 775 780
Lys Val Ser Glu Ala Asn Ile Pro Asn Leu Arg His Gln Ala Val Arg
785 790 795 800
Arg Tyr Lys Thr Phe Ile Ser Phe Ala Leu Pro Asn Asp Lys Asp Gly
805 810 815
Asn Lys Thr Pro Met Ala Phe Leu Val His Lys Leu Gln Ser Ala Leu
820 825 830
Ser Ser Leu Glu Arg Phe Pro Val Val Leu Ser His Ser Gly Arg Ala
835 840 845
Ser Thr Leu Gly Gly Ser Arg Leu Thr Thr Gly Leu Gly Ser Leu Ser
850 855 860
Gln Pro Ile Lys Leu Arg Leu Cys Arg Ala Pro Gly Glu Lys Ser Leu
865 870 875 880
Lys Asp Phe Ser Ser Asn Val Val Leu Ile Asp Ser Leu Ala Ser Leu
885 890 895
Ala Ala Val Glu Asp Phe Leu Trp Pro Arg Val Gln Arg Thr Glu Pro
900 905 910
Val Leu Lys Pro Pro Met Ser Ser Ala Asn Asn Ser Gly Ser Gly Ala
915 920 925
Ala Ser Ser Thr Ala Cys Ala Pro Ser Ile Pro Ser Glu Thr Gln Ser
930 935 940
Val Arg Arg Thr Ser Leu Arg Ser Lys Ser Ser Ala Ala Thr Ser Gly
945 950 955 960
Ala Ile Lys Lys Asp Tyr Gln Glu Gly Ser Ile Asn Thr Ser Lys Gly
965 970 975
Lys Gly Lys Ala Val Leu Lys Leu Ser Leu Asp Glu Pro Lys Gly Pro
980 985 990
His Thr Arg Asn Ala Ala Arg Arg Lys Ala Thr Ser Glu Lys Asp Val
995 1000 1005
Glu Leu Lys Pro Ser His Gly His Ile Thr Ser Glu Asp Glu Asp
1010 1015 1020
Leu Asp Ala Ser Pro Val Glu Ile Asp Asp Ala Leu Ile Leu Asp
1025 1030 1035
Asp Asp Asp Glu Asp Val Pro Asp Asp Glu Asp Asp Asp His Glu
1040 1045 1050
Ala Val Leu Arg Gly Ser Leu Pro Ser Cys Val Pro Glu Arg Val
1055 1060 1065
His Asp Val Lys Leu Gly Asp Ala Asp Asp Ser Ser Val Ala Ser
1070 1075 1080
Leu Ala Asn Asp Asn Gln Ala Gln Pro Ser Ser Gly Ser Ser Thr
1085 1090 1095
Lys Asn Thr Ser Ser Arg Gly Leu Asp Thr Ala Glu Phe Arg Ser
1100 1105 1110
Pro Ala Thr Phe Gly Ser Arg Gly Ala Met Ser Phe Ala Ala Ala
1115 1120 1125
Ala Met Ala Gly Leu Thr Pro Val Gly Gly Arg Gly Ile Arg Gly
1130 1135 1140
Ser Arg Asp Arg Asn Gly Leu Pro Leu Gly Ala Arg Ala Thr Glu
1145 1150 1155
His Tyr Asn Lys Leu Ile Phe Thr Ala Ala Gly Lys Gln Leu Asn
1160 1165 1170
Lys His Leu Thr Val Tyr Gln Ala Val Gln Arg Gln Val Val His
1175 1180 1185
Ala Glu Asp Asp Glu Asp Arg Phe Gly Gly Ser Asp Leu Pro Asp
1190 1195 1200
Asp Gly Asn His Phe Trp Asp Asp Ile Arg Gly Asp Val Phe Thr
1205 1210 1215
Ile Thr Tyr Gln Lys Ala Asp Asn Thr Ala Glu Lys Gly Ser Val
1220 1225 1230
Gly Gly Ser Ala Ser Val Pro Lys Ser Ser Lys Ser Asp Ser Cys
1235 1240 1245
Arg Thr Leu Ser Glu Lys Gln Cys Thr Ser Leu Leu Asp Ser Ile
1250 1255 1260
Leu Gln Gly Glu Leu Pro Cys Asp Leu Glu Lys Ser Asn Gln Thr
1265 1270 1275
Tyr Asn Ile Leu Ser Leu Leu His Val Leu Glu Gly Leu Asn Gln
1280 1285 1290
Leu Ser Pro Arg Leu Arg Leu Gln Ser Ala Cys Asp Asp Phe Ala
1295 1300 1305
Glu Gly Lys Val Ala Thr Leu Asn Gly Leu Tyr Asp Val Gly Ala
1310 1315 1320
Lys Val Pro Ser Lys Glu Phe Ile Asn Ser Lys Met Thr Pro Lys
1325 1330 1335
Leu Ala Arg Gln Ile Gln Asp Val Leu Ala Leu Cys Ser Gly Ser
1340 1345 1350
Leu Pro Ser Trp Cys Tyr Gln Leu Thr Lys Ala Cys Pro Phe Leu
1355 1360 1365
Phe Pro Phe Glu Thr Arg Arg Gln Tyr Phe Tyr Ser Thr Ala Phe
1370 1375 1380
Gly Leu Ser Arg Ala Leu His Arg Leu Gln Gln Gln Pro Gly Asn
1385 1390 1395
Asp Asn Asn Thr Ala Phe Glu Arg Glu Val Arg Ile Gly Arg Leu
1400 1405 1410
Gln Arg Gln Lys Val Arg Val Ser Arg Asn Arg Ile Leu Asp Ser
1415 1420 1425
Ala Ala Lys Val Met Glu Met Phe Ser Asn Gln Lys Ala Val Leu
1430 1435 1440
Glu Val Glu Tyr Phe Gly Glu Val Gly Thr Gly Leu Gly Pro Thr
1445 1450 1455
Leu Glu Phe Tyr Thr Leu Leu Ser Arg Glu Leu Gln Arg Val Asp
1460 1465 1470
Leu Gly Leu Trp Arg Ser His Ser Ser Asp Asn Ser Gly Met Gln
1475 1480 1485
Ile Asp Ala Asn Ala Asp Asp Leu Ile Arg Ser Lys Asn His Glu
1490 1495 1500
Ser Glu Ser Leu Thr Glu Ser Arg Asn Ile Val Gln Ser Pro Leu
1505 1510 1515
Gly Leu Phe Pro Gln Pro Trp Pro Pro Thr Ala Ala Ala Ser Glu
1520 1525 1530
Gly Ser Lys Phe Phe Lys Val Val Glu Tyr Phe Arg Leu Val Gly
1535 1540 1545
Arg Val Met Ala Lys Ala Leu Gln Asp Gly Arg Leu Leu Asp Leu
1550 1555 1560
Pro Leu Ser Thr Ala Phe Tyr Lys Leu Leu Leu Gly Gln Glu Leu
1565 1570 1575
Asp Leu Tyr Asp Ile Leu Ser Phe Asp Thr Glu Phe Gly Lys Thr
1580 1585 1590
Leu Gln Glu Leu Gln Ile Leu Val Ala Arg Lys Gln Phe Leu Asp
1595 1600 1605
Ser Cys Ser Ser Glu Ser Gln Lys Ile Asp Leu Cys Phe Arg Gly
1610 1615 1620
Ala Pro Val Glu Asp Leu Tyr Leu Asp Phe Thr Leu Pro Gly Tyr
1625 1630 1635
Pro Glu Tyr Val Leu Lys Glu Gly Gly Glu Asn Ala Glu Val Asn
1640 1645 1650
Ile Cys Asn Leu Glu Glu Tyr Ile Ser Leu Val Val Asp Ala Thr
1655 1660 1665
Val Lys Thr Gly Ile Met Arg Gln Val Glu Ala Phe Lys Ala Gly
1670 1675 1680
Phe Asn Gln Val Phe Asp Ile Ser Ser Leu Gln Ile Phe Ser Pro
1685 1690 1695
Gln Glu Leu Asp Tyr Leu Ile Cys Gly Arg Cys Glu Leu Trp Glu
1700 1705 1710
Pro Glu Thr Leu Pro Glu His Ile Lys Phe Asp His Gly Tyr Thr
1715 1720 1725
Ser Lys Ser Pro Ala Ile Ile Asn Phe Leu Glu Ile Met Ala Glu
1730 1735 1740
Phe Thr Pro Glu Gln Gln His Ala Phe Cys Gln Phe Val Thr Gly
1745 1750 1755
Ala Pro Arg Leu Pro Pro Gly Gly Leu Ala Ala Leu Asn Pro Lys
1760 1765 1770
Leu Thr Ile Val Arg Lys His Ser Ser Val Ala Asn Asn Asn Ser
1775 1780 1785
Asn Ala Thr Gly Ala Thr Glu Ser Ala Asp Asp Asp Leu Pro Ser
1790 1795 1800
Val Met Thr Cys Ala Asn Tyr Leu Lys Leu Pro Pro Tyr Ser Thr
1805 1810 1815
Lys Ala Ile Met Leu Lys Lys Leu Leu Tyr Ala Ile Asn Glu Gly
1820 1825 1830
Gln Gly Ser Phe Asp Leu Ser
1835 1840
<210> 83
<211> 1833
<212> PRT
<213> 玉米
<400> 83
Met Glu Thr Arg Ser Ser Ser Arg Lys Arg Ala Ala Ala Ala Ala Ser
1 5 10 15
Ser Ser Ala Ser Lys Arg Pro Arg Arg Asn Ser Ser Ser Ser Ser Ser
20 25 30
Ala Pro Pro Arg Gln Gln Pro Pro Met Glu Pro Ser Pro Ser Ser Arg
35 40 45
Arg Arg Ser Arg Ala Ala Ala Ala Asp Lys Gly Lys Asp Pro Asp Pro
50 55 60
Ala Ser Ser Asp His Pro Ser Pro Pro Ala Pro Asp Asp Asp Ala Asp
65 70 75 80
Val Thr Phe Pro Gln Pro Phe Thr Ser Ala Ser Thr Ala Leu Gln Gly
85 90 95
Leu Leu Arg Arg Leu Gly Ala Gly Leu Asp Asp Leu Leu Pro Ser Ser
100 105 110
Ser Ser Ala Pro Ser Ser Ala Thr Ser Gly His Leu Lys Arg Ile Leu
115 120 125
Ala Gly Leu Gln Ala His Gly Asp Glu Ser Arg Gln Leu Gln Ser Leu
130 135 140
Met Gln Leu Cys Glu Met Leu Ser Ile Gly Thr Glu Asp Ser Leu Ala
145 150 155 160
Ala Phe Pro Val Asp Ala Phe Val Pro Ile Leu Val Gly Met Leu Gly
165 170 175
Arg Glu Asp Glu Pro Ala Thr Ala Gly Ala Ser Pro Asp Val Met Leu
180 185 190
Leu Ala Ala Arg Ala Leu Ser Asn Leu Val Asp Val Leu Pro Ser Ser
195 200 205
Cys Ser Ala Val Val His Tyr Gly Ala Ile Gln Cys Phe Cys Ala Arg
210 215 220
Leu Leu Thr Ile Glu Tyr Met Asp Leu Ala Glu Gln Ser Leu Gln Ala
225 230 235 240
Leu Lys Lys Ile Ser Leu Glu His Pro Thr Ala Cys Leu Arg Ala Gly
245 250 255
Ala Leu Met Ala Val Leu Ser Tyr Leu Asp Phe Phe Ser Thr Gly Val
260 265 270
Gln Arg Val Ala Leu Ser Thr Ala Ala Asn Ile Cys Arg Lys Leu Pro
275 280 285
Ser Asp Ala Ser Glu Phe Val Met Glu Ala Val Pro Leu Leu Thr Asn
290 295 300
Leu Leu Asn His His Asp Ser Lys Val Leu Glu His Ala Ser Val Cys
305 310 315 320
Leu Thr Arg Ile Ala Glu Ala Phe Ala Tyr Tyr Pro Asp Lys Leu Asp
325 330 335
Glu Leu Cys Asn His Gly Leu Val Ala Gln Ala Ala Ser Leu Ile Ala
340 345 350
Val Gly Asn Ser Ser Gly Gln Thr Ser Leu Ser Thr Ser Thr Tyr Thr
355 360 365
Gly Leu Ile Arg Leu Leu Ser Ile Cys Ala Ser Gly Ser Leu Leu Ala
370 375 380
Val Lys Thr Leu Leu Leu Leu Gly Ile Ser Gly Thr Leu Lys Asp Ile
385 390 395 400
Leu Ser Gly Ser Ala Leu Ile Ser Gly Thr Ser Val Ser Pro Ala Val
405 410 415
Thr Arg Pro Ala Asp Gln Met Phe Glu Ile Val Ser Leu Ala Asp Asp
420 425 430
Leu Leu Pro His Ile Pro Ala Arg Ile Ile Lys Leu Pro Thr Tyr Tyr
435 440 445
His Thr Tyr Lys Gly Ser Ser Thr Lys Lys Ser Ala Ser Ile Lys Gln
450 455 460
Glu Gly Gly Gly Ser Thr Gly Asn Glu Arg Ser Gly Arg Glu Arg Leu
465 470 475 480
Leu Arg Glu Gln Pro Glu Leu Leu Gln Gln Phe Gly Met Asp Leu Leu
485 490 495
Pro Thr Met Thr Gln Val Tyr Gly Ser Ser Val Asn Ala Pro Ile Arg
500 505 510
His Lys Cys Leu Ser Ile Ile Gly Lys Leu Met Tyr Tyr Ser Ser Ala
515 520 525
Glu Thr Ile Gln Ser Leu Leu Gly Thr Thr Asn Ile Ser Ser Phe Leu
530 535 540
Ala Gly Ile Leu Ala Trp Lys Asp Pro Gln Val Leu Ile Pro Ala Leu
545 550 555 560
Gln Ile Ala Glu Ile Met Met Glu Lys Leu Pro Glu Thr Phe Ser Lys
565 570 575
Leu Phe Val Arg Glu Gly Val Val His Ala Val Glu Ser Leu Ile Cys
580 585 590
Ser Glu Ser Ser Asn Lys Met Pro Ser Gln Val Pro Pro Gln Asp Lys
595 600 605
Asp Lys Asp Ser Ala Met Pro Ser Arg Ser Arg Arg Gln Arg Arg Arg
610 615 620
Ala Gly Ala Val Ala Ala Glu Asn Ser Ser Leu Asp Glu Ser Asn Ser
625 630 635 640
Ser Asn Leu Gly Val Met Cys Ser Thr Ala Thr Ala Ser Glu Ala Pro
645 650 655
Asn Thr Ser Leu Arg Phe Thr Val Ser Asp His Ala Lys Ser Phe Lys
660 665 670
Asp Lys Tyr Phe Pro Ala Asp Thr Asp Ser Ser Asp Ile Gly Phe Thr
675 680 685
Asp Asp Leu Leu Lys Leu Arg Ala Leu Cys Ala Lys Leu Asn Thr Val
690 695 700
Ser Glu Asn Val Lys Thr Lys Ala Lys Gly Lys Ser Lys Ala Ile Ser
705 710 715 720
Thr Asn Phe Leu Asp Ile Ser Ile Asp Val Glu Glu Gln Leu Asp Lys
725 730 735
Ile Ile Ser Glu Met Leu Ser Glu Leu Ser Lys Val Asn Gly Val Ser
740 745 750
Thr Phe Glu Phe Ile Arg Ser Gly Val Val Ile Ala Leu Leu Asp Tyr
755 760 765
Leu Ser Cys Gly Thr Phe Gly Lys Glu Lys Val Ser Glu Gly Asn Leu
770 775 780
Ser Gln Leu Arg Gln Gln Ala Leu Arg Arg Tyr Lys Thr Phe Ile Ser
785 790 795 800
Val Ala Leu Ser Ile Asp His Gly Arg Asp Glu Thr Pro Met Ala Leu
805 810 815
Leu Val Gln Lys Leu Gln Ser Ala Leu Ser Ser Leu Glu Arg Phe Pro
820 825 830
Val Val Leu Ser Gln Ser Ser Arg Ile Gly Ile Gly Gly Ser Arg Leu
835 840 845
Thr Ser Gly Leu Ser Ala Leu Ala Gln Pro Phe Lys Leu Arg Leu Ser
850 855 860
Arg Ala Gln Gly Glu Lys Ser Leu Arg Asp Tyr Ser Ser Asn Ile Val
865 870 875 880
Leu Ile Asp Pro Phe Ala Ser Leu Ala Ser Val Glu Glu Phe Leu Trp
885 890 895
Pro Arg Val Gln Arg Ser Glu Val Ala Ser Lys Pro Ile Ile Pro Ser
900 905 910
Gly Asn Asn Ser Glu Ser Gly Val Pro Gly Thr Thr Ala Gly Ala Ser
915 920 925
Leu Thr Ala Ala Met Ala Gln Ser Gly Arg Arg Pro Thr Thr Arg Ser
930 935 940
Lys Ser Ser Ala Ala Gly Gly Leu Thr Ser Lys Lys Asp Ser His Asp
945 950 955 960
Glu Ser Thr Ser Thr Ala Lys Gly Lys Gly Lys Ala Ile Val Lys Pro
965 970 975
Asn Ser Asp Glu Ser Lys Gly Pro Asn Thr Arg Asn Ala Ala His Gln
980 985 990
Lys Ser Ala Ser Glu Lys Asp Leu Glu Met Lys Gln Ala His Gly His
995 1000 1005
Ser Ser Ser Glu Asp Glu Glu Leu Asp Thr Ser Pro Val Glu Ile
1010 1015 1020
Asp Asp Ala Leu Met Ile Asp Asp Asp Asp Ile Ser Asp Asp Asp
1025 1030 1035
Asp Asp Asp Asp Asp Asp His Glu Val Leu Gln Glu Gly Ser Leu
1040 1045 1050
Pro Ile Cys Ser Gln Asp Gly Val His Asp Val Lys Leu Gly Asp
1055 1060 1065
Ala Asp Glu Ser Asn Ile Gly Ser Ala Thr Phe Ser His Ala Gln
1070 1075 1080
Pro Ser Ser Gly Ser Gly Ala Arg Asn Thr Ser Ser Arg Gly Pro
1085 1090 1095
Asp Ser Thr Glu Phe Arg Ser Met Gly Thr Phe Gly Ser Arg Gly
1100 1105 1110
Ala Met Ser Phe Val Ala Ala Thr Met Ala Gly Leu Ala Ser Val
1115 1120 1125
Gly Gly Arg Ser Val Arg Gly Val Arg Asp Arg Arg Gly Leu Ser
1130 1135 1140
Leu Gly Gly Ser Met Asn Asp His Asn Lys Leu Ile Phe Thr Ala
1145 1150 1155
Gly Gly Lys Gln Leu Asn Lys His Leu Thr Val Tyr Gln Ala Ile
1160 1165 1170
Gln Arg Gln Leu Met Leu Asp Glu Asp Asp Glu Glu Arg Phe Asn
1175 1180 1185
Gly Ser Asp Ile Pro Asn Asp Gly Asn Arg Phe Trp Gly Asp Val
1190 1195 1200
Phe Thr Ile Thr Tyr Gln Lys Ala Asp Asn Gln Val Glu Lys Gly
1205 1210 1215
Cys Gln Gly Thr Ser Thr Ser Leu Asn Ile Lys Ser Asp Ser Tyr
1220 1225 1230
Arg Ser Ile Ser Glu Ala Gln Gly Val Ser Leu Leu Asp Ser Ile
1235 1240 1245
Leu Gln Gly Glu Leu Pro Cys Asp Leu Glu Arg Thr Asn Ser Thr
1250 1255 1260
Tyr Asn Ile Leu Ala Leu Leu Arg Ile Leu Glu Gly Leu Asn Gln
1265 1270 1275
Leu Ser Pro Arg Leu Arg Val Gln Gly Ala Ser Asp Asp Phe Ala
1280 1285 1290
Glu Gly Lys Ile Asn Thr Leu Asp Glu Leu Tyr Arg Thr Gly Ala
1295 1300 1305
Lys Val Pro Ser Glu Val Phe Val Asn Ser Lys Leu Thr Pro Lys
1310 1315 1320
Leu Ala Arg Gln Met Gln Asp Val Leu Ala Leu Cys Ser Gly Ser
1325 1330 1335
Leu Pro Ser Trp Cys Tyr Gln Met Thr Lys Ala Cys Pro Phe Leu
1340 1345 1350
Phe Pro Phe Glu Thr Arg Arg Gln Tyr Phe Tyr Ser Thr Ala Phe
1355 1360 1365
Gly Leu Ser Arg Ala Leu Asn Arg Leu Gln Gln Gln Gln Ser Asp
1370 1375 1380
Asn His Ser Ser Gly Gly Glu Arg Glu Val Arg Phe Gly Arg Leu
1385 1390 1395
Gln Arg Gln Lys Val Arg Val Ser Arg Asn Arg Ile Leu Asp Ser
1400 1405 1410
Ala Ala Lys Val Met Glu Met Phe Ser Ser Gln Arg Ala Val Leu
1415 1420 1425
Glu Val Glu Tyr Phe Gly Glu Val Gly Thr Gly Leu Gly Pro Thr
1430 1435 1440
Leu Glu Phe Tyr Thr Leu Leu Ser His Glu Leu Gln Cys Ala Gln
1445 1450 1455
Leu Gly Leu Trp Arg Ala Thr Ser Ser Tyr Asp Ser Gly Leu His
1460 1465 1470
Thr Asp Met Lys Asp Val Ile Arg Leu Asp Pro Glu Asp Gly Ser
1475 1480 1485
Ser Gly Lys Glu Leu Asn Thr Asp Leu Pro Gly Asp Gly Met Tyr
1490 1495 1500
Leu Ile Gln Ala Pro Leu Gly Leu Phe Pro Arg Pro Trp Pro Pro
1505 1510 1515
Lys Val Asp Thr Ser Glu Gly Ser Arg Phe Phe Lys Val Leu Glu
1520 1525 1530
Tyr Phe Arg Leu Ile Gly Gln Val Ile Ala Lys Val Leu Gln Asp
1535 1540 1545
Gly Arg Leu Leu Asp Leu Pro Leu Ser Thr Ala Phe Tyr Lys Leu
1550 1555 1560
Ile Leu Gly Gln Glu Leu Asp Leu Phe Asp Ile Val Ser Phe Asp
1565 1570 1575
Ser Glu Phe Gly Lys Thr Leu Gln Glu Leu Gln Val Leu Val Glu
1580 1585 1590
Arg Lys Gln Phe Leu Glu Ser Thr Ser Gly Lys Asn Gln Leu Gln
1595 1600 1605
Val Ala Asp Leu Cys Phe His Gly Ala Ser Ile Glu Asp Leu Cys
1610 1615 1620
Leu Asp Phe Thr Leu Pro Gly Tyr Pro Asp Tyr Val Leu Lys Glu
1625 1630 1635
Gly Glu Gly Gly Thr Ile Val Asn Ile Tyr Asn Leu Glu Glu Tyr
1640 1645 1650
Ile Thr Leu Val Val Asp Ala Thr Val Lys Ser Gly Ile Lys Arg
1655 1660 1665
Gln Val Glu Ala Phe Arg Ser Gly Phe Asn Gln Val Phe Asp Met
1670 1675 1680
Ser Ser Leu His Ile Phe Ser Pro Gln Glu Leu Asp Tyr Leu Ile
1685 1690 1695
Cys Gly Arg Gln Glu Ile Trp Glu Ser Glu Ser Leu Val Asp Asn
1700 1705 1710
Ile Lys Phe Asp His Gly Tyr Thr Ala Lys Ser Pro Ala Ile Val
1715 1720 1725
Asn Leu Leu Glu Ile Leu Ser Glu Phe Thr Pro Glu Gln Gln His
1730 1735 1740
Ala Phe Cys Gln Phe Val Thr Gly Ala Pro Arg Leu Pro His Gly
1745 1750 1755
Gly Leu Ala Ala Leu Ser Pro Lys Leu Thr Ile Val Arg Lys His
1760 1765 1770
Pro Ser Ser Ala Val Asn Thr Ser Asn Ser Thr Gly Ala Val Glu
1775 1780 1785
Ser Ala Asp Asp Asp Leu Pro Ser Val Met Thr Cys Ala Asn Tyr
1790 1795 1800
Leu Lys Leu Pro Pro Tyr Ser Thr Lys Glu Ile Met Arg Lys Lys
1805 1810 1815
Leu Leu Tyr Ala Ile Leu Glu Gly Arg Gly Ser Phe Asp Leu Ser
1820 1825 1830
<210> 84
<211> 74
<212> PRT
<213> 大豆
<400> 84
Ser Ser Ser Ala Ser Ala Ala Val Asn Thr Arg Ser Arg Ala Ser Asn
1 5 10 15
Thr Lys Glu Pro Leu Pro Pro Lys Asn Pro Pro Pro Pro Leu Pro Pro
20 25 30
Met Asp Ser Ala Asn Glu Ser Ser Gly Ser Arg Arg Asp Arg Arg Asn
35 40 45
Asn Lys Glu Asn Ser Ser Asp Lys Gly Lys Glu Lys Glu His Asp Val
50 55 60
Arg Ile Arg Asp Arg Asp Ala Ala Leu Asn
65 70
<210> 85
<211> 68
<212> PRT
<213> 大豆
<400> 85
Ser Ser Ser Ser Ser Ala Ser Ala Ala Ala Ala Ala Ala Ser Val Ser
1 5 10 15
Ser Val Asn Thr Arg Ser Arg Ser Ser Arg Thr Lys Glu Pro Leu Pro
20 25 30
Pro Lys Asn Pro Pro Pro Met Asp Ser Ala Asn Glu Ser Ser Gly Ser
35 40 45
Arg Arg Asp Arg Arg Asn Lys Asp Asn Ser Asp Lys Gly Lys Glu Lys
50 55 60
Glu His Asp Val
65
<210> 86
<211> 84
<212> PRT
<213> 大豆
<400> 86
Asp Asp Asn Asp Ser Glu Gly Gly Val Gly Ile Leu His Gln Asn Leu
1 5 10 15
Ile Ser Ala Ser Thr Phe Arg Gly Leu Leu Arg Lys Leu Gly Ala Gly
20 25 30
Leu Asp Asp Leu Leu Pro Ala Thr Ala Met Gly Gly Ser Val Pro Ser
35 40 45
Ser His Gln Thr Gly Gly Leu Lys Gln Ile Leu Ser Gly Leu Arg Ala
50 55 60
Asp Gly Glu Glu Gly Arg Gln Val Glu Ala Leu Thr His Leu Cys Asp
65 70 75 80
Met Leu Ser Ile
<210> 87
<211> 71
<212> PRT
<213> 大豆
<400> 87
Gly Lys Glu Lys Glu Gln Asp Val Arg Ile Arg Asp Ala Glu Arg Glu
1 5 10 15
Arg Glu Arg Ala Leu Ala Leu Asn Met Glu Ser Glu Asp Val Gly Asp
20 25 30
Asp Asp Asp Asn Asp Ser Asp Gly Gly Val Gly Ile Leu His Gln Asn
35 40 45
Leu Thr Ser Ala Ser Ser Ala Leu Gln Gly Leu Leu Arg Lys Leu Gly
50 55 60
Ala Gly Leu Asp Asp Leu Leu
65 70
<210> 88
<211> 58
<212> PRT
<213> 大豆
<400> 88
Ala Ala Asn Ala Ser Ser Asn Gly Asn Gly Pro Ser Glu Leu Ala Asp
1 5 10 15
Asp Asp Leu Pro Ser Val Met Thr Cys Ala Asn Tyr Leu Lys Leu Pro
20 25 30
Pro Tyr Ser Thr Lys Glu Ile Met Tyr Lys Lys Leu Leu Tyr Ala Ile
35 40 45
Ser Glu Gly Gln Gly Ser Phe Asp Leu Ser
50 55
<210> 89
<211> 51
<212> PRT
<213> 大豆
<400> 89
Ala Val Asn Thr Ser Ser Asn Gly Asn Gly Pro Ser Glu Ser Ala Asp
1 5 10 15
Asp Asp Leu Pro Ser Val Met Thr Cys Ala Asn Tyr Leu Lys Leu Pro
20 25 30
Pro Tyr Ser Thr Lys Glu Ile Met Tyr Lys Lys Leu Leu Tyr Ala Ile
35 40 45
Asn Glu Gly
50
<210> 90
<211> 51
<212> PRT
<213> 大豆
<400> 90
Ala Val Asn Asn Ser Ser Asn Gly Asn Gly Pro Ser Glu Ser Ala Asp
1 5 10 15
Asp Asp Leu Pro Ser Val Met Thr Cys Ala Asn Tyr Leu Lys Leu Pro
20 25 30
Pro Tyr Ser Thr Lys Glu Ile Met Tyr Lys Lys Leu Leu Tyr Ala Ile
35 40 45
Ser Glu Gly
50
<210> 91
<211> 51
<212> PRT
<213> 玉米
<400> 91
Val Ala Asn Asn Asn Ser Asn Ala Thr Gly Ala Thr Glu Ser Ala Asp
1 5 10 15
Asp Asp Leu Pro Ser Val Met Thr Cys Ala Asn Tyr Leu Lys Leu Pro
20 25 30
Pro Tyr Ser Thr Lys Ala Ile Met Leu Lys Lys Leu Leu Tyr Ala Ile
35 40 45
Asn Glu Gly
50
<210> 92
<211> 51
<212> PRT
<213> 玉米
<400> 92
Ser Ala Val Asn Thr Ser Asn Ser Thr Gly Ala Val Glu Ser Ala Asp
1 5 10 15
Asp Asp Leu Pro Ser Val Met Thr Cys Ala Asn Tyr Leu Lys Leu Pro
20 25 30
Pro Tyr Ser Thr Lys Glu Ile Met Arg Lys Lys Leu Leu Tyr Ala Ile
35 40 45
Leu Glu Gly
50
<210> 93
<211> 22
<212> PRT
<213> 大豆
<400> 93
Asp Asp Asp Leu Pro Ser Val Met Thr Cys Ala Asn Tyr Leu Lys Leu
1 5 10 15
Pro Pro Tyr Ser Thr Lys
20
<210> 94
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 94
tccttcccct tgtcggagga gtt 23
<210> 95
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 95
gggggcaagg gttccttagt att 23
<210> 96
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 96
ttccgtgaac actcgctccc gtt 23
<210> 97
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 97
ttgcggcgat cgcggcggga gcc 23
<210> 98
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 98
atttctgcca gtacctttcg agg 23
<210> 99
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 99
cgtgccgatg gggaagaagg tcg 23
<210> 100
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 100
acgccaaggc tcgctctcgc tcc 23
<210> 101
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 101
acgtctgcca gtagtgccct tca 23
<210> 102
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 102
ccaagtgtga tgacgtgtgc aaa 23
<210> 103
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 103
cctagtgtga tgacatgtgc taa 23
<210> 104
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 104
atggcatgaa cagtcccttc aag 23
<210> 105
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 105
tttcagcggg ttgcattgtc tac 23
<210> 106
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 106
agcgggttgc attgtctact gct 23
<210> 107
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 107
ttgatgtgag cagtctcttc aag 23
<210> 108
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 108
atgtgagcag tctcttcaag cac 23
<210> 109
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 109
cacatatttg cagcagtaga caa 23
<210> 110
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 110
ttgcacatat ttgcagcagt aga 23
<210> 111
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 111
ttgatgtgaa cagtctcttc aag 23
<210> 112
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 112
atgtgaacag tctcttcaag cac 23
<210> 113
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 113
gagagttgca ttatctacag ctg 23
<210> 114
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 114
aggtactgga gcatgcttct gtc 23
<210> 115
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 115
tgcaaatatt tgcagcggta gac 23
<210> 116
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 116
gcaatgcgtg tcaaacaaac aga 23
<210> 117
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 117
cctagtgtca tgacttgcgc caa 23
<210> 118
<211> 23
<212> DNA
<213> 人工序列
<220>
<223> 合成寡核甘酸
<400> 118
cctagtgtga tgacgtgcgc caa 23
<210> 119
<211> 11119
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 119
gaggagcgta gctagttcag ttccgtttga tgaacgaaaa tgaccaatcc accgagcgaa 60
attgtaattt tcaacaataa aaaaggcaaa gaaagttaca tatgaaagcc tgttttgttc 120
tgtccctttt tattcttccc tttctttctt acttacccct ttccctggct tagggttttc 180
tgcccccctg aatcctcatc gctattcatg attacactcc ttacaattct cactcagcga 240
ctcgttcgaa atacgtgaaa tccccttatc tccaatttct agggtttcga ttcattcaac 300
tcaccacaaa gattggatcc atggtagcgt gattggcgct cggtggcgag cgagttgata 360
attatcgggt tgggtggatt ttgtatggaa actcggagcc ggaagcgggc ggaggcttcc 420
tcagctgccc cttcatcccc ttcctctggt cccaccactc gttccagcaa gcgtgcacgc 480
ctctcttctt cttcctccgc ttccgccgcc gctgccgccg catccgtttc ttccgtgaac 540
actcgctccc gttcatcccg gactaaggaa cccttgcccc ctaagaatcc tcctcccatg 600
gactctgcca atgaatcctc tggctcccgc cgcgatcgcc gcaacaaagt ccgacaaggg 660
gaaggagaaa gagcatgatg ttaggattag ggacagggat gctgacagag ggctggcatt 720
gaacatggat ggtggcggag acgatgatga caatgacagc gaaggcggtg tggggatttt 780
gcatcaaaac ttgacctctg cgagtagtgc acttcagggg cttcttcgga aacttggtgc 840
tggtttggat gatctgcttc catcatcggc tatgggttct gcgtcttcct ctcaccagag 900
tgggaggctg aaaaagatcc tcttcggatt gcgggcggat ggcgaagaag ggcggcaggt 960
tgaagcattg acacagctct gtgagatgct ttcgattggg actgaagagt cacttagtac 1020
attctctgtt gattcgtttg ttcccgtgct tgtggggttg ttgaatcacg agagcaaccc 1080
tgacatcatg cttcttgctg ccagagcatt gacccatctc tgtgatgtgc tcccttcatc 1140
ttgtgctgct gttgtgcatt atggtgccgt gtcaatcttt tgtgcgaggt tgctcaccat 1200
agaatacatg gacctggctg agcaggttgt tccttggttc atgactattg tggaattaat 1260
gaaaatggtg ctagaaagtt ataaaataaa tatttgtgtt tcagtgttga accctggtgc 1320
tgcctaattg tttttaaaac tatatatttc gatgtttctg ggtcaaattt tcagttgtag 1380
aagaattatg cctttttttt ttaatctaag acttagcatt ttgaaaatga aattcatata 1440
gtttgattac tgtatttgat ggcatgaaca gtcccttcaa gctctgaaga agatatctca 1500
ggagcatccc actgcctgcc tacgagcagg tgctcttatg gccgtgcttt cctatttgga 1560
cttcttttca acaggagttc aggtaaattg tcagtgaagt acctacattt agaggatgat 1620
tgcatcccac tggtcctttt ggacagtcat aaatcatttg gccacaggtg ccattagtga 1680
gcaagtgtat gttgacatta ttctttgttt cagcgggttg cattgtctac tgctgcaaat 1740
atgtgcaaaa agcttccttc agatgcagct gattttgtga tggaagctgt tcctcttctg 1800
acaaaccttc ttcagtacca tgattccaag gtaaggtcat gttttgttgc aagtcttgtc 1860
acataatgga aactgtattt ttccttttgc atccataaac ttgcctttta aggaagttta 1920
gcttgagaag agggaaattt tgatccctat atcccatggt ataaaatatt tatctatagc 1980
ttcaccttat gcattttcaa tttttgtagg ttctggagca cgcctctgtt tgtttgactc 2040
gaatagctga agcttttgcg tcatctccag acaaattaga tgaattgtgc aaccatggac 2100
ttgtaacaca agctgcctcc ctcatttcta acagcagttc tgggggtggt caggcttctc 2160
tcagcacgcc gacatatact gtaagtgcaa tttttacttt tagttagatg cattttgcct 2220
atagtttggt ccttgacctg ggtatatgca gggtttaatc cgacttcttt caacttgtgc 2280
gagtggatct cctcttggag ctaaaacctt actacttctt ggaattagtg gtattcttaa 2340
agatattcta tctggttctg gagtttcttc taaggcctct gtttctcctg cattaagtag 2400
gccgccagaa caggtatagt ataacatcag aacttttctt ttggtcattc atgtgtagtt 2460
ttatctcgta atgttcatta aacagacact gaccttaaat ccatcattat tctaattctt 2520
ggtttcaaat atatagatat ttgagattgt aaacctcacg aatgagcttc tgcctccatt 2580
gccacatgga acaatttctc tccctatcat ctccaacatg tttttgaaag ggcccattgt 2640
aaagaagtct cctgctggta gctctggaaa acaagaagac acaaatggaa atgttcctga 2700
gatatcggct cgtgagaaac tattaaatga tcagcctgaa ctacttaagc aatttgcgat 2760
ggatctcctt ccagttttaa tacaggttga tatttgtgca tcaattgctt aaactttgct 2820
tgataaattt gttaaattga aaaaaatgtt ctgataaatt gctcctttgc ttccttattt 2880
gtcttctttg gttaattgat gatattggct tgctgttgat atagatatat ggttctagtg 2940
tcaatggtcc tgttcggcac aaatgccttt ctgtcattgg aaaattgatg tatttcagca 3000
cagcagagat gatccagtct ttgttgagtg tgacaaatat atcaaggtat gttgaaattt 3060
aattgagttg atattgcttg ataccctcac tgattttatg ggtttagaaa atttatgcat 3120
tgttgctttg atcatatagt ttcttagctg gcgtcttagc atggaaagat ccacatgttt 3180
tgcttcctgc cttgaaaatt gctgaaattc ttatggaaaa gcttcctggg acattctcca 3240
agatgttcat tagagaaggt gttgtgcatg cagtggacca acttatttta actggaaatt 3300
cgaccaatat ctctacacag gcatcttctg ccgagaagga taatgattct atatctggag 3360
catcatctcg ctctaggcgt tatcggcgac gcagtgggaa ttccaatcct gatggaaatc 3420
ctttggacga tttgaaaact ccagtttcag taaatgttgg ttcacctcct aattctgtgg 3480
atatgccaac agtaaattcc agtattcggt tatctgttag tacagctgcc aaagctttta 3540
aagataagta ttttccttca gatcctgggg ctgctgaagt gggtattact gatgatcttt 3600
tgcatctgaa aaatctttgc atgaagttaa atgctggtgc tgatgaacaa aggaccaatg 3660
gaaaggggaa atctaaaact tctggatttg gtctggaaga gtatttaatt gggatcatag 3720
ctgacatgct aaaggaactt ggcaaaggag atggggtatc tacttttgaa ttcattggta 3780
gtggtgttgt tgcagctctg ttgaattatt tttcttgtgg gtatttctct aaagatcgac 3840
cattagaaac ccaccttccc aagcttcgcc aacaagcact tacaaggttt aagttattta 3900
tagctgttgc actaccttct actactgaag atgggactgt ggctcctatg actgtcttgg 3960
tccagaagct tcaaaatgcc ttgtcctcct tggagcgttt ccctgttgtg ctgagtcatt 4020
catctaggtc atctagtggg agtgcacgcc tctcctctgg actaagtgca ttatctcagc 4080
ccttcaagtt gcggctttgt cgagcccagg gtgaaaggtc acttagggat tattcatcca 4140
atgttgtact ggttgatcca ttagcaagtt tagcagcaat tgaggaattt gtttggcctc 4200
gtatccaacg aagtgaattg ggtcagaagt ccactgtacc tgctgggaat tctgaatctg 4260
ggacaactcc tacaggagct ggtgtatcct ctccaactac ccatcgccat tctactagat 4320
ccagatcatc tgttaatata ggtgatacat ctagaaagga aataagtcaa gataaaagca 4380
caagctcttc caagggtaag ggaaaagctg tattaaagcc tgcacaagag gaggcaagag 4440
gacctcagac cagaaatgct actcgcagaa gagaagctct tgataaagat gctcaaataa 4500
agcctgtaaa cggtgactct acttctgagg tatactgatg gaaaccgagt taggggcaga 4560
tatgacagtt atctgtagaa aataactgct ttagaaatca attctgtata agaaactcaa 4620
ataactgtct tagaaaacag tactgtatta gaaactgcag gatggtagtt atgtgtagaa 4680
aataactgct ggtagttatg tattaggagt atatgtctgt tagtatgtgg tgtaattggg 4740
cagtgtaaag tcatttttaa gtttgtggat agagaatact ctgtaggaga cttctgtctc 4800
tgggacagct agtgctgttt tttgtatatg ataggagact ttagtctcta ggacggcagg 4860
tgtattgctg tatttgtatg tgataggaga cttttgtctc tgggacagca agtgtattgc 4920
tgtattgtgt gatttcagtg ttctatatat tttaggtaaa tgctagggct ctggttgagg 4980
aagtaaaaag agggatgcgt aaaaatgtac tcctcctatg atttccaaaa aaaaactttt 5040
cttttttaat tccttaacca gtgcctggca ctggttagca agaccctata ttttattact 5100
ttatcatttg gtgttctatc atatactgtg aaatcctagc gatgacttta caatgcttca 5160
acttttttct tctgttaatt tataacttcc ttctggctat gtatgacctg actatgaatg 5220
ctctgttttc atgttggctt ataaagtgaa atacgaagag aatacttgat aatgccaata 5280
taagatgtaa agatgcatta ttacattttt cattgcaagc ttgaaaagac atcttacatt 5340
tctctgtatc tgtaactttg gacatgctgg attttgttgt ctgtagatcc ttaaaatgtt 5400
acctctgcca tttagtttta taaatggttt ttgattataa ttatctattt aagatatcat 5460
ctctaatatg agaaacactg cctaggtttt ctttggatta tgttgaacag ttgctttttt 5520
gcaccattga tttctttaca tacattaaaa ttaagctgtt gtaaggtttc cctgcaaaag 5580
tgttgattta ctaaaaaatt gaggcagagc tagcactgag aggataactt atctattttg 5640
cgttgaagta acattgctgt aagtaatagt gtatttggat gtaatcagat ggatactttg 5700
tatatgctga tcctcatcct tctttcatta agtgcatgtg tcagaatttt tagtatggta 5760
ccactcaaaa agtcaaaatc gattcctatt ttcagtttag cttagttttt aattattttc 5820
ttataacatt aaacttgtct gtcatgaagt tgtagtttgt atgactctca ttggggaata 5880
ttttagatta tttgtgggcg ggttacatat tttttccgtt taggtgaagc tttcttgttt 5940
tgttttttgt tgcttttgat tatggcagtg tttacatgca tgttgtttag ctaggttctt 6000
gtggttgtta aactggtgga agcatctttg agtataaatt ttttttttgg aaggcgtctt 6060
tgagtataaa tattagtcat ttgtttttcc tgctatgctt tggactaaat catgaaccta 6120
atccaagtat cttgaagtag tcatttgttt ttccttttct ttttttaccc ttctagtgac 6180
attttccaat gtctacattg taggatgaag atttggatat atcccctgtt gagattgatg 6240
aggcattggt gattgaagat gatgatattt ctgatgatga agatgatgac catgatgatg 6300
tatgttatct actgtttctt cttctttggc taggattttc ttacttcctt ggtgatgagt 6360
atctcattta agtaattact gtgtttctgt cttttttttt tttttgcttt tctgacatca 6420
tttccttttt taacatggct tttaaatatg ttaaaatctg tgtgcatctt tattttttta 6480
tttccagtgc tggtgtcctt caatcttgtt tacaattttt tcaaaatgag ttgctgctgt 6540
ctttctatca tttctttgtt ttctattttt cttttctggg tataatgatg caatgaagtt 6600
ttggtttatg accttatgca aactatccat aatccaagtc tctgaccaat agctcaatcc 6660
tggtgtttta tttccaaatt tttaatttta gctaccatca ttattgttaa ccaagattga 6720
acattataat taaattcagt tgtatctatt aaaatttttg tgcttgtttt gttttttctg 6780
atgcacaggt actgagggat gattctcttc ctgtttgttc acctgacaaa gtacatgatg 6840
tgaaattggg cgacatagtg gaggagagta atgttgctcc tgcaactagt gatggtggcc 6900
agactaatgc tgcctcaggt tctagtagca aagctggtac agtcagggga tcagactctg 6960
ctgattttag gagtggctat acctcaagct caagaggtgc aatgtcattt gctgctgctg 7020
ctatggctgg acttggatct gccaatagca gaggtatcag gggtggaaga gatcgactag 7080
ggcgtccatt gtttggtagt tctaatgatc ctccaaagtt gatatttact gctggtggga 7140
agcagcttaa taggcatttg actatttatc aggcaattca aaggcagctt gtgctagatg 7200
aagatgatga ggagagattt gctggcagta gtgactatgt atccagtgat ggaagcaggt 7260
tgtggggtga tatttatact ataacttatc agagggcaga gaaccagaca gataggactc 7320
cccctggagg ttcaacctct aatgcttcaa aatctggcaa atctgggtct gtattgaatt 7380
ctagttctga agacaagcta aatcagacat ctgtattaga tagtattttg cagggagaat 7440
tgccctgtga actggagaaa tctaatccta catacaatat attggcatta ttgcgggtgc 7500
ttgagggttt gaaccaactt gcatctcgtt tgagggccca agtggttact gatagctttg 7560
cagagggaaa aattttggat ttagttgagc taagttttac cagtggtgct agggttccta 7620
cagaggaatt tataagcagc aaacttactc caaaattagc taggcaaata caagatgccc 7680
ttgccttatg tagtgggagt cttccctcat ggtgttacca gttatctaaa gcgtgccctt 7740
ttttgtttcc ttttgagacc cggcgacagt atttttattc aactgccttt gggttatctc 7800
gtgcattgta tcgccttcag cagcagcagg gtgctgatgg tcatggatca acaaatgaaa 7860
gagaggtcag ggttgggaga ttgcagcgtc aaaaggttcg tgtctctcga aatcgcattt 7920
tggattctgc tgctaaggtg atggagttgt attctagtca aaaggctgta cttgaagtag 7980
aatattttgg tgaagttggc accggtctgg gtcccactct ggagttctac acacttctca 8040
gtcatgactt acaaaaagtt gtacttcaaa tgtggagatc aggttcttca gagaaatatc 8100
aaatggaaat tgatggagat gaaaagaaaa tgaaaaatag tgaaggctct tttgttggag 8160
atggagaact cgttcaagct cctcttgggc tgtttcctcg accttggcct gcaaatgctg 8220
atgcatcaga gggtacccaa attttcaaag tgattgaata tttccgatta ttaggccgtg 8280
taatggctaa agctcttcaa gatggacgct tattggattt accattgtca gtggcatttt 8340
ataagcttgt tcttggtcaa gtaagttatg aaatgttgat gtcttgtctg atttcatgtg 8400
tatcttaagg ttgattttta gtctctatat atttagcctt tgatatattg caggagcttg 8460
atttgcatga cattcttttc attgatgctg aacttgggaa aactttgcaa gagttaaatg 8520
cccttgtttg ccggaaatgt tttatagaat ctattggtgg tagctacact gatacctttg 8580
ctaatttgca ttttcgtggg gccccaatag aagatctctg cttggacttc acacttcctg 8640
gttatccaga gtacatcttg aaacctggag atgaaattgt atgtattcag tctgtttttt 8700
ttacctggtt tttgttttgg ttctgattct gtctgtaata aaaattgctt tgaacttact 8760
gtcaaacttt caggttgaca tcaataatct agaggagtac atatccatgg tggttgaggc 8820
aacggttaag actggaatca tgcgtcaaat ggaagctttt agagcagggt ttaatcaggt 8880
tatatgttgt ctcaataaat tcatgtaact ttgtctttga ctgtgcatct tgtttggtga 8940
tgctgagtat aaaaaatatc atgtattttt taactgatta atggttcatt ctttttggta 9000
ttccttttct agtttctctc aaacaatttt attgaaaact aaacttgact ggggtttaat 9060
ttgaaaatat tgagtatgga tttttcagct tttagattct taaggggcat tgttttctac 9120
taaaaattgt tacttttggt tatgtcttga gcagtgaact gtatatatat cctgaatctc 9180
ggatgtatca attaagaaat tactaaatgt ttgtttctga cttttactta tgtttgctac 9240
caaccttgta tcccccttcc cctgcagatg aagaatggaa gttagaaaat acacattttt 9300
tttgtattga tgtcaaatat tcagttattt aatgtcaaaa ttttacaagt gaaatgagct 9360
actagcctaa cttatatgga agagatgggc tgggcaatag tttgaacttg gaacaactag 9420
ttggataatt tgttcatttg cttgagtcca aaaactaaac atttgtcact ttcccacttg 9480
ttcttgtcaa ttcaggtttt tgacatctca tctttacaaa ttttttctcc ccaagaactg 9540
gattacttgc tttgcggccg gagagaattg tggaaggtat tcttttttat acaaaagtat 9600
tactgctgct tacaacaatc ttttagatgt taccatggat aatgtagtta taattttttt 9660
tctcctatct gcagactgag acactagctg atcatataaa atttgaccat ggttatactg 9720
ccaagagccc tgccatagtt aatgtatgtt tttttattcc tgtagaagga caattgtgtt 9780
tttggaaatt taggcttgtt atatttggtg ctgaacctgt atgatgctat tttcagttac 9840
tcgaaattat gggagaattc acaccagagc agcagcgtgc cttctgtcaa tttgttactg 9900
gtgcacctag gctgcctcct ggtggactgg cagttctaaa tccaaaatta acgattgtga 9960
ggaaggtatt gaaaaatatt tttgatcact tgcaacctgt gttattcatt catgccttca 10020
tgcaattttg tacttgatat cttgaatgtt aaagtttttt ggggggcgag gatctatttg 10080
aacttcggta gcaagatgtg tctggattta ctgcctgaca tatgttgctc cactattcct 10140
ttacctcttg aaggggggtt ttcaaaatgc aatgttagta agtgattaca tttacatgtc 10200
tgggtgcagc tttcgtcaag tgcagctaat gcttcatcta acgggaatgg gccttcagaa 10260
ttagcagatg atgacttgcc aagtgtgatg acgtgtgcaa attacctgaa gcttcctcct 10320
tattctacca aggtagaaca ctgcaaagca ttgttggtta tatgatcatg catgtcaaag 10380
tgtcttttga tctttgattt ccattttaaa acaggaaatt atgtacaaga agctactcta 10440
tgcaatcagt gaaggccagg gatcctttga tttatcatga gttctggaac tacccaaccc 10500
ggccctgtat gttaatatga tgatgaatca gggtttactc gagacttaat tattttttgg 10560
ttgcatgtac tttcctccct gcttctcaaa ataatggcct ctctcgggtc acgcacatca 10620
aaggtggtga catggttcag gtgagtttac cattgttgtg aaacctcact atgattcttg 10680
acattactag accattgttg tgagcttacc aagagcctct tattattttg ttcattttca 10740
tttgttggta aactaaatta atattaattc tttgattatt ggcagttgaa tcagatcgcc 10800
tcgtttttaa aggtgtaaag gaatttgcgg agctcgtttt agctgttaca tgtaaataga 10860
ataggaatgt tgagtaattc cctcccaaat ttttcatcat tctgttttca actcaaagta 10920
ttggcagtta gttatcatac ctgtatggaa gatggtaaaa ggaatagaaa taatatcgta 10980
tactatgctt tatgatttca tgttagatct gttacgagct tttgttgagg taaaatgcga 11040
attattctat gcttgtttgc tatgattgtc ccaaaatgag gcgttctagc ataaataagt 11100
actgtttgag ttaccgatt 11119
<210> 120
<211> 11114
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 120
gaggagcgta gctagttcag ttccgtttga tgaacgaaaa tgaccaatcc accgagcgaa 60
attgtaattt tcaacaataa aaaaggcaaa gaaagttaca tatgaaagcc tgttttgttc 120
tgtccctttt tattcttccc tttctttctt acttacccct ttccctggct tagggttttc 180
tgcccccctg aatcctcatc gctattcatg attacactcc ttacaattct cactcagcga 240
ctcgttcgaa atacgtgaaa tccccttatc tccaatttct agggtttcga ttcattcaac 300
tcaccacaaa gattggatcc atggtagcgt gattggcgct cggtggcgag cgagttgata 360
attatcgggt tgggtggatt ttgtatggaa actcggagcc ggaagcgggc ggaggcttcc 420
tcagctgccc cttcatcccc ttcctctggt cccaccactc gttccagcaa gcgtgcacgc 480
ctctcttctt cttcctccgc ttccgccgcc gctgccgccg catccgtttc ttccgtgaac 540
actcatcccg gactaaggaa cccttgcccc ctaagaatcc tcctcccatg gactctgcca 600
atgaatcctc tggctcccgc cgcgatcgcc gcaacaaaga caactccgac aaggggaagg 660
agaaagagca tgatgttagg attagggaca gggatgctga cagagggctg gcattgaaca 720
tggatggtgg cggagacgat gatgacaatg acagcgaagg cggtgtgggg attttgcatc 780
aaaacttgac ctctgcgagt agtgcacttc aggggcttct tcggaaactt ggtgctggtt 840
tggatgatct gcttccatca tcggctatgg gttctgcgtc ttcctctcac cagagtggga 900
ggctgaaaaa gatcctcttc ggattgcggg cggatggcga agaagggcgg caggttgaag 960
cattgacaca gctctgtgag atgctttcga ttgggactga agagtcactt agtacattct 1020
ctgttgattc gtttgttccc gtgcttgtgg ggttgttgaa tcacgagagc aaccctgaca 1080
tcatgcttct tgctgccaga gcattgaccc atctctgtga tgtgctccct tcatcttgtg 1140
ctgctgttgt gcattatggt gccgtgtcaa tcttttgtgc gaggttgctc accatagaat 1200
acatggacct ggctgagcag gttgttcctt ggttcatgac tattgtggaa ttaatgaaaa 1260
tggtgctaga aagttataaa ataaatattt gtgtttcagt gttgaaccct ggtgctgcct 1320
aattgttttt aaaactatat atttcgatgt ttctgggtca aattttcagt tgtagaagaa 1380
ttatgccttt tttttttaat ctaagactta gcattttgaa aatgaaattc atatagtttg 1440
attactgtat ttgatggcat gaacagtccc ttcaagctct gaagaagata tctcaggagc 1500
atcccactgc ctgcctacga gcaggtgctc ttatggccgt gctttcctat ttggacttct 1560
tttcaacagg agttcaggta aattgtcagt gaagtaccta catttagagg atgattgcat 1620
cccactggtc cttttggaca gtcataaatc atttggccac aggtgccatt agtgagcaag 1680
tgtatgttga cattattctt tgtttcagcg ggttgcattg tctactgctg caaatatgtg 1740
caaaaagctt ccttcagatg cagctgattt tgtgatggaa gctgttcctc ttctgacaaa 1800
ccttcttcag taccatgatt ccaaggtaag gtcatgtttt gttgcaagtc ttgtcacata 1860
atggaaactg tatttttcct tttgcatcca taaacttgcc ttttaaggaa gtttagcttg 1920
agaagaggga aattttgatc cctatatccc atggtataaa atatttatct atagcttcac 1980
cttatgcatt ttcaattttt gtaggttctg gagcacgcct ctgtttgttt gactcgaata 2040
gctgaagctt ttgcgtcatc tccagacaaa ttagatgaat tgtgcaacca tggacttgta 2100
acacaagctg cctccctcat ttctaacagc agttctgggg gtggtcaggc ttctctcagc 2160
acgccgacat atactgtaag tgcaattttt acttttagtt agatgcattt tgcctatagt 2220
ttggtccttg acctgggtat atgcagggtt taatccgact tctttcaact tgtgcgagtg 2280
gatctcctct tggagctaaa accttactac ttcttggaat tagtggtatt cttaaagata 2340
ttctatctgg ttctggagtt tcttctaagg cctctgtttc tcctgcatta agtaggccgc 2400
cagaacaggt atagtataac atcagaactt ttcttttggt cattcatgtg tagttttatc 2460
tcgtaatgtt cattaaacag acactgacct taaatccatc attattctaa ttcttggttt 2520
caaatatata gatatttgag attgtaaacc tcacgaatga gcttctgcct ccattgccac 2580
atggaacaat ttctctccct atcatctcca acatgttttt gaaagggccc attgtaaaga 2640
agtctcctgc tggtagctct ggaaaacaag aagacacaaa tggaaatgtt cctgagatat 2700
cggctcgtga gaaactatta aatgatcagc ctgaactact taagcaattt gcgatggatc 2760
tccttccagt tttaatacag gttgatattt gtgcatcaat tgcttaaact ttgcttgata 2820
aatttgttaa attgaaaaaa atgttctgat aaattgctcc tttgcttcct tatttgtctt 2880
ctttggttaa ttgatgatat tggcttgctg ttgatataga tatatggttc tagtgtcaat 2940
ggtcctgttc ggcacaaatg cctttctgtc attggaaaat tgatgtattt cagcacagca 3000
gagatgatcc agtctttgtt gagtgtgaca aatatatcaa ggtatgttga aatttaattg 3060
agttgatatt gcttgatacc ctcactgatt ttatgggttt agaaaattta tgcattgttg 3120
ctttgatcat atagtttctt agctggcgtc ttagcatgga aagatccaca tgttttgctt 3180
cctgccttga aaattgctga aattcttatg gaaaagcttc ctgggacatt ctccaagatg 3240
ttcattagag aaggtgttgt gcatgcagtg gaccaactta ttttaactgg aaattcgacc 3300
aatatctcta cacaggcatc ttctgccgag aaggataatg attctatatc tggagcatca 3360
tctcgctcta ggcgttatcg gcgacgcagt gggaattcca atcctgatgg aaatcctttg 3420
gacgatttga aaactccagt ttcagtaaat gttggttcac ctcctaattc tgtggatatg 3480
ccaacagtaa attccagtat tcggttatct gttagtacag ctgccaaagc ttttaaagat 3540
aagtattttc cttcagatcc tggggctgct gaagtgggta ttactgatga tcttttgcat 3600
ctgaaaaatc tttgcatgaa gttaaatgct ggtgctgatg aacaaaggac caatggaaag 3660
gggaaatcta aaacttctgg atttggtctg gaagagtatt taattgggat catagctgac 3720
atgctaaagg aacttggcaa aggagatggg gtatctactt ttgaattcat tggtagtggt 3780
gttgttgcag ctctgttgaa ttatttttct tgtgggtatt tctctaaaga tcgaccatta 3840
gaaacccacc ttcccaagct tcgccaacaa gcacttacaa ggtttaagtt atttatagct 3900
gttgcactac cttctactac tgaagatggg actgtggctc ctatgactgt cttggtccag 3960
aagcttcaaa atgccttgtc ctccttggag cgtttccctg ttgtgctgag tcattcatct 4020
aggtcatcta gtgggagtgc acgcctctcc tctggactaa gtgcattatc tcagcccttc 4080
aagttgcggc tttgtcgagc ccagggtgaa aggtcactta gggattattc atccaatgtt 4140
gtactggttg atccattagc aagtttagca gcaattgagg aatttgtttg gcctcgtatc 4200
caacgaagtg aattgggtca gaagtccact gtacctgctg ggaattctga atctgggaca 4260
actcctacag gagctggtgt atcctctcca actacccatc gccattctac tagatccaga 4320
tcatctgtta atataggtga tacatctaga aaggaaataa gtcaagataa aagcacaagc 4380
tcttccaagg gtaagggaaa agctgtatta aagcctgcac aagaggaggc aagaggacct 4440
cagaccagaa atgctactcg cagaagagaa gctcttgata aagatgctca aataaagcct 4500
gtaaacggtg actctacttc tgaggtatac tgatggaaac cgagttaggg gcagatatga 4560
cagttatctg tagaaaataa ctgctttaga aatcaattct gtataagaaa ctcaaataac 4620
tgtcttagaa aacagtactg tattagaaac tgcaggatgg tagttatgtg tagaaaataa 4680
ctgctggtag ttatgtatta ggagtatatg tctgttagta tgtggtgtaa ttgggcagtg 4740
taaagtcatt tttaagtttg tggatagaga atactctgta ggagacttct gtctctggga 4800
cagctagtgc tgttttttgt atatgatagg agactttagt ctctaggacg gcaggtgtat 4860
tgctgtattt gtatgtgata ggagactttt gtctctggga cagcaagtgt attgctgtat 4920
tgtgtgattt cagtgttcta tatattttag gtaaatgcta gggctctggt tgaggaagta 4980
aaaagaggga tgcgtaaaaa tgtactcctc ctatgatttc caaaaaaaaa cttttctttt 5040
ttaattcctt aaccagtgcc tggcactggt tagcaagacc ctatatttta ttactttatc 5100
atttggtgtt ctatcatata ctgtgaaatc ctagcgatga ctttacaatg cttcaacttt 5160
tttcttctgt taatttataa cttccttctg gctatgtatg acctgactat gaatgctctg 5220
ttttcatgtt ggcttataaa gtgaaatacg aagagaatac ttgataatgc caatataaga 5280
tgtaaagatg cattattaca tttttcattg caagcttgaa aagacatctt acatttctct 5340
gtatctgtaa ctttggacat gctggatttt gttgtctgta gatccttaaa atgttacctc 5400
tgccatttag ttttataaat ggtttttgat tataattatc tatttaagat atcatctcta 5460
atatgagaaa cactgcctag gttttctttg gattatgttg aacagttgct tttttgcacc 5520
attgatttct ttacatacat taaaattaag ctgttgtaag gtttccctgc aaaagtgttg 5580
atttactaaa aaattgaggc agagctagca ctgagaggat aacttatcta ttttgcgttg 5640
aagtaacatt gctgtaagta atagtgtatt tggatgtaat cagatggata ctttgtatat 5700
gctgatcctc atccttcttt cattaagtgc atgtgtcaga atttttagta tggtaccact 5760
caaaaagtca aaatcgattc ctattttcag tttagcttag tttttaatta ttttcttata 5820
acattaaact tgtctgtcat gaagttgtag tttgtatgac tctcattggg gaatatttta 5880
gattatttgt gggcgggtta catatttttt ccgtttaggt gaagctttct tgttttgttt 5940
tttgttgctt ttgattatgg cagtgtttac atgcatgttg tttagctagg ttcttgtggt 6000
tgttaaactg gtggaagcat ctttgagtat aaattttttt tttggaaggc gtctttgagt 6060
ataaatatta gtcatttgtt tttcctgcta tgctttggac taaatcatga acctaatcca 6120
agtatcttga agtagtcatt tgtttttcct tttctttttt tacccttcta gtgacatttt 6180
ccaatgtcta cattgtagga tgaagatttg gatatatccc ctgttgagat tgatgaggca 6240
ttggtgattg aagatgatga tatttctgat gatgaagatg atgaccatga tgatgtatgt 6300
tatctactgt ttcttcttct ttggctagga ttttcttact tccttggtga tgagtatctc 6360
atttaagtaa ttactgtgtt tctgtctttt tttttttttt gcttttctga catcatttcc 6420
ttttttaaca tggcttttaa atatgttaaa atctgtgtgc atctttattt ttttatttcc 6480
agtgctggtg tccttcaatc ttgtttacaa ttttttcaaa atgagttgct gctgtctttc 6540
tatcatttct ttgttttcta tttttctttt ctgggtataa tgatgcaatg aagttttggt 6600
ttatgacctt atgcaaacta tccataatcc aagtctctga ccaatagctc aatcctggtg 6660
ttttatttcc aaatttttaa ttttagctac catcattatt gttaaccaag attgaacatt 6720
ataattaaat tcagttgtat ctattaaaat ttttgtgctt gttttgtttt ttctgatgca 6780
caggtactga gggatgattc tcttcctgtt tgttcacctg acaaagtaca tgatgtgaaa 6840
ttgggcgaca tagtggagga gagtaatgtt gctcctgcaa ctagtgatgg tggccagact 6900
aatgctgcct caggttctag tagcaaagct ggtacagtca ggggatcaga ctctgctgat 6960
tttaggagtg gctatacctc aagctcaaga ggtgcaatgt catttgctgc tgctgctatg 7020
gctggacttg gatctgccaa tagcagaggt atcaggggtg gaagagatcg actagggcgt 7080
ccattgtttg gtagttctaa tgatcctcca aagttgatat ttactgctgg tgggaagcag 7140
cttaataggc atttgactat ttatcaggca attcaaaggc agcttgtgct agatgaagat 7200
gatgaggaga gatttgctgg cagtagtgac tatgtatcca gtgatggaag caggttgtgg 7260
ggtgatattt atactataac ttatcagagg gcagagaacc agacagatag gactccccct 7320
ggaggttcaa cctctaatgc ttcaaaatct ggcaaatctg ggtctgtatt gaattctagt 7380
tctgaagaca agctaaatca gacatctgta ttagatagta ttttgcaggg agaattgccc 7440
tgtgaactgg agaaatctaa tcctacatac aatatattgg cattattgcg ggtgcttgag 7500
ggtttgaacc aacttgcatc tcgtttgagg gcccaagtgg ttactgatag ctttgcagag 7560
ggaaaaattt tggatttagt tgagctaagt tttaccagtg gtgctagggt tcctacagag 7620
gaatttataa gcagcaaact tactccaaaa ttagctaggc aaatacaaga tgcccttgcc 7680
ttatgtagtg ggagtcttcc ctcatggtgt taccagttat ctaaagcgtg cccttttttg 7740
tttccttttg agacccggcg acagtatttt tattcaactg cctttgggtt atctcgtgca 7800
ttgtatcgcc ttcagcagca gcagggtgct gatggtcatg gatcaacaaa tgaaagagag 7860
gtcagggttg ggagattgca gcgtcaaaag gttcgtgtct ctcgaaatcg cattttggat 7920
tctgctgcta aggtgatgga gttgtattct agtcaaaagg ctgtacttga agtagaatat 7980
tttggtgaag ttggcaccgg tctgggtccc actctggagt tctacacact tctcagtcat 8040
gacttacaaa aagttgtact tcaaatgtgg agatcaggtt cttcagagaa atatcaaatg 8100
gaaattgatg gagatgaaaa gaaaatgaaa aatagtgaag gctcttttgt tggagatgga 8160
gaactcgttc aagctcctct tgggctgttt cctcgacctt ggcctgcaaa tgctgatgca 8220
tcagagggta cccaaatttt caaagtgatt gaatatttcc gattattagg ccgtgtaatg 8280
gctaaagctc ttcaagatgg acgcttattg gatttaccat tgtcagtggc attttataag 8340
cttgttcttg gtcaagtaag ttatgaaatg ttgatgtctt gtctgatttc atgtgtatct 8400
taaggttgat ttttagtctc tatatattta gcctttgata tattgcagga gcttgatttg 8460
catgacattc ttttcattga tgctgaactt gggaaaactt tgcaagagtt aaatgccctt 8520
gtttgccgga aatgttttat agaatctatt ggtggtagct acactgatac ctttgctaat 8580
ttgcattttc gtggggcccc aatagaagat ctctgcttgg acttcacact tcctggttat 8640
ccagagtaca tcttgaaacc tggagatgaa attgtatgta ttcagtctgt tttttttacc 8700
tggtttttgt tttggttctg attctgtctg taataaaaat tgctttgaac ttactgtcaa 8760
actttcaggt tgacatcaat aatctagagg agtacatatc catggtggtt gaggcaacgg 8820
ttaagactgg aatcatgcgt caaatggaag cttttagagc agggtttaat caggttatat 8880
gttgtctcaa taaattcatg taactttgtc tttgactgtg catcttgttt ggtgatgctg 8940
agtataaaaa atatcatgta ttttttaact gattaatggt tcattctttt tggtattcct 9000
tttctagttt ctctcaaaca attttattga aaactaaact tgactggggt ttaatttgaa 9060
aatattgagt atggattttt cagcttttag attcttaagg ggcattgttt tctactaaaa 9120
attgttactt ttggttatgt cttgagcagt gaactgtata tatatcctga atctcggatg 9180
tatcaattaa gaaattacta aatgtttgtt tctgactttt acttatgttt gctaccaacc 9240
ttgtatcccc cttcccctgc agatgaagaa tggaagttag aaaatacaca ttttttttgt 9300
attgatgtca aatattcagt tatttaatgt caaaatttta caagtgaaat gagctactag 9360
cctaacttat atggaagaga tgggctgggc aatagtttga acttggaaca actagttgga 9420
taatttgttc atttgcttga gtccaaaaac taaacatttg tcactttccc acttgttctt 9480
gtcaattcag gtttttgaca tctcatcttt acaaattttt tctccccaag aactggatta 9540
cttgctttgc ggccggagag aattgtggaa ggtattcttt tttatacaaa agtattactg 9600
ctgcttacaa caatctttta gatgttacca tggataatgt agttataatt ttttttctcc 9660
tatctgcaga ctgagacact agctgatcat ataaaatttg accatggtta tactgccaag 9720
agccctgcca tagttaatgt atgttttttt attcctgtag aaggacaatt gtgtttttgg 9780
aaatttaggc ttgttatatt tggtgctgaa cctgtatgat gctattttca gttactcgaa 9840
attatgggag aattcacacc agagcagcag cgtgccttct gtcaatttgt tactggtgca 9900
cctaggctgc ctcctggtgg actggcagtt ctaaatccaa aattaacgat tgtgaggaag 9960
gtattgaaaa atatttttga tcacttgcaa cctgtgttat tcattcatgc cttcatgcaa 10020
ttttgtactt gatatcttga atgttaaagt tttttggggg gcgaggatct atttgaactt 10080
cggtagcaag atgtgtctgg atttactgcc tgacatatgt tgctccacta ttcctttacc 10140
tcttgaaggg gggttttcaa aatgcaatgt tagtaagtga ttacatttac atgtctgggt 10200
gcagctttcg tcaagtgcag ctaatgcttc atctaacggg aatgggcctt cagaattagc 10260
agatgatgac ttgccaagtg tgatgacgtg tgcaaattac ctgaagcttc ctccttattc 10320
taccaaggta gaacactgca aagcattgtt ggttatatga tcatgcatgt caaagtgtct 10380
tttgatcttt gatttccatt ttaaaacagg aaattatgta caagaagcta ctctatgcaa 10440
tcagtgaagg ccagggatcc tttgatttat catgagttct ggaactaccc aacccggccc 10500
tgtatgttaa tatgatgatg aatcagggtt tactcgagac ttaattattt tttggttgca 10560
tgtactttcc tccctgcttc tcaaaataat ggcctctctc gggtcacgca catcaaaggt 10620
ggtgacatgg ttcaggtgag tttaccattg ttgtgaaacc tcactatgat tcttgacatt 10680
actagaccat tgttgtgagc ttaccaagag cctcttatta ttttgttcat tttcatttgt 10740
tggtaaacta aattaatatt aattctttga ttattggcag ttgaatcaga tcgcctcgtt 10800
tttaaaggtg taaaggaatt tgcggagctc gttttagctg ttacatgtaa atagaatagg 10860
aatgttgagt aattccctcc caaatttttc atcattctgt tttcaactca aagtattggc 10920
agttagttat catacctgta tggaagatgg taaaaggaat agaaataata tcgtatacta 10980
tgctttatga tttcatgtta gatctgttac gagcttttgt tgaggtaaaa tgcgaattat 11040
tctatgcttg tttgctatga ttgtcccaaa atgaggcgtt ctagcataaa taagtactgt 11100
ttgagttacc gatt 11114
<210> 121
<211> 10728
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 121
atggaaactc ggagccggaa gcgggcggag gcttcctcag ctgccccttc atccccttcc 60
tctggtccca ccactcgttc aagcaagcgc gcacgtctct cttcttcttc ttccgcttcc 120
gccgccgtga acactcggtc ccgtgaagga acccttgccc cctaaaaatc cccctcctcc 180
tcttcctccc atggactctg ccaacgaatc ctctggctcc cgccgtgatc gccgcaacaa 240
caaagaaaac tcctccgaca aggggaagga gaaagagcat gatgtcagga ttagggacag 300
ggatgcggca ttgaacatgg atggcagtgg tggagacgag gacgatgaca atgacaatga 360
cagtgaaggc ggtgtgggga ttttgcatca aaacttgacc tctgcgagta gtgcacttca 420
ggggcttctt cggaaactcg gtgctggttt ggatgatctg cttccatcgt cggctatggg 480
ctctgcatct tcctctcacc agagtgggag gctgaaaaag atcctctttg gattgcgggc 540
tgacggcgaa gaagggcggc aggtcgaagc attgacacag ctctgtgaga tgctttcgat 600
tgggactgaa gaatcgctta gtacattctc tgttgattcg tttgttcctg tgctcgtggg 660
gttgttgaat cacgagagca accctgacat catgcttctt gctgccagag cattgaccca 720
tctctgcgat gtgctccctt catcttgtgc tgcggttgtg cattatggtg cagtctcaat 780
cttttgtgcc aggttgctca caatagaata catggacttg gctgagcagg ttgttcttta 840
atgggaatgg tgttagaaga aacttatgaa atataaatat ttgtcttgaa ctctggtgct 900
gcctaattgt tttttaaaac tatgtatatt gcaatgtttc tgggttcagt tgtagaagaa 960
tcatgcattt tgaaaatgaa attcttatag tttgattact gtgtttgatg gcatgaacag 1020
tcccttcaag ctttgaagaa gatctctctg gagcatccca ctgcctgcct acgagcaggt 1080
gctcttatgg ccgtgctttc ttatttggac ttcttttcaa caggagttca ggtaaattgt 1140
cagtgaagta cctaaattta gaggatgatt atattatatg gtccttttgg atagtcataa 1200
atcatttggc cacagatgcc tttagtgagc aagtgtgtgt atgttgacat tattctttgt 1260
ttcagcgggt tgcattgtct actgctgcaa atatgtgcaa gaagcttcct tcagatgcag 1320
ctgattttgt gatggaagct gttcctcttc tgacaaacct tcttcagtac catgattcca 1380
aggtaaggtc atgttttttg ttttaagtct tgtcacataa tggaaactgt atttttcctt 1440
ttgcatctat aacttttccc gttaaggaag ttttttttag cttgagagga gggagatttt 1500
gatccctgta tcccatagta taaaatattt ttctatagct tcaccttatg cattatcaat 1560
ttttgtaggt tctggagcac gcctctgttt gtttgactcg aatagctgaa gcttttgcat 1620
catctccaga caaattagat gagttgtgca accatggact tgtaacacaa gctacctccc 1680
tcatttctaa cagcagttct ggaggtggtc aggcttctct cagcacaccg acatataccg 1740
tgagtgcaat ttttacttta agtcggatgc attttgcatg tagttttggt ccttgacttg 1800
ggcatatgca gggtttaatc cgacttcttt caacttgtgc gagtggatcc ctcttggagc 1860
taaaacctta ttacttcttg gaattagtgg tattcttaaa gatattctat ctggttctgg 1920
agtttcttct aatgcctctg tttctctgca ttaagtaggc caccagaaca ggtatagtat 1980
aacaccagaa cttttctttt agtcattcat gtgtagtttt atctcgtaat gttcattaag 2040
cagcctttga ccttaaatac accattattt agatccttga tatttccagt tctcttacat 2100
ttgtgaagga atgaaattat tctaattctc tgtttcaaat atatagatat ttgagattgt 2160
aaacctggcg aatgagcttc tgcccccatt gccacatgga acaatttctc tccctatcat 2220
ctccaacatg tttttgaaag ggcccattgt aaagaagtct ccttctggta gctctgggaa 2280
acaagaagac actaatggaa atgttcctga gatatcggct cgtgagaaac tattaaatga 2340
tcagcctgaa ctacttaagc aatttgcgat ggatctcctc ccagttttaa tacaggttga 2400
catttgtgca ttgatcattt gcttaaattt tgcttgataa atttgttaaa ttaaaaaaga 2460
agttctgata ttgctccttt gcttccttat ttatcttctt tggttaattg atggtattgg 2520
cttgcttttg atacagatat atggttctag tgtcaatggt cctgttcggc acaaatgcct 2580
ttctgtcatt gggaaattga tgtatttcag cacggcagag atgatccagt ctttgttgag 2640
tgtgacaaat atatcaaggt atgtagaagt ttaattgggt tgatattgct tgatatccta 2700
ctgattttat gggtttagaa aatttatgca ttgttgcttt gatcatatag tttcttagct 2760
ggcgtcctag catggaaaga tccacatgtt ttgcttcctg ccttgaaaat tgctgaaatt 2820
cttatggaaa agcttcccgg gacattctcc aagatgttca ttagagaagg tgttgtgcat 2880
gcagtggacc aacttatttt agcctcaaat tcaaccaata tctctacaca ggcatctcct 2940
gctgagaagg ataatgattc tatatctgga gcatcatctc gctctaggcg ttatcggcga 3000
cggagtggga attccaatcc cgatggaaat cctttggacg atttgaaaac tccagtttca 3060
gtaaatgttg gttcacctcc aagttctgtg gatatgccaa cattaaattc cagtattcgg 3120
ttatctgtta gtacagctgc caaagctttt aaagataagt actttccttc agatcctggg 3180
gctgctgaag tgggtattac tgatgatctt ttgcatctga aaaatctttg catgaagtta 3240
aatgctggtg atgatgaaca aaggactaat ggaaaggggg aatctaaaac ttctggattt 3300
ggtccggaag agtatttaat tgggatcata gctaacatgc taaaggaact tggcaaagga 3360
gatggtgtat ctacttttga attcattggt agtggtgttg ttgcagctct gttgaattat 3420
ttttcttgtg ggtatttctc taaagatcga ccattagaaa cccaccttcc caagcttcgc 3480
caacaagcac ttacaaggtt taagttattt atagcagttg cactaccttc tactattgaa 3540
gttgggactg tggctcctat gactgtcttg gtccagaagc ttcaaaatgc cttgtcctcc 3600
ttggagcgtt tccctgttgt gctgagtcat tcatctaggt catctagtgg gagtgcacgc 3660
ctctcctctg gactaagtgc tttatctcag cccttcaagt tgcggctttg tcgagcccag 3720
ggtgaaaagt cacttaggga ttattcatcc aacgttgtac tggttgatcc attagcaagt 3780
ttagcagcga ttgaggaatt tgtttggcct cgtatccaac gtagtgaatc tggtcagaag 3840
tccactgtag ccactgggaa ttctgaatct gggacaactc ctgcaggagc tggtgtatcc 3900
tctcctacta cccgtcgcca ttctactaga tccagatcat ctgttaatat aggtgataca 3960
tctagaaagg aaataactca agataaaagt acaagttctt ccaagggtaa gggaaaagtt 4020
gtattaaagc ctgcacaaga ggaggcaaga ggacctcaga ccagaaatgc tactcgcaga 4080
agagcagctc ttgataaaga tgctcaaatg aagcctgtaa acgctgactc tacttctgag 4140
gtataccgat gtaaactgag ttagggactg ttatgacagt tatctgtaga gaataactgc 4200
tttagaaatc aatgttagaa actcaaatat ctgtcttaga aaacagtact gtattagaaa 4260
cttcaggata gtagttatgt atttggagta tatgtctgtt agtatgtggt tttaattagg 4320
cagtttaaag ttatttttaa gttcgtggac agagaatact ctgtaggaga cttctgtctc 4380
tgggacagta agtgctgttt ttgtatgtga taggagactc tagtctttag tacagcaaat 4440
gtattgctgt atttgtatgt gataggaaac ttttgtctct gggacagcaa gtgtattgct 4500
gtattgtgtg tgatttaagt gttctatata ttttaggcaa atgctagggc gctgtttgag 4560
gaagtaaaaa gggaaagttt tttattggga tgcgtaaaat gcactccccc tatattttat 4620
ttctttactt tatcatttgg tgttctatca tatactttga aatcctagcg atgagtttac 4680
aatgctacaa cttttttctt ctgttaattt ataactttca gttatttagt ttttgttgga 4740
tggttttctt cttttgctct ccacttcctt ctggctatgt atgatctgac tatgaatgtt 4800
gctgttttca tgttggttta tacagtgaaa tatgaggaga atacttaata atgccaatat 4860
aaggtgtaaa gatgcattat tagattttgc atcgcaagct tgaaaagtca tctggaattt 4920
ctctgcaact ttggacacac tggattttgt tgtctgtaga tccttgaaat gttttacctc 4980
tgccatttag ttttataaat ggcttttggt tacaattatc tatttagata tatctcttat 5040
gtgagaaaca ctacctaagt gttcttttga ttattatgtt gaacagttgc ttttttgcac 5100
cattgatttc tttacataca ttaaaattaa gctgttataa gatttccctg caaaagtgtt 5160
gagttactac aaaagtgagg aacattgcca agcctcatcc ttctagcatt aagtgtatgt 5220
gtcaggattg ttttagatta tttgtgggtg ggttgcatat tttttctatt taggttatac 5280
ttgttttgtt tttcattgct tttgattatg gcagtgttta catgcatgtt gtttagctag 5340
gttcttgtgg ttgttaaact ggtggaagga tctttagagt ataaaataca acacacattc 5400
tatatgcagt tgatagggct caacttggct gctaaactca tctttttact gctatgcttt 5460
ggactaaatc atgaacctaa tccaagtatc ctgaagtatc taaatattag tctcttgttt 5520
tttcttttct ttttttaccc ttctattgac atttttcatt gtctacattg taggatgaag 5580
atttggatat atcccctgtt gagattgatg aggcattggt gattgaagat gacgatattt 5640
ctgatgatga agatgatgac cacgaagatg tatgttatct actgttttgt ctacttttct 5700
tacttggtga tgagtatcac atttaagtaa ttactgtgtt actgtaattt ttttttttgc 5760
ttttctgaca acccccagct ttttctacgt ttaacacggc ttttaatata ttaaaatcta 5820
tttttggcta ccaacgctgg tgtccttcaa tcttgtttac atttttttca aaatgatatg 5880
ctgctgtctt tgtatcattt ctttgctttc tattttttct tctgggtata atgatgcaat 5940
gaagttttag tttatgatct tatgcgaact atccaagcct ctgaccaata gctcaatcta 6000
ggtgttttat ttccaaaatt ttaattttag ctaccatcat tattgttaac caagattgaa 6060
tgttatagtt aaattcagtt gtatctattc ttaaaatatt ggtgcttgtt ttatttttat 6120
tgatgcaggt actgagggat gattctcttc ctgtttgttc acctgacaaa gtacacgatg 6180
tgaaattagg cgacttagcg gaggagagta atgttgctcc tgcaacaagt gatggccagg 6240
ctaatgctgc ctcaggttct agtagcaaag ctggtacagt caggggatca gactctactg 6300
attttaggag tggctataac tcaagctcaa gaggtgcaat gtcatttgct gctgctgcta 6360
tggctggact tggatctgcc aatagcagag gtatcagggg tggaagagat cgactagggc 6420
gtccattgtt tggtagttct aatgatcctc caaagttgat atttactgct ggtgggaagc 6480
agcttaatag gcatttgact atttatcagg caattcaaag gcagcttgtg ctagatgatg 6540
atgaaagatt tgctggcagt agtgactatg tatccagtga cggaagcagg ttgtggggtg 6600
atatttatac tataacttat catagggctg agaaccagac agataggact ccccctggag 6660
gttcaacctc taatgcttca aaatcttgca aatctgggtc tgtatcgaat tctagttccg 6720
aagccaagct gcatcagaca tctgtactag atagtatctt gcagggagaa ttgccctgtg 6780
aactggagaa atctaatcct acatacaata tattggcatt attgcgggtg cttgagggtt 6840
tgaaccaact tgcatctcgt ttgagggccc aagtggttac tgatagcttt gctgagggaa 6900
aaattttgga tttagatgag ctaagtgtta ccagcggtgc tagggttcct acagaggaat 6960
ttataagcag caaacttact ccaaaattag ctaggcaaat acaagatgcc cttgccttat 7020
gcagtgggag tcttccctcg tggtgttacc agttatctaa agcgtgccct tttttgtttc 7080
cttttgagac ccggcgacag tatttttatt caactgcctt tgggttatct cgtgcattgt 7140
atcgccttca gcagcagcag ggtgctgatg gtcatggatc aacaaatgaa agagaggtca 7200
gggttgggag attgcagcgt caaaaggttc gtgtctctcg aaatcgcatt ttggattctg 7260
ctgctaaagt gatggagttg tattctagtc aaaaggctgt acttgaagta gaatattttg 7320
gtgaagttgg caccggtctg ggtcccactc tggagttcta cacacttctc agtcatgact 7380
tacaaaaaat tatacttgaa atgtggagat caggttcttc agagaaatat caaatgaaaa 7440
ttgatggaga tgaaaagaaa atgaaacgta gtgaaggctc ttttgttgga gatggagaac 7500
ttgttcaagc tcctcttggg ctgtttcctc gaccttggcc tgcaaatgct gatgcatcag 7560
agggtaccca attttttaaa gtgattgaat atttccgact attaggccgt gtaatggcta 7620
aggctcttca agatggacgc ctattggatt taccaatgtc agtggcattt tataagcttg 7680
ttcttggtca agtaagttat gaaatgttga tgtcttatct gatttgatgt gcatcttaag 7740
gttgattttt agtatgtata tatttaaccc ttgatatatt gcaggagctt gatttgcatg 7800
acattctttt cattgatgct gagcttggga aaactttgca agagttaaat gcccttgttt 7860
gccgaaaaca ttatatacaa tctactggtg gtagctacac cgataccttt gctaatttgc 7920
attttcgtgg ggccccaata gaagatctct gcttggactt cacacttcct ggttatccag 7980
agtacatctt gaaacctgga gatgaaattg tatgtattca gtctgtttct attacctggt 8040
ttttgttttg gttctgattc tgtctgttat gaaaattgct ttgaacttac tgtcaaactt 8100
ccaggttgac atcaataatc tagaggagta catatccatg gtggttgagg caactgttaa 8160
gactggaatc atgcgtcaaa tggaagcatt tagagcaggg ttcaatcagg ttatatgctg 8220
tctcaattaa ttcatgtaac ttttgtcttt gactgtgcac cttgtttggt gatgctgtgt 8280
aagctcttat ctaaaatgtc atgtattttt taactgtgat cagaaagttc tgtaatatca 8340
ttgttgtctg tcttttactt gcacattgtc aattcttatt taaaacatca tgtatttttt 8400
aattgtgatc agaaagttct gtaatatcat tgtctgtctt ttacttgcac attgtccatt 8460
ctttttggtg ttccttctct caaacaattt tattgaaaag taaacttgac tggggtttaa 8520
tttgataata ttgagcatgg attcttcagc ttttagattc ttaaggggca ttgttttttc 8580
taataaaaat tattactttt ggttatatct tgagcagtga actgcatata tagcctgaat 8640
atcggatgta ttaattaaga aattacaaaa tgtttgtttt tgacttttac ttatgtttgc 8700
taccaacctt gtgtccccct tcccctggcc cctgcacatc aagaatggaa gatagaaaat 8760
acacattttt ttttatattg atgtcaaata ttcagttatt taatgtcaaa attttacaag 8820
tgaaataagc tactggctga ggaagagatg ggctgggctg ggcaatagtt ggaacttgga 8880
aaaactagtt tgatgatttg ttcatttgct tgagtccaaa aactaaacat ttgtcacttg 8940
cccacttgtt cttttcaatt caggtttttg acatctcatc tttacaaatt ttttctcccc 9000
aagaactgga ttacttgctt tgtggccgga gagaattgtg gaaggtattc ttttttgtgc 9060
aaaagtatta ctgctgctta caacaatatt ttaaatgtta ccatgaataa tgtagttata 9120
aatttatttt tctcttttct gcagactgag acactagccg atcatataaa atttgaccat 9180
ggttatactg ccaagagccc tgccatagtt aatgtatggt tcttttcccc atagaaggac 9240
atttgtgttt ttggaaattt aggcttgcta tatttgatgc tgaacctcta tgatgctatt 9300
ttcagttact cggaattatg ggagaattca caccagagca acagcgtgcc ttctgtcaat 9360
ttgttactgg tgcacctagg ttgcctcctg gtggactggc agttctaaat ccaaaattaa 9420
cgattgtgag gaaggtattg aaaaaatatt tttgatcact tgcaacctgt gctatttatt 9480
catgcggcct tcacgcaatt ttgtacttga atgttaaagt ggctttcctc tacttttcct 9540
ttttacctct taaaaggggg cttacaaaat gcaagtttgt aagtgattac atttacatgt 9600
ctgggtgcag ctttcatcaa gtgcagctaa tgcttcatct aatgggaacg gtccttcaga 9660
attagcggat gatgacttgc caagtgtgat gacgtgtgca aattacctta agcttcctcc 9720
ttattctacc aaggtagaac actgcaaagc attgttggta atatgatcat gcatgtcaaa 9780
gtgtcttttg gtgttgaact gttgatattt gatttccatt ttaaaacagg aaattatgta 9840
caagaagcta ctctatgcaa tcagtgaagg ccagggatcc tttgatttat cgtgagtttc 9900
tggaactaac caacccggcc ctgtatgtta atatgatgat gaatcagggt ttactcttga 9960
gacttgatta ttttttggtt atatgtactt tcctccctgc ttctcaaata atggcctctc 10020
ccgggtcaca catcaaaggt ggtgacatgg ttcaggtgag tttaccattg ttgtgaaacc 10080
tcactatgat tcttgacact actagaccat tgttgtgagc ttaccaagag cctcttatta 10140
ttttgttcat tttcatttgt tggtaaacta attaatatca tttctttgat tactggcagt 10200
tgaatcggat cgcctcgttc taaaggtgta aaggaatttg cggagctcgt tttagctgtt 10260
acaacatgta aatagaatag gaatgttgag taattccctc cccctctaaa ttttccatca 10320
ttctgttttc aactcaaatt attggcagtt agttattata catgcatgga agatggtaaa 10380
aggaatagaa ataatatcaa atactatgct ttattatttc atgttagatc tgttacgagc 10440
ttttgttgag gtaaaatgtg gatatcactc tatgctgatg ctatttgcta tgattgcccc 10500
aaaatgaggc gctctatagc ataaataagt gctgtttggg ttgccgattg gttagtcgct 10560
aatgtacttt tgctaaagca tctcttttgt tacttctccg ttgaaatgca ttgaaattcc 10620
cgcaaccaag gtttgaatgg gaattcaaac attccctaaa tttcatgtac tagctagggc 10680
tatctaaact gtgatagcgc caattctaat gcactttaaa ttttcccc 10728
<210> 122
<211> 16846
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 122
catggtacac cttgttctgg agaaatgcaa caattacctt tttattttct gtttacacat 60
taccctgtct tgaatgctac agagtttcta ctgattagtt cagtttatga ctatatggtc 120
ttacttattc tctgatacat ttggcaaagg taaaacttca gggccaacat aggccatgct 180
ctcccctgat cattttagca actctgcaga atatctgaca gtattttgag ctatattgct 240
aatgaaacat gtaaatatgc ttttggaaac actgaattta tcttcaaatc acaaccctct 300
ttgcctccta acatcggctg caagctctgc cactggtttt tgatagaaca tttttctgct 360
ttctattgtt gtttggatct ctagaaataa tatcttcacg tttgaaatac tatgcataca 420
gataatagac gcatacgtgt accatctctt tgtatttgta tgcatgctct tgatggtggc 480
ctaacatata tgctacgttt ttttatttag tcattgcaag ctcttaaaaa aatatccctg 540
gagcatccaa ctgcgtgctt gcgagctggt gcactaatgg cagtcctatc ataccttgac 600
tttttctcca caggtgttca ggtaatcact ctctgcctat tatagacttt tgcatcacaa 660
aatactttgt ttgtttcagc tcatttatct catagcttat tcttatgcat tacttgtcat 720
tatattgtga cagatgaact gtattattta ttcttgttat ttatttttat tactaactgt 780
tgtttttgtt gatggaatgt tttcagagag ttgcattatc tacagctgct aatatgtgta 840
ggaaacttcc ttctgatgca tcagattttg taatggaagc ggttccactt cttactaatc 900
tactgaacta tcatgattca aaagtacaat tgccattcca aatgctgata tcctctatga 960
taccatttat gtttcagatg ctgatattcc gttttgctta ttccaggtac tggagcatgc 1020
ttctgtctgc cttacccgta ttgcggaatc tttttcacca tttccagaaa aattggatga 1080
gttgtgcagt catggattgg ttgcacaagc tgctagctta gtgtctgtta gcaactcagc 1140
aggacaggca tccttgagta catcaacata tacagtacgc tccattttgg ctttttgggt 1200
acaattattt gttttacgtt tgtgaagttt atgccatgtt tttatagggt gtgattcgtc 1260
ttctctcaat atgcgcaagt ggatcacctc tggcagctaa aacacttctc cttcttggaa 1320
ttagtggcat acttaaagat atcctttcag gctctgggtt ggttgctggc acgactgtgt 1380
cccctgcttt aacaagacca gctgatcagg tgattttgtc ttttcatagg cacttcaagg 1440
ctctttgggt atcatgtgtg cttgatgtct acatttgttg tttatcttaa caagctaagg 1500
ccccgtttgg cactgctcca cttcacaaaa acaagcgcaa cttcatgaac ttcaaaatat 1560
agcagcccag ctttgaagtt gcagagtttt tgaagagttt ggcttgtaga ccagctttag 1620
tttcataaaa aaatataatt aaaaaaaact tgtttcttct tcctttggcc gtggcggcca 1680
gaggcgcacg tgcgcgactt gggaagacag acacaacgct gcgatggcct ggcgtggctc 1740
tggcggccca gcgcgctgca cgcagcctgg cggcacgttg tcggccgatg cggccctgct 1800
cgctagcgca ggcggtggca cgcgcgctgc cagagtttgg ttcaggcgtg gctctggcag 1860
cggcacatgg cctggcggtg cggctgccgg tcggcacggc cctgctcgct ggcgtaactg 1920
tgtgcggcct gagttccctt gagaacagaa cgaaagaaaa catttcgtga gggattgcgt 1980
aactgatggc aaaggtgggt aatttcaccc caacttaatg agggagttaa aaatacccat 2040
tcgtgaagta tacttttggt tgtttcatgg atttggtgaa gttagggtca acttcacctt 2100
ttttggtgaa gctgggagtg tttggctaga gttttataaa gctaagctag attttgtgaa 2160
gtgaagcagt cccaaacatt ccctaagtgt aaattaatgg agctaggctt actctatgct 2220
tcagatagta cttcaaccat tgattccaat ttccaagcct caggatatca tctctattaa 2280
ggcccccttt ggcacggctc tggctctagc ttcttgcagc cccttggaag gagccctgcc 2340
aaacagccat ctaaaacaca gctcctggca aggagccagg gccagaaaaa tggtttctcc 2400
cagctcctta caaacttgat acaaattatt acaaaactgc cactggacct gttttgacca 2460
aaccttttcc aaaacggctt caactggccc aaattataaa gccagcacaa ttaatcccat 2520
acacttccca accttacaag ccccttaata cagcccatct cttccttcat tctccataca 2580
tatctgaaag tcttccatgt tctctctctt ctaccacttc gtggcgcagt gaggggggct 2640
tacttctagg agcgcctaac atgtgctata taacatcttt acgcacaatg acataggaac 2700
aagtgtcgtc aaagaaaaca aataatattt gtgtatatta aaacattttt tgatgaattt 2760
gatgaaatga aaccaaatcg tcccataaca tgtttttctc ataatgcatt ttgtttttta 2820
atcgccacca ataatatact ttgagcttgt agtcaatgtt cgtttatggt atgacattgt 2880
ttttctttaa tgtgagcata ctcatactac ttcccatgtc gagcatttgt gaacatgatg 2940
ttatcctatg ttctattcat gatagcacaa tttatatttg tgatgaattg tcatctatgt 3000
taaattttaa tgggtactgt aacttgtaaa gctactgact ttttggtagt aatagattat 3060
caccacacaa ttccatatgc aaatgatgcc attatgtgtt ttttggtatt acacagatga 3120
atgagattgt gaagcttgca gatgagttac ttccttctct gcctgttggg accatttctt 3180
taccagtgta ttctggtgtt cacatgaaag gttgttctgt aaagaaatcc acttctagta 3240
agcagggtga acatggttca acagcaaatg aactatcagg tcgggagaag ttattgcgtg 3300
atcagcctga acttctgcag caatttggca tggacctgtt acctaccatg acacaggtca 3360
gtcttgtctt attaatgttc tctttgatgt tgttaatggt aatgctgatg tggtattacg 3420
ggacattgtt gctatcatgt tggtgtgata cttaaggact tcaatacctt gaacaggtgt 3480
atggctctag tgtaagtgga ccaatacggc acaggtgctt atctgtcatt gggaaattaa 3540
tgtactatag ctcagctgag atgatccagt ctctccttag cacaacaaac atttccaggt 3600
gcttaacaca atatatttca tcgagttatc tcatgttatt agaaactatc ttatcacatg 3660
aattccaaat atttgtttca agctattaac ttctatatca accattgcag ttttttggct 3720
ggcattttag cttggaaaga tccacaagtg ttgatccctg ctcttcagat agcagaagtt 3780
ttaatggaaa agcttccaga gatttttttg aagatgtttg tgagggaagg cgttgttcat 3840
gctgttgagt cacttatatg cccagaatta tctggtcagg tgactcctca tgtggattcc 3900
attacatctt cacataatag gcgcaaccgc cgtagaaata atgctgtgaa cactggaaat 3960
aacttgcctg atggaccaaa aggttccaat tctatgattg ccaattcacc cccaagcatg 4020
gctgaagttc caaataatag tcttcgtgct ttagttagta atcatgcaaa gtcatttaag 4080
gataaatatt tcccttccga gcctggctca agtgacattg cagttaccga tgaccttctt 4140
aaactgaggg cactttgtgc aaaactgaat acaacagctg acactatcaa aacaaaagct 4200
aaaggaaaat caaaggtagt gtctgacaat agttttgatg tcttgtgcaa tattgaggag 4260
caattggatg atataatagc tgaaatgttg tctgagctta gcaagggtga tggtgtttcg 4320
acatttgagt tcatcggaag tggagttgtt acggctttgc taacctatct gtcatgtgga 4380
acatttggga gggaaaaggt gtctgaggca aacataccaa atttgcgtca tcaggcagtc 4440
agacgataca aagcatttat atcttttgca cttccaaatg ataaagatgg gaataaaact 4500
cctatggcat tcctagtcca taaactgcaa agtgccctat cttcgttgga acgtttccca 4560
gttgtgctca gccattctgg aagggcatcg actttgggag gatcccgcct gacaacaggt 4620
ttaggatctc tgtcgcagcc catcaaattg cgtctttgtc gagcgcctgg tgagaagtca 4680
cttaaggatt tttcgtccaa tgttgttctt attgactcat tagctagtct ggcagctgtt 4740
gaagattttc tttggcctag agtccagcgc actgaaccag tattgaagcc tccaatgtca 4800
tctgcaaata attctggatc tggagcagca agctccacag cttgtgcgcc ttcaattcca 4860
tcagaaactc aatctgtccg tcgcacatcg ttaagatcaa agtcatcagc tgccaccagt 4920
ggtgcaatta agaaggatta tcaggaggga agcataaaca cctcaaaggg aaaaggaaaa 4980
gctgttctta aattgagttt ggatgaacct aaaggcccac atactaggaa tgctgcgcgc 5040
agaaaagcta cttcagagaa agatgttgaa ctgaagccat cacatggtca catcacttca 5100
gaggttagat actcgtgcat tcaatcttat ttgttatttg cataatatat gcgaaatggc 5160
ctctagccta gtggttaaag gcttccgaat agcacctcca agtctcgggt tcgattcccc 5220
tcaggggcga attttccggc ttcctgtgcg ccaccctccg gttggaccgc tgtagagggg 5280
acggttgacg tcagctcgtt agtgatgggg aggggggggg gcagagtttg gggattttct 5340
tggccgagac cattgtttta gtctatcttg atataatacc gggatggcgg tcattccttc 5400
cctggctgag ttttttttat ttgcataata tatcaattga gccctcctaa gtgagctccg 5460
ttgggaagtt tagctttaat ttctggctct aggacacatt gattgcaaat gtttttttaa 5520
caatgccttt tgtttactca gctgacaaat gttatcatag taatgataca acagtaatgc 5580
agctgattta ttttgcctta agtgtatcta gtttgatcag tactttgttt tagtatgtaa 5640
gcaaaggatt tcaaagggat gttcttcata gacatgatgt acagattgta taactataaa 5700
aggagggggg gcagtgctgc aggctccgac atgaggagtc tggggaaggg ataaaccgag 5760
gcaacccttc tcccgtaaat acggagagac actgctttga acccacaacc tggtgactcc 5820
gggagacaat tcacaccact gacgtacaga ttgtataact ataagtctat aatatcataa 5880
atgtataaca atgaaaatgc actgaactgt ggcatttctg tgtatcatgc atttataata 5940
ttatatctga gataggatat aaaaactgaa gcatttttta taagagcaac aaagattaaa 6000
gtagaaatat gcctgtttca ctaatatttt ataatttagt catattatgt aatgacctga 6060
ctacgctttc gtaacttagt atactggact atattgtgga tactcacaaa tatgaaatat 6120
cagtgtaatc ttttactcta attctctaaa taaaagatat tgttgtattt tgattttaaa 6180
tatctaagtg aagttttcag gggaaacaac tttatgaatt attatttgga atagtacgtg 6240
catgttcttt tataagtttg atatctgaat tttagaaatt aaaaagtttg aaagccctgc 6300
aacatatatt tatattatta tacctgcagg atgaagatct tgatgcttct cctgttgaga 6360
ttgatgatgc tttaattctt gatgatgatg acgaggatgt cccagatgat gaagatgatg 6420
atcatgaggc ggtaattatt ttttaaatct tgcttattac tacaaaggtt tgtattgagg 6480
ggacatgatt tggaactcag tagattttgt tgaaatgatc aggttctccg aggttctctt 6540
ccttcctgtg ttcctgagag agtgcatgat gtgaaattag gagatgctga tgattctagt 6600
gttgcctcat tggcaaatga taaccaggca cagccctcat ctggttctag tacaaaaaat 6660
acttctagca ggggattgga cactgctgaa tttagaagtc cagccacatt tggttcacga 6720
ggtgcaatgt cgtttgctgc agctgcaatg gctggattaa ctccagtagg tggtcgtgga 6780
attagaggta gccgagatcg gaatggcctt ccattgggtg ctcgtgcaac tgagcattac 6840
aacaaattga tatttacagc tgctgggaag cagctgaaca aacatttgac tgtatatcaa 6900
gctgttcaaa gacaagtagt tcatgccgag gatgatgaag atcggttcgg tggatctgat 6960
ttacctgatg atggtaacca cttctgggat gatataaggg gtgatgtgtt cactataacg 7020
tatcagaagg ctgataacac agcggagaag gggtctgttg gaggttcagc ttcagtgcca 7080
aaatcttcca aatcagattc ttgcagaact ttgtctgaaa aacagcgtac ttctcttctt 7140
gatagtattt tgcaaggaga gcttccctgt gatttagaga aatcaaacca aacctacaat 7200
atcttatcac tattacatgt gttggagggt ttgaatcagt tatcacctcg tctgagactg 7260
cagtcagcct gtgatgattt tgctgaagga aaagttgcta cattaaatgg gctatacgat 7320
gttggagcta aggtaccctc aaaggagttt atcaacagta agatgacccc aaaacttgct 7380
cggcaaattc aggatgttct tgcactgtgt agtggcagtt taccatcttg gtgttatcag 7440
ctgacgaaag cttgtccttt tctgtttcct ttcgaaacac gaaggcaata cttctattcc 7500
acagcttttg ggttgtctcg tgcacttcat cgtcttcagc aacaaccggg caatgataat 7560
aacactgctt ttgaaagaga agtcaggatt ggtagattgc aacgccagaa agtccgtgtt 7620
tctcgtaacc gtatcttgga ttctgcagct aaagttatgg agatgttctc taatcaaaag 7680
gctgtcctag aagttgaata ctttggtgaa gttggaactg gtcttggtcc aactttggag 7740
ttttatactc tcttaagccg tgagctgcaa agggttgact taggattgtg gagatctcat 7800
tcttcagata attctgggat gcaaattgat gcgaatactg atgatttaat aagaagtaaa 7860
aatcatgaat cagaatcact tactgagagc aggaacatag tacaatcacc tcttggatta 7920
tttcctcagc cttggccacc tactgctgct gcatcagaag gtagcaaatt cttcaaagtt 7980
gtcgagtatt tccgcttagt tggtcgagtg atggcaaaag cattgcaaga tggaaggctt 8040
ctcgatttgc ctttgtcgac agcattttac aaacttttgc ttggacaagt aagcatgagg 8100
gcctccttga gctagtttta tcttatgttg tcttctaaaa ccttcctctt ttcggttggc 8160
acttaacttt ctgtgccttt tccttattaa tttaccaaat ttgctttaat aggaacttga 8220
tttatatgat atattatctt ttgataccga gttcggaaag acattgcaag aactgcaaat 8280
tctcgttgca cgtaaacaat ttttggactc ctgctctagt gagagccaaa agatagatct 8340
atgtttccgt ggtgctcccg ttgaagattt atatttggac ttcactcttc cgggctatcc 8400
tgaatatgtt ctcaaggaag gtggagagaa tgcagaggta agttatatga taccttagtt 8460
tgtttttttc ttctagtgtt gctgcagatg actcattgtt atcctgtctt ataggtcaac 8520
atttgtaact tagaagagta tatttctttg gttgtggatg ctactgttaa gaccggcata 8580
atgcggcaag tagaggcatt taaggcagga ttcaatcagg tcttctttct tattttgtga 8640
tttgtgggca atttctatta ataattgatc ctagaactaa ctagcacatt tatttattta 8700
ttcagaaaaa aatactcatt gtatatttct cacttcaggt gtttgacata tcatcactgc 8760
aaatattttc tcctcaagaa cttgattatc tgatttgtgg tcgctgtgaa ctttgggagg 8820
tattgctttt gtggtcattc ttgatgttgt acttcctctg tcccagaatg ataagcatag 8880
tttgtatagg aaaaagtcaa atttagaatt ttatttccca gaaattgctt gtattatact 8940
agtttatgtg gctttgtgtc ccgctgccat aggtcaatgt tagctattga aagctagttg 9000
ctctagtcta ctgcttccct ttttttttcg ggcggggggg gggggggggg gggtattctc 9060
cctcctttgc agttcacctt ttttctctct taatgaaatg atacacaatt ctcttttgtg 9120
ttcgagaatt ctttttacaa actttggcca acaattaatc aaattatata catgttttgg 9180
acaaacttgc atcaatagat tcatattcag agtgcatttg agatgtgatt aattgtttag 9240
caaacataaa catattgtac gagaaattaa tggtcaattt ttttaataga ttatatgagt 9300
tattgtactc agtctaagtg ttatggaccc tggtcccgcc catgtaactc cttttccaat 9360
atatgcaacg tagcccaccc tgattggagt taacatggta gcaagcttgg tcttttctct 9420
ccctcccttc cctagccgcc acctcctggc tgccatcact accgtcgggt cgctattgtc 9480
gtctgcgcct ccatctcctc cctagcgcct ttcttgctca tcggccaccc atatttatta 9540
aggcagtgtt aataaccatg gggccaccac cttcacttta gctttctagg cgcgactggt 9600
ggccttgttg gccctacggt tggcgccttg cccaccggtg ctttagtccc ggtcacctcg 9660
attgctgccc tagtcgctcc cgtcgccctt gcctttgctg tggccatcta tggcatgggt 9720
gccgccagcc agggcccaga cctagctgac tctggcctag cttatctttc ggtcgcgtta 9780
cttccccctg atgtcgcttt cgctcactcc accttggcca tagcccccat tgctgccaag 9840
acggtgtttg ctgctgctcg ggatcgggaa catgttgctg cccttgctta ggggcacaaa 9900
cgcaccacga cggatgcact cgctcgatag cagaccaatg ctgagggtca cctgctcggc 9960
tcctcctgca tcgagaccct cccagcgctc tagtggtcgc ccctttgtcg gagtacgagg 10020
tcgagatcat caccaacctc tatgccgagg tggcgtgtgt ccagaacatt tgctctatga 10080
ttcccgtcat ccttgacaca acttcctcca actacgcctt ctggcgggcg atcttgtctt 10140
cactctccat cgctacgcct tggactacca cgtcctcacc aacactgtct ccctcactaa 10200
cctatcctgg cggcagatgg atagtgtggt cctctcgtgg atccccagga ccgccatcgt 10260
tgcgctgcat gacgtggttt gcgagcgtgg tggcactatc cgtcttggac gtcgtcgagt 10320
agttccttgg caatcgtgag gcttgcactc tctaccttga tgtcgccttt cagacttttg 10380
tctaggggaa cctctttgtc actgagtact tttgtcagag gaagggcatg caagcctccc 10440
tctatgacct tggggagccc gtctcccata gcatgctcat cctcaacctc ctgcatggcc 10500
ttcacctcca cacttcgatc acttcctcag ttccgttcca caaggtcaag gtcaacaacg 10560
acctcatttc cgaggagctc accaaggggc tgctaccacc ctctacaact ccaccactgg 10620
ggggccagcg catgcaccct cttctactac tatcttggga ggatcgtcgt tgcgctcgtc 10680
tgttcatcaa atctcaagca ttgcacatcc tccccacttg ccttggtctg aggggagggg 10740
tctagggtgg cggtcgacga aaaggcggtc gtggtggtgg ccatggggac actccttggc 10800
cttccatcta caacctgtgg actgatcgca tctccatgtg gcctggtcct tctccgagag 10860
tcccttctca gcgtatcact ccaccgtagc cagctcactt gacattgctc ggtctggggt 10920
atcaccttgg ccatcctacg ccattagcac ctgagtcacc acttctgtca ctgccacccc 10980
acttcgcttt gtcctggaac ccgtggccag gtgggtggga tcaacagtct cttgctagct 11040
ccttcaacac gatgaccttg actcctccca ctgtcaccta ctaggtggct gattcctggt 11100
gcctcctatc acaccacttt ggacgcgagt atgctatatt cttatccacc tcttcgtcta 11160
ctcctaccat tgtgggtaac gagaacattc tctcggtcac ctctgtcagt gattcggttc 11220
ttcctaggcc ctttcacctt cacaatgttc ttgttgcccc acatatcctt taatatttta 11280
tccgttcacc tattgagcat cggtaattct tgttccatag attttgatcc ttttggcttg 11340
tctgtgaagg atcttgctac ctagagtctt cttgcttgtt ttgatagatc taggcccctg 11400
tacacccatg caagcctgcg tctacctccc cacatactgg tttcgctaca tcatcacatt 11460
cgtcctcgct acagtcgcct actttgacta cttccactat ttgtagtact tggcatcgtc 11520
gacttgtcca tcccaacctt gtagcgtagt ccaagctatg tagtacttta gtaatttctt 11580
gtagtagggg cacccttgag catctatgcc agttgtgtaa gttaggtcgt cattttcgac 11640
ttcctttctc tagcttctca caacatacgc tgtgatttgt ggacctctcc agctctcctg 11700
ttaccaatgt tttgggatat aaatattatt tggtgattct tgatgattgc tctcattgtt 11760
tgtggatttt tccattgagt ttaaagtctg acacctttac gactctgctc ggttacttcg 11820
cttgggttta cacttagttt ggttgcacta tgaagatcgt ccagtgtgac aatggtcgtg 11880
agtttgataa cttctctcga tcctttacct cacttggagt ctaccttcga atgtcttgcc 11940
cctacacttc ccatcagagt ggtaaggctg agtgtatgat ttgcaccacg aataatgtaa 12000
tgtgttcctt gatatttcaa tcatctgttt ccgctcgcta gtgggatggg agcctccaca 12060
ctgctaccta cctccttaat tgtcttccta ctacgacaac tcttatgtca tgccccccac 12120
cccactttgc tcgaatggca aggcttagcg catgaattaa tgtcatgttt ttcttgctat 12180
tttactattt taggcatccg ttccttctcc ttactggtct gagagcctcc acaccaccac 12240
ctacctcttt aattgttttc ctattacagc gactcgtgcc cccacttccc actttgctct 12300
ttttgtcaac actccaatga ccatcttcgt gttttgggta tgtgtgccac cctaaccttt 12360
ccaccactac tctccatctt gttcttcgtt ctgctcgctg tgtctttctc gggtactcat 12420
ctgatcacaa agggtactgg tgtcttgacc tcaccactca tcgccatttg atctctcatc 12480
acgtcatatt cgttgggatg gatgtcccct tctcaactaa cacctagccc accaccgcct 12540
cccctttcga gttgtatttt cttcatgatc ctgactccgt ggtgcccttt gtccaaccac 12600
tgtttttcta gttgctgttt tcctctccac tgtttcagcc ggtggcccct gtgtgcccat 12660
ggtggatcat acacccttga attcctttcg ggttgcaccg ttgcccctcg tcgacctgtg 12720
tgtcacctcg tcaacccatg ttgggtccct aggccccggt ggccccactt ttgacattgc 12780
cttcgttgct agggccctct ttattgccct cgcctgtgcc ctcgccttgt tgctatgccc 12840
aacctgtgca tttctatcag cgacatgccc gagtgggtac accgcctcgc tcttgcgacg 12900
agccgacaat gtatcacccc ttcatcgtga ctctcaccac atccacttga tggcgaattg 12960
ccaggccact ggggtcctta gccctgttga ttgcctcatc ctctccgtga cctcctcatt 13020
gtcggtttct catgtgtcgt cttgcgttga ttgcaccctt tctaatactc attggcgcca 13080
cgctatgaag aaagagtatc tgactatcct tgccaatcac acctgggttc tggtattctg 13140
gtgccccatc tttcgtgcga caatgtggtg attgacaagt gggtctggac acataagtgg 13200
caggcggctg gctcgctggg ctactcagtg tcgtgctgtt gactacgatg agacgttaat 13260
tctcgttgtg aagcctgcta ccgaccgtgc catcctcact ttggccctct cccattttcg 13320
gccaattcat cacttggatg tcaaaaatgc ttttctgcat gggactctca cttagacttg 13380
tctactgctg tcagcctact agctgtgttg atcatgcaca ccccggtatg gtatgcaagc 13440
tcaacaaagt atgcaaactc aacaagtccc tctatggttc gaggcaagcc ccacacgttt 13500
ggtacaaccg cttcaccata tatctgctct ctttgttttg ttgaggccaa gttagacaca 13560
tttctgttca tctatcggtg tggtagtgac actgtatacc tactcctata tgttgatgac 13620
atcgtgctca ctacctcctt tctcatgcac tagatcacca ccgctcttca gcatgagttc 13680
accatgaagg atttaggccc ccttcaccac tttatgggga ttgctgttga gcatcgcttt 13740
gatggcctct tttttcagca gcgacgatac accctggaca tcctcgagcg tgctggcatt 13800
ctggattgca agccttgtgt ggcgtcagtg gacatgtagg ccaagctctc tggcgtcagt 13860
gctccagtca gtggccccat cacctaccat agcctcatcg gtgctcttca ccatctcacc 13920
ttcaccagac ctgacatcgt gtatgccatc caacaagtat gtctttatat gcatgatccc 13980
tgtgacctgt acttgggtgt ggtcaagtgg atccttattg ttgactatgg tctcctcctt 14040
tggcgctcct tcatccatga gctaatcatc tatactgacg ccgatgggcc gagtgtcctg 14100
gacacctgtc ggtatacctc ggcgaacctc atctcctggt cctccaagcg gtagccagtg 14160
gtctcccgtt ccagtgctga cgttgagtac tggctgttgc taatggggtg gctgaggcga 14220
gcaagctgca ccaacttctc cagtagcttc acaacccact caccatgagt accctggtct 14280
actacgacaa cgtcagcatc gtctacctct ctgtcaaccc ttttcaacac caccgtattg 14340
accgttttac tattggggac gtccgcgtat gtttcaacga gctcgtagtt tgatgacacc 14400
ttcactaagg gttttccctt gttcttggag tttcggttgt tggggcggga gacagaaccg 14460
gtcctcaact accaattgta ctcaccactc accaagattc ctaatgcaga gtaaccgact 14520
aggaggtgcg aaggcctatg gcggaagggg caagccgcgg aggccgaccc caacttcggt 14580
ccttcctaac aatcgcgaag gctacttgac tacgcaggga ccttggccaa acctcgacga 14640
gggacaaaca cgttcagcga gggcaggcgg gggctaggac gcgtcgaaga cccccgaagc 14700
accgaacgaa gacccaactg ctaccagccg atgttgaccg aggcgagatc actcctcttt 14760
ggtaggccgt cgtgaggcag tttttctcta aggcccccac gcagaggcca taagacgagg 14820
aatgttggat tccgccaagc ggcggctcag ttgtgggcgt agactcgcct actccacacg 14880
tcagcctttg atgtgggtga gaaagatggt gtaatatgga atgtagccag gggacctgta 14940
attaccccgt tgcacacctg ttgcacggta tatttgtggc atgtagtagg taaccaaggg 15000
cattacgata ttttaggcct tgggcccttg gccgccctat agatagcccc atcctgtagc 15060
tggatgggac acacttgaca agacatttgt gctcccagcc aattgttttg tcgtgccacc 15120
tgtaacaccg ctctcgagtg cttgggcaca ctgttctaag tcccaacatc ggtccagtct 15180
caacatcttc cactactaga gtttcatctg cgggggtggg ggtaggggtg ttagattctt 15240
gtgcgttatt gtactcaatt taagtgttat gggccttggc ccagcccatg taacccatgc 15300
aacacagttt accctaattt gggttaggtt tccaacaatt ttgtttgact tttccattat 15360
caaaacacac ttaggccttg ttcgtttacg tcggattgca cccggaaacg ttccagctaa 15420
tcaaagttta tataaattag agaagcaatc cggctaggaa tcgttccgac ccaccaatcc 15480
gacacaaacg aacaagacct taatatttca cgatgaatac tcaatttaag tgttatgggc 15540
cttggcccag cccatgtaac ccatgcaaca cagttcaccc taatttgggt taggtttcca 15600
acaattttgt ttgacttttc cattatcaaa acacacttaa taagttaata tttcacgatg 15660
aatgtagttg ttttggctta ttctgaacaa tttcacatca tgcagccgga aacactgcct 15720
gaacatataa aattcgacca tggttatacc tctaagagtc ctgcaattgt taatgtaagt 15780
ttccctgctt ctttgtaatt catttgtagg tttcgtaagt ttatttggat gtttctaatg 15840
ttgtgttttg cagtttcttg agatcatggc agaatttact cctgagcagc aacatgcttt 15900
ctgccagttt gtgactggtg ctcctcggct tccacctggg ggtttggctg ctctaaatcc 15960
taagttgacc attgttagga aggtaaattg gctatctttg ttcttatttg tactttatgg 16020
ttgacatgcc tgcccacatt gttgaagttt tgaattcttc ttaaattcca gcactcttct 16080
gtggcaaata atagttcaaa tgcaactgga gcgacagagt ctgcagatga tgatttgcct 16140
agtgtcatga ccaactatct taaactacca ccatactcca ccaaggtatg cttctttctg 16200
ctttttggct aactgtggtt atatctcctg tattgtctta taaattgagg attcagaaac 16260
ccagtcacca aagaattact tcatatagcc ttatcgtaac aggtaactgg acaaattttc 16320
aactaaggac gtggaaacta aaatttaatg tgggcagcac cttccagcca ctcattagtt 16380
agaattatta tttcgttagt ttaaatcaaa tagcatattc cacattgctt gaacccttta 16440
tattgacgca tgctggtttt ttttctggaa ggaaatattt acagtgctac tcgtgctagt 16500
gtgtggcttg ttatctgtgt ctgaattttg ttgaaaactt cttgcaggct atcatgctaa 16560
agaaactgct ttatgcgatc aacgaaggcc aagggtcatt tgatctttcg tgaatctcaa 16620
cactaacata ggtattggtc cacctagaaa tctgcgtcat tgttacccag agttagtttc 16680
tacctcattc atgtatgaca taggttaaac tcagctctcc ggagtcccac cgaaggtttg 16740
gagcccgtac ctttgggtgt ggatgtctat actctctttt cttggttgta tattcttgcg 16800
gatctttata gtgaataata gtaataaatt gttttgcgct tcttac 16846
<210> 123
<211> 10181
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 123
aggcctggag ttgccgacca cgagagatcc tgaccagcag ttgctgggct gtgtgtgtgt 60
ctgcgtctgt cactctcctc tctcgcgtct tggtcctcct cctccccttc atcgaacgaa 120
tccatccaag gcaaaaggga acgagggaag caagcaaaaa atccccaatt cctcctagat 180
cgatccaccg ccgacgggac gaatccaatc caggagctat cggctggaat tggattcacg 240
cgcacccgcc cgctgccgct ggccgccgcc tagtctcggc cggccggccg accggccgcc 300
ctccccgcgc gcgcgccttg ccttgtatgg aaacacgcag cagcagtcgc aagcgcgcag 360
cggccgccgc ctcctcgtcc gcctccaagc gcccccgccg caactcctct tcctcctcct 420
ctgcgccgcc gcgccagcag ccaccgatgg aaccctcgcc ctcctcccgc cgccgctccc 480
gcgccgccgc cgccgacaag ggcaaggacc ccgaccccgc ctcctccgac cacccttccc 540
cacccgcccc tgacgatgac gccgacgtca ccttccccca gcccttcacc tccgccagca 600
ccgccctcca gggcctcctc cgcaggctcg gcgccggcct cgacgacctc ctcccatcct 660
cctcctcagc gccctcctcc gccacgtccg gtcacctcaa gaggatcctc gccggcctgc 720
aggcgcacgg cgacgagtcg cgccagctcc agtccctcat gcagctctgc gagatgctct 780
ccatcggcac cgaggactcg ctcgccgcct tcccggtcga tgcttttgtc ccaatcctcg 840
tcgggatgct cggccgcgag gacgagcctg ccaccgctgg tgccagcccc gacgtcatgc 900
tgctcgcggc gcgggcactc tccaacctcg ttgatgtgct cccctcctcc tgctccgcgg 960
tcgtgcacta cggggccata cagtgcttct gtgctcgact gctcaccatc gagtacatgg 1020
acctcgctga gcaggtggtg ccttcccttt gtcttcttct tcctcgtttt atcctgtatc 1080
atcgctcatc ctcactatca tctaaaaatg aaatatgatc agtcatcctc tgcatgcatg 1140
cccttagccc cttaccaagg tgctgctaac acatttactt tactcgcctg ctcagtctct 1200
acaagccctt aagaagatct ctctggagca cccaactgcc tgtttgcgag ctggtgccct 1260
aatggctgtt ctatcgtatc tcgacttctt ctccactggg gttcaggtcg ggtcctctga 1320
tctccttaca accgttctta tcttgtgcac tgtttgcatc tacttacacc tcttcccatg 1380
ggggacgtac ttgttgtaga gggtagcctt gtctaccgct gcaaatattt gcaggaagct 1440
gccgtccgat gcttccgagt ttgtcatgga agctgtcccc ttgctcacga atcttctaaa 1500
tcaccatgat tccaaggtac tttgcaagct cagacaacca tacctctgga ttatgatata 1560
ttgaggtata tcatgtattt cccaaccctt catctatgtg ccttacaggt actagagcat 1620
gcttctgttt gtttgacacg cattgctgaa gcttttgcct attaccccga caaacttgat 1680
gaactctgca atcatggatt agttgctcaa gctgctagct tgatagctgt tggcaactca 1740
tctggccaga catcactgag cacatccaca tatacagtat gggccatttt attttgatac 1800
ccttttattg tttgctgtgg cccctctttc tcacacttat gctaaaatta ggttgcacgc 1860
ttttcagggc ttaatccgtc ttctctcaat atgtgcaagt ggatccctgc tggcagttaa 1920
aacactactt cttcttggga tcagcggcac tcttaaagat atcctttctg gctctgcttt 1980
gatttctgga acatcagtct cccctgctgt aacaaggcct gcagatcagg tgaatatttc 2040
tatcactgac ttaggacttg actattctgc tagtgctggc cttttaaaaa aaaatcaatt 2100
tagccacaat ttctctctgt gattggattc aataaaattc cgatttatca gtgaaaaatt 2160
aggctttggc atctcacact ttctgaaatt acatctacca cacgtatcat gcagactttt 2220
agaactctaa aggcgacatc acatgttcat aattatatgt accattgagc aattgtttgg 2280
gtgatttttt tgagcattga acatagatgc taaaatgtcg ggaattgtgc tcgtcttctt 2340
aaaccatgca aataatcatt caaaagagca gtgaactagt attttcaaag gcctgcatac 2400
tttcgttgtg cttttatatt gcctgagtaa gtcttttgaa atgttttaat gaagcagaaa 2460
atgttgattt atggcacaca ggggaacaaa agtctcactt gtaatatgat agagcaaaat 2520
ctcatttgga ttaagtagct gctgtgattc ttgtcgtctt attttcttcc tagtgtggtg 2580
caagtaaatt gtgcttttgg tcaatcgcat ttggtatggt acagatcgaa gagttttttt 2640
tccatgctat acaattgagt atgaatatat aatgccatct cagcttctgc ttgttgctgt 2700
cttgcttaat tgctaaaatc atttgtggta catacacaca cgagtctggc attttttgag 2760
tatgatcgtc tgcaacagct gctgtatctt ctcacctagc tgctgaaaca ttaattcttg 2820
aaacccacac tgtttgactc atcagtttca tgcacctgta tgttttttgt tgtacaaatg 2880
tttttcatat gtttacccca aatattgcag atgtttgaga ttgtgagcct tgccgatgat 2940
ttgcttccgc atatccctgc gagaatcatt aagttaccaa catattacca tacatataaa 3000
ggctcttcaa caaagaaatc tgcttccatc aaacaagagg ggggtggttc aacaggaaat 3060
gaaaggtctg gccgtgagag gctattgcgt gagcaacctg agcttttaca acagtttggt 3120
atggatttgt tacccacaat gacacaggtt agtgctggaa ttcttttcct gagtcctgac 3180
agccatttca attgctattt gtgttttgtc tttgtgtact ttaatagcct gtgtgcttgt 3240
tcaggtgtat ggttctagtg taaatgcacc aatacgtcac aagtgtttat ctatcattgg 3300
aaaattaatg tactacagct ccgccgaaac aatccagtct ctccttggca caacaaacat 3360
atccaggttt tctctctctc tcacctggaa cacacaaaag agatgcatgt ttcattaata 3420
gaaccgaaaa aattctcagt tcatttctag agaatgattt cagaacaatg gatgttaaaa 3480
gacaaaaaga aagtctctca agtctcaatg tacatacttc ttttccaatc ccttctaagt 3540
tatttcattt ctatctgcag cttccttgcg ggcattcttg catggaaaga tcctcaagtg 3600
ttgattcctg ctcttcagat agcagaaata atgatggaaa agcttccaga gacattttcc 3660
aagttatttg tgagggaagg tgtcgttcat gctgtggagt cactcatatg ttcagaatct 3720
tccaacaaaa tgccttctca ggtgccacct caggataaag ataaagattc tgccatgcca 3780
tcacgttcta gacgtcaacg ccggcgtgcg ggtgctgttg cagcagaaaa tagttcattg 3840
gatgaatcaa atagttccaa ccttggtgtt atgtgctcaa cagcaaccgc ttcagaagct 3900
ccaaacacca gccttcgttt cacagttagt gaccatgcaa agtctttcaa agataaatac 3960
tttccggcgg atactgattc aagtgatatc gggtttactg atgaccttct taaactgagg 4020
gcactctgtg caaagttgaa tactgtctct gagaacgtca aaacaaaagc aaaagggaaa 4080
tcaaaagcta taagcactaa ttttttggac atctcaattg atgttgaaga acaattagac 4140
aaaataatat ctgaaatgct ctctgagctt agcaaagtta atggtgtttc cacatttgaa 4200
ttcattagaa gtggtgttgt catagcattg cttgactatt tgtcctgtgg gacatttggg 4260
aaggaaaagg tatctgaagg aaacctatct cagcttcgtc agcaggccct tagacgatac 4320
aaaaccttca tatctgttgc cctttctatt gatcatggac gagatgaaac tcccatggca 4380
ctgttggttc agaaactgca aagtgcattg tcttcgctgg aacgtttccc tgttgtgctc 4440
agccagtcta gtaggattgg tattggaggg tctcgtctga cttcaggtct tagtgctctg 4500
gctcaaccct tcaagcttcg tctttcccga gctcaaggcg aaaaatcact tcgggattat 4560
tcatcaaata ttgtgcttat cgacccattt gctagtcttg catctgttga agaattcctt 4620
tggcctagag ttcaacgcag tgaggttgca tcgaagccta taattccatc tggtaataat 4680
tctgaatctg gggttcctgg caccacggct ggtgcatcat taacagctgc aatggctcaa 4740
tctggtcggc gcccaacaac aagatcaaag tcatctgctg caggtggtct tacatctaag 4800
aaggattctc atgatgaaag cactagtact gccaaaggaa aaggaaaagc tattgtaaag 4860
ccaaactcag atgaatcaaa aggacctaat acaaggaatg ctgcccacca aaaatctgct 4920
tcagagaaag atttggaaat gaagcaggca catggccaca gtagctctga ggtatttttt 4980
ttgtttacat gcctcattcc tggttagttt ggcttgcaaa gaaaagttac tgtttctatt 5040
caagttgggt agcagctctt ctcaaaagtc gcattgtcag ggcctaagga ggcccacgga 5100
ggggctgcgg cagcagcagc catgggccct ccaaggggga ttagataatg atagttagag 5160
ataagattag aagattagtt gagattattt aggagattag ttgagattat ctaggaaggt 5220
tatcagttag tttgttgaga gtttttagga gagttttaag gagataaagg tctcaagcta 5280
gacaagggcg gccacccggt ctatttatgt aatcaatgga taagaaataa attaggcaag 5340
aattagaagg gagaaaccct cctcttgctc ggccgtgggc agaggccctc ggccgacgtt 5400
cctgcccgtc gatacccctc tctaatccct caccgattta gcgtccataa cacacatcca 5460
cttctctggt gtcatatttg gtgccaccta taaagctcat ggcttgggcg ccaaatgaac 5520
atgataattt tagccgcgga tctagttctt aatttatatt accctcataa tatgtaaaca 5580
tgattttgtg agttcaaaac tgtgccaatc taaaaaatat taggataatg atcaagaagt 5640
aggctcattt tctagttgag agtgtaagca tggtaactga taagtaaata cattctatgt 5700
tggtatatgt aggatgaaga gctcgacaca tctcctgttg agatcgatga tgctttgatg 5760
attgatgatg atgacatttc agatgatgat gatgatgacg acgatgatca tgaggtaact 5820
gtttgaaaat tgaatgccca ctcttttttt tgcttcgttg caaccatccg tagttgctta 5880
ggttcatatt caaggtacta aatagcatgg gttcatagca gtttgatgtt gcggatgtca 5940
ggtgctatta cggaccgcta agttccaata gcagtatata ataatcaaac ataaatttat 6000
gtatatatat atcattaata ttaatttaat agacgtttaa ttaattatac ttagataagt 6060
gatgtctcac atcattatta tcaacaaagc cttttagccc caagcaagtt ggggtaggct 6120
agagttgaaa cccatcggaa gccacaaatt aaggttgagg catgtggaaa tcgatgtctc 6180
actctcacat actttaaatt aaataaagga acatgatatt caacatatta catataagtc 6240
atcacacatt ggactggagt ctacaaagaa atatattttg atagccgtaa aatagtcata 6300
aaatcatgtt tcattgtcat gtcagaagta gtaacctcaa tagtttcatg atagttaaaa 6360
tgggataggg ttatttgttg tatatcatac aacactattg tggacactat gaagaagtgc 6420
ggagcactat aacagcctgc tacctcaaat agcagccgct aagttccaac ctcaaatagt 6480
ggtagcggta agagaccgct accgtttttt gcggacgcca tagcggttgc tactaacgct 6540
atttagtacc atgcttatat ttatttactt tatgacaaaa ataacctttt agttggtcca 6600
agatgatcat ttgtgccctt acttgactaa tttaataacc tctccaaatt cttaggttct 6660
ccaagaaggg tctcttccaa tctgttctca agatggtgtg catgacgtga aactgggtga 6720
tgccgatgaa tctaacattg gatcagctac ttttagccat gcacagccat catctggttc 6780
tggggctaga aacacttcta gcaggggacc ggattctact gaatttcgaa gtatgggcac 6840
attcggttca cgaggtgcaa tgtcatttgt tgctgcgacg atggctggtc tggcttctgt 6900
tggtggtcgc agtgttagag gtgtccgaga tcggcgtggt ctgtcacttg gaggtagcat 6960
gaatgatcac aataaactta tattcactgc tggtggcaag cagcttaaca aacatttgac 7020
tgtatatcaa gctatccaac gtcaacttat gctcgacgag gatgatgaag aaaggtttaa 7080
tggatctgat ataccaaatg atggaaaccg cttctggggt gatgtgttca caataacgta 7140
ccagaaggct gataaccaag ttgagaaggg ttgccaaggg acttccacct cattaaacat 7200
aaaatcagat tcttacaggt ccatttctga agcacaaggg gtttctcttc ttgatagcat 7260
cttacaagga gaacttcctt gtgatcttga gaggacaaac tcaacgtaca acattttagc 7320
attattgcgt atacttgagg ggcttaatca gttatcccct cgtttaagag tacagggagc 7380
atctgatgat tttgctgagg gaaaaatcaa taccctggat gagttatata gaactggagc 7440
caaggtacct tctgaagtgt ttgtcaatag caagttaaca ccaaagcttg ctcggcaaat 7500
gcaggacgtt ctcgcactat gtagtggcag tttaccttct tggtgttatc aaatgaccaa 7560
ggcctgccca ttcctgtttc cttttgagac aaggaggcag tacttctatt ccacagcctt 7620
tgggttgtca cgagcattga atcgactcca gcaacagcag agtgataatc acagctctgg 7680
tggtgaaaga gaggtcaggt ttggcaggtt acaacgccag aaagttcgtg tttcccgtaa 7740
ccgtattcta gattctgctg ctaaagttat ggagatgttc tccagtcaga gggctgttct 7800
tgaggtggaa tactttggcg aagttggaac tgggctgggg cctactctag agttttatac 7860
tctcttaagc catgaactac aatgtgctca actgggatta tggagagcta cttcttcata 7920
tgattcagga ttgcacactg atatgaagga tgtgatccgt ttagatcctg aagatgggtc 7980
gtcaggaaaa gaactcaaca cagatctacc aggtgatggc atgtatttga tacaagctcc 8040
tcttggattg ttccctcgac cttggccacc taaagttgat acttcagaag gtagcagatt 8100
cttcaaagtt ttggagtatt tccgcttaat aggtcaagtg atcgcaaaag tcctgcaaga 8160
tggcagactt ttagatttgc ctttatcaac agcgttttat aagcttatac ttggacaggt 8220
atgtacagag aagctctatt tccaagtgct ttgctttcat tttgttttgc tttgttttat 8280
gtattaatta gttgtttgta ttcacttttc acgacttaat gtgttgcaat tggccaggaa 8340
cttgatttgt ttgacatagt gtcatttgat tctgagtttg ggaagacatt gcaagaactg 8400
caagttcttg ttgaacgaaa gcagttcctt gaatccactt ctggcaagaa ccagctacaa 8460
gttgcagatt tgtgtttcca tggcgcttct attgaagatt tgtgtttaga ttttactctt 8520
ccaggttatc ctgattatgt gcttaaagag ggtgaaggag gcacaattgt aagtgatgaa 8580
gttccatatt agcgtgtatt tatcttggtg atttcacttt gtatgtttta atctgttagt 8640
atcatctgca ggttaatatt tataatttag aagagtatat tactttggtg gtggatgcta 8700
cagttaagtc agggataaag cgacaggttg aggcatttag atcagggttt aatcaggtac 8760
agcctcattg gcattgctct aacacaatct tataataatt ctacttgttc tgattttgtg 8820
tagttcagtt tactctaata gttcttgttt attttaatgt ttcgtaggtg tttgacatgt 8880
catccctcca catattttca cctcaagaac ttgactatct aatatgtggt cgacaagaaa 8940
tttgggaggt aatgttctgt cctttttatc caagttaatg tatgtaaaag tgtggcacta 9000
aataaacctt ggaatctgca gtcggaatcc ctggtggata acataaaatt tgatcatgga 9060
tatactgcta aaagtcctgc gatcgtgaat gtaagctcat tcgtcctcac atgctacatc 9120
tcttacatac atccagttgt ggtttttgag aagtgtggtt ccttgattct tgctcttgtt 9180
tgtagctact cgagattttg tcagaattca cccctgagca gcagcatgcg ttctgccagt 9240
ttgtaaccgg tgctcctcgg cttccacatg gtggattggc tgcccttagc cccaagctta 9300
ctatagttag gaaggtacct tttcatggtc tatctatcgt ggcatttgct ataatgacat 9360
gtgcacacct attctgcttg atgttatttt tggggtaact cggtgttttg attattgcca 9420
gcatccttca agtgctgtga atacttcaaa ttcaactggg gcagtagagt ccgctgatga 9480
tgatttgcct agtgtgatga cgtgttatct gaagctgccc ccctactcca caaaagtaca 9540
gtgtccttgc tgttttatca ctaaaaagtc agcttgacat tctgatggtt cctgctgacg 9600
ggctgttgct attgtttgga caggaaatca tgcgcaagaa gttgctttat gcgatcctgg 9660
aaggccgagg atcatttgat ctttcataag aaatatacaa tatccgaact aacatacagg 9720
ggtcagtttt attctggaga tttagtttct tggtccatgt ataacactga gttgaggtta 9780
ccagctcact cagcaaacgg cttcaagggc gagagcccca gtcctcactc ggatgcaggt 9840
gctgctgtcg atttctccct tcttttacat ttgttcagct actgtattct cttcgagggt 9900
tgcccttgtc acctgcaatt atgaatctac agcttttgtc atcgaggaag tttaacagat 9960
tttggtttct ctaccatgat ctgtttccat tgggcaggct aagctatagt aattgaggat 10020
gtgcgccttg agtgaggatg tgcgccaaca agaacccccg tttgtttcag tacatgtaaa 10080
taaaagggat tttttgttcc agaattgtta tctgtaacaa gaacatctac atctgtgatg 10140
actgaccatt aataaatgtg tcccggtgta ccatttgttt t 10181
<210> 124
<211> 20826
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<220>
<221> misc_feature
<222> (1870)..(1969)
<223> n是a, c, g或t
<400> 124
atgagccgag gccaggggta gcggtgtcag cacggacaga ggcagagttg gctcgaaaag 60
aagactgggt cggccggagc ctggccgggt cgtccgttag cgggaccgac gccagagtta 120
atctgccgag gcctcgggct gggctgatgt cttcggggga cagctggtcg aggcctcggg 180
gtgaccagct gagccgcctg ctcgagccgg attcttggag aagaccctgg cggcgatggc 240
ccgggcgtga tgatgatgtc gtccttcaga gtggagatcc tcggaccgcg tcgccgtccg 300
aggctaggtc ggacctctcc gaaggtgtag ttgacgccga gggtgctgct gctccctcca 360
accgtcaaga tccgagcctg caggatcgga ttatcttgtt gtgtgtgttt tctgcggccg 420
ccgaggccca aacacaccct cgccgtgttg taaagctgcg cttcttttcc tcttgtttcg 480
agtatctgga ctttttcgtc ggtaacagaa ttgtctgtgc gagcgagagt tgcttttcat 540
ggaaggtgat gagtgaggta tccgtatccc ggaggcgtag gaatccttcg gctcggtcgg 600
ccttgccgct tacgcgcact cttgcccgtc catagggttc tgtcaccgac gcagtcaaga 660
aggcccgaag aatcgcttca gcagaggagc tttcgagcgt gaagacttgt tcggtccgcg 720
gaatcactta tccgaacgtg agttacttat cgcagaaggt gatgagtgag gtatctgtat 780
cccggaggcg taggagtccc tcggctcggt cagccttggc tgcttacgtg tactccgtcg 840
ttttcgggat ccactttcga agtagtcgaa aaggacgaaa gacattctag cggaaaagat 900
cttttttcga ggaaaaattt cgacgcagag ggggttcccc ccttttagcc cccgagggag 960
ggtcgggctt tgccgaggca aggctgaccc ttccttgatg actaaacttt gcgtgggaat 1020
gaggtatgcg aacaaattga aagcatctta agggtagaag cgacgtagct gttggatgtt 1080
ccaagcgttg ctgtagacct cgccttgact gttggccagc ttgtacgttc cgagcttcag 1140
aactttggcg atgacgaacg tcacttccca gggaggcgtg agcttgtgcc gccctcgggc 1200
gtcctgtcgc agccgaagca ctaggtcgcc cacctggagg tctcgggacc ggacccctcg 1260
ggcgtggtag catcgcaggg actgctggta ccgcgccgag tgtagtaagg ccatgtcccg 1320
agcctcttcc agctggtcca gcgagtcttc tcgactagct tggttgcttt ggtcggcata 1380
ggccctcgtc ctcgaggaac cgtattctaa gtctgtgggc aagatggcct cggccccata 1440
gactagaaag aacgacgtga agcccgtggc tcggctcggc gtcgtcctca ggctccagac 1500
caccgagggg agttccttca tccatcgctt gccgaacttg ttgaggtcgt tgtagatccg 1560
aggcttgagt ccttgtagaa tcatgtcgtt ggcacgctct acttgcccat tcgtcatggg 1620
gtgagccatg gcggcccagt ccacccggat gtggtgatcc tcgcagaagt ctaggaactt 1680
tctgccgatg aactgggtgc cgttgtcggt gatgatggag ttcgggaccc caaaacgatg 1740
gatgatgttg gtgaagaacg ccacctcctg ttcggacctg atgcgtttag gggtcggacc 1800
tcgatccact tggagaattt gtcgatggcg accaacaggt gccgtgtagc cccgggtgcc 1860
ttctgcaagn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1920
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnna agacaaggag 1980
ttaaggaaca caactttgtt catggtacac cttgttctgg agaaatgcaa caattacctt 2040
tttattttct gtttacacat taccctgtct tgaatgctac agagtttcta ctgattagtt 2100
cagtttatga ctatatggtc ttacttattc tctgatacat ttggcaaagg taaaacttca 2160
gggccaacat aggccatgct ctcccctgat cattttagca actctgcaga atatctgaca 2220
gtattttgag ctatattgct aatgaaacat gtaaatatgc ttttggaaac actgaattta 2280
tcttcaaatc acaaccctct ttgcctccta acatcggctg caagctctgc cactggtttt 2340
tgatagaaca tttttctgct ttctattgtt gtttggatct ctagaaataa tatcttcacg 2400
tttgaaatac tatgcataca gataatagac gcatacgtgt accatctctt tgtatttgta 2460
tgcatgctct tgatggtggc ctaacatata tgctacgttt ttttatttag tcattgcaag 2520
ctcttaaaaa aatatccctg gagcatccaa ctgcgtgctt gcgagctggt gcactaatgg 2580
cagtcctatc ataccttgac tttttctcca caggtgttca ggtaatcact ctctgcctat 2640
tatagacttt tgcatcacaa aatactttgt ttgtttcagc tcatttatct catagcttat 2700
tcttatgcat tacttgtcat tatattgtga cagatgaact gtattattta ttcttgttat 2760
ttatttttat tactaactgt tgtttttgtt gatggaatgt tttcagagag ttgcattatc 2820
tacagctgct aatatgtgta ggaaacttcc ttctgatgca tcagattttg taatggaagc 2880
ggttccactt cttactaatc tactgaacta tcatgattca aaagtacaat tgccattcca 2940
aatgctgata tcctctatga taccatttat gtttcagatg ctgatattcc gttttgctta 3000
ttccaggtac tggagcatgc ttctgtctgc cttacccgta ttgcggaatc tttttcacca 3060
tttccagaaa aattggatga gttgtgcagt catggattgg ttgcacaagc tgctagctta 3120
gtgtctgtta gcaactcagc aggacaggca tccttgagta catcaacata tacagtacgc 3180
tccattttgg ctttttgggt acaattattt gttttacgtt tgtgaagttt atgccatgtt 3240
tttatagggt gtgattcgtc ttctctcaat atgcgcaagt ggatcacctc tggcagctaa 3300
aacacttctc cttcttggaa ttagtggcat acttaaagat atcctttcag gctctgggtt 3360
ggttgctggc acgactgtgt cccctgcttt aacaagacca gctgatcagg tgattttgtc 3420
ttttcatagg cacttcaagg ctctttgggt atcatgtgtg cttgatgtct acatttgttg 3480
tttatcttaa caagctaagg ccccgtttgg cactgctcca cttcacaaaa acaagcgcaa 3540
cttcatgaac ttcaaaatat agcagcccag ctttgaagtt gcagagtttt tgaagagttt 3600
ggcttgtaga ccagctttag tttcataaaa aaatataatt aaaaaaaact tgtttcttct 3660
tcctttggcc gtggcggcca gaggcgcacg tgcgcgactt gggaagacag acacaacgct 3720
gcgatggcct ggcgtggctc tggcggccca gcgcgctgca cgcagcctgg cggcacgttg 3780
tcggccgatg cggccctgct cgctagcgca ggcggtggca cgcgcgctgc cagagtttgg 3840
ttcaggcgtg gctctggcag cggcacatgg cctggcggtg cggctgccgg tcggcacggc 3900
cctgctcgct ggcgtaactg tgtgcggcct gagttccctt gagaacagaa cgaaagaaaa 3960
catttcgtga gggattgcgt aactgatggc aaaggtgggt aatttcaccc caacttaatg 4020
agggagttaa aaatacccat tcgtgaagta tacttttggt tgtttcatgg atttggtgaa 4080
gttagggtca acttcacctt ttttggtgaa gctgggagtg tttggctaga gttttataaa 4140
gctaagctag attttgtgaa gtgaagcagt cccaaacatt ccctaagtgt aaattaatgg 4200
agctaggctt actctatgct tcagatagta cttcaaccat tgattccaat ttccaagcct 4260
caggatatca tctctattaa ggcccccttt ggcacggctc tggctctagc ttcttgcagc 4320
cccttggaag gagccctgcc aaacagccat ctaaaacaca gctcctggca aggagccagg 4380
gccagaaaaa tggtttctcc cagctcctta caaacttgat acaaattatt acaaaactgc 4440
cactggacct gttttgacca aaccttttcc aaaacggctt caactggccc aaattataaa 4500
gccagcacaa ttaatcccat acacttccca accttacaag ccccttaata cagcccatct 4560
cttccttcat tctccataca tatctgaaag tcttccatgt tctctctctt ctaccacttc 4620
gtggcgcagt gaggggggct tacttctagg agcgcctaac atgtgctata taacatcttt 4680
acgcacaatg acataggaac aagtgtcgtc aaagaaaaca aataatattt gtgtatatta 4740
aaacattttt tgatgaattt gatgaaatga aaccaaatcg tcccataaca tgtttttctc 4800
ataatgcatt ttgtttttta atcgccacca ataatatact ttgagcttgt agtcaatgtt 4860
cgtttatggt atgacattgt ttttctttaa tgtgagcata ctcatactac ttcccatgtc 4920
gagcatttgt gaacatgatg ttatcctatg ttctattcat gatagcacaa tttatatttg 4980
tgatgaattg tcatctatgt taaattttaa tgggtactgt aacttgtaaa gctactgact 5040
ttttggtagt aatagattat caccacacaa ttccatatgc aaatgatgcc attatgtgtt 5100
ttttggtatt acacagatga atgagattgt gaagcttgca gatgagttac ttccttctct 5160
gcctgttggg accatttctt taccagtgta ttctggtgtt cacatgaaag gttgttctgt 5220
aaagaaatcc acttctagta agcagggtga acatggttca acagcaaatg aactatcagg 5280
tcgggagaag ttattgcgtg atcagcctga acttctgcag caatttggca tggacctgtt 5340
acctaccatg acacaggtca gtcttgtctt attaatgttc tctttgatgt tgttaatggt 5400
aatgctgatg tggtattacg ggacattgtt gctatcatgt tggtgtgata cttaaggact 5460
tcaatacctt gaacaggtgt atggctctag tgtaagtgga ccaatacggc acaggtgctt 5520
atctgtcatt gggaaattaa tgtactatag ctcagctgag atgatccagt ctctccttag 5580
cacaacaaac atttccaggt gcttaacaca atatatttca tcgagttatc tcatgttatt 5640
agaaactatc ttatcacatg aattccaaat atttgtttca agctattaac ttctatatca 5700
accattgcag ttttttggct ggcattttag cttggaaaga tccacaagtg ttgatccctg 5760
ctcttcagat agcagaagtt ttaatggaaa agcttccaga gatttttttg aagatgtttg 5820
tgagggaagg cgttgttcat gctgttgagt cacttatatg cccagaatta tctggtcagg 5880
tgactcctca tgtggattcc attacatctt cacataatag gcgcaaccgc cgtagaaata 5940
atgctgtgaa cactggaaat aacttgcctg atggaccaaa aggttccaat tctatgattg 6000
ccaattcacc cccaagcatg gctgaagttc caaataatag tcttcgtgct ttagttagta 6060
atcatgcaaa gtcatttaag gataaatatt tcccttccga gcctggctca agtgacattg 6120
cagttaccga tgaccttctt aaactgaggg cactttgtgc aaaactgaat acaacagctg 6180
acactatcaa aacaaaagct aaaggaaaat caaaggtagt gtctgacaat agttttgatg 6240
tcttgtgcaa tattgaggag caattggatg atataatagc tgaaatgttg tctgagctta 6300
gcaagggtga tggtgtttcg acatttgagt tcatcggaag tggagttgtt acggctttgc 6360
taacctatct gtcatgtgga acatttggga gggaaaaggt gtctgaggca aacataccaa 6420
atttgcgtca tcaggcagtc agacgataca aagcatttat atcttttgca cttccaaatg 6480
ataaagatgg gaataaaact cctatggcat tcctagtcca taaactgcaa agtgccctat 6540
cttcgttgga acgtttccca gttgtgctca gccattctgg aagggcatcg actttgggag 6600
gatcccgcct gacaacaggt ttaggatctc tgtcgcagcc catcaaattg cgtctttgtc 6660
gagcgcctgg tgagaagtca cttaaggatt tttcgtccaa tgttgttctt attgactcat 6720
tagctagtct ggcagctgtt gaagattttc tttggcctag agtccagcgc actgaaccag 6780
tattgaagcc tccaatgtca tctgcaaata attctggatc tggagcagca agctccacag 6840
cttgtgcgcc ttcaattcca tcagaaactc aatctgtccg tcgcacatcg ttaagatcaa 6900
agtcatcagc tgccaccagt ggtgcaatta agaaggatta tcaggaggga agcataaaca 6960
cctcaaaggg aaaaggaaaa gctgttctta aattgagttt ggatgaacct aaaggcccac 7020
atactaggaa tgctgcgcgc agaaaagcta cttcagagaa agatgttgaa ctgaagccat 7080
cacatggtca catcacttca gaggttagat actcgtgcat tcaatcttat ttgttatttg 7140
cataatatat gcgaaatggc ctctagccta gtggttaaag gcttccgaat agcacctcca 7200
agtctcgggt tcgattcccc tcaggggcga attttccggc ttcctgtgcg ccaccctccg 7260
gttggaccgc tgtagagggg acggttgacg tcagctcgtt agtgatgggg aggggggggg 7320
gcagagtttg gggattttct tggccgagac cattgtttta gtctatcttg atataatacc 7380
gggatggcgg tcattccttc cctggctgag ttttttttat ttgcataata tatcaattga 7440
gccctcctaa gtgagctccg ttgggaagtt tagctttaat ttctggctct aggacacatt 7500
gattgcaaat gtttttttaa caatgccttt tgtttactca gctgacaaat gttatcatag 7560
taatgataca acagtaatgc agctgattta ttttgcctta agtgtatcta gtttgatcag 7620
tactttgttt tagtatgtaa gcaaaggatt tcaaagggat gttcttcata gacatgatgt 7680
acagattgta taactataaa aggagggggg gcagtgctgc aggctccgac atgaggagtc 7740
tggggaaggg ataaaccgag gcaacccttc tcccgtaaat acggagagac actgctttga 7800
acccacaacc tggtgactcc gggagacaat tcacaccact gacgtacaga ttgtataact 7860
ataagtctat aatatcataa atgtataaca atgaaaatgc actgaactgt ggcatttctg 7920
tgtatcatgc atttataata ttatatctga gataggatat aaaaactgaa gcatttttta 7980
taagagcaac aaagattaaa gtagaaatat gcctgtttca ctaatatttt ataatttagt 8040
catattatgt aatgacctga ctacgctttc gtaacttagt atactggact atattgtgga 8100
tactcacaaa tatgaaatat cagtgtaatc ttttactcta attctctaaa taaaagatat 8160
tgttgtattt tgattttaaa tatctaagtg aagttttcag gggaaacaac tttatgaatt 8220
attatttgga atagtacgtg catgttcttt tataagtttg atatctgaat tttagaaatt 8280
aaaaagtttg aaagccctgc aacatatatt tatattatta tacctgcagg atgaagatct 8340
tgatgcttct cctgttgaga ttgatgatgc tttaattctt gatgatgatg acgaggatgt 8400
cccagatgat gaagatgatg atcatgaggc ggtaattatt ttttaaatct tgcttattac 8460
tacaaaggtt tgtattgagg ggacatgatt tggaactcag tagattttgt tgaaatgatc 8520
aggttctccg aggttctctt ccttcctgtg ttcctgagag agtgcatgat gtgaaattag 8580
gagatgctga tgattctagt gttgcctcat tggcaaatga taaccaggca cagccctcat 8640
ctggttctag tacaaaaaat acttctagca ggggattgga cactgctgaa tttagaagtc 8700
cagccacatt tggttcacga ggtgcaatgt cgtttgctgc agctgcaatg gctggattaa 8760
ctccagtagg tggtcgtgga attagaggta gccgagatcg gaatggcctt ccattgggtg 8820
ctcgtgcaac tgagcattac aacaaattga tatttacagc tgctgggaag cagctgaaca 8880
aacatttgac tgtatatcaa gctgttcaaa gacaagtagt tcatgccgag gatgatgaag 8940
atcggttcgg tggatctgat ttacctgatg atggtaacca cttctgggat gatataaggg 9000
gtgatgtgtt cactataacg tatcagaagg ctgataacac agcggagaag gggtctgttg 9060
gaggttcagc ttcagtgcca aaatcttcca aatcagattc ttgcagaact ttgtctgaaa 9120
aacagcgtac ttctcttctt gatagtattt tgcaaggaga gcttccctgt gatttagaga 9180
aatcaaacca aacctacaat atcttatcac tattacatgt gttggagggt ttgaatcagt 9240
tatcacctcg tctgagactg cagtcagcct gtgatgattt tgctgaagga aaagttgcta 9300
cattaaatgg gctatacgat gttggagcta aggtaccctc aaaggagttt atcaacagta 9360
agatgacccc aaaacttgct cggcaaattc aggatgttct tgcactgtgt agtggcagtt 9420
taccatcttg gtgttatcag ctgacgaaag cttgtccttt tctgtttcct ttcgaaacac 9480
gaaggcaata cttctattcc acagcttttg ggttgtctcg tgcacttcat cgtcttcagc 9540
aacaaccggg caatgataat aacactgctt ttgaaagaga agtcaggatt ggtagattgc 9600
aacgccagaa agtccgtgtt tctcgtaacc gtatcttgga ttctgcagct aaagttatgg 9660
agatgttctc taatcaaaag gctgtcctag aagttgaata ctttggtgaa gttggaactg 9720
gtcttggtcc aactttggag ttttatactc tcttaagccg tgagctgcaa agggttgact 9780
taggattgtg gagatctcat tcttcagata attctgggat gcaaattgat gcgaatactg 9840
atgatttaat aagaagtaaa aatcatgaat cagaatcact tactgagagc aggaacatag 9900
tacaatcacc tcttggatta tttcctcagc cttggccacc tactgctgct gcatcagaag 9960
gtagcaaatt cttcaaagtt gtcgagtatt tccgcttagt tggtcgagtg atggcaaaag 10020
cattgcaaga tggaaggctt ctcgatttgc ctttgtcgac agcattttac aaacttttgc 10080
ttggacaagt aagcatgagg gcctccttga gctagtttta tcttatgttg tcttctaaaa 10140
ccttcctctt ttcggttggc acttaacttt ctgtgccttt tccttattaa tttaccaaat 10200
ttgctttaat aggaacttga tttatatgat atattatctt ttgataccga gttcggaaag 10260
acattgcaag aactgcaaat tctcgttgca cgtaaacaat ttttggactc ctgctctagt 10320
gagagccaaa agatagatct atgtttccgt ggtgctcccg ttgaagattt atatttggac 10380
ttcactcttc cgggctatcc tgaatatgtt ctcaaggaag gtggagagaa tgcagaggta 10440
agttatatga taccttagtt tgtttttttc ttctagtgtt gctgcagatg actcattgtt 10500
atcctgtctt ataggtcaac atttgtaact tagaagagta tatttctttg gttgtggatg 10560
ctactgttaa gaccggcata atgcggcaag tagaggcatt taaggcagga ttcaatcagg 10620
tcttctttct tattttgtga tttgtgggca atttctatta ataattgatc ctagaactaa 10680
ctagcacatt tatttattta ttcagaaaaa aatactcatt gtatatttct cacttcaggt 10740
gtttgacata tcatcactgc aaatattttc tcctcaagaa cttgattatc tgatttgtgg 10800
tcgctgtgaa ctttgggagg tattgctttt gtggtcattc ttgatgttgt acttcctctg 10860
tcccagaatg ataagcatag tttgtatagg aaaaagtcaa atttagaatt ttatttccca 10920
gaaattgctt gtattatact agtttatgtg gctttgtgtc ccgctgccat aggtcaatgt 10980
tagctattga aagctagttg ctctagtcta ctgcttccct ttttttttcg ggcggggggg 11040
gggggggggg gggtattctc cctcctttgc agttcacctt ttttctctct taatgaaatg 11100
atacacaatt ctcttttgtg ttcgagaatt ctttttacaa actttggcca acaattaatc 11160
aaattatata catgttttgg acaaacttgc atcaatagat tcatattcag agtgcatttg 11220
agatgtgatt aattgtttag caaacataaa catattgtac gagaaattaa tggtcaattt 11280
ttttaataga ttatatgagt tattgtactc agtctaagtg ttatggaccc tggtcccgcc 11340
catgtaactc cttttccaat atatgcaacg tagcccaccc tgattggagt taacatggta 11400
gcaagcttgg tcttttctct ccctcccttc cctagccgcc acctcctggc tgccatcact 11460
accgtcgggt cgctattgtc gtctgcgcct ccatctcctc cctagcgcct ttcttgctca 11520
tcggccaccc atatttatta aggcagtgtt aataaccatg gggccaccac cttcacttta 11580
gctttctagg cgcgactggt ggccttgttg gccctacggt tggcgccttg cccaccggtg 11640
ctttagtccc ggtcacctcg attgctgccc tagtcgctcc cgtcgccctt gcctttgctg 11700
tggccatcta tggcatgggt gccgccagcc agggcccaga cctagctgac tctggcctag 11760
cttatctttc ggtcgcgtta cttccccctg atgtcgcttt cgctcactcc accttggcca 11820
tagcccccat tgctgccaag acggtgtttg ctgctgctcg ggatcgggaa catgttgctg 11880
cccttgctta ggggcacaaa cgcaccacga cggatgcact cgctcgatag cagaccaatg 11940
ctgagggtca cctgctcggc tcctcctgca tcgagaccct cccagcgctc tagtggtcgc 12000
ccctttgtcg gagtacgagg tcgagatcat caccaacctc tatgccgagg tggcgtgtgt 12060
ccagaacatt tgctctatga ttcccgtcat ccttgacaca acttcctcca actacgcctt 12120
ctggcgggcg atcttgtctt cactctccat cgctacgcct tggactacca cgtcctcacc 12180
aacactgtct ccctcactaa cctatcctgg cggcagatgg atagtgtggt cctctcgtgg 12240
atccccagga ccgccatcgt tgcgctgcat gacgtggttt gcgagcgtgg tggcactatc 12300
cgtcttggac gtcgtcgagt agttccttgg caatcgtgag gcttgcactc tctaccttga 12360
tgtcgccttt cagacttttg tctaggggaa cctctttgtc actgagtact tttgtcagag 12420
gaagggcatg caagcctccc tctatgacct tggggagccc gtctcccata gcatgctcat 12480
cctcaacctc ctgcatggcc ttcacctcca cacttcgatc acttcctcag ttccgttcca 12540
caaggtcaag gtcaacaacg acctcatttc cgaggagctc accaaggggc tgctaccacc 12600
ctctacaact ccaccactgg ggggccagcg catgcaccct cttctactac tatcttggga 12660
ggatcgtcgt tgcgctcgtc tgttcatcaa atctcaagca ttgcacatcc tccccacttg 12720
ccttggtctg aggggagggg tctagggtgg cggtcgacga aaaggcggtc gtggtggtgg 12780
ccatggggac actccttggc cttccatcta caacctgtgg actgatcgca tctccatgtg 12840
gcctggtcct tctccgagag tcccttctca gcgtatcact ccaccgtagc cagctcactt 12900
gacattgctc ggtctggggt atcaccttgg ccatcctacg ccattagcac ctgagtcacc 12960
acttctgtca ctgccacccc acttcgcttt gtcctggaac ccgtggccag gtgggtggga 13020
tcaacagtct cttgctagct ccttcaacac gatgaccttg actcctccca ctgtcaccta 13080
ctaggtggct gattcctggt gcctcctatc acaccacttt ggacgcgagt atgctatatt 13140
cttatccacc tcttcgtcta ctcctaccat tgtgggtaac gagaacattc tctcggtcac 13200
ctctgtcagt gattcggttc ttcctaggcc ctttcacctt cacaatgttc ttgttgcccc 13260
acatatcctt taatatttta tccgttcacc tattgagcat cggtaattct tgttccatag 13320
attttgatcc ttttggcttg tctgtgaagg atcttgctac ctagagtctt cttgcttgtt 13380
ttgatagatc taggcccctg tacacccatg caagcctgcg tctacctccc cacatactgg 13440
tttcgctaca tcatcacatt cgtcctcgct acagtcgcct actttgacta cttccactat 13500
ttgtagtact tggcatcgtc gacttgtcca tcccaacctt gtagcgtagt ccaagctatg 13560
tagtacttta gtaatttctt gtagtagggg cacccttgag catctatgcc agttgtgtaa 13620
gttaggtcgt cattttcgac ttcctttctc tagcttctca caacatacgc tgtgatttgt 13680
ggacctctcc agctctcctg ttaccaatgt tttgggatat aaatattatt tggtgattct 13740
tgatgattgc tctcattgtt tgtggatttt tccattgagt ttaaagtctg acacctttac 13800
gactctgctc ggttacttcg cttgggttta cacttagttt ggttgcacta tgaagatcgt 13860
ccagtgtgac aatggtcgtg agtttgataa cttctctcga tcctttacct cacttggagt 13920
ctaccttcga atgtcttgcc cctacacttc ccatcagagt ggtaaggctg agtgtatgat 13980
ttgcaccacg aataatgtaa tgtgttcctt gatatttcaa tcatctgttt ccgctcgcta 14040
gtgggatggg agcctccaca ctgctaccta cctccttaat tgtcttccta ctacgacaac 14100
tcttatgtca tgccccccac cccactttgc tcgaatggca aggcttagcg catgaattaa 14160
tgtcatgttt ttcttgctat tttactattt taggcatccg ttccttctcc ttactggtct 14220
gagagcctcc acaccaccac ctacctcttt aattgttttc ctattacagc gactcgtgcc 14280
cccacttccc actttgctct ttttgtcaac actccaatga ccatcttcgt gttttgggta 14340
tgtgtgccac cctaaccttt ccaccactac tctccatctt gttcttcgtt ctgctcgctg 14400
tgtctttctc gggtactcat ctgatcacaa agggtactgg tgtcttgacc tcaccactca 14460
tcgccatttg atctctcatc acgtcatatt cgttgggatg gatgtcccct tctcaactaa 14520
cacctagccc accaccgcct cccctttcga gttgtatttt cttcatgatc ctgactccgt 14580
ggtgcccttt gtccaaccac tgtttttcta gttgctgttt tcctctccac tgtttcagcc 14640
ggtggcccct gtgtgcccat ggtggatcat acacccttga attcctttcg ggttgcaccg 14700
ttgcccctcg tcgacctgtg tgtcacctcg tcaacccatg ttgggtccct aggccccggt 14760
ggccccactt ttgacattgc cttcgttgct agggccctct ttattgccct cgcctgtgcc 14820
ctcgccttgt tgctatgccc aacctgtgca tttctatcag cgacatgccc gagtgggtac 14880
accgcctcgc tcttgcgacg agccgacaat gtatcacccc ttcatcgtga ctctcaccac 14940
atccacttga tggcgaattg ccaggccact ggggtcctta gccctgttga ttgcctcatc 15000
ctctccgtga cctcctcatt gtcggtttct catgtgtcgt cttgcgttga ttgcaccctt 15060
tctaatactc attggcgcca cgctatgaag aaagagtatc tgactatcct tgccaatcac 15120
acctgggttc tggtattctg gtgccccatc tttcgtgcga caatgtggtg attgacaagt 15180
gggtctggac acataagtgg caggcggctg gctcgctggg ctactcagtg tcgtgctgtt 15240
gactacgatg agacgttaat tctcgttgtg aagcctgcta ccgaccgtgc catcctcact 15300
ttggccctct cccattttcg gccaattcat cacttggatg tcaaaaatgc ttttctgcat 15360
gggactctca cttagacttg tctactgctg tcagcctact agctgtgttg atcatgcaca 15420
ccccggtatg gtatgcaagc tcaacaaagt atgcaaactc aacaagtccc tctatggttc 15480
gaggcaagcc ccacacgttt ggtacaaccg cttcaccata tatctgctct ctttgttttg 15540
ttgaggccaa gttagacaca tttctgttca tctatcggtg tggtagtgac actgtatacc 15600
tactcctata tgttgatgac atcgtgctca ctacctcctt tctcatgcac tagatcacca 15660
ccgctcttca gcatgagttc accatgaagg atttaggccc ccttcaccac tttatgggga 15720
ttgctgttga gcatcgcttt gatggcctct tttttcagca gcgacgatac accctggaca 15780
tcctcgagcg tgctggcatt ctggattgca agccttgtgt ggcgtcagtg gacatgtagg 15840
ccaagctctc tggcgtcagt gctccagtca gtggccccat cacctaccat agcctcatcg 15900
gtgctcttca ccatctcacc ttcaccagac ctgacatcgt gtatgccatc caacaagtat 15960
gtctttatat gcatgatccc tgtgacctgt acttgggtgt ggtcaagtgg atccttattg 16020
ttgactatgg tctcctcctt tggcgctcct tcatccatga gctaatcatc tatactgacg 16080
ccgatgggcc gagtgtcctg gacacctgtc ggtatacctc ggcgaacctc atctcctggt 16140
cctccaagcg gtagccagtg gtctcccgtt ccagtgctga cgttgagtac tggctgttgc 16200
taatggggtg gctgaggcga gcaagctgca ccaacttctc cagtagcttc acaacccact 16260
caccatgagt accctggtct actacgacaa cgtcagcatc gtctacctct ctgtcaaccc 16320
ttttcaacac caccgtattg accgttttac tattggggac gtccgcgtat gtttcaacga 16380
gctcgtagtt tgatgacacc ttcactaagg gttttccctt gttcttggag tttcggttgt 16440
tggggcggga gacagaaccg gtcctcaact accaattgta ctcaccactc accaagattc 16500
ctaatgcaga gtaaccgact aggaggtgcg aaggcctatg gcggaagggg caagccgcgg 16560
aggccgaccc caacttcggt ccttcctaac aatcgcgaag gctacttgac tacgcaggga 16620
ccttggccaa acctcgacga gggacaaaca cgttcagcga gggcaggcgg gggctaggac 16680
gcgtcgaaga cccccgaagc accgaacgaa gacccaactg ctaccagccg atgttgaccg 16740
aggcgagatc actcctcttt ggtaggccgt cgtgaggcag tttttctcta aggcccccac 16800
gcagaggcca taagacgagg aatgttggat tccgccaagc ggcggctcag ttgtgggcgt 16860
agactcgcct actccacacg tcagcctttg atgtgggtga gaaagatggt gtaatatgga 16920
atgtagccag gggacctgta attaccccgt tgcacacctg ttgcacggta tatttgtggc 16980
atgtagtagg taaccaaggg cattacgata ttttaggcct tgggcccttg gccgccctat 17040
agatagcccc atcctgtagc tggatgggac acacttgaca agacatttgt gctcccagcc 17100
aattgttttg tcgtgccacc tgtaacaccg ctctcgagtg cttgggcaca ctgttctaag 17160
tcccaacatc ggtccagtct caacatcttc cactactaga gtttcatctg cgggggtggg 17220
ggtaggggtg ttagattctt gtgcgttatt gtactcaatt taagtgttat gggccttggc 17280
ccagcccatg taacccatgc aacacagttt accctaattt gggttaggtt tccaacaatt 17340
ttgtttgact tttccattat caaaacacac ttaggccttg ttcgtttacg tcggattgca 17400
cccggaaacg ttccagctaa tcaaagttta tataaattag agaagcaatc cggctaggaa 17460
tcgttccgac ccaccaatcc gacacaaacg aacaagacct taatatttca cgatgaatac 17520
tcaatttaag tgttatgggc cttggcccag cccatgtaac ccatgcaaca cagttcaccc 17580
taatttgggt taggtttcca acaattttgt ttgacttttc cattatcaaa acacacttaa 17640
taagttaata tttcacgatg aatgtagttg ttttggctta ttctgaacaa tttcacatca 17700
tgcagccgga aacactgcct gaacatataa aattcgacca tggttatacc tctaagagtc 17760
ctgcaattgt taatgtaagt ttccctgctt ctttgtaatt catttgtagg tttcgtaagt 17820
ttatttggat gtttctaatg ttgtgttttg cagtttcttg agatcatggc agaatttact 17880
cctgagcagc aacatgcttt ctgccagttt gtgactggtg ctcctcggct tccacctggg 17940
ggtttggctg ctctaaatcc taagttgacc attgttagga aggtaaattg gctatctttg 18000
ttcttatttg tactttatgg ttgacatgcc tgcccacatt gttgaagttt tgaattcttc 18060
ttaaattcca gcactcttct gtggcaaata atagttcaaa tgcaactgga gcgacagagt 18120
ctgcagatga tgaactatct taaactacca ccatactcca ccaaggtatg cttctttctg 18180
ctttttggct aactgtggtt atatctcctg tattgtctta taaattgagg attcagaaac 18240
ccagtcacca aagaattact tcatatagcc ttatcgtaac aggtaactgg acaaattttc 18300
aactaaggac gtggaaacta aaatttaatg tgggcagcac cttccagcca ctcattagtt 18360
agaattatta tttcgttagt ttaaatcaaa tagcatattc cacattgctt gaacccttta 18420
tattgacgca tgctggtttt ttttctggaa ggaaatattt acagtgctac tcgtgctagt 18480
gtgtggcttg ttatctgtgt ctgaattttg ttgaaaactt cttgcaggct atcatgctaa 18540
agaaactgct ttatgcgatc aacgaaggcc aagggtcatt tgatctttcg tgaatctcaa 18600
cactaacata ggtattggtc cacctagaaa tctgcgtcat tgttacccag agttagtttc 18660
tacctcattc atgtatgaca taggttaaac tcagctctcc ggagtcccac cgaaggtttg 18720
gagcccgtac ctttgggtgt ggatgtctat actctctttt cttggttgta tattcttgcg 18780
gatctttata gtgaataata gtaataaatt gttttgcgct tcttactgtg ctaatcatca 18840
gtgtcccacc cgaagcgtca gccgtcagtc gtacaaattt tgctcgatgg tttcgctgcc 18900
cacaaatcgg atggatgggg gcccatgaac aagggcacgg gatttccggg gctatctgaa 18960
atagtgacgg gcatgcaaac acacccaagg tccacggcct gcttttggtc gacacagtgc 19020
cacgcgaccg tgctggatct tatcactggc ctgtcagagg catctgagga tgtcaagttg 19080
tcaaccgagg cgcctatgtg ggcacgggac tgatcacttt cactcgggtc acagcgttcg 19140
gtcttggcgc ttcagccaca accccaagga tgacgatagc tgggcacaag gccacgacag 19200
ttgcacagat cccctcgtct cgtttgcagg taccagcgaa attgctaacg tgcgatgcga 19260
cccgctgcga aaactacgga tcacgtatca gcggtcgttg tcatatatga tcagtcggcc 19320
gtgccttggc actgcacaag ccaataaaac tccgccagaa ctgaggaaag atggaaccgt 19380
ccaggagttg ctaagccatc gattgcacgg atagtatggc aagcggtaca tccagtaggt 19440
ggtttgacct aacattagtg tctgatcccg ttagaacatc tctaactccg tcttaaatta 19500
atgcactatc ataaaatata ggacttataa aaaacagttc caccagtgcc ttaatctaaa 19560
atttttcatc aaaaaatata ggccaaacta tgaaatgatc taaatatact acacccccgg 19620
ctagtgtcct atctttatgt tgtacattat tttcaatatt ttcttttcta ataatgtaat 19680
ttactttcca aattatatga tttaaggctt aactgttgga gtataattta tttttagatc 19740
cctaaacatt ttaaacgatg tagtatttta atttttaggg cactattttg agccaacttg 19800
ttggagatgc tcttaggtta cctacggatt ccggataaaa aaatcttgaa atttctgcca 19860
taagattcta gaggaatcta acacacaggc acaaaattca atgaatacta gagattattt 19920
ttatccagaa tccagttgat gttggctatt attttattct tctattttgg atccaaagaa 19980
aagaagaaga gtaaatccat agaaattcct aactaaaaat gtgatttcta aagatttcat 20040
tgtaattctt cgaattatcg aacaaatcca atccaacata atacaatttt tttgaaatta 20100
gtaaattaag taataaagca tagagaaata attaaagaat acaatcgtta tccatctttt 20160
ctttcttttt tgcttcatat actaaaaaga attcaaaaag gcaaaatttt cgtcatgtca 20220
tgaagaattc tatctcttgt atacgaataa ctaggattta aagaaggaaa tgacaaagca 20280
tatgtgaata aacgattggt ttctatcata aacctgctaa agcctcaaac cactgagtac 20340
ttagcacggg tgctacaaag agatatgttt ttacatatat aatcagatac tcttgtccaa 20400
gatcatttgt atttcttttg tttaggctaa tttcctaact aaaaaagaaa aataaatatc 20460
gtatatatag aatgaaccat atagatgaga tttttctatc aagtgagatg tgaaaaaatg 20520
ttgttggctg tgagctgtaa aaagctaaaa atagtttagt ggaagctgct agaatccact 20580
aaaatttctt ctattatatt tttacacggt tccatctgaa agttgctaaa aacaagttca 20640
tagtgcttcc aattttgtaa accgaggcaa tcgttttttt aaaaagctga ttcttgaatt 20700
gagtccttcg gtttggcttt tggctcttga gaggcaaaag ccaaagccaa aagccaaacc 20760
aaacatacca tttgttattc tacacttaaa acaaactaat atataactcg ttaatatcag 20820
ttccaa 20826
<210> 125
<211> 10181
<212> DNA
<213> 人工序列
<220>
<223> 合成多核苷酸
<400> 125
aggcctggag ttgccgacca cgagagatcc tgaccagcag ttgctgggct gtgtgtgtgt 60
ctgcgtctgt cactctcctc tctcgcgtct tggtcctcct cctccccttc atcgaacgaa 120
tccatccaag gcaaaaggga acgagggaag caagcaaaaa atccccaatt cctcctagat 180
cgatccaccg ccgacgggac gaatccaatc caggagctat cggctggaat tggattcacg 240
cgcacccgcc cgctgccgct ggccgccgcc tagtctcggc cggccggccg accggccgcc 300
ctccccgcgc gcgcgccttg ccttgtatgg aaacacgcag cagcagtcgc aagcgcgcag 360
cggccgccgc ctcctcgtcc gcctccaagc gcccccgccg caactcctct tcctcctcct 420
ctgcgccgcc gcgccagcag ccaccgatgg aaccctcgcc ctcctcccgc cgccgctccc 480
gcgccgccgc cgccgacaag ggcaaggacc ccgaccccgc ctcctccgac cacccttccc 540
cacccgcccc tgacgatgac gccgacgtca ccttccccca gcccttcacc tccgccagca 600
ccgccctcca gggcctcctc cgcaggctcg gcgccggcct cgacgacctc ctcccatcct 660
cctcctcagc gccctcctcc gccacgtccg gtcacctcaa gaggatcctc gccggcctgc 720
aggcgcacgg cgacgagtcg cgccagctcc agtccctcat gcagctctgc gagatgctct 780
ccatcggcac cgaggactcg ctcgccgcct tcccggtcga tgcttttgtc ccaatcctcg 840
tcgggatgct cggccgcgag gacgagcctg ccaccgctgg tgccagcccc gacgtcatgc 900
tgctcgcggc gcgggcactc tccaacctcg ttgatgtgct cccctcctcc tgctccgcgg 960
tcgtgcacta cggggccata cagtgcttct gtgctcgact gctcaccatc gagtacatgg 1020
acctcgctga gcaggtggtg ccttcccttt gtcttcttct tcctcgtttt atcctgtatc 1080
atcgctcatc ctcactatca tctaaaaatg aaatatgatc agtcatcctc tgcatgcatg 1140
cccttagccc cttaccaagg tgctgctaac acatttactt tactcgcctg ctcagtctct 1200
acaagccctt aagaagatct ctctggagca cccaactgcc tgtttgcgag ctggtgccct 1260
aatggctgtt ctatcgtatc tcgacttctt ctccactggg gttcaggtcg ggtcctctga 1320
tctccttaca accgttctta tcttgtgcac tgtttgcatc tacttacacc tcttcccatg 1380
ggggacgtac ttgttgtaga gggtagcctt gtctaccgct gcaaatattt gcaggaagct 1440
gccgtccgat gcttccgagt ttgtcatgga agctgtcccc ttgctcacga atcttctaaa 1500
tcaccatgat tccaaggtac tttgcaagct cagacaacca tacctctgga ttatgatata 1560
ttgaggtata tcatgtattt cccaaccctt catctatgtg ccttacaggt actagagcat 1620
gcttctgttt gtttgacacg cattgctgaa gcttttgcct attaccccga caaacttgat 1680
gaactctgca atcatggatt agttgctcaa gctgctagct tgatagctgt tggcaactca 1740
tctggccaga catcactgag cacatccaca tatacagtat gggccatttt attttgatac 1800
ccttttattg tttgctgtgg cccctctttc tcacacttat gctaaaatta ggttgcacgc 1860
ttttcagggc ttaatccgtc ttctctcaat atgtgcaagt ggatccctgc tggcagttaa 1920
aacactactt cttcttggga tcagcggcac tcttaaagat atcctttctg gctctgcttt 1980
gatttctgga acatcagtct cccctgctgt aacaaggcct gcagatcagg tgaatatttc 2040
tatcactgac ttaggacttg actattctgc tagtgctggc cttttaaaaa aaaatcaatt 2100
tagccacaat ttctctctgt gattggattc aataaaattc cgatttatca gtgaaaaatt 2160
aggctttggc atctcacact ttctgaaatt acatctacca cacgtatcat gcagactttt 2220
agaactctaa aggcgacatc acatgttcat aattatatgt accattgagc aattgtttgg 2280
gtgatttttt tgagcattga acatagatgc taaaatgtcg ggaattgtgc tcgtcttctt 2340
aaaccatgca aataatcatt caaaagagca gtgaactagt attttcaaag gcctgcatac 2400
tttcgttgtg cttttatatt gcctgagtaa gtcttttgaa atgttttaat gaagcagaaa 2460
atgttgattt atggcacaca ggggaacaaa agtctcactt gtaatatgat agagcaaaat 2520
ctcatttgga ttaagtagct gctgtgattc ttgtcgtctt attttcttcc tagtgtggtg 2580
caagtaaatt gtgcttttgg tcaatcgcat ttggtatggt acagatcgaa gagttttttt 2640
tccatgctat acaattgagt atgaatatat aatgccatct cagcttctgc ttgttgctgt 2700
cttgcttaat tgctaaaatc atttgtggta catacacaca cgagtctggc attttttgag 2760
tatgatcgtc tgcaacagct gctgtatctt ctcacctagc tgctgaaaca ttaattcttg 2820
aaacccacac tgtttgactc atcagtttca tgcacctgta tgttttttgt tgtacaaatg 2880
tttttcatat gtttacccca aatattgcag atgtttgaga ttgtgagcct tgccgatgat 2940
ttgcttccgc atatccctgc gagaatcatt aagttaccaa catattacca tacatataaa 3000
ggctcttcaa caaagaaatc tgcttccatc aaacaagagg ggggtggttc aacaggaaat 3060
gaaaggtctg gccgtgagag gctattgcgt gagcaacctg agcttttaca acagtttggt 3120
atggatttgt tacccacaat gacacaggtt agtgctggaa ttcttttcct gagtcctgac 3180
agccatttca attgctattt gtgttttgtc tttgtgtact ttaatagcct gtgtgcttgt 3240
tcaggtgtat ggttctagtg taaatgcacc aatacgtcac aagtgtttat ctatcattgg 3300
aaaattaatg tactacagct ccgccgaaac aatccagtct ctccttggca caacaaacat 3360
atccaggttt tctctctctc tcacctggaa cacacaaaag agatgcatgt ttcattaata 3420
gaaccgaaaa aattctcagt tcatttctag agaatgattt cagaacaatg gatgttaaaa 3480
gacaaaaaga aagtctctca agtctcaatg tacatacttc ttttccaatc ccttctaagt 3540
tatttcattt ctatctgcag cttccttgcg ggcattcttg catggaaaga tcctcaagtg 3600
ttgattcctg ctcttcagat agcagaaata atgatggaaa agcttccaga gacattttcc 3660
aagttatttg tgagggaagg tgtcgttcat gctgtggagt cactcatatg ttcagaatct 3720
tccaacaaaa tgccttctca ggtgccacct caggataaag ataaagattc tgccatgcca 3780
tcacgttcta gacgtcaacg ccggcgtgcg ggtgctgttg cagcagaaaa tagttcattg 3840
gatgaatcaa atagttccaa ccttggtgtt atgtgctcaa cagcaaccgc ttcagaagct 3900
ccaaacacca gccttcgttt cacagttagt gaccatgcaa agtctttcaa agataaatac 3960
tttccggcgg atactgattc aagtgatatc gggtttactg atgaccttct taaactgagg 4020
gcactctgtg caaagttgaa tactgtctct gagaacgtca aaacaaaagc aaaagggaaa 4080
tcaaaagcta taagcactaa ttttttggac atctcaattg atgttgaaga acaattagac 4140
aaaataatat ctgaaatgct ctctgagctt agcaaagtta atggtgtttc cacatttgaa 4200
ttcattagaa gtggtgttgt catagcattg cttgactatt tgtcctgtgg gacatttggg 4260
aaggaaaagg tatctgaagg aaacctatct cagcttcgtc agcaggccct tagacgatac 4320
aaaaccttca tatctgttgc cctttctatt gatcatggac gagatgaaac tcccatggca 4380
ctgttggttc agaaactgca aagtgcattg tcttcgctgg aacgtttccc tgttgtgctc 4440
agccagtcta gtaggattgg tattggaggg tctcgtctga cttcaggtct tagtgctctg 4500
gctcaaccct tcaagcttcg tctttcccga gctcaaggcg aaaaatcact tcgggattat 4560
tcatcaaata ttgtgcttat cgacccattt gctagtcttg catctgttga agaattcctt 4620
tggcctagag ttcaacgcag tgaggttgca tcgaagccta taattccatc tggtaataat 4680
tctgaatctg gggttcctgg caccacggct ggtgcatcat taacagctgc aatggctcaa 4740
tctggtcggc gcccaacaac aagatcaaag tcatctgctg caggtggtct tacatctaag 4800
aaggattctc atgatgaaag cactagtact gccaaaggaa aaggaaaagc tattgtaaag 4860
ccaaactcag atgaatcaaa aggacctaat acaaggaatg ctgcccacca aaaatctgct 4920
tcagagaaag atttggaaat gaagcaggca catggccaca gtagctctga ggtatttttt 4980
ttgtttacat gcctcattcc tggttagttt ggcttgcaaa gaaaagttac tgtttctatt 5040
caagttgggt agcagctctt ctcaaaagtc gcattgtcag ggcctaagga ggcccacgga 5100
ggggctgcgg cagcagcagc catgggccct ccaaggggga ttagataatg atagttagag 5160
ataagattag aagattagtt gagattattt aggagattag ttgagattat ctaggaaggt 5220
tatcagttag tttgttgaga gtttttagga gagttttaag gagataaagg tctcaagcta 5280
gacaagggcg gccacccggt ctatttatgt aatcaatgga taagaaataa attaggcaag 5340
aattagaagg gagaaaccct cctcttgctc ggccgtgggc agaggccctc ggccgacgtt 5400
cctgcccgtc gatacccctc tctaatccct caccgattta gcgtccataa cacacatcca 5460
cttctctggt gtcatatttg gtgccaccta taaagctcat ggcttgggcg ccaaatgaac 5520
atgataattt tagccgcgga tctagttctt aatttatatt accctcataa tatgtaaaca 5580
tgattttgtg agttcaaaac tgtgccaatc taaaaaatat taggataatg atcaagaagt 5640
aggctcattt tctagttgag agtgtaagca tggtaactga taagtaaata cattctatgt 5700
tggtatatgt aggatgaaga gctcgacaca tctcctgttg agatcgatga tgctttgatg 5760
attgatgatg atgacatttc agatgatgat gatgatgacg acgatgatca tgaggtaact 5820
gtttgaaaat tgaatgccca ctcttttttt tgcttcgttg caaccatccg tagttgctta 5880
ggttcatatt caaggtacta aatagcatgg gttcatagca gtttgatgtt gcggatgtca 5940
ggtgctatta cggaccgcta agttccaata gcagtatata ataatcaaac ataaatttat 6000
gtatatatat atcattaata ttaatttaat agacgtttaa ttaattatac ttagataagt 6060
gatgtctcac atcattatta tcaacaaagc cttttagccc caagcaagtt ggggtaggct 6120
agagttgaaa cccatcggaa gccacaaatt aaggttgagg catgtggaaa tcgatgtctc 6180
actctcacat actttaaatt aaataaagga acatgatatt caacatatta catataagtc 6240
atcacacatt ggactggagt ctacaaagaa atatattttg atagccgtaa aatagtcata 6300
aaatcatgtt tcattgtcat gtcagaagta gtaacctcaa tagtttcatg atagttaaaa 6360
tgggataggg ttatttgttg tatatcatac aacactattg tggacactat gaagaagtgc 6420
ggagcactat aacagcctgc tacctcaaat agcagccgct aagttccaac ctcaaatagt 6480
ggtagcggta agagaccgct accgtttttt gcggacgcca tagcggttgc tactaacgct 6540
atttagtacc atgcttatat ttatttactt tatgacaaaa ataacctttt agttggtcca 6600
agatgatcat ttgtgccctt acttgactaa tttaataacc tctccaaatt cttaggttct 6660
ccaagaaggg tctcttccaa tctgttctca agatggtgtg catgacgtga aactgggtga 6720
tgccgatgaa tctaacattg gatcagctac ttttagccat gcacagccat catctggttc 6780
tggggctaga aacacttcta gcaggggacc ggattctact gaatttcgaa gtatgggcac 6840
attcggttca cgaggtgcaa tgtcatttgt tgctgcgacg atggctggtc tggcttctgt 6900
tggtggtcgc agtgttagag gtgtccgaga tcggcgtggt ctgtcacttg gaggtagcat 6960
gaatgatcac aataaactta tattcactgc tggtggcaag cagcttaaca aacatttgac 7020
tgtatatcaa gctatccaac gtcaacttat gctcgacgag gatgatgaag aaaggtttaa 7080
tggatctgat ataccaaatg atggaaaccg cttctggggt gatgtgttca caataacgta 7140
ccagaaggct gataaccaag ttgagaaggg ttgccaaggg acttccacct cattaaacat 7200
aaaatcagat tcttacaggt ccatttctga agcacaaggg gtttctcttc ttgatagcat 7260
cttacaagga gaacttcctt gtgatcttga gaggacaaac tcaacgtaca acattttagc 7320
attattgcgt atacttgagg ggcttaatca gttatcccct cgtttaagag tacagggagc 7380
atctgatgat tttgctgagg gaaaaatcaa taccctggat gagttatata gaactggagc 7440
caaggtacct tctgaagtgt ttgtcaatag caagttaaca ccaaagcttg ctcggcaaat 7500
gcaggacgtt ctcgcactat gtagtggcag tttaccttct tggtgttatc aaatgaccaa 7560
ggcctgccca ttcctgtttc cttttgagac aaggaggcag tacttctatt ccacagcctt 7620
tgggttgtca cgagcattga atcgactcca gcaacagcag agtgataatc acagctctgg 7680
tggtgaaaga gaggtcaggt ttggcaggtt acaacgccag aaagttcgtg tttcccgtaa 7740
ccgtattcta gattctgctg ctaaagttat ggagatgttc tccagtcaga gggctgttct 7800
tgaggtggaa tactttggcg aagttggaac tgggctgggg cctactctag agttttatac 7860
tctcttaagc catgaactac aatgtgctca actgggatta tggagagcta cttcttcata 7920
tgattcagga ttgcacactg atatgaagga tgtgatccgt ttagatcctg aagatgggtc 7980
gtcaggaaaa gaactcaaca cagatctacc aggtgatggc atgtatttga tacaagctcc 8040
tcttggattg ttccctcgac cttggccacc taaagttgat acttcagaag gtagcagatt 8100
cttcaaagtt ttggagtatt tccgcttaat aggtcaagtg atcgcaaaag tcctgcaaga 8160
tggcagactt ttagatttgc ctttatcaac agcgttttat aagcttatac ttggacaggt 8220
atgtacagag aagctctatt tccaagtgct ttgctttcat tttgttttgc tttgttttat 8280
gtattaatta gttgtttgta ttcacttttc acgacttaat gtgttgcaat tggccaggaa 8340
cttgatttgt ttgacatagt gtcatttgat tctgagtttg ggaagacatt gcaagaactg 8400
caagttcttg ttgaacgaaa gcagttcctt gaatccactt ctggcaagaa ccagctacaa 8460
gttgcagatt tgtgtttcca tggcgcttct attgaagatt tgtgtttaga ttttactctt 8520
ccaggttatc ctgattatgt gcttaaagag ggtgaaggag gcacaattgt aagtgatgaa 8580
gttccatatt agcgtgtatt tatcttggtg atttcacttt gtatgtttta atctgttagt 8640
atcatctgca ggttaatatt tataatttag aagagtatat tactttggtg gtggatgcta 8700
cagttaagtc agggataaag cgacaggttg aggcatttag atcagggttt aatcaggtac 8760
agcctcattg gcattgctct aacacaatct tataataatt ctacttgttc tgattttgtg 8820
tagttcagtt tactctaata gttcttgttt attttaatgt ttcgtaggtg tttgacatgt 8880
catccctcca catattttca cctcaagaac ttgactatct aatatgtggt cgacaagaaa 8940
tttgggaggt aatgttctgt cctttttatc caagttaatg tatgtaaaag tgtggcacta 9000
aataaacctt ggaatctgca gtcggaatcc ctggtggata acataaaatt tgatcatgga 9060
tatactgcta aaagtcctgc gatcgtgaat gtaagctcat tcgtcctcac atgctacatc 9120
tcttacatac atccagttgt ggtttttgag aagtgtggtt ccttgattct tgctcttgtt 9180
tgtagctact cgagattttg tcagaattca cccctgagca gcagcatgcg ttctgccagt 9240
ttgtaaccgg tgctcctcgg cttccacatg gtggattggc tgcccttagc cccaagctta 9300
ctatagttag gaaggtacct tttcatggtc tatctatcgt ggcatttgct ataatgacat 9360
gtgcacacct attctgcttg atgttatttt tggggtaact cggtgttttg attattgcca 9420
gcatccttca agtgctgtga atacttcaaa ttcaactggg gcagtagagt ccgctgatga 9480
tgatttgcct agtgtgatga ccaattatct gaagctgccc ccctactcca caaaagtaca 9540
gtgtccttgc tgttttatca ctaaaaagtc agcttgacat tctgatggtt cctgctgacg 9600
ggctgttgct attgtttgga caggaaatca tgcgcaagaa gttgctttat gcgatcctgg 9660
aaggccgagg atcatttgat ctttcataag aaatatacaa tatccgaact aacatacagg 9720
ggtcagtttt attctggaga tttagtttct tggtccatgt ataacactga gttgaggtta 9780
ccagctcact cagcaaacgg cttcaagggc gagagcccca gtcctcactc ggatgcaggt 9840
gctgctgtcg atttctccct tcttttacat ttgttcagct actgtattct cttcgagggt 9900
tgcccttgtc acctgcaatt atgaatctac agcttttgtc atcgaggaag tttaacagat 9960
tttggtttct ctaccatgat ctgtttccat tgggcaggct aagctatagt aattgaggat 10020
gtgcgccttg agtgaggatg tgcgccaaca agaacccccg tttgtttcag tacatgtaaa 10080
taaaagggat tttttgttcc agaattgtta tctgtaacaa gaacatctac atctgtgatg 10140
actgaccatt aataaatgtg tcccggtgta ccatttgttt t 10181
<210> 126
<211> 10
<212> DNA
<213> 大豆
<400> 126
cgctcccgtt 10
<210> 127
<211> 11
<212> DNA
<213> 大豆
<400> 127
catccaatac t 11
<210> 128
<211> 26
<212> DNA
<213> 大豆
<400> 128
tttgcctagt gtcatgactt gcgcca 26

Claims (95)

1.一种植物或其植物部分,其包含编码HECT E3 UPL多肽的内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的至少一个非天然突变。
2.权利要求1的植物或其部分,其中所述内源HECT E3 UPL基因是编码HECT E3 UPL3多肽的内源HECT E3 UPL3基因。
3.权利要求2的植物或其部分,其中所述内源HECT E3 UPL3基因是内源HECT E3 UPL3a基因、内源HECT E3 UPL3b基因、内源HECT E3 UPL3b基因或内源HECT E3 UPL3d基因。
4.权利要求2或权利要求3的植物或其部分,其中所述植物或植物部分包含二至四个拷贝的内源HECT E3 UPL基因(例如,HECT E3UPL3a基因、HECT E3 UPL3b基因、HECT E3 UPL3b基因和/或HECT E3 UPL3d基因),并且所述二至四个拷贝的内源HECT E3 UPL基因包含一个或多个非天然突变。
5.权利要求1-4中任一项的植物或其部分,其中所述至少一个非天然突变是无效突变或显性失活突变。
6.权利要求1-5中任一项的植物或其部分,其中所述至少一个非天然突变是碱基取代、碱基缺失、碱基插入和/或倒位。
7.权利要求1-6中任一项的植物或其部分,其中所述至少一个非天然突变包括对A、T、G或C的碱基取代。
8.权利要求1-7中任一项的植物或其部分,其中所述至少一个非天然突变包括G至C的碱基取代。
9.权利要求1-6中任一项的植物或其部分,其中所述至少一个非天然突变是至少一个碱基对的缺失,任选地约1个碱基对至约100个连续碱基对的缺失。
10.权利要求1-6中任一项的植物或其部分,其中所述至少一个非天然突变是至少一个碱基对的插入。
11.权利要求9或权利要求10的植物或其部分,其中所述插入或缺失是框外插入或缺失或框内插入或缺失,任选地其中所述插入或缺失是产生提前终止密码子的框外插入或缺失,任选地其中所述缺失导致与SEQ ID NO:119-125的核苷酸序列中的任一个具有至少90%的序列同一性的突变的HECT E3 UPL基因。
12.权利要求2-11中任一项的植物或其部分,其中所述至少一个非天然突变位于由内源HECT E3 UPL3基因编码的HECT E3 UPL3多肽的N端区域中。
13.权利要求12的植物或其部分,其中HECT E3 UPL3多肽的N端区域包含:(a)与SEQ IDNO:84-87中的任一个具有至少80%序列同一性的序列,和/或(b)与以下的连续氨基酸残基区域具有至少80%序列同一性的氨基酸序列:参考SEQ ID NO:78的残基编号,从残基50到残基95、从残基43到残基102和/或从残基36到残基109,参考SEQ ID NO:79的残基编号,从残基48到残基87、从残基41到残基94和/或从残基34到残基101,参考SEQ ID NO:80的残基编号,从残基139到残基194、从残基132到残基201和/或从残基125到残基208,和/或参考SEQ ID NO:81的残基编号,从残基104到残基146、从残基97到残基153和/或从残基90到残基160。
14.权利要求2-11中任一项的植物或其部分,其中所述至少一个非天然突变位于由内源HECT E3 UPL3基因编码的HECT E3 UPL3多肽的活性位点中。
15.权利要求14的植物或其部分,其中所述活性位点位于HECT E3 UPL3多肽的区域中,所述区域包含:(a)与SEQ ID NO:88-93中的任一个具有至少80%同一性的氨基酸序列,和/或(b)与以下的连续氨基酸残基区域具有至少80%序列同一性的氨基酸序列:参考SEQ IDNO:78的残基编号,从残基1814到残基1865、从残基1822到残基1857和/或从残基1829到残基1851,参考SEQ ID NO:79的残基编号,从残基1820到残基1870、从残基1827到残基1863和/或从残基1834到残基1856,参考SEQ ID NO:80的残基编号,从残基1838到残基1888、从残基1845到残基1881和/或从残基1852到残基1874,参考SEQ ID NO:81的残基编号,从残基1834到残基1884、从残基1841到残基1877和/或从残基1848到残基1870,参考SEQ ID NO:82的残基编号,从残基1783到残基1833、从残基1790到残基1826和/或从残基1797到残基1818,或参考SEQ ID NO:83的残基编号、从残基1776到残基1826、从残基1783到残基1819和/或从残基1790到残基1812。
16.权利要求14或权利要求15的植物或其部分,其中所述HECT E3 UPL3基因的活性位点中的至少一个非天然突变导致取代的氨基酸残基。
17.权利要求2-8或14-16中任一项的植物或其部分,其中所述至少一个非天然突变是位于以下位置处的氨基酸残基的取代:参考SEQ ID NO:78的氨基酸位置编号的位置1839,参考SEQ ID NO:79的氨基酸位置编号的位置1844,参考SEQ ID NO:80的氨基酸位置编号的位置1862,参考SEQ ID NO:81的氨基酸位置编号的位置1858,参考SEQ ID NO:82的氨基酸位置编号的位置1807,或者参考SEQ ID NO:83的氨基酸位置编号的位置1800。
18.权利要求6-8或14-17中任一项的植物或其部分,其中所述取代是半胱氨酸至丝氨酸。
19.前述权利要求中任一项的植物或其部分,其中所述内源HECT E3 UPL基因编码多肽,所述多肽包含与SEQ ID NO:78-83的氨基酸序列中的任一个具有至少80%序列同一性的序列,包含与SEQ ID NO:84-92的氨基酸序列中的任一个具有至少80%序列同一性的区域,和/或包含与SEQ ID NO:93具有至少90%同一性的区域。
20.前述权利要求中任一项的植物或其部分,其中所述HECT E3UPL基因包含与SEQ IDNO:72-77的核苷酸序列中的任一个具有至少80%序列同一性的序列。
21.前述权利要求中任一项的植物或其植物部分,其中所述植物是玉米、大豆、油菜、小麦、水稻、棉花、甘蔗、甜菜、大麦、燕麦、苜蓿、向日葵、红花、油棕、芝麻、椰子、烟草、土豆、甘薯、木薯、咖啡、苹果、李子、杏、桃子、樱桃、梨、无花果、香蕉、柑橘、可可、鳄梨、橄榄、杏仁、核桃、草莓、西瓜、胡椒、葡萄、番茄、黄瓜或芸苔属植物物种。
22.前述权利要求中任一项的植物或其部分,其中所述植物是大豆、油菜或玉米。
23.前述权利要求中任一项的植物或其部分,其中包含至少一个非天然突变的植物与缺乏所述至少一个突变的植物(例如,同基因植物(例如,野生型未编辑植物或无效分离子)相比具有一种或多种改善的产量性状的表型。
24.权利要求23的植物或其部分,其中所述一种或多种改善的产量性状包括与缺乏所述至少一个非天然突变的对照植物相比更高的产量(蒲式耳/英亩)、增加的籽粒行数,任选地其中穗长度基本上没有减少、增加的籽粒数量、增加的籽粒大小、增加的穗长度、减少的分蘖数、减少的雄穗分枝数、每个节增加的豆荚数、每株植物增加的豆荚数和/或增加的种子重量,任选地,其中所述一种或多种改善的产量性状是增加的种子大小和/或增加的种子油含量。
25.一种植物细胞,其包含编辑系统,所述编辑系统包含:
(a)CRISPR-Cas效应蛋白;和
(b)引导核酸,其包含与编码HECT E3 UPL多肽的内源靶基因互补的间隔序列。
26.权利要求25的植物细胞,其中所述内源靶基因:(a)包含与SEQ ID NO:72-77的核苷酸序列中的任一个具有至少80%序列同一性的序列,(b)编码与SEQ ID NO:78-83中任一个的氨基酸序列具有至少80%序列同一性的序列,(c)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的区域,和/或(d)编码与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域。
27.权利要求25或权利要求26的植物细胞,其中所述引导核酸包含SEQ ID NO:94-118的核苷酸序列中任一个的序列。
28.权利要求25-27中任一项的植物细胞,其中所述植物细胞是玉米细胞、油菜细胞或大豆细胞。
29.由权利要求1-24中任一项的植物部分或权利要求25-28中任一项的植物细胞再生的植物,任选地其中所述植物包含突变的HECT E3UPL3基因,所述突变的HECT E3 UPL3基因包含与SEQ ID NO:119-125中的任一个具有至少90%序列同一性的核苷酸序列。
30.权利要求29的植物,其中所述植物包含一种或多种改善的产量性状的表型,任选地所述一种或多种改善的产量性状包括与缺乏所述至少一个非天然突变的对照植物相比更高的产量(蒲式耳/英亩)、增加的籽粒行数,任选地其中穗长度基本上没有减少、增加的籽粒数量、增加的籽粒大小、增加的穗长度、减少的分蘖数、减少的雄穗分枝数、每个节增加的豆荚数、每株植物增加的豆荚数和/或增加的种子重量,任选地,其中所述一种或多种改善的产量性状是增加的种子大小和/或增加的种子油含量。
31.一种植物细胞,其包含内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因内的至少一个非天然突变,其中所述突变是使用编辑系统引入的取代、插入、缺失或倒位,所述编辑系统包含与内源HECT E3 UPL基因中的靶位点结合的核酸结合结构域。
32.权利要求31的植物细胞,其中所述内源HECT E3 UPL基因是内源HECT E3 UPL3基因。
33.权利要求32的植物细胞,其中所述内源HECT E3 UPL3基因是内源HECT E3 UPL3a基因、内源HECT E3 UPL3b基因、内源HECT E3 UPL3b基因或内源HECT E3 UPL3d基因。
34.权利要求31-33中任一项的植物细胞,其中所述至少一个非天然突变是显性失活等位基因或无效等位基因。
35.权利要求31-34中任一项的植物细胞,其中所述靶位点在HECT E3 UPL基因的区域内,所述区域
(a)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的序列,
(b)编码与以下的连续氨基酸残基的区域具有至少80%序列同一性的序列:参考SEQID NO:78的残基编号,从残基50到残基95、从残基43到残基102、从残基36到残基109、从残基1814到残基1865、从残基1822到残基1857和/或从残基1829到残基1851,参考SEQ ID NO:79的残基编号,从残基48到残基87、从残基41到残基94、从残基34到残基101、从残基1820到残基1870、从残基1827到残基1863和/或从残基1834到残基1856,参考SEQ ID NO:80的残基编号,从残基139到残基194、从残基132到残基201、从残基125到残基208、从残基1838到残基1888、从残基1845到残基1881和/或从残基1852到残基1874,参考SEQ ID NO:81的残基编号,从残基104到残基146、从残基97到残基153、从残基90到残基160、从残基1834到残基1884、从残基1841到残基1877和/或从残基1848到残基1870,参考SEQ ID NO:82的残基编号,从残基1783到残基1833、从残基1790到残基1826和/或从残基1797到残基1818,和/或参考SEQ ID NO:83的残基编号,从残基1776到残基1826、从残基1783到残基1819和/或从残基1790到残基1812;和/或
(c)编码与SEQ ID NO:93的氨基酸序列具有至少90%序列同一性的序列。
36.权利要求31-35中任一项的植物细胞,其中所述编辑系统还包含核酸酶,所述核酸结合结构域结合与编码SEQ ID NO:84-92中任一个的序列具有至少80%序列同一性或与编码SEQ ID NO:93的序列具有至少90%序列同一性的序列内的靶位点,并且所述HECT E3UPL基因内的至少一个非天然突变是在核酸酶切割后产生的。
37.权利要求36的植物细胞,其中所述核酸酶是锌指核酸酶、转录激活因子样效应核酸酶(TALEN)、核酸内切酶(例如Fok1)或CRISPR-Cas效应蛋白。
38.权利要求31-37中任一项的植物细胞,其中所述核酸结合结构域是锌指、转录激活因子样DNA结合结构域(TAL)、argonaute或CRISPR-Cas效应子核酸结合结构域。
39.权利要求31-38中任一项的植物细胞,其中所述HECT E3 UPL基因内的至少一个非天然突变是插入和/或缺失,任选地所述至少一个非天然突变是框外插入或缺失或者框内插入或缺失,任选地其中所述框外插入或缺失导致提前终止密码子。
40.权利要求39的植物细胞,其中所述HECT E3 UPL基因内的至少一个非天然突变是缺失,任选地其中包含所述缺失的突变的HECT E3 UPL基因包含与SEQ ID NO:119-125中的任一个具有至少90%序列同一性的核苷酸序列。
41.权利要求31-40中任一项的植物细胞,其中所述HECT E3 UPL基因内的至少一个非天然突变包括点突变。
42.权利要求31-41中任一项的植物细胞,所述至少一个非天然突变导致位于以下位置处的修饰的氨基酸残基:参考SEQ ID NO:78的氨基酸位置编号的位置1839,参考SEQ IDNO:79的氨基酸位置编号的位置1844,参考SEQ ID NO:80的氨基酸位置编号的位置1862,参考SEQ ID NO:81的氨基酸位置编号的位置1858,参考SEQ ID NO:82的氨基酸位置编号的位置1807,或者参考SEQ ID NO:83的氨基酸位置编号的位置1800。
43.一种由权利要求31-42中任一项的植物细胞再生的植物,其包含HECT E3 UPL基因内的至少一个非天然突变,任选地其中突变的HECT E3 UPL基因包含与SEQ ID NO:119-125中的任一个具有至少90%序列同一性的核苷酸序列。
44.权利要求43的植物,其中与缺乏所述至少一个突变的植物(例如,同基因植物(例如,野生型未编辑植物或无效分离子)相比,所述植物表现出一种或多种改善的产量性状的表型。
45.权利要求44的植物,其中所述一种或多种改善的产量性状包括与缺乏所述至少一个非天然突变的对照植物相比更高的产量(蒲式耳/英亩)、增加的籽粒行数,任选地其中穗长度基本上没有减少、增加的籽粒数量、增加的籽粒大小、增加的穗长度、减少的分蘖数、减少的雄穗分枝数、每个节增加的豆荚数、每株植物增加的豆荚数、增加的种子大小(例如,面积、重量)和/或增加的种子油含量,任选地,其中所述一种或多种改善的产量性状是增加的种子大小和/或增加的种子油含量。
46.一种产生/培育无转基因的经编辑的植物的方法,包括:
将权利要求1-24、29、30或43-45中任一项的植物与无转基因的植物杂交,从而将所述至少一个非天然突变引入到所述无转基因的植物中;和
选择包含所述至少一个非天然突变并且无转基因的后代植物,从而产生无转基因的经编辑的植物。
47.一种提供具有一种或多种改善的产量性状的多个植物的方法,所述方法包括在生长区域中种植权利要求1-24、29、30或43-45中任一项的两个或更多个植物,从而提供与不包含所述至少一个非天然突变的多个对照植物相比具有一种或多种改善的产量性状的多个植物。
48.一种在E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)多肽的区域中产生变异的方法,包括:
将编辑系统引入到植物细胞中,其中所述编辑系统靶向编码HECT E3 UPL多肽的区域的HECT E3 UPL基因的区域,其中HECT E3 UPL多肽的区域包含
(a)与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的序列,
(b)与以下的连续氨基酸残基的区域具有至少80%序列同一性的序列:参考SEQ IDNO:78的残基编号,从残基50到残基95、从残基43到残基102、从残基36到残基109、从残基1814到残基1865、从残基1822到残基1857和/或从残基1829到残基1851,参考SEQ ID NO:79的残基编号,从残基48到残基87、从残基41到残基94、从残基34到残基101、从残基1820到残基1870、从残基1827到残基1863和/或从残基1834到残基1856,参考SEQ ID NO:80的残基编号,从残基139到残基194、从残基132到残基201、从残基125到残基208、从残基1838到残基1888、从残基1845到残基1881和/或从残基1852到残基1874,参考SEQ ID NO:81的残基编号,从残基104到残基146、从残基97到残基153、从残基90到残基160、从残基1834到残基1884、从残基1841到残基1877和/或从残基1848到残基1870,参考SEQ ID NO:82的残基编号,从残基1783到残基1833、从残基1790到残基1826和/或从残基1797到残基1818,和/或参考SEQ ID NO:83的残基编号,从残基1776到残基1826、从残基1783到残基1819和/或从残基1790到残基1812;或者
(c)包含与SEQ ID NO:93的氨基酸序列具有至少90%序列同一性的序列;
将HECT E3 UPL基因的区域与所述编辑系统接触,从而将突变引入到植物细胞中的HECT E3 UPL多肽的区域;和
在HECT E3 UPL多肽的区域中产生变异。
49.一种用于编辑植物细胞基因组中的特定位点的方法,所述方法包括:以位点特异性方式切割植物细胞中内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因内的靶位点,所述内源HECT E3 UPL基因:
(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列,
(b)编码与SEQ ID NO:78-83中任一个的氨基酸序列具有至少80%序列同一性的序列,
(c)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的区域,和/或
(d)编码与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域,从而在植物细胞的内源HECT E3 UPL基因中产生编辑并产生包含内源HECT E3 UPL基因中的编辑的植物细胞。
50.权利要求49的方法,进一步包括从包含所述内源HECT E3UPL基因中的编辑的植物细胞再生植物,以产生在其内源HECT E3UPL基因中包含编辑的植物。
51.权利要求49或权利要求50的方法,其中所述编辑导致非天然突变,任选地其中所述编辑导致与SEQ ID NO:119-125中任一个的核苷酸序列具有至少90%序列同一性的突变的HECT E3 UPL基因。
52.权利要求50或权利要求51的方法,其中在其内源HECT E3UPL基因中包含编辑的植物与缺乏所述至少一个突变的植物(例如,同基因植物(例如野生型未编辑植物或无效分离子)相比表现出一种或多种改善的产量性状的表型,任选地其中一种或多种改善的产量性状包括与缺乏所述至少一个非天然突变的对照植物相比更高的产量(蒲式耳/英亩)、增加的籽粒行数,任选地其中穗长度基本上没有减少、增加的籽粒数量、增加的籽粒大小、增加的穗长度、减少的分蘖数、减少的雄穗分枝数、每个节增加的豆荚数、每株植物增加的豆荚数、增加的种子重量/大小和/或增加的种子油含量。
53.权利要求49-52中任一项的方法,其中所述内源HECT E3 UPL基因编码HECT E3 UPL多肽,并且所述编辑导致位于以下位置处的氨基酸残基的变异:参考SEQ ID NO:78的氨基酸位置编号的位置1839,参考SEQ ID NO:79的氨基酸位置编号的位置1844,参考SEQ IDNO:80的氨基酸位置编号的位置1862,参考SEQ ID NO:81的氨基酸位置编号的位置1858,参考SEQ ID NO:82的氨基酸位置编号的位置1807,或者参考SEQ ID NO:83的氨基酸位置编号的位置1800。
54.一种制备植物的方法,包括:
(a)使包含内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的植物细胞群体与连接至核酸结合结构域(例如,编辑系统)的核酸酶接触,所述核酸结合结构域结合(a)与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性,(b)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列,(c)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列,和/或(d)编码包含与SEQ IDNO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列的序列;
(b)从所述群体中选择其中内源HECT E3 UPL基因已突变的植物细胞,从而产生包含内源HECT E3 UPL基因中的突变的植物细胞;和
(c)将选择的植物细胞生长成植物。
55.一种用于改善植物中的一种或多种产量性状的方法,包括
(a)使包含内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的植物细胞与靶向内源HECT E3 UPL基因的核酸酶接触,其中所述核酸酶连接至结合内源HECT E3 UPL基因中的靶位点的核酸结合结构域(例如,编辑系统),其中所述内源HECT E3 UPL基因:
(i)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列;
(ii)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;
(iii)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或
(iv)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,以产生包含内源HECT E3 UPL基因中的突变的植物细胞;和
(b)使植物细胞生长成包含内源HECT E3 UPL基因中的突变的植物,从而产生具有突变的内源HECT E3 UPL基因和一种或多种改善的产量性状的植物,任选地其中所述一种或多种改善的产量性状是增加的种子大小和/或增加的种子油含量。
56.一种用于产生植物或其部分的方法,所述植物或其部分包含至少一个具有突变的内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因的细胞,所述方法包括
将植物或植物部分中的内源HECT E3 UPL基因中的靶位点与包含切割结构域和核酸结合结构域的核酸酶接触,其中核酸结合结构域结合内源HECT E3 UPL基因中的靶位点,
其中所述内源HECT E3 UPL基因
(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列;
(b)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;
(c)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或
(d)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,从而产生包含至少一个具有内源HECT E3UPL基因中的突变的细胞的植物或其部分。
57.一种用于产生植物或其部分的方法,所述植物或其部分包含突变的内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因并表现出一种或多种改善的产量性状,所述方法包括
将植物或植物部分中的内源HECT E3 UPL基因中的靶位点与包含切割结构域和核酸结合结构域的核酸酶接触,其中核酸结合结构域结合内源HECT E3 UPL基因中的靶位点,
其中所述内源HECT E3 UPL基因:
(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列;
(b)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;
(c)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或
(d)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,从而产生包含具有突变的内源HECT E3 UPL基因并表现出一种或多种改善的产量性状的植物或其部分。
58.权利要求55或权利要求57的方法,其中所述一种或多种改善的产量性状包括与缺乏所述至少一个非天然突变的对照植物相比更高的产量(蒲式耳/英亩)、增加的籽粒行数,任选地其中穗长度基本上没有减少、增加的籽粒数量、增加的籽粒大小、增加的穗长度、减少的分蘖数、减少的雄穗分枝数、每个节增加的豆荚数、每株植物增加的豆荚数、增加的种子大小(例如,面积、重量)和/或增加的种子油含量,任选地,其中一种或多种改善的产量性状是增加的种子大小和/或增加的种子油含量。
59.权利要求48-58中任一项的方法,其中所述内源HECT E3 UPL基因是内源HECT E3UPL3基因。
60.权利要求59的方法,其中所述内源HECT E3 UPL3基因是内源HECT E3 UPL3a基因、内源HECT E3 UPL3b基因、内源HECT E3UPL3b基因或内源HECT E3 UPL3d基因。
61.权利要求54-60中任一项的方法,其中所述核酸酶切割所述内源HECT E3 UPL基因,从而将突变引入到内源HECT E3 UPL基因中。
62.权利要求54-61中任一项的方法,其中所述突变是非天然突变。
63.权利要求54-62中任一项的方法,其中所述突变是显性失活突变或无效突变。
64.权利要求54-63中任一项的方法,其中所述突变是取代、插入、倒位和/或缺失,任选地其中所述突变是框外插入或缺失或框内插入或缺失,任选地其中框外插入或缺失导致提前终止密码子。
65.权利要求54-64中任一项的方法,其中所述突变包括点突变。
66.权利要求54-64中任一项的方法,其中所述突变是一个碱基对至约100个碱基对的缺失,任选地其中所述缺失导致与SEQ ID NO:119-125中任一个的核苷酸序列具有至少90%序列同一性的突变的HECT E3 UPL基因。
67.权利要求54-66中任一项的方法,其中所述突变导致位于以下位置处的修饰的氨基酸残基:参考SEQ ID NO:78的氨基酸位置编号的位置1839,参考SEQ ID NO:79的氨基酸位置编号的位置1844,参考SEQ ID NO:80的氨基酸位置编号的位置1862,参考SEQ ID NO:81的氨基酸位置编号的位置1858,参考SEQ ID NO:82的氨基酸位置编号的位置1807,或者参考SEQ ID NO:83的氨基酸位置编号的位置1800。
68.权利要求54-67中任一项的方法,其中所述核酸酶是锌指核酸酶、转录激活因子样效应核酸酶(TALEN)、核酸内切酶或CRISPR-Cas效应蛋白。
69.权利要求54-68中任一项的方法,其中所述核酸结合结构域是锌指、转录激活因子样DNA结合结构域(TAL)、argonaute或CRISPR-Cas效应子DNA结合结构域。
70.一种通过权利要求46或54-69中任一项的方法产生的植物,任选地其中所述植物包含与SEQ ID NO:119-125中任一个的核苷酸序列具有至少90%序列同一性的突变的HECTE3 UPL基因。
71.一种引导核酸,其结合E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的靶位点,该靶位点位于HECT E3 UPL3基因的区域中,所述区域:
(a)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的序列,
(b)编码与以下的连续氨基酸残基的区域具有至少80%序列同一性的序列:参考SEQID NO:78的残基编号,从残基50到残基95、从残基43到残基102、从残基36到残基109、从残基1814到残基1865、从残基1822到残基1857和/或从残基1829到残基1851,参考SEQ ID NO:79的残基编号,从残基48到残基87、从残基41到残基94、从残基34到残基101、从残基1820到残基1870、从残基1827到残基1863和/或从残基1834到残基1856,参考SEQ ID NO:80的残基编号,从残基139到残基194、从残基132到残基201、从残基125到残基208、从残基1838到残基1888、从残基1845到残基1881和/或从残基1852到残基1874,参考SEQ ID NO:81的残基编号,从残基104到残基146、从残基97到残基153、从残基90到残基160、从残基1834到残基1884、从残基1841到残基1877和/或从残基1848到残基1870,参考SEQ ID NO:82的残基编号,从残基1783到残基1833、从残基1790到残基1826和/或从残基1797到残基1818,和/或参考SEQ ID NO:83的残基编号,从残基1776到残基1826、从残基1783到残基1819和/或从残基1790到残基1812;和/或
(c)编码与SEQ ID NO:93的氨基酸序列具有至少90%序列同一性的序列。
72.权利要求71的引导核酸,其中所述引导核酸包含间隔区,所述间隔区包含SEQ IDNO:94-118的核苷酸序列中的任一个。
73.一种系统,其包含权利要求71或权利要求72的引导核酸和与所述引导核酸缔合的CRISPR-Cas效应蛋白。
74.权利要求73的系统,还包含与所述引导核酸和CRISPR-Cas效应蛋白缔合的tracr核酸,任选地其中所述tracr核酸和所述引导核酸共价连接。
75.一种基因编辑系统,其包含与引导核酸缔合的CRISPR-Cas效应蛋白,其中所述引导核酸包含与内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因结合的间隔序列。
76.权利要求74的基因编辑系统,其中所述E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因:
(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列,
(b)编码与SEQ ID NO:78-83中任一个的氨基酸序列具有至少80%序列同一性的序列,
(c)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的区域,和/或
(d)编码与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域。
77.权利要求75或权利要求76的基因编辑系统,其中所述引导核酸包含间隔序列,所述间隔序列包含SEQ ID NO:94-118的核苷酸序列中的任一个。
78.权利要求75-77中任一项的基因编辑系统,还包含与所述引导核酸和CRISPR-Cas效应蛋白缔合的tracr核酸,任选地其中所述tracr核酸和所述引导核酸共价连接。
79.一种复合物,其包含引导核酸和包含切割结构域的CRISPR-Cas效应蛋白,其中所述引导核酸结合内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的靶位点,其中所述内源HECT E3 UPL3基因:
(a)包含与SEQ ID NO:72-77中任一个的核苷酸序列具有至少80%序列同一性的序列,
(b)编码与SEQ ID NO:78-83中任一个的氨基酸序列具有至少80%序列同一性的序列,
(c)编码与SEQ ID NO:84-92中任一个的氨基酸序列具有至少80%序列同一性的区域,和/或
(d)编码与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域,其中所述切割结构域切割HECT E3 UPL3基因中的靶标链。
80.一种表达盒,其包含(a)编码包含切割结构域的CRISPR-Cas效应蛋白的多核苷酸,和(b)结合内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的靶位点的引导核酸,其中所述引导核酸包含间隔序列,所述间隔序列互补于并结合
(i)与SEQ ID NO:72-77的核苷酸序列中的任一个具有至少80%序列同一性的序列的一部分;
(ii)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列的核酸的一部分;
(iii)编码与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的氨基酸序列的核酸的一部分;和/或
(iv)编码与SEQ ID NO:93具有至少90%序列同一性的氨基酸序列的核酸的一部分。
81.一种编码E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)多肽的显性失活突变或无效突变的核酸,任选地其中所述核酸包含与SEQ ID NO:119-125中的任一个具有至少90%序列同一性的核苷酸序列。
82.一种修饰的E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)多肽,其包含位于以下位置处的氨基酸残基的突变:参考SEQ ID NO:78的氨基酸位置编号的位置1839,参考SEQ IDNO:79的氨基酸位置编号的位置1844,参考SEQ ID NO:80的氨基酸位置编号的位置1862,参考SEQ ID NO:81的氨基酸位置编号的位置1858,参考SEQ ID NO:82的氨基酸位置编号的位置1807,和/或参考SEQ ID NO:83的氨基酸位置编号的位置1800。
83.一种植物或其部分,其包含权利要求80的核酸或权利要求82的修饰的HECT E3 UPL多肽。
84.一种在植物中产生内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的突变的方法,包括:
(a)将基因编辑系统靶向至HECT E3 UPL基因的一部分,其
(i)编码对应于参考SEQ ID NO:78的氨基酸位置编号的位置1839、参考SEQ ID NO:79的氨基酸位置编号的位置1844、参考SEQ ID NO:80的氨基酸位置编号的位置1862、参考SEQID NO:81的氨基酸位置编号的位置1858、参考SEQ ID NO:82的氨基酸位置编号的位置1807和/或参考SEQ ID NO:83的氨基酸位置编号的位置1800的氨基酸;
(ii)编码与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列;
(iii)编码包含与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域的氨基酸序列;和/或
(iv)编码包含与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域的氨基酸序列,并且
(b)选择植物,其包含(i)参考SEQ ID NO:78的氨基酸位置编号的位置1839、参考SEQID NO:79的氨基酸位置编号的位置1844、参考SEQ ID NO:80的氨基酸位置编号的位置1862、参考SEQ ID NO:81的氨基酸位置编号的位置1858、参考SEQ ID NO:82的氨基酸位置编号的位置1807和/或参考SEQ ID NO:83的氨基酸位置编号的位置1800处的氨基酸残基的取代;(ii)与SEQ ID NO:78-83中的任一个具有至少80%序列同一性的氨基酸序列中的突变;(iii)与SEQ ID NO:84-92中的任一个具有至少80%序列同一性的区域中的突变;和/或(iv)与SEQ ID NO:93的氨基酸序列具有至少90%同一性的区域中的突变。
85.一种玉米植物或其植物部分,其包含在至少一个内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的至少一个非天然突变,所述基因具有Zm00001d014920和/或Zm00001d004139的基因识别号(基因ID),任选地其中所述非天然突变包含与SEQ ID NO:122-125中任一个的核苷酸序列具有至少90%序列同一性的突变的HECT E3 UPL基因。
86.一种大豆植物或其植物部分,其包含在至少一个内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的至少一个非天然突变,所述基因具有Glyma.11G107500、Glyma.12G032500、Glyma.06G003600和/或Glyma.04G004000的基因识别号(基因ID),任选地其中所述非天然突变包含与SEQ ID NO:119-121中任一个的核苷酸序列具有至少90%序列同一性的突变的HECT E3 UPL基因。
87.一种引导核酸,其结合E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的靶核酸,所述基因具有Zm00001d014920、Zm00001d004139、Glyma.11G107500、Glyma.12G032500、Glyma.06G003600和/或Glyma.04G004000的基因识别号(基因ID)。
88.一种产生植物的方法,所述植物包含内源E6AP C端同源(HECT)E3泛素蛋白连接酶(UPL)基因中的突变和至少一种感兴趣的多核苷酸,所述方法包括
将第一植物与包含至少一种感兴趣的多核苷酸的第二植物杂交以产生后代植物,所述第一植物为权利要求1-24、29-30、43-45或70中任一项的植物;和
选择包含所述HECT E3 UPL基因中的突变和所述至少一种感兴趣的多核苷酸的后代植物,从而产生包含内源HECT E3 UPL基因中的突变和至少一种感兴趣的多核苷酸的植物。
89.一种产生植物的方法,所述植物包含内源HECT E3 UPL基因中的突变和至少一种感兴趣的多核苷酸,所述方法包括
将至少一种感兴趣的多核苷酸引入到权利要求1-24、29-30、43-45或70中任一项的植物中,从而产生包含HECT E3 UPL基因中的突变和至少一种感兴趣的多核苷酸的植物。
90.一种产生植物的方法,所述植物包含内源HECT E3 UPL基因中的突变并且表现出改善的产量性状、改善的植物结构和/或改善的防御性状的表型,所述方法包括
将第一HECT E3 UPL基因植物与表现出改善的产量性状、改善的植物结构和/或改善防御性状的表型的第二植物杂交,所述第一HECT E3 UPL基因植物为权利要求1-24、29-30、43-45或70中任一项的植物;和
选择包含所述HECT E3 UPL基因中的突变和改善的产量性状、改善的植物结构和/或改善的防御性状的表型的后代植物,从而产生包含内源HECT E3 UPL基因中的突变并与对照植物相比表现出改善的产量性状、改善的植物结构和/或改善的防御性状的表型的植物。
91.一种控制容器(例如花盆或种子盘等)、生长室、温室、田地、休闲区、草坪或路边中的杂草的方法,包括
将除草剂施用于生长在容器、生长室、温室、田地、休闲区、草坪或路边中的权利要求1-24、29-30、43-45或70中任一项的一种或多种(多个)植物,从而控制一种或多种植物生长在其中的容器、生长室、温室、田地、休闲区、草坪或路边中的杂草。
92.一种减少植物上的昆虫捕食的方法,包括
将杀虫剂施用于权利要求1-24、29-30、43-45或70中任一项的一种或多种植物,从而减少所述一种或多种植物上的昆虫捕食。
93.一种减少植物上的真菌病害的方法,包括
将杀真菌剂施用于权利要求1-24、29-30、43-45或70中任一项的一种或多种植物,从而减少所述一种或多种植物上的真菌病害。
94.权利要求92或权利要求93的方法,其中一种或多种大豆植物生长在容器、生长室、温室、田地、休闲区、草坪或路边中。
95.权利要求88-94中任一项的方法,其中所述感兴趣的多核苷酸是赋予除草剂耐受性、抗虫性、抗病性、增加的产量、增加的养分利用效率或非生物胁迫抗性的多核苷酸。
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