CN101939434A - 用于在大豆中提高种子贮藏油脂的生成和改变脂肪酸谱的来自解脂耶氏酵母的dgat基因 - Google Patents
用于在大豆中提高种子贮藏油脂的生成和改变脂肪酸谱的来自解脂耶氏酵母的dgat基因 Download PDFInfo
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Abstract
本文描述了总脂肪酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量相比高出至少10%的转基因大豆种子。来自解脂耶氏酵母的DGAT基因被用于实现种子贮藏油脂的提高。
Description
本申请要求申请日为2007年5月24日、申请号为60/939872和申请日为2007年12月13日、申请号为61/013406的美国临时专利申请的优先权,上述两个申请的全部公开内容特此并入本申请中。
发明领域
本发明属于生物技术领域,尤其涉及编码二酰基甘油酰基转移酶基因的多核苷酸序列以及利用此类酰基转移酶在大豆中提高种子贮藏油脂的生成和改变脂肪酸谱。
发明背景
植物油脂在工业上和营养学方面具有多种用途,并且对植物膜功能的实现和气候适应性起着重要作用。这些油脂表现出大量的化学结构,这些结构决定着上述油脂的生理学和工业特性。大量的上述结构直接或间接地产生自令上述油脂的不饱和度改变的代谢过程。不同植物的不同代谢模式产生出这些改变的油脂,而要大量、经济地生产所期望的油脂,则通常需要对外来植物物种进行驯化或者对已适应的农作物加以改良。
利用诱变方式来改变脂肪酸的组成和含量存在严重的局限性。筛选法很难发现以下情形的突变:a)导致显性(功能获得性)表型,b)位于植物生长所必需的基因中,以及c)位于非限速酶和由一种以上基因编码的酶的编码基因中。在获得了具有所期望表型的突变玉米品系的实例中,由于所期望的油脂成分很可能是若干隐性基因的产物,通过传统育种技术对原种品系进行的基因渗入既耗时又昂贵。
近来的分子和细胞生物学技术提供了克服诱变方法的某些局限的潜在可能,包括能满足大范围育种的需要。一些特别有用的技术包括在转基因植物中对外来基因进行种子特异性的表达(参见Goldberg等人(1989)Cell 56:149-160),以及利用反义RNA以显性或组织特异性方式对植物靶基因进行抑制(参见van der Krol等人(1988)Gene72:45-50)。其他进展包括向工业油料作物的商品化品种原种中转入外来基因,例如大豆(Chee等人(1989)Plant Physiol.91:1212-1218;Christou等人(1989)Proc.Natl.Acad.Sci.U.S.A.86:7500-7504;Hinchee等人(1988)Bio/Technology 6:915-922;欧洲专利公布0 301749 A2),卡诺拉(De Block等人(1989)Plant Physiol.91:694-701),和向日葵(Everett等人(1987)Bio/Technology 5:1201-1204),以及在育种程序中利用一些基因作为限制性片段长度多态性(RFLP)标记,从而令隐性性状向原种品系的基因渗入既快速又花费较少(Tanksley等人(1989)Bio/Technology 7:257-264)。然而,上述各种技术的应用,需要对具有重要商业意义的基因进行鉴定和分离。
大多数的游离脂肪酸被酯化为辅酶A(CoA),以产生酰基辅酶A。然后这些分子在细胞内质网内作为甘油脂类合成的底物,产生磷脂酸和二酰甘油(DAG)。这些代谢中间产物中任何一种都可被用于形成膜磷脂(例如磷脂酰甘油、磷脂酰乙醇胺和磷脂酰胆碱),或者DAG可被用于形成三酰基甘油(TAG),后者是真核细胞中所储存的类脂类主要储备物。
在植物、真菌和哺乳动物中,二酰基甘油酰基转移酶(“DGAT”)是一种在三酰基甘油生成反应中催化最后一个酶促步骤的整合膜蛋白。该酶负责将一个酰基基团从酰基-辅酶A上转移到1,2-二酰基甘油(“DAG”)的sn-3位上,以形成三酰基甘油(“TAG”)。在植物和真菌中,DGAT与膜和脂质体片段相联,尤其在油料种子中,其参与组成用作能量储备的碳贮藏库。TAG被认为是细胞中用于储存能量的一种重要化学物质。已知DGAT可调控TAG的结构并指导TAG的合成。此外,已知DGAT反应为油脂合成所特有。
在植物中,TAG是植物油的基本成分,种子将其作为供萌芽期使用的能量的一种贮藏形式。
已鉴定出的DGAT蛋白包括两个不同的家族。DGAT蛋白的第一个家族(“DGAT1”)与酰基辅酶A:胆固醇酰基转移酶(“ACAT”)相关,描述于美国专利6,100,077和6,344,548。DGAT蛋白的第二个家族(“DGAT2”)与上述DGAT1家族不相关,描述于2004年2月5日公布的PCT专利公布WO 2004/011671。关于DGAT基因及其在植物中的使用的其他参考文献包括PCT专利公布WO2004/011,671、WO1998/055,631和WO2000/001,713,以及美国专利公布20030115632。
申请人之受让人的已公布的共同待决专利申请US 2006-0094088描述了植物和真菌中的DGAT基因,并且描述了它们的用途是在食用油中调节多不饱和脂肪酸(“PUFA”)的水平。
申请人之受让人的已公布的国际专利申请WO 2005/003322描述了对用于改变含油酵母中PUFA和油脂含量的磷脂酰胆碱二酰基甘油酰基转移酶以及DGAT2的克隆。
发明概述
本发明涉及一种转基因大豆种子,所述转基因大豆种子的总脂肪酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量相比高出至少10%。
在第二种实施方案中,本发明涉及一种提高大豆种子的总脂肪酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量相比高出至少10%的步骤(a)中的转化的大豆细胞。
在第三种实施方案中,本发明涉及转基因玉米谷粒,所述转基因玉米谷粒的总脂肪酸含量与非转基因的、无转基因分离子玉米谷粒的总脂肪酸含量相比高出至少10%。
在第四种实施方案中,本发明涉及提高玉米谷粒的总脂肪酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个玉米谷粒;
(b)选择总脂肪酸含量与非转基因的、无转基因分离子玉米谷粒的总脂肪酸含量相比高出至少10%的步骤(a)中的转化的玉米谷粒。
在第五种实施方案中,本发明涉及转基因大豆种子,与非转基因的、无转基因分离子大豆种子的对应含量相比,所述转基因大豆种子的总脂肪酸含量高出至少10%并且油酸含量高出至少25%。
在另一个实施方案中,本发明涉及转基因大豆种子,与非转基因的、无转基因分离子大豆种子的对应含量相比,所述转基因大豆种子的总脂肪酸含量高出至少10%,并且具有至少一种下列特征:i)油酸含量高出至少25%;ii)亚麻酸含量降低至少25%;iii)亚油酸含量降低至少4%;iv)棕榈酸含量降低至少8%;以及v)硬脂酸含量高出至少14%。
在第六种实施方案中,本发明涉及提高大豆种子的总脂肪酸含量和油酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和油酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和油酸含量相比分别高出至少10%和至少25%的步骤(a)中的转化的大豆细胞。
在第七种实施方案中,本发明涉及提高大豆种子的总脂肪酸含量并降低其亚麻酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和亚麻酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和亚麻酸含量相比分别高出至少10%和降低至少25%的步骤(a)中的转化的大豆细胞。
在第八种实施方案中,本发明涉及提高大豆种子的总脂肪酸含量并降低其亚油酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和亚油酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和亚油酸含量相比分别高出至少10%和降低至少4%的步骤(a)中的转化的大豆细胞。
在第九种实施方案中,本发明涉及提高大豆种子的总脂肪酸含量并降低其棕榈酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和棕榈酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和棕榈酸含量相比分别高出至少10%和降低至少8%的步骤(a)中的转化的大豆细胞。
在第十种实施方案中,本发明涉及提高大豆种子的总脂肪酸含量和硬脂酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和硬脂酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和硬脂酸含量相比分别高出至少10%和至少14%的步骤(a)中的转化的大豆细胞。
本发明的任何一种转基因种子可以包含具有至少一种DGAT序列的重组构建体,所述DGAT序列可以选自DGAT1、DGAT2和DGAT1与DGAT2的组合。此外,所述DGAT序列可以是一种耶氏酵母的序列。
从本发明的转基因大豆种子获得的产品和/或副产品同样属于本发明的范围。
耶氏酵母DGAT基因保持了其功能的突变型也同样属于本发明的范围。特定的例子包括但不限于一种分离的多核苷酸,所述多核苷酸包含编码具有二酰基甘油酰基转移酶活性的多肽的序列,其中所述多肽如SEQ ID:83、88或93所示。所述核苷酸序列的互补序列,包含所述核苷酸序列的重组DNA构建体,以及掺入了所述重组DNA构建体的细胞也同样属于本发明的范围。产生油料种子的植物适用于包含所述重组构建体。
附图和序列表的简要说明
根据以下的详细描述和附图以及序列清单,可以更全面地理解本发明,以下的详细描述和附图以及序列清单形成本申请的一部分。
图1显示了pFBAIn-YLDGAT1、pFBAIn-YLDGAT2和pFBAIn-MOD1的质粒图谱。
图2显示了KS352和KS332的质粒图谱。
图3显示了KS349、KS362和KS364的质粒图谱。
图4显示了携带KS349的体细胞胚油酸含量和总酯化脂肪酸含量之间的强相关性(R2≥0.59)。
图5显示了单独携带KS362或同时携带KS362与KS349和KS364其中之一的体细胞胚油酸含量和总酯化脂肪酸含量之间的强相关性(R2≥0.67)。
图6显示了通过共转化质粒KS349和KS362得到的转基因大豆种子(T1世代)油酸含量和油脂含量之间的相关性(R2≥0.45)。
图7显示了通过共转化质粒KS349和KS362(A)和单独转化质粒KS362(B)得到的T1种子的油脂含量和种子重量。
图8显示了来自基因组DNA印迹的杂交结果。基因组DNA分离自来自事件AFS4818.1.2、AFS4818.1.3、AFS4818.1.5、AFS48182.6和AFS4818.1.9(参见实施例6)的转基因大豆。用限制酶EcoRI或HindIII消化DNA,然后在一块凝胶上跑出并与尼龙滤膜杂交(泳道1和2为AFS4818.1.2,泳道3和4为AFS4818.1.3,泳道5和6为AFS4818.1.5,泳道7和8为AFS48182.6,泳道9和10为AFS4818.1.9,泳道11和12为同样用EcoRI和HindIII消化后的非转基因的野生型DNA)。上方的印迹(A)所用的杂交探针为一种耶氏酵母DGAT1特异性探针,下方的印迹所用的则为一种耶氏酵母DGAT2特异性探针。
图9显示了来自基因组DNA印迹的杂交结果。图中的印迹与图8中所描述的相似,不同之处在于DNA全部用限制酶BstXI消化并且印迹利用一种耶氏酵母DGAT2特异性探针进行探测。
序列描述概要说明了后附的序列列表。此序列列表中以单个字母表示核苷酸,以单独的3字母表示氨基酸,如Nucleic Acids Research13:3021-3030(1985)和Biochemical Journal 219(2):345-373(1984)所描述的IUPAC-IUB标准中所规定的。
核酸和蛋白质SEQ ID Number摘要
SEQ ID NO:7-8为PCR引物oYLDGAT2-1(SEQ IOD NO:7)和oYLDGAT2-2(SEQ IOD NO:8)的对应序列,上述引物用于从一份酵母溶菌液中扩增解脂耶氏酵母二酰基甘油酰基转移酶2(YL DGAT2)基因(详见实施例1)。
SEQ ID NO:12-15为用于从解脂耶氏酵母基因组DNA中扩增YL DGAT1(YDGAT1-F和YDGAT1-R,SEQ ID NO分别为12和13)和YL DGAT2(YDGAT2-F和YDGAT2-R,SEQ ID NO分别为14和15)编码区域的寡核苷酸引物的对应序列。
SEQ ID NO:23(oCal-15)和SEQ ID NO:24(oCal-6)为用于从如PCT专利公布WO 02/008269中所述的模板质粒CalFad2-2中扩增DNA片段ca1 a24-4(SEQ ID NO:22)的寡核苷酸引物的对应序列。
SEQ ID NO:28(oKR85-1)和SEQ ID NO:29(oKR85-2)为用于从质粒pKR85(SEQ ID NO:27)中扩增β-大豆伴球蛋白启动子-(NotI克隆位点)-云扁豆蛋白3′终止子区域的引物的对应序列。
SEQ ID NO:37(oGy1-1)和SEQ ID NO:38(oGy1-2)为用于扩增大豆的大豆球蛋白1启动子(SEQ ID NO:36),并在5′和3′端分别插入了BamHI和NcoI位点的引物的对应序列。
SEQ ID NO:54(oP34-1)和SEQ ID NO:55(oP34-2)为用于扩增大豆P34启动子(SEQ ID NO:53),并在5′和3′端分别插入了BamHI和NcoI位点的引物的对应序列。
SEQ ID NO:69(oSGly-2)和SEQ ID NO:70(oSGly-3)为用于扩增大豆球蛋白GY1启动子的引物的对应序列。
SEQ ID Nos:76(oYDG2-1)和SEQ ID NO:77(oYDG2-2)为用于扩增解脂耶氏酵母DGAT2(SEQ ID NO:10)的引物的对应序列,所述解脂耶氏酵母DGAT2随后被插入pKR1254(SEQ ID NO:78)。
SEQ ID NO:79(Y1D2_Y326F-5)和SEQ ID NO:80(Y1D2_Y326F-3)为用于使解脂耶氏酵母DGAT2(SEQ ID NO:10)326位点的氨基酸由酪氨酸突变为苯丙氨酸的引物的对应序列。
SEQ ID NO:84(Y1D2_Y326L-5)和SEQ ID NO:85(Y1D2_Y326L-3)为用于使解脂耶氏酵母DGAT2(SEQ ID NO:10)326位点的氨基酸由酪氨酸突变为亮氨酸的引物的对应序列。
SEQ ID NO:89(Y1D2_Y327K-5)和SEQ ID NO:90(Y1D2_Y327K-3)为用于使解脂耶氏酵母DGAT2(SEQ ID NO:10)327位点的氨基酸由精氨酸突变为赖氨酸的引物的对应序列。
SEQ ID NO:108至111(分别为GmTE25-1、GmTE23-1、GmTE25-2和GmTE23-2)为用于扩增大豆硫酯酶2基因的引物的对应序列。
SEQ ID NO:115至118(分别为GmFad2-15-1、GmFad2-13-1、GmFad2-1 5-2和GmFad2-1 3-2)为用于扩增大豆脂肪酸去饱和酶2-1基因的引物的对应序列。
发明详述
藉此将本文中所列出的每篇参考文献的公开内容的全文以引用的方式并入本文。
如本文所用的并在所附权利要求书中的单数形式“一个”和“所述”包括复数涵义,除非上下文中清楚地另有指明。因此,例如,“一株植物”的涵义包括多株此类植物,“一个细胞”的涵义包括一个或多个细胞及其本领域的技术人员已知的等同物,等等。
在本公开内容的上下文中,使用了若干术语和缩写。给出了如下定义。
“开放阅读框”缩写为ORF。
“聚合酶链式反应”缩写为PCR。
“美国典型培养物保藏中心”缩写为ATCC。
“酰基-辅酶A:固醇-酰基转移酶”缩写为ARE2。
“磷脂:二酰基甘油酰基转移酶”缩写为PDAT。
“二酰基甘油酰基转移酶”缩写为DAG AT或DGAT。
“二酰基甘油”缩写为DAG。
“三酰基甘油”缩写为TAG。
“辅酶A”缩写为CoA。
术语“脂肪酸”指不同链长的长链脂族酸(链烷酸),链长为约C12至C22(尽管更长和更短链长的酸两者都是已知的)。主要的链长介于C16和C22之间。脂肪酸的结构可用简单的记号系统“X:Y”来表示,其中X表示具体脂肪酸中碳(C)原子的总数,而Y表示双键的数目。
总的来说,脂肪酸被分为饱和的和不饱和的两类。术语“饱和脂肪酸”是指其碳主链不含“双键”的脂肪酸。相对的,“不饱和脂肪酸”的碳主链(通常为顺式构型)则含有“双键”。“单不饱和脂肪酸”的碳主链含有一个“双键”(例如,在棕榈油酸(16:1)和油酸(18:1)中通常位于第9号和第10号碳原子之间。),而“多不饱和脂肪酸”(或者叫“PUFA”)的碳主链则含有至少两个双键(例如,在亚油酸(18:2)中位于第9号和第10号以及第12号和第13号碳原子之间;在α-亚麻酸(18:3)中位于第9号和第10号、第12号和第13号以及第15号和第16号碳原子之间)。
“微生物油脂”或者“单细胞油脂”是由微生物(例如藻类、含油酵母和丝状真菌)在其生命周期中自然产生的油脂。术语“油脂”是指在25℃时为液体且通常为多不饱和的脂类物质。相对的,术语“脂肪”是指在25℃时为固体且通常为饱和的脂类物质。
“脂质体”是指通常以特异性蛋白质和一层磷脂单分子层为界的脂类小滴。此类细胞器是多数生物转运/贮藏中性脂类的位点。脂质体被认为来源于包含TAG-生物合成酶类的内质网的微结构域,并且它们的合成和大小似乎受特异性蛋白质组分的控制。
“中性脂类”是指通常发现于细胞的脂质体内,作为储存性脂肪或油脂的那些脂类,其之所以得名是由于在胞内的pH值下,此类脂类不含带电基团。总而言之,它们完全没有极性,与水没有亲和力。中性脂类通常是指与脂肪酸相连的甘油单酯、二酯和/或三酯,分别也被称为单酰基甘油、二酰基甘油或者TAG(或者总称为酰基甘油)。要将脂肪酸从酰基甘油上释放出来,必须发生水解反应。
术语“三酰基甘油”、“油”和“TAG”指由与甘油分子酯化的三个脂肪酰残基组成的中性脂质(并且这些术语在本公开内容中将可以互换使用)。这些油可含有长链PUFA以及较短的饱和的和不饱和的脂肪酸以及较长链的饱和脂肪酸。因此,“油的生物合成”一般指细胞中TAG的合成。
术语“DAG AT”是指一种二酰基甘油酰基转移酶(也叫酰基-辅酶A-二酰基甘油酰基转移酶或二酰基甘油O-酰基转移酶)(EC2.3.1.20)。该酶负责酰基-辅酶A和1,2-二酰基甘油向TAG和CoA的转化(由此涉及TAG生物合成的最后步骤)。存在两个家族的DAGAT酶:DGAT1和DGAT2。前一家族同酰基-辅酶A:胆固醇酰基转移酶(ACAT)基因家族具有同源性,而后一家族与之无关(Lardizabal等人,J.Biol.Chem.276(42):38862-28869(2001))。
术语“PDAT”是指一种磷脂:二酰基甘油酰基转移酶(EC2.3.1.158)。该酶负责将一个酰基基团从磷脂的sn-2位上转移到1,2-二酰基甘油的sn-3位上,从而产生溶血磷脂和TAG(由此涉及TAG生物合成的最后步骤)。该酶与DGAT(EC 2.3.1.20)不同的是,其合成TAG时通过的是一种不依赖于酰基-辅酶A的机制。
术语“ARE2”是指一种酰基-辅酶A:固醇-酰基转移酶(EC2.3.1.26;也叫固醇-酯合酶2),该酶催化下列反应:酰基-辅酶A+胆固醇=辅酶A+胆固醇酯。
本文所用的“核酸”意指多核苷酸,并包括单链或双链的脱氧核糖核苷酸或核糖核苷酸碱基聚合物。核酸也可以包括片段和经修饰的核苷酸。因此,术语“多核苷酸”、“核酸序列”、“核苷酸序列”或“核酸片段”可互换使用,并且是作为单链或双链的RNA或DNA聚合物,任选含有合成的、非天然的或改变的核苷酸碱基。核苷酸(通常以它们的5′-单磷酸形式存在)可以用如下它们的单字母名称指代:“A”表示腺苷酸或脱氧腺苷酸(分别针对RNA或DNA),“C”表示胞苷酸或脱氧胞苷酸,“G”表示鸟苷酸或脱氧鸟苷酸,“U”表示尿苷酸,“T”表示脱氧胸苷酸,“R”表示嘌呤(A或G),“Y”表示嘧啶(C或T),“K”表示G或T,“H”表示A或C或T,“I”表示肌苷,“N”表示任何核苷酸。
术语“具有相当功能的亚片段”和“功能上相当的亚片段”在本文中可互换使用。这些术语是指一种分离的核酸片段的一个部分或亚序列,其中无论所述片段或亚片段是否编码有活性的酶,其都保留着改变基因表达或导致某种表型的能力。例如,所述片段或亚片段可以被用于设计嵌合基因,以在一种转化的植物中产生所期望的表型。可以通过将一种核酸片段或亚片段以相对于一种植物启动子序列有义或反义的方向与所属启动子序列连接,从而设计出用于抑制的嵌合基因,其中所述核酸片段或亚片段无论是否编码有活性的酶均可。
术语“保守结构域”或“基序”指进化上相关的蛋白质的比对序列中在特定位置保守的一组氨基酸。虽然同源蛋白质之间在其它位置的氨基酸可以发生变化,但在特定位置高度保守的氨基酸表示对蛋白质的结构、稳定性或活性来说是必需的氨基酸。因为它们可通过它们在蛋白质同源物家族的比对序列中的高度保守来鉴定,所以它们可用作识别标记或“签名”来确定具有新的测定序列的蛋白质是否属于以前鉴定的蛋白质家族。
术语“同源性”、“同源的”、“基本上类似的”和“基本上对应的”本文可互换使用。它们指其中一个或多个核苷酸碱基的变化不会影响核酸片段介导基因表达或产生某种表型的能力的核酸片段。这些术语也指本发明的核酸片段的修饰(例如缺失或插入一个或多个核苷酸),相对于初始的未经修饰的核酸片段,基本上不会改变所得核酸片段的功能特性。因此,正如本领域技术人员应该理解的,本发明不仅仅涵盖这些具体的示例性序列。
此外,技术人员认识到,本发明所涵盖的基本上类似的核苷酸序列也由它们在中等严格条件(如0.5×SSC,0.1%SDS,60℃)下,与本文所示例的序列杂交的能力,或杂交至本文所公开的核苷酸序列的任何部分以及杂交至与本文所公开的任何核苷酸序列功能相当的序列的能力所限定。可以调节严格性条件以筛选中度相似的片段例如来自远缘生物的同源序列,到筛选高度相似的片段例如从近缘生物复制功能性酶的基因。杂交后的洗涤确定严格性条件。
术语“选择性杂交”包括指在严格杂交条件下,核酸序列与特定核酸靶序列以比其与非靶核酸序列的杂交更高的可检测程度(例如至少两倍于背景)杂交,并指基本排除了非靶核酸。选择性杂交的序列通常彼此具有约至少80%的序列同一性,或90%的序列同一性,最多并包括100%的序列同一性(即完全互补)。
术语“严格条件”或“严格杂交条件”包括指探针将选择性杂交至其靶序列的条件。严格条件是序列依赖性的,并将因不同的环境而异。通过控制杂交和/或洗涤条件的严格性,可鉴定与探针100%互补的靶序列(同源探测)。作为另一种选择,可调节严格条件以允许序列中的一些错配,以便检测到更低程度的相似性(异源探测)。通常,探针的长度少于约1000个核苷酸,任选长度少于500个核苷酸。
一般地,严格条件将是如下那些条件:在pH7.0至8.3下盐浓度低于约1.5M钠离子,通常约0.01至1.0M钠离子浓度(或其它盐),并且对于短探针(例如10至50个核苷酸)温度为至少约30℃,而对于长的探针(例如多于50个核苷酸)温度为至少约60℃。严格条件也可以通过加入诸如甲酰胺之类的去稳定剂来实现。示例性的低严格条件包括在37℃于含有30至35%甲酰胺、1M NaCl、1%SDS(十二烷基硫酸钠)的缓冲液中杂交,以及在50至55℃用1×至2×SSC(20×SSC=3.0M NaCl/0.3M柠檬酸三钠)洗涤。示例性的中等严格条件包括在37℃于40至45%甲酰胺、1M NaCl、1%SDS中杂交,以及在55至60℃下用0.5×至1×SSC洗涤。示例性的高严格条件包括在37℃于50%甲酰胺、1M NaCl、1%SDS中杂交,以及在60至65℃用0.1×SSC洗涤。
特异性通常取决于杂交后洗涤,最后的洗涤溶液的离子强度和温度是关键因素。对于DNA-DNA杂交,可以从Meinkoth等人,Anal.Biochem.138:267-284(1984)中的公计算出近似的Tm∶Tm=81.5℃+16.6(log M)+0.41(%GC)-0.61(%form)-500/L;其中M是单价阳离子的体积摩尔浓度,%GC是DNA中鸟苷和胞嘧啶核苷酸的百分比,%form是杂交溶液中甲酰胺的百分比,而L是以碱基对表示的杂交体的长度。Tm是(在确定的离子强度和pH下)50%的互补靶序列与完全匹配的探针杂交时的温度。每出现1%的错配,Tm降低约1℃;因此,可调节Tm杂交和/或洗涤条件以与具有期望同一性的序列杂交。例如,如果要寻求具有≥90%同一性的序列,则可将Tm降低10℃。通常,在确定的离子强度和pH下,严格条件选择为比特定序列及其互补序列的熔解温度(Tm)低约5℃。然而,极严格条件可采用在比熔解温度(Tm)低1、2、3或4℃的温度下杂交和/或洗涤;中等严格条件可采用在比熔解温度(Tm)低6、7、8、9或10℃的温度下杂交和/或洗涤;低严格条件可采用在比熔解温度(Tm)低11、12、13、14、15或20℃温度下杂交和/或洗涤。利用所述等式、杂交和洗涤组合物以及理想的Tm,本领域技术人员将会理解,杂交和/或洗涤溶液的严格性的变化已经在本质上得以描述。如果所需的错配程度导致Tm低于45℃(水溶液)或32℃(甲酰胺溶液),则优选增加SSC的浓度以便能使用更高的温度。对于核酸杂交的详尽指导可在以下文献中找到:Tijssen,Laboratory Techniques in Biochemistry and Molecular Biology--Hybridization with Nucleic Acid Probes,第I部分,第2章“Overview of principles of hybridization and the strategy of nucleic acid probe assays”,Elsevier,New York(1993);and Current Protocols in Molecular Biology,第2章,Ausubel等人,编辑,Greene Publishing and Wiley-Interscience,New York(1995)。杂交和/或洗涤条件可进行至少10、30、60、90、120或240分钟。
“序列一致性”或“一致性”当涉及核酸或多肽序列时,是指两段序列中的核酸碱基或氨基酸残基当在一个指定的比对窗口上进行最大对应性匹配时是相同的。
因此,“序列一致性百分比”是指通过在一个比对窗口上比较两段最大化匹配的序列所决定的值,其中在所述比对窗口中的多核苷酸或多肽序列的部分与(不包含附加或缺失)的参考序列相比可以包含附加或缺失(即间隙),以获得两段序列之间的最大匹配。所述百分比的计算方法是,统计相同的核酸碱基或氨基酸残基在两段序列中同时出现的位点的数量以得到匹配位点数,将此匹配位点数除以比对窗口中的总位点数,再将结果乘以100,从而得到序列一致性百分比。序列一致性百分比的可用的例子包括但不限于50%、55%、60%、65%、70%、75%、80%、85%、90%和95%,或者从50%至100%的任意整数百分比。上述一致性可以利用本文所述的任意程序来确定。
序列对比和百分比同一性或类似性可以用设计用于检测同源序列的多种比较方法来确定,这些方法包括(但不限于)LASERGENE生物信息计算包(DNASTAR Inc.,Madison,WI)的MegAligTM程序。在本专利申请案的上下文中应当理解,使用序列分析软件进行分析时,除非另外指明,否则分析结果将基于所参考程序的“默认值”。在此所用的“默认值”是指在首次初始化软件时软件最初加载的任何值或参数集。
“Clustal V序列比对方法”对应于Clustal V标记的比对方法(描述于Higgins and Sharp,CABIOS.5:151-153(1989);Higgins,D.G.等人(1992)Comput.Appl.Biosci.8:189-191)并且存在于the MegAlignTM program of the LASERGENE bioinformatics computing suite(DNASTAR Inc.,Madison,WI).对于多重比对,默认值对应于缺口罚分(GAP PENALTY)=10和缺口长度罚分(GAP LENGTH PENALTY)=10。用Clustal方法进行成对比对和蛋白质序列的百分比同一性计算的默认参数为KTUPLE=1、空位罚分=3、窗口(WINDOW)=5和DIAGONALS SAVED=5。而对于核酸,这些参数为KTUPLE=2,空位罚分=5,窗口=4和DIAGONALS SAVED=4。用Clustal V程序比对序列后,可通过查看同一程序中的“序列距离”表来获得“百分比同一性”。
“BLASTN比对方法”是由国家生物技术信息中心(National Center for Biotechnology Information,NCBI)提供的用以采用默认参数比较核苷酸序列的算法。
本领域的技术人员非常清楚,多种程度的序列同一性可用于从其它物种中鉴定多肽,其中这类多肽具有相同或相似的功能或活性。同一性百分比的可用的实例包括但不限于50%、55%、60%、65%、70%、75%、80%、85%、90%或95%、或50%至100%之间的任何整数百分比。实际上,50%至100%之间的任何整数氨基酸同一性可用于描述本发明,诸如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%或99%。此分离核苷酸片段的任何全长或部分互补的序列也是有用的。
“基因”是指能够表达特定蛋白质的核酸片段,其包括编码序列前的调控序列(5′非编码序列)和编码序列后的调控序列(3′非编码序列)。“天然基因”是指天然存在的具有其自己的调控序列的基因。“嵌合基因”是指不是天然基因的任何基因,包含在天然情况下不是一起存在的调节序列和编码序列。因此,嵌合基因可包括源于不同来源的调控序列和编码序列,或者包括源于同一来源但以不同于天然存在的方式排列的调控序列和编码序列。“外来基因”指正常情况下不存在于宿主生物中的基因,它通过基因转移导入宿主生物。外来基因可以包含插入到非天然生物体内的天然基因,或嵌合基因。“转基因”是通过转化方法导入基因组内的基因。
在应用于植物细胞时,术语“基因组”不仅包括在细胞核内发现的染色体DNA,并且还包括在细胞的亚细胞成分(例如线粒体、质体)内发现的细胞器DNA。
“经密码子最优化的基因”是其密码子使用频率经设计用以模仿宿主细胞优选的密码子使用频率的基因。
“等位基因”是染色体上占据给定位点的基因的几种供选择替代形式的其中一种。当染色体上给定位点处存在的所有等位基因相同时,植物在该位点是纯合子。如果染色体上给定位点处存在的等位基因不同的话,植物在该位点是杂合子。
“编码序列”指编码特定氨基酸序列的DNA序列。“调控序列”指位于编码序列的上游(5′非编码序列)、中间或下游(3′非编码序列),并且影响相关编码序列的转录、RNA加工或稳定性或者翻译的核苷酸序列。调控序列可包括,但不限于:启动子、翻译前导序列、内含子、多腺苷酸化识别序列、RNA加工位点、效应子结合位点和茎环结构。
“启动子”指能够控制编码序列或功能性RNA表达的DNA序列。启动子序列由近侧和较远端上游元件组成,后者经常指增强子。因此,“增强子”是能刺激启动子活性的DNA序列,它可以是启动子的天然元件,或插入以增强启动子水平或组织特异性的异源元件。启动子可以整个源于天然基因,或者由源于不同的天然存在的启动子的不同元件组成,或者甚至包括合成的DNA片段。本领域内的技术人员应当理解,不同的启动子可以在不同的组织或细胞类型中,或者在不同的发育阶段,或者响应不同的环境条件而引导基因的表达。还应认识到,由于在大多数情况下还不能完全确定调控序列的确切范围,一些变型的DNA片段可能具有相同的启动子活性。在多数情况下引起基因在大多数细胞型中表达的启动子通常称为“组成型启动子”。目前不断在发现可用于植物细胞中的不同类型的新启动子;在以下文献中可以发现多个实例:Okamuro,J.K.and Goldberg,R.B.编辑的Biochemistry of Plants,15:1-82(1989).
“翻译前导序列”指位于基因启动子序列和编码序列之间的多核苷酸序列。翻译前导序列存在于翻译起始序列的经完全加工后的mRNA上游。翻译前导序列可影响mRNA的初级转录过程、mRNA稳定性或翻译效率。翻译前导序列的实例已经有所描述(Turner,R.and Foster,G.D.,Mol.Biotechnol.3:225-236(1995))。
“3′非编码序列”,“转录终止子”或“终止序列”是指位于编码序列下游,且包括能够影响mRNA加工或基因表达的多腺苷酸化识别序列和其他序列编码调节信号的DNA序列。多腺苷酸化信号通常表征为影响多腺苷酸片添加到mRNA前体的3′末端。Ingelbrecht,I.L.等人例示了不同3′非编码序列的用途Plant Cell 1:671-680(1989)。
“RNA转录物”指由RNA聚合酶催化的DNA序列的转录所产生的产物。当RNA转录物与DNA序列拷贝完全互补时,它指初级转录物。当RNA转录物是来源于转录后加工的初级转录物的RNA序列时,它指成熟的RNA。“信使RNA”或“mRNA”指无内含子并且可以由细胞翻译成蛋白质的RNA。“cDNA”指与mRNA模板互补并用反转录酶从mRNA模板合成的DNA。cDNA可以是单链,或使用DNA聚合酶I的Klenow片段转化成双链。“有义”RNA指包括mRNA和在细胞内或体外能被翻译成蛋白质的RNA在内的RNA转录物。“反义RNA”指与全部或部分靶初级转录物或mRNA互补,并阻止靶基因表达的RNA(美国专利5,107,065)。反义RNA可以与特定基因转录物的任何部分,即5′非编码序列、3′非编码序列、内含子、或编码序列互补。“功能性RNA”指反义RNA、核酶RNA或其它不能翻译但是对细胞过程有影响作用的RNA。术语“互补的”和“反向互补的”对mRNA转录来说可互换使用,并且用以定义信使反义RNA。
术语“可操作地连接”指单个核酸片段上的核酸序列的关联,使得其中一个核酸序列的功能受到另一个核酸序列的调控。例如,当启动子能够调节编码序列的表达(即,该编码序列受到该启动子的转录控制)时,则该启动子与该编码序列可操作地连接。编码序列可以以有义或反义的取向可操作地连接至调控序列。在另一个例子中,本发明的互补RNA区域,无论直接或间接连接、5′端与目标mRNA或3′端与目标mRNA连接、或者位于目标mRNA内、或者第一互补区域以5′端而其互补序列以3′端与目标mRNA连接,均可构成可用的连接。
本文使用的标准重组DNA和分子克隆技术是本领域所熟知的并且在如下文献中有更全面的描述:Sambrook,J.,Fritsch,E.F.and Maniatis,T.,Molecular Cloning:A Laboratory Manual;Cold Spring Harbor Laboratory:Cold Spring Harbor,NY(1989)。转化方法为本领域的技术人员所熟知,并描述于下文。
“PCR”或“聚合酶链式反应”是一种用于合成大量的特定DNA片段的技术,其包括一系列重复的循环(Perkin Elmer Cetus Instruments,Norwalk,CT)。通常,双链DNA被热变性,两种与目标片段的3′端区域互补的引物在较低温度下与之退火,然后在中间温度下得到延伸。一组的上述三个连续的步骤被称为一个“循环”。
术语“重组”指例如通过化学合成或者通过用基因工程技术操纵分离的核酸片段来实现的两个原本分离的序列片段的人工组合。
术语“质粒”、“载体”和“框”指通常携带有不属于细胞中心代谢的部分的基因的染色体外元件,并且常常是环状双链DNA片段的形式。这类元件可以是源自任何来源的自主复制序列、基因组整合序列、噬菌体或单链或双链DNA或RNA的核苷酸序列(线性或环状),其中多个核苷酸序列已连接或重组进入一种独特构建体中,该独特构建体能够将所选基因产物的启动子片段和DNA序列与相应的3′末端非翻译序列一起引入细胞中。“转化框”指含有外来基因并且除了该外来基因外还含有有利于转化特定宿主细胞转化的元件的特定载体。“表达框”是指包含一种外来基因,并具有能令所述基因在一种宿主中增强表达的所述基因以外的元件的一种特定载体(即指可将一种核酸序列或片段移入其中的不连续的核酸片段)。
术语“重组构建体”、“表达构建体”、“嵌合构建体”、“构建体”和“重组DNA构建体”本文可互换使用。重组构建体包括人造的核酸片段组合,例如天然条件下不一起存在的调控序列和编码序列。例如嵌合构造可包括源于不同来源的调控序列和编码序列,或者包括源于同一来源但以不同于天然存在的方式排列的调控序列和编码序列。此类构建体可独自使用或与载体组合使用。如果使用载体,则载体的选择取决于用以转化宿主细胞的方法,该宿主细胞是本领域的技术人员所熟知的。例如可以使用质粒载体。技术人员熟知为了成功地转化、筛选和繁殖包括本发明任何分离的核酸片段的宿主细胞,必须存在于载体上的遗传元件。技术人员也将认识到不同的独立转化事件将导致不同水平和不同模式的表达(Jones等人,EMBO J.4:2411-2418(1985);De Almeida等人,Mol.Gen.Genetics 218:78-86(1989)),因此,必须筛选多次事件以获得显示期望表达水平和模式的品系。这样的筛选可以通过DNA的Southern印迹分析、mRNA表达的Northern印迹分析、蛋白表达的免疫印迹分析或表型分析等完成。
本文所用术语“表达”是指功能性终产物(例如,一种mRNA或一种蛋白质(既可以是前体也可以是成熟形式))的生产。
术语“导入”是指将核酸(例如表达构建体)或蛋白提供至细胞中。导入包括指将核酸整合进真核或原核细胞中,在该细胞中核酸可以整合进细胞的基因组内,以及包括指将核酸或蛋白暂时提供给细胞。导入包括指稳定的或暂时性的转化方法以及有性杂交。因此,将核酸片段(例如重组DNA构建体/表达构建体)插入细胞内的情况下的“导入”意指“转染”或“转化”或“转导”,并且包括指将核酸片段整合进真核或原核细胞中,在该细胞中核酸片段可以整合进细胞的基因组(如染色体、质粒、质体或线粒体DNA)内、转变成自主的复制子或瞬时表达(例如转染的mRNA)。
“成熟”蛋白质指经翻译后加工的多肽(即已经去除了存在于初始翻译产物中的任何前肽或肽原的多肽)。“前体”蛋白质指mRNA的初级翻译产物,即前肽和肽原仍然存在。前肽和肽原可以是(但不限于)细胞内定位信号。
“稳定转化”指将核酸片段转入宿主生物的基因组以产生稳定的遗传特性,该基因组包括核基因组和细胞器基因组。相反“瞬时转化”指将核酸片段转入宿主生物的核中或包含DNA的细胞器中,在无整合或无稳定的遗传特性的情况下引起基因表达。含有转化核酸片段的宿主生物被称为“转基因”生物体。
如本文所用的“转基因”是指其基因组内含有异源多核苷酸的植物或细胞。优选的是,异源多核苷酸被稳定地整合进基因组中,使得该多核苷酸能传递至连续的世代。异源多核苷酸可以单独地或作为表达构建体的部分整合进基因组中。本文所用的转基因包括因异源核酸存在而已经改变了基因型的任何细胞、细胞系、愈伤组织、组织、植物部分或植物,包括初始进行此类改变的转基因以及从初始的转基因通过有性杂交或无性繁殖产生的那些转基因。如本文所用的术语“转基因”不涵盖通过常规植物育种方法或通过诸如随机异花受精、非重组病毒感染、非重组细菌转化、非重组转座或自发突变之类的自然发生事件导致的基因组(染色体基因组或染色体外基因组)改变。
“反义抑制”指产生能够抑制靶蛋白表达的反义RNA转录物。“共抑制”是指产生能抑制相同的或充分相似的外来或内源性基因表达的有义RNA转录本(美国专利5,231,020)。以前已经通过集中正向过表达具有与内源mRNA同源性的核酸序列设计出了植物中的共抑制构建体,它导致所有与过表达序列具有同源性的RNA减少(Vaucheret等人,Plant J.16:651-659(1998);Gura,Nature 404:804-808(2000))。该现象的总体效率较低,并且RNA减少的程度变化较大。更为近期的工作描述了以互补的方向与一种mRNA编码序列的部分或全部结合,从而使表达的RNA产生一种潜在的“茎-环”结构的“发夹”结构的使用(PCT专利公布WO 99/53050,公布于1999年10月21日;PCT专利公布WO 02/00904,公布于2002年1月3日)。这在获得的转基因植物中提高了共抑制的频率。另一种变型描述了将植物病毒序列用于引导对近端mRNA编码序列的抑制,或“沉默”,(于1998年8月20日公开的PCT专利公布WO 98/36083)。这些共抑制现象均未从机制上得到阐明,但遗传证据已经开始揭示这种复杂的情况(Elmayan等人,Plant Cell,10:1747-1757(1998))。
术语“含油的”指那些趋于以脂质形式贮存它们的能源的生物(Weete,Fungal Lipid Biochemistry,第二版,Plenum,1980)。FungalLipid Biochemistry,第二版,Plenum,1980)。一类被鉴定为含油的植物通常被称为“油料种子”植物。油料种子植物的例子包括但不限于:大豆(大豆属)、亚麻(亚麻属)、卡诺拉(芸苔属)、玉米、棉花、红花(红花属)和向日葵(向日葵属)。
通常,含油微生物的细胞油脂或TAG含量符合S形曲线,其中脂质浓度增加直至在晚对数培育生长期或早稳定培育生长期时它达到最高浓度,随后在晚稳定培育生长期和死亡期期间逐渐下降(Yongrmanitchai and Ward,Appl.Environ.Microbiol.57:419-25(1991))。
术语“含油酵母”指那些产油并被归类为酵母的微生物。含油微生物积累油脂超过它们的干细胞重量的约25%的情形并不鲜见。含油酵母的实例包括但不限于如下属:解脂耶氏酵母属(Yarrowia)、假丝酵母属(Candida)、红酵母属(Rhodotorula)、红冬孢酵母属(Rhodosporidium)、隐球酵母属(Cryptococcus)、丝孢酵母属(Trichosporon)和油脂酵母属(Lipomyces)。
术语“植物”包括整个植株、植物器官、植物组织、种子和植物细胞,种子以及同一植株的子代。植物细胞包括但不限于得自下列物质的细胞:种子、悬浮培养物、胚、分生区域、愈伤组织、叶、根、芽、配子体、孢子体、花粉和小孢子。
“子代”包括植物的任何后续世代。
“非转基因的、无转基因分离子(null segregant)大豆种子”是指缺少所转基因的近等基因植物或种子,和/或用于转化过程以获得转基因事件的父母本植物。无转基因分离子可以是由杂合的转基因植物在繁殖中经正常的遗传分离而产生的不含转基因性状的植物或种子。
“谷粒”是玉米的颖果,由一个成熟的胚胎和作为双受精产物的胚乳组成。术语“玉米”指玉米(Zea mays L.)的任何变种、栽培种或种群。
“谷物”包括由商业种植者生产、出于除栽种或繁殖该物种以外的目的而用于农用或售予消费者的成熟的玉米谷粒。“种子”是出于繁殖该物种的目的并用于售予商业种植者而生产的成熟的玉米谷粒。本文所用术语种子、谷粒和谷物可以互换使用。术语“胚胎”或者“胚”是指快速生长期(种子萌芽)开始之前的孢子体植物幼体。玉米的胚胎(胚)包含了谷粒中所发现的绝大多数油脂。谷物类胚胎的结构包括胚轴和盾片。“盾片”是谷物类胚胎中专门吸收胚乳的单个子叶。“糊粉”是蛋白质性质的物质,通常以小颗粒的形式存在,产生于玉米以及其他谷物的胚乳的最外层细胞。
本发明涉及一种转基因大豆种子,所述转基因大豆种子的总脂肪酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量相比高出至少10%。当然,转基因相对于非转基因的、无转基因分离子在总脂肪酸含量方面的任何可测量的提高都是有用的。所述总脂肪酸含量方面的提高包括但不限于至少1%、2%、3%、4%、5%、6%、7%、8%、9%、11%、12%、13%、14%、15%、16%、17%、18%、19%、或20%。
本发明的一种转基因油料种子可以包含具有至少一种DGAT序列的重组构建体。所述DGAT序列可以选自DGAT1、DGAT2和DGAT1与DGAT2的组合。此外,至少一种DGAT序列可以来自解脂耶氏酵母。可用于实施本发明的适当的DGAT序列的例子论述于下文的实施例中。包括本发明中以下SEQ ID NO的序列:1、9、64、66、82、87和92。本领域的技术人员将认识到,本发明包括但不限于本文所公开的DGAT序列。
所述重组构建体启动子包括不同的元件,例如一个是一种DNA序列的启动子,其能指导一种植物的胞内装置从该启动子下游(3′)临近的编码序列产生RNA。所述启动子区域影响着基因的RNA转录本合成的速率、合成时的发育阶段和细胞类型。所述RNA转录物被加工为mRNA,后者作为用于将RNA序列翻译为所编码多肽的氨基酸序列的模板。5′端非翻译前导序列是mRNA上位于蛋白质编码区域上游的区域,其可能对mRNA翻译的起始和进行发挥作用。3′转录终止/多聚腺苷酸化信号是位于蛋白质编码区域下游的非翻译区域,其在植物细胞中的作用是导致RNA转录的终止和多聚腺苷酸向RNA3′端的添加。
用于驱动DGAT编码序列表达的启动子的来源并不重要,只要它具有足够的转录活性以通过在适当的时间、在所期望的宿主组织内表达所期望的核酸片段的可翻译的mRNA,从而达到本发明的目的即可。异种的或非异种的(即内源的)启动子均可用于实施本发明。例如,适当的启动子包括但不限于:β大豆伴球蛋白启动子的α引发亚单位、库尼兹(Kunitz)胰蛋白酶抑制剂3启动子、膜联蛋白启动子、大豆球蛋白Gy1启动子、β大豆伴球蛋白启动子的β亚单位、P34/Gly Bd m 30K启动子、白蛋白启动子、Leg A1启动子和Leg A2启动子。
上述膜联蛋白或P34启动子描述于PCT专利公布WO2004/071178(公布于2004年8月26日)。上述膜联蛋白启动子的活性水平与任何已知的强启动子相当,所述强启动子如:(1)CaMV35S启动子(Atanassova等人,Plant Mol.Biol.37:275-285(1998);Battraw and Hall,Plant Mol.Biol.15:527-538(1990);Holtorf等人,Plant Mol.Biol.29:637-646(1995);Jefferson等人,EMBO J.6:3901-3907(1987);Wilmink等人,Plant Mol.Biol.28:949-955(1995));(2)拟南芥油质蛋白(oleosin)启动子(Plant等人,Plant Mol.Biol.25:193-205(1994);Li,Texas A&M University Ph.D.dissertation,第107-128页(1997));(3)拟南芥泛素延伸蛋白启动子(Callis等人,J Biol.265(21):12486-93(1990));(4)一种番茄泛素基因启动子(Rollfinke等人,Gene.211(2):267-76(1998));(5)一种大豆热激蛋白启动子(Schoffl等人,Mol Gen Genet.217(2-3):246-53(1989));以及(6)一种玉米H3组蛋白基因启动子(Atanassova等人,Plant Mol Biol.37(2):275-85(1989))。
上述膜联蛋白启动子的另一有用特性是其在发育种子中的表达谱。上述膜联蛋白启动子在早期(授粉后10天以内)的发育中种子中最为活跃,而在随后的时期基本保持沉默。上述膜联蛋白启动子的表达谱与许多种子特异性启动子的都不同,后者如通常在发育后期表现出最高活性的种子贮藏蛋白启动子(Chen等人,Dev.Genet.10:112-122(1989);Ellerstrom等人,Plant Mol.Biol.32:1019-1027(1996);Keddie等人,Plant Mol.Biol.24:327-340(1994);Plant等人,(supra);Li,(supra))。上述膜联蛋白启动子具有更加常规的表达谱,不过仍与其他已知的种子特异性启动子不同。因此,当希望在早期发育阶段过表达或抑制一个基因时,上述膜联蛋白启动子是一个很有吸引力的选择。例如,可能希望过表达一个调控早期胚胎发育的基因或一个在种子成熟之前参与代谢的基因。
在鉴定出适合表达特异性的DGAT编码序列的适当启动子之后,利用为本领域的技术人员所熟知的传统方法,将所述启动子以有义方向进行可操作地连接。
本文所用的标准的重组DNA和分子克隆技术为本领域所熟知,并且Sambrook,J.等人在Molecular Cloning:A Laboratory Manual;2nded.;Cold Spring Harbor Laboratory Press:Cold Spring Harbor,New York,1989(以下简称“Sambrook等人,1989”)或Ausubel,F.M.,Brent,R.,Kingston,R.E.,Moore,D.D.,Seidman,J.G.,Smith,J.A.and Struhl,K.,编辑,;在Current Protocols in Molecular Biology;John Wiley and Sons:New York,1990(以下简称“Ausubel等人,1990”)中有更加详尽的描述。
另一方面,本发明涉及一种提高大豆种子的总脂肪酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量相比高出至少10%的步骤(a)中的转化的大豆细胞。
在构建了重组构建体之后,可以通过为本领域的普通技术人员所熟知的方法(例如转染、转化和电穿孔),将其导入一种植物细胞。优选的植物细胞为油料种子植物细胞。然后在允许对总脂肪酸含量比非转基因的、无转基因分离子大豆种子的总脂肪酸含量高出至少10%的转化的大豆细胞进行选择的适当条件下,对转化的植物细胞进行人工培养和再生。
此类重组构建体可以被导入至一个植物细胞;或者也可将每一重组构建体导入至不同的植物细胞。
如前所述,在植物细胞中的表达方式可以是暂时性的也可以是稳定的。
从此类转化的植物获得的种子或植物部分也属于本发明的范围。
植物部分包括分化和未分化的组织,包括但不限于下列部分:根、茎、芽、叶、花粉、种子、肿瘤组织和多种形式的细胞和培养物(例如单个细胞、原生质体、胚和愈伤组织)。植物组织可存在于植株或植物器官、组织或细胞培养物中。
术语“植物器官”是指构成植株的形态和功能不同部分的植物组织或组织群。术语“基因组”是指:(1)存在于生物体的每一个细胞或者病毒或细胞器中的全套遗传物质(基因和非编码序列);和/或(2)作为(单倍体)单位从一个亲本遗传的全套染色体。
用于转化双子叶植物(主要利用根癌农杆菌)并获得转基因植物的方法,已与其他方法一起公布于下列文献:棉花(美国专利5,004,863;美国专利5,159,135);大豆(美国专利5,569,834;美国专利5,416,011);芸苔属植物(美国专利5,463,174);花生(Cheng等人.Plant Cell Rep.15:653-657(1996);McKently等人.Plant Cell Rep.14:699-703(1995));番木瓜果(Ling,K.等人Bio/technology 9:752-758(1991));以及豌豆(Grant等人.Plant Cell Rep.15:254-258(1995))。对植物转化的其他常用方法的综述参见Newell,C.A.(Mol.Biotechnol.16:53-65(2000))。这些转化方法之一利用了发根农杆菌(Tepfler,M.and Casse-Delbart,F.Microbiol.Sci.4:24-28(1987))。已公布的利用直接输送DNA进行的大豆转化,有利用PEG融合(PCT专利公布WO 92/17598)、利用电穿孔(Chowrira,G.M.等人,Mol.Biotechnol.3:17-23(1995);Christou,P.等人,Proc.Natl.Acad.Sci.U.S.A.84:3962-3966(1987))、利用显微注射和利用基因枪轰击(McCabe,D.E.等人,Bio/Technology 6:923(1988);Christou等人,Plant Physiol.87:671-674(1988))进行的。
存在多种用于从植物组织再生植物的方法。再生的具体方法将取决于起始植物组织以及待再生的具体植物物种。从单植物原生质体转化体或从多种经转化的外植体再生、发育和培育植物是本领域所熟知的(Weissbach和Weissbach(编辑),载于:Methods for Plant Molecular Biology,(Eds.),Academic:San Diego,CA(1988))。该再生和生长方法通常包括如下步骤:选择转化的细胞和培养这些单独化的细胞通过胚发育的通常阶段以及通过生根小植株阶段。转基因胚以及种子以类似的方式再生。随后将所得的转基因的生根小苗种植在诸如土壤之类的合适植物生长培养基中。优选地,将再生的植物进行自花授粉以产生纯合的转基因植物。或者,将得自再生植物的花粉与农学上重要的品系的产生种子的植株进行杂交。相反,将来自这些重要品系的植物用于给再生植物授粉。利用本领域技术人员所熟知的方法培育含有所需多肽的本发明的转基因植物。
除以上所讨论的程序以外,专业人员亦熟悉描述了用于下列目的特定条件和程序的标准源资料:大分子的构建、操作和分离(例如DNA分子、质粒等等);重组DNA片段和重组表达体的构建;以及克隆的筛选和分离。例如参见:Sambrook等人,Molecular Cloning:A Laboratory Manual,Cold Spring Harbor:NY(1989);Maliga等人,Methods in Plant Molecular Biology,Cold Spring Harbor:NY(1995);Birren等人,Genome Analysis:Detecting Genes,第1卷,Cold Spring Harbor:NY(1998);Birren等人,Genome Analysis:Analyzing DNA,第2卷,Cold Spring Harbor:NY(1998);Plant Molecular Biology:A Laboratory Manual,编辑,Clark,Springer:NY(1997)。
油料种子植物的例子包括但不限于:大豆、芸苔属植物、向日葵、玉米、棉花、亚麻和红花。
另一方面,本发明涉及一种转基因玉米谷粒,所述转基因玉米谷粒的总脂肪酸含量与非转基因的、无转基因分离子玉米谷粒的总脂肪酸含量相比高出至少10%。所述转基因玉米谷粒可以包含具有至少一种DGAT序列的重组构建体。所述DGAT序列可以选自DGAT1、DGAT2和DGAT1与DGAT2的组合。
在另一方面,本发明涉及一种提高玉米谷粒的总脂肪酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个玉米谷粒;
(b)选择总脂肪酸含量与非转基因的、无转基因分离子玉米谷粒的总脂肪酸含量相比高出至少10%的步骤(a)中的转化的玉米谷粒。
本发明还涉及一种转基因大豆种子,与非转基因的、无转基因分离子大豆种子的对应含量相比,所述转基因大豆种子的总脂肪酸含量高出至少10%并且油酸含量高出至少25%。并且,本发明还涉及一种转基因大豆种子,与非转基因的、无转基因分离子大豆种子的对应含量相比,所述转基因大豆种子的总脂肪酸含量高出至少10%,并且具有至少一种下列特征:i)油酸含量高出至少25%;ii)亚麻酸含量降低至少25%;iii)亚油酸含量降低至少4%;iv)棕榈酸含量降低至少8%;以及v)硬脂酸含量高出至少14%。
此外的另一方面,本发明还涉及一种提高大豆种子的总脂肪酸含量和油酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和油酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和油酸含量相比分别高出至少10%和至少25%的步骤(a)中的转化的大豆细胞。
另一方面,本发明还涉及一种提高大豆种子的总脂肪酸含量并降低其亚麻酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和亚麻酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和亚麻酸含量相比分别高出至少10%和降低至少25%的步骤(a)中的转化的大豆细胞。
再一方面,本发明还涉及一种提高大豆种子的总脂肪酸含量并降低其亚油酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和亚油酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和亚油酸含量相比分别高出至少10%和降低至少4%的步骤(a)中的转化的大豆细胞。
再一方面,本发明还涉及一种提高大豆种子的总脂肪酸含量并降低其棕榈酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和棕榈酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和棕榈酸含量相比分别高出至少10%和降低至少8%的步骤(a)中的转化的大豆细胞。
再一方面,本发明涉及一种提高大豆种子的总脂肪酸含量和硬脂酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和硬脂酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和硬脂酸含量相比分别高出至少10%和至少14%的步骤(a)中的转化的大豆细胞。
如前文所论及,本文所述的任何一种转基因油料种子均可包含具有至少一种DGAT序列的重组构建体。所述DGAT序列可以选自DGAT1、DGAT2和DGAT1与DGAT2的组合。此外,至少一种DGAT序列来自解脂耶氏酵母。
利用电穿孔、基因枪轰击和农杆菌对单子叶植物进行的转化已有报道。已实现转化和植物再生的包括:芦笋(Bytebier等人,Proc.Natl.Acad.Sci.(USA)84:5354,(1987));大麦(Wan and Lemaux,Plant Physiol 104:37(1994));玉米(Rhodes等人,Science 240:204(1988),Gordon-Kamm等人,Plant Cell 2:603-618(1990),Fromm等人,BiolTechnology 8:833(1990),Koziel等人,BiolTechnology 11:194,(1993),Armstrong等人,Crop Science 35:550-557(1995));燕麦(Somers等人,BiolTechnology 10:1589(1992));鸭茅(Horn等人,Plant Cell Rep.7:469(1988));水稻(Toriyama等人,TheorAppl.Genet.205:34,(1986);Part等人,Plant Mol.Biol.32:1135-1148,(1996);Abedinia等人,Aust.J.Plant Physiol.24:133-141(1997);Zhang和Wu,Theor.Appl.Genet.76:835(1988);Zhang等人Plant Cell Rep.7:379,(1988);Battraw和Hall,Plant Sci.86:191-202(1992);Christou等人,Bio/Technology9:957(1991));裸麦(De la Pena等人,Nature 325:274(1987));甘蔗(Bower和Birch,Plant J.2:409(1992));高羊茅(Wang等人,BiolTechnology 10:691(1992)),和小麦(Vasil等人,Bio/Technology10:667(1992);美国专利5,631,152)。
通过利用聚乙二醇处理、电穿孔或基因枪轰击将核酸分子导入植物细胞,已发展出基于克隆核酸构造的短暂表达的基因表达分析法(Marcotte等人,Nature 335:454-457(1988);Marcotte等人,Plant Cell1:523-532(1989);McCarty等人,Cell 66:895-905(1991);Hattori等人,Genes Dev.6:609-618(1992);Goff等人,EMBO J.9:2517-2522(1990))。
短暂表达体系可用于对基因结构进行功能性剖析(参见Maliga等人,Methods in Plant Molecular Biology,Cold Spring Harbor Press(1995))。当然,本发明的任何核酸分子都能以永久性或暂时性的方式与其他遗传元件,如载体、启动子、增强子等一起,被导入至植物细胞。
除以上所讨论的程序以外,专业人员亦熟悉描述了用于大分子的构建、操作和分离(例如DNA分子、质粒等等)、重组生物体的构建以及克隆的筛选和分离的特定条件和程序的标准源资料(例如参见Sambrook等人,Molecular Cloning:A Laboratory Manual,Cold Spring Harbor Press(1989);Maliga等人,Methods in Plant Molecular Biology,Cold Spring Harbor Press(1995);Birren等人,Genome Analysis:Detecting Genes,1,Cold Spring Harbor,New York(1998);Birren等人,Genome Analysis:Analyzing DNA,2,Cold Spring Harbor,New York(1998);Plant Molecular Biology:A Laboratory Manual,编辑,Clark,Springer,New York(1997))。
本发明的转基因大豆种子可用于加工生产大豆油脂、大豆产品和/或大豆副产品。
“大豆产品”可以包括但不限于表1A所列的项目。
表1A
由大豆种子获得的大豆蛋白产品
a
a 参见Soy蛋白质Products:Characteristics,Nutritional Aspects and Utilization(1987).大豆蛋白委员会。
“加工”是指用以获得表1A中所列产品的任何物理和化学方法,所述方法包括但不限于对完整种子或种子局部进行的热处理、去皮与碾磨、挤压、溶剂萃取或水浸萃取。此外,“加工”还包括用以从完整种子或种子局部浓缩和提取大豆蛋白质的方法,以及东方国家制作大豆发酵食物产品的各种传统方法。关于建立起多种上述产品的贸易标准和规格(参见National油脂种子Processors Association Yearbook and Trading Rules 1991-1992)。当指某种产品为“高蛋白的”或“低蛋白的”时,此产品如上述标准规格所述。“NSI”是指如如美国油脂化学家协会方法Ac4 41所定义的氮溶解指数。“KOH氮溶解度”是豆粕质量的一个指标,其是指在如Araba和Dale((1990)Poult.Sci.69:76-83)所述的条件下,溶解于0.036M KOH中的氮的量。“低变性”豆粕是指在控制湿热以令NSI为约85至90的条件下经过处理并脱脂的、脱壳去皮的大豆子叶。此术语亦可指被碾磨至能通过100号美国标准筛尺寸的、具有相似NSI的大豆粉。“煮熟的”是指NSI为约20至60的大豆蛋白产品,典型地为一种大豆粉。“烤的”是指NSI低于20的大豆蛋白产品,典型地为一种大豆粉。“粗磨粉”是指在10号和80号美国标准筛尺寸之间的脱脂去壳的子叶。“大豆蛋白浓缩物”是指脱脂去壳的大豆经以下三个基本步骤所获得的产品:酸浸(pH约为4.5),酒精(约55%至80%)萃取,以及在水萃取之前用湿热处理令蛋白质变性。用于制备大豆蛋白浓缩物的典型条件描述于Pass((1975)美国专利3,897,574和Campbell等人,(1985)in New Protein Foods,由Altschul and Wilcke编辑,Academic Press,第5卷,第10章,Seed Storage Proteins,第302-338页)。“挤压”是指令原料(粗磨粉、粉或浓缩物)在高温高压下通过一种带外壳的螺旋输送机,作为一种改变所述原料构造的方法的过程。“质构化”和“构造化”是指用以改变所述原料物理性状的挤压过程。包括热塑挤压在内的上述过程的特征此前已有描述(Atkinson(1970)美国专利3,488,770,Horan(1985)In New Protein Foods,ed.by Altschul and Wilcke,Academic Press,第1A卷,第八章,第367-414页]。此外,包含大豆蛋白产品的复合食品混合物的挤压过程所使用的条件此前也已有描述(Rokey(1983)Feed Manufacturing Technology III,222-237;McCulloch,美国专利4,454,804)。
表1B
大豆油脂和副产品生产的一般步骤
更准确地说,大豆种子经清洁、回火、脱壳和去皮从而提高了油脂萃取的效率。油脂的萃取通常利用溶剂(例如己烷)萃取来完成,但也可以通过联合使用物理压榨和/或溶剂萃取而实现。所得到的油脂称为原油。可以通过使磷脂以及其他极性和中性的脂类复合物发生水合反应从而促进它们与非水合的甘油三酯部分(大豆油脂)分离,而令上述原油脱胶。所得到的卵磷脂胶可以被深加工为作为乳化剂和脱模(即抗粘结)剂用于多种食品和工业品中的具有重要商业意义的卵磷脂产品。脱胶油可以进一步精炼以除去杂质(主要是游离脂肪酸、色素和残留胶)。通过添加一种与游离脂肪酸反应形成皂并使上述原油中的磷脂和蛋白质发生水合反应的腐蚀剂而实现精炼。用水洗去精炼过程所形成的皂类痕迹。上述皂脚副产品可直接用于动物饲料或将其酸化以回收游离脂肪酸。利用一种漂白土的吸附作用除去大部分的叶绿素和类胡萝卜素复合物而实现脱色。精炼的油脂可以是氢化的,从而得到具有不同熔解性质和构造的脂肪。可以通过在精细控制的冷却条件下结晶,从而利用冷凝(分馏)从氢化油脂中除去硬脂。脱臭(主要通过真空蒸汽蒸馏)是最后一个步骤,目的是除去那些令油脂具有臭味或气味的复合物。脱臭过程中还可以除去其他有价值的副产品,如生育酚和甾醇类。包含这些副产品的脱臭馏出液可以出售,其可用于生产天然维生素E以及其他高价值的药品。经精炼、漂白、(氢化、分馏)以及脱臭的油脂和脂肪可以直接包装出售,或深加工为更加专门化的产品。关于大豆种子加工、大豆油脂生产以及副产品利用的更加详尽的参考文献参见Erickson,Practical Handbook of Soybean Processing and Utilization,The American 油脂Chemists’Society and United Soybean Board(1995)。大豆油脂同椰子、棕榈、棕榈仁以及可可等的油脂相比饱和脂肪酸含量相对较低,因此其在室温下为液态。
包含经精炼和/或纯化的PUFA的植物和微生物油脂可以被氢化,从而得到具有不同熔解性质和构造的脂肪。多种经加工的脂肪(包括涂抹油、糖果脂肪、硬质脂肪、人造黄油、烤酥油等)要求在室温下具有的硬度各不相同,而它们只能通过改变原料油脂的物理特性而制得。通常通过催化氢化实现这点。
氢化作用是在催化剂,如镍的催化下,将氢加到不饱和脂肪酸双键上的化学反应。例如,高油酸大豆油脂包含不饱和的油酸、亚油酸和亚麻酸,而上述每种脂肪酸都能被氢化。氢化作用有两个主要的效应。第一,由于不饱和脂肪酸含量的减少,油脂的氧化稳定性得到提高。第二,由于脂肪酸的修饰提高了熔点,改变了油脂的物理性质,从而得到在室温下为半液态或固态的脂肪。
影响氢化反应的参数有很多,它们进而又影响着终产品的组成。包括压力、温度、催化剂的类型和浓度、搅拌以及反应器设计在内的操作条件属于可控的重要参数范畴。可以利用选择性的氢化反应条件来实现对不饱和程度较高的脂肪酸相对于不饱和程度较低的脂肪酸的优先氢化。轻微或涂刷氢化(brush hydrogenation)常被用于提高液态油脂的稳定性。进一步的氢化则使得液态油脂转化为一种物理上的固态脂肪。氢化的程度依赖于针对特定的终产物所期望的性能和熔解性质。通过氢化可以获得无数种可能的油脂和脂肪产品,包括液态酥油(用于制作焙烤产品,用于商业化的油炸和烘烤操作的固态脂肪和酥油)以及用于制造人造黄油的基础油。对氢化作用和氢化产品的更加详尽的描述参见Patterson,H.B.W.,Hydrogenation of Fats and油脂s:Theory and Practice.The American油脂Chemists’Society(1994)。
由于氢化过程中会产生反式的脂肪酸同分异构体,关于氢化油脂已存在一些争议。摄食大量的反式同分异构体被认为与一些有害健康的效应存在联系,包括血浆中低密度脂蛋白对高密度脂蛋白的比率的升高,以及罹患冠心病的风险升高。
本发明的另一方面涉及一种分离的多核苷酸,所述多核苷酸包括:
(a)编码具有二酰基甘油酰基转移酶活性的多肽的核苷酸序列,其中所述多肽如SEQ ID:83、88或93所示;
(b)(a)中的核苷酸序列的互补序列,其中所述互补序列与所述核苷酸序列由相同数目的核苷酸组成并且100%互补。
以及,包含与至少一种调控序列可操作地连接的所述分离的核酸片段的重组DNA构建体。以及,在其基因组中包含所述重组DNA构建体的细胞。
所述细胞可以是一种油料种子植物的一部分,例如但不限于,大豆、玉米、卡诺拉(canola)、向日葵、亚麻、棉花和红花。
最后,本发明涉及一种提高油料种子的总脂肪酸含量的方法,所述方法包括:
(a)用上述重组构建体转化至少一个油料种子细胞;
(b)选择总脂肪酸含量高于非转基因的、无转基因分离子油料种子的总脂肪酸含量的步骤(a)中的转化的油料种子细胞。
实施例
本发明将在下面的实施例中进一步限定,其中份数和百分比是以重量计并且度数是摄氏度,除非另外说明。应该理解,尽管这些实施例说明了本发明的优选实施方案,但仅是以例证的方式给出的。根据上面的论述和这些实施例,本领域的技术人员能确定本发明的基本特征,并在不脱离本发明的精神和范围的情况下,能够对本发明做出多种改变和修饰,以使其适用于多种用法和条件。因此,除了那些本文所示和描述的那些之外,根据前文所述,本发明的各种修改形式对本领域的技术人员来说将是显而易见的。这些修改形式也旨在属于附加的权利要求书的范围内。
缩写的含义如下:“sec”表示秒,“min”表示分钟,“h”表示小时,“d”表示天,“μL”表示微升,“mL”表示毫升,“L”表示升,“μM”表示微摩尔浓度,“mM”表示毫摩尔浓度,“M”表示摩尔浓度,“mmol”表示毫摩尔,“μmole”表示微摩尔,“g”表示克,“μg”表示微克,“ng”表示纳克,“U”表示单位,“bp”表示碱基对而“kB”表示千碱基对。
实施例1
解脂耶氏酵母DGAT基因在酿酒酵母中的表达
通过限制酶NcoI和NotI的消化,将解脂耶氏酵母DGAT1基因(SEQ ID NO:1)自质粒载体pYDA1(SEQ ID NO:2)上切下。用T4DNA聚合酶(Promega,Madison,WI,USA)将DNA片段的末端补平,并连接到pY75(SEQ ID NO:3)上惟一的NotI位点。在用于克隆之前,先将pY75载体用NotI线性化,用T4DNA聚合酶补平,并用虾碱性磷酸酶((NEB,Beverly,MA,USA))去磷酸化。利用标准方法分离质粒DNA,并用EcoRI进行限制性消化,以鉴定出起始密码子邻近pY75中GPD启动子3′端的质粒克隆(DGAT1基因的有义方向)。鉴定出的阳性克隆质粒称为pY75 YL DGAT1(SEQ ID NO:4)。pYDA1的构建描述于PCT专利公布WO 2006/052914,此公布在此引入本文以供参考。
酵母附加体质粒(YEp)型载体pRS425(SEQ ID NO:5)(Christianson等人,Gene 110:119-122(1992))包含来自酿酒酵母2微米内源性质粒的序列、一个LEU2选择性标记以及基于一种多功能噬菌粒,pBluescript II SK(+)的序列。酿酒酵母组成型的甘油醛-3-磷酸脱氢酶(GPD)强启动子按照如Jia等人(Physiol.Genomics 3:83-92(2000))to produce pGPD-425(SEQ ID NO:6)所述的相同方式被克隆至pRS425的SacII和SpeI位点之间,从而得到pGPD-425(SEQ ID NO:6)。一个NotI位点被引入至pGPD-425的BamHI位点,从而得到两侧为BamHI位点的一个NotI位点,该质粒称为pY75(SEQ ID NO:3)。
DGAT2基因按下述方法PCR扩增至解脂耶氏酵母(ATCC登录号20362)基因组。酵母细胞在固体YPD培养基上生长72h。将细胞悬浮于200μL的DNA提取缓冲液(100mM Tris pH 7.5,10mMEDTA,100mM NaCl,0.1%Triton X-100)并加入2至5个玻璃珠(直径3mm)和约0.1g的玻璃珠(直径0.5mm)。用旋涡混合器将酵母细胞悬液剧烈振荡混匀,75℃孵育25min。将溶菌液冷却至室温并离心澄清。
利用下列两种寡核苷酸引物以产生一种约1600bp的PCR片段:
oYLDGAT2-1:
GCGGCCGCATGACTATCGACTCACAATACTACAAGT(SEQ IDNO:7),和
oYLDGAT2-2:
GCGGCCGCTTACTCAATCATTCGGAACTCTGGGGCT(SEQ IDNO:8)。
简言之,制备了一种包含2.5mM MgCl2、2mM dNTPs、10mMTris/HCl(pH 8.8)、50mM KCl、0.08%Nonidet P40、1μM oYLDGAT2-1(SEQ ID NO:7)和oYLDGAT2-2(SEQ ID NO:8)、10U Taq聚合酶(Fermentas,Hanover,MD)以及2μL酵母溶菌液的PCR反应混合液(100μL)。将上述PCR混合液分为四份25μL的等份,进行35个循环的扩增,每一循环包括94℃45sec,各退火温度45sec,以及72℃1min。将PCR产物进行凝胶纯化,并按生产商的说明书克隆至pGEM T-easy(Promega)。
对十个独立的质粒克隆进行了完整测序。该分析的共有序列如SEQ ID NO:9所示。该DNA序列在两个不同的核苷酸位点与PCT专利公布WO 2005/003322所公开的DGAT2序列不一致。DNA序列的差异影响了DGAT2开放阅读框的nt448和nt672。前一个nt差异改变了预测的DGAT蛋白氨基酸序列。它将PCT专利公布WO2005/003322所公开的DGAT序列中的一个丝氨酸替换为苏氨酸残基。第二个序列差异并未改变PCT专利公布WO 2005/003322所公开的氨基酸序列。本文所公开的解脂耶氏酵母DGAT2序列和PCT专利公布WO 2005/003322所公开的解脂耶氏酵母DGAT2序列之间的差异可以归因于用于分离DGAT2基因的解脂耶氏酵母菌株之间的差异。菌株ATCC登录号20362的DGAT蛋白所预测的氨基酸序列如SEQ ID NO:10所示。
将DGAT基因(SEQ ID NO:9)作为NotI限制性片段自pGEMT-easy载体上切下,并连接到经NotI线性化并去磷酸化的pY75(SEQ ID NO:3)DNA上。从重组构建体克隆分离质粒DNA,并用SacI和PacI进行限制性消化,以鉴定出DGAT2的起始密码子邻近pY75中GPD启动子3′端的克隆(DGAT2基因的有义方向)。鉴定出的阳性克隆质粒称为pY75 YL DGAT2(SEQ ID NO:11)。
利用标准方法(Gietz,R.Daniel;Woods,Robin A.,Meth.Enzymol.350:87-96(2002))将pY75YL DGAT1(SEQ ID NO:4)质粒DNA和pY75空载体转入酿酒酵母菌株INVSC1(Invitrogen,USA)。在添加了CSM-leu(Qbiogene,Carlsbad,CA)的DOBA培养基上对重组酵母克隆进行选择。将五个独立生长的克隆分别接种到五份50mL的添加了CSM-leu的DOBA培养基中,并在30℃培养72h。离心收集细胞,并重悬于除去除了作为氮源的硫酸铵以外同上述DOBA培养基相同的培养基中。将培养物再培养60h,离心收集细胞。将细胞在干冰上冷冻并冻干。
将每一酵母细胞样品分为一式三份测定其总脂肪酸含量如下。约5至15毫克的酵母粉末被称重并置于一个带有旋盖和特氟隆密封件的13x 100mm的玻璃培养管底部。加入5μL 17:0TAG(10mg/mL,溶于甲苯)储备液,再加入500μL溶于(无水)甲醇的5%硫酸。将样品在95℃孵育1.5h。待玻璃管冷却至室温后加入1mL 1M的氯化钠并混匀。加入1mL庚烷,混匀玻璃管内容物并将样品短暂旋转以实现相分离。将约500μL的有机相转入一个气相色谱小瓶。用气相色谱法分析脂肪酸甲基酯。在配套了Omegawax 320熔融石英毛细管柱(Supelco Inc.,Catalog No.24152)的Hewlett-Packard 6890气相色谱仪上分析4μL的庚烷萃取液。设定炉温程序如下:220℃保持2.7分钟,然后以20℃/分钟的速度升温至240℃并再保持2.3分钟。载气通过Whatman氢气发生器供应。保留时间与那些可商购获得的标准甲酯比较(Nu-Chek Prep,Inc.catalog#U-99-A)。
如前所述,将pY75YL DGAT2(SEQ ID NO:11)质粒DNA和pY75空载体转入酿酒酵母菌株INVSC1,并分析重组酵母培养物的总脂肪酸含量。与在酵母中过表达两种DGAT基因相关的结果总结于表2中。
表2
酿酒酵母培养物的总脂肪酸含量
表2显示,同仅携带pY75空质粒的细胞相比,携带pY75YLDGAT1(SEQ ID NO:4)的酵母细胞中总脂肪酸含量有显著提高。在相同的培养条件下,五个独立培养物平均的脂肪酸细胞干重(DCW)百分比为11.9%,而空载体对照组为9.3%。总而言之,总脂肪酸产量提高了28%。此外,脂肪酸谱还发生了同YL DGAT1的表达相关的轻微变化,表现为棕榈酸的增加。
与相同条件下培养的空载体对照组相比,在氮饥饿培养条件下YL DGAT2的组成型表达令总脂肪酸含量提高了110%。空载体对照组的总脂肪酸含量为6.6%,而YL DGAT2转化体的平均脂肪酸含量为14.2%。YL DGAT2的表达导致了脂肪酸谱的改变。棕榈酸含量显著提高,与之相伴的是棕榈油酸含量发生中等程度的降低。此外,伴随着油酸含量的明显降低,硬脂酸含量发生了显著提高。上述结果总体上显示,在提高了碳/氮比率的条件下,YL DGAT在酵母中的过表达导致了脂肪酸累积的增加。YL DGAT酶类的过表达能够在酿酒酵母中提高内源性DGAT活性。两种DGAT基因的类似实验均重复了两次以上。每次均能在YL DGAT培养物和空载体对照组之间观察到脂肪酸含量的差异,所述差异就YL DGAT1(SEQ ID NO:1)而言为至少8%,就YL DGAT2(SEQ ID NO:9)而言为至少14.3%。
实施例2
解脂耶氏酵母DGAT1和DGAT2基因向解脂耶氏酵母表达载体中
的克隆
本实施例描述了分别包含FBAINm::YDGAT1::PEX20嵌合基因和FBAINm::YDGAT2::PEX20嵌合基因的pFBAIN-YDG1和pFBAIN-YDG2的制备。上述质粒被设计用于DGAT1和DGAT2在解脂耶氏酵母中的过表达。
设计并合成用以从解脂耶氏酵母基因组DNA(分离自ATCC登录号20362菌株,购自American Type Culture Collection(Rockville,MD))扩增DGAT1ORF的寡核苷酸YDGAT1-F(SEQ ID NO:12)和YDGAT-R(SEQ ID NO:13),同时设计并合成用以扩增DGAT2ORF的寡核苷酸YDGAT2-F(SEQ ID NO:14)和YDGAT2-R(SEQ IDNO:15)。
用解脂耶氏酵母基因组DNA作为模板的PCR反应,每一反应总体积为50μL,包括:20μM的正向和反向引物各1μL,1μL基因组DNA(100ng),10μL 5X PCR缓冲液,1μL dNTP混合液(每种10μM),35μL水以及1μL Phusion聚合酶(New England Biolabs,Inc.,Ipswich,MA)。扩增程序为:98℃1min;98℃10sec、55℃10sec、72℃30sec共30个循环;最终延伸步骤为72℃5min。得到分别包含DGAT1 ORF和DGAT2ORF的1603bp DNA片段(SEQ ID NO:16)和1567bp DNA片段(SEQ ID NO:17)。
按生产商的说明书用Qiagen PCR Purification试剂盒纯化上述PCR片段。用NcoI和NotI消化纯化后的DNA样品,继而用Qiagen reaction clean-up试剂盒纯化,然后定向连接至NcoI/NotI消化后的pFBAIN-MOD-1(图1C;SEQ ID NO:18)。具体而言,上述连接反应包含:10μL 2X连接缓冲液,1μL T4DNA连接酶(Promega),1600bp片段(即DGAT1;SEQ ID NO:16)或1564bp片段(即DGAT2;SEQ ID NO:17)4μL(约300ng)以及1μL pFBAIN-MOD-1(约150ng)。将反应混合液在室温孵育2h,并用其转化E.coli Top10感受态细胞(Invitrogen)。用Qiagen Miniprep试剂盒回收转化体中的质粒DNA。通过限制梅切作图鉴定出正确的克隆,最终的质粒构造分别命名为“pFBAIN-YDG 1”和“pFBAIN-YDG2”。
由此,pFBAIN-YDG1(图1A;SEQ ID NO:19)包含下述组分:
表3
质粒pFBAIN-YDG1(SEO ID NO:19)的组分
质粒pFBAIN-YDG2(图1B;SEQ ID NO:20)所包含的组分与pFBAIN-YDG1所包含的基本一致,不同之处在于解脂耶氏酵母DGAT2ORF(SEQ ID NO:17;图1B中标记为YDG2)取代了pFBAIN-YDG1中的解脂耶氏酵母DGAT1ORF。
术语“FBAINm启动子”或“FBAINm启动子区域”是指FBAIN启动子(参见下文)的一个经修饰的版本,其中FBAINm在ATG翻译起始密码子和FBAIN启动子的内含子之间有52bp的缺失(从而仅包括N-端的22个氨基酸),并且在上述内含子之后有一种新的翻译共有基序。此外,当上述FBAIN启动子与一个待表达的基因的编码区融合时会产生一种融合蛋白质,而上述FBAINm启动子则并不产生这样的融合蛋白质。上述FBAIN启动子是指为表达所必需的、位于由fba1基因编码的解脂耶氏酵母果糖-二磷酸醛缩酶(E.C.4.1.2.13)“ATG”翻译起始密码子之前的5′上游非翻译区域,加上含有fba1基因一个内含子的5′编码区的一个部分。对上述启动子的描述详见PCT专利公布WO 2005/049805和美国专利7,202,356,上述专利均全文引入本文以供参考。
实施例3
解脂耶氏酵母DGAT1和DGAT2基因在解脂耶氏酵母菌株
Y2224中的过表达
本实施例描述了在经过转化从而共表达解脂耶氏酵母DGAT1(SEQ ID NO:16)或解脂耶氏酵母DGAT2(SEQ ID NO:17)的解脂耶氏酵母菌株Y2224中,脂肪酸含量的提高和每种脂肪酸相对丰度的改变。Y2224菌株是由ATCC登录号为20362的野生型解脂耶氏酵母菌株经Ura3基因自发突变而来的一种FOA抗性突变体。
菌株Y2224的产生:以如下方式分离菌株Y2224:将来自一块YPD琼脂平板(1%酵母抽提物,2%细菌用蛋白胨,2%葡萄糖,2%琼脂)的ATCC登录号为20362的解脂耶氏酵母细胞,划线接种至一块包含250mg/L 5-FOA(5-氟乳清酸;Zymo Research)基本培养基平板(含尿嘧啶和尿苷各75mg/L,含硫酸铵不含氨基酸的酵母氮源(YNB)6.7g/L以及葡萄糖20g/L)。将平板在28℃下孵育并将四个得到的菌落分别贴在含有200mg/mL 5-FOA的基本培养基(MM)平板上和无尿嘧啶和尿苷的MM平板上以确定尿嘧啶Ura3营养缺陷型。
Y2224菌株的转化:如下所述,将一个pFBAIn-YDG1克隆、一个pFBAIn-YDG2克隆和对照组质粒pFBAIN-MOD-1转入解脂耶氏酵母Y2224菌株。
除非另外说明,否则解脂解脂耶氏酵母的转化根据Chen,D.C.等人(Appl.Microbiol Biotechnol.48(2):232-235(1997))的方法。简而言之,将解脂耶氏酵母划线到YPD琼脂平板上,并在30℃生长大约18小时。从平板上刮掉几大环量的细胞并将其重悬浮于含有下面成分的1mL转化缓冲液中:2.25mL 50%PEG,平均分子量为3350;0.125mL 2M醋酸锂,pH6.0;0.125mL 2M DTT,以及50μg经剪切的鲑精DNA。然后,将大约500ng线性化的质粒DNA在100μL重悬浮的细胞中孵育,并将其在39℃下维持1小时,每间隔15分钟进行涡旋混合。
将每种转化的细胞接种至缺乏尿嘧啶的基本培养基(MM)平板(0.17%不含硫酸铵和氨基酸的酵母氮源(DIFCO Laboratories,Detroit,MI),2%葡萄糖,0.1%脯氨酸,pH 6.1,20g/L琼脂),并在30℃培养2天。将来自各转化平板的三种转化体分别接种至25mL的MM培养基(0.17%不含硫酸铵和氨基酸的酵母氮源(DIFCOLaboratories),2%葡萄糖,0.1%脯氨酸,pH 6.1)。每一培养物在30℃培养2天,然后转入25mL的高葡萄糖培养基(“HG培养基”,包含80g/L葡萄糖,27g/L K2HPO4,6.3g/L KH2PO4,pH约7.5)在30℃继续培养5天。
脂质分析:提取总脂质,通过酯交换反应制备脂肪酸甲基酯(FAMEs),并随后用Hewlett-Packard 6890GC分析。
更具体地讲,为了进行脂肪酸分析,将细胞通过离心收集并提取脂质,如Bligh,E.G.&Dyer,W.J.(Can.J.Biochem.Physiol.37:911-917(1959))中描述的那样。脂肪酸甲基酯通过脂质提取物和甲醇钠之间的酯交换反应进行制备(Roughan,G.,和Nishida I.,Arch.Biochem Biophys.276(1):38-46(1990)),然后用配备30-m X 0.25mm(i.d.)HP-INNOWAX(Hewlett-Packard)柱的Hewlett-Packard6890GC进行分析。炉温以3.5℃/分钟从170℃(保持25分钟)升到185℃。
为了进行直接的碱催化酯交换反应,收获解脂耶氏酵母培养物(3mL),在蒸馏水中洗涤一次,并在Speed-Vac中在真空下干燥5至10分钟。将甲醇钠(100μL,浓度为1%)加入样本中,然后将样本涡旋振荡20分钟。在加入3滴1M氯化钠和400μL己烷后,涡旋并离心样本。移出上层并如上所述的用GC进行分析。
基于上述分析,测定了分别包含pFBAIn-YDG1、pFBAIn-YDG2和pFBAIN-MOD-1(对照组)的Y2224转化体菌株的脂质含量和组分,显示于下列表4中。对每种菌株的三个独立的转化体进行了分析,平均结果显示于突出表示为灰色的行中。各种脂肪酸分别表示为16:0(棕榈酸)、16:1(棕榈油酸)、18:0、18:1(油酸)和18:2(LA);并且每种组分用占总脂肪酸的百分比表示。
“%FAME/DCW”表示脂肪酸甲基酯/细胞干重的百分比。通过从10mL培养物中离心收集细胞、用水清洗上述细胞一次以除去残留的培养基、在真空烘箱中80℃过夜烘干上述细胞以及称量烘干后细胞的重量,测定出细胞干重。通过将GC谱中所有峰的面积与加入的一种已知量的C15:0脂肪酸内标物的峰的面积进行比较,测定出一份样品中脂肪酸甲基酯的总量。
表4
过表达YDGAT1和YDGAT2的解脂耶氏酵母Y2224菌株中的
脂质含量A\和组成
GC分析显示,与携带pFBAIn-MOD-1的细胞相比,携带pFBAIn-YDG1的细胞中的总脂肪酸有显著的增加。平均脂肪酸由对照组的21.39%FAME/DCW增加至表达YDGAT1的细胞的23.70%FAME/DCW(即提高了10.8%)。此外,还发生了C16:1和C18:1脂肪酸量的提高和C18:2脂肪酸量的降低(表4)。
与对照组相比,携带pFBAIn-YDG2的细胞的总脂肪酸含量同样有了大幅增加,其平均为27.32%FAME/DCW(即提高了27.7%)。脂肪酸重量的分布也同样发生了显著的改变。具体而言,C18:0和C18:1的量提高了,而C16:0、C16:1和C18:2的量降低了。
总而言之,上述结果说明解脂耶氏酵母DGAT1或DGAT2的过表达既影响总脂肪酸含量,又影响各种脂肪酸的相对丰度。
实施例4
解脂耶氏酵母DGAT基因在拟南芥种子中的表达
如下所述,制备了一种适用于农杆菌介导的转化的二元载体。通过若干克隆步骤,将多个限制性位点添加到来自KS123(SEQ ID NO:21)的Bcon/NotI/Phas3′框的末端,其中所述KS123质粒已描述于PCT专利公布WO 02/008269中(该专利的内容特此引入本文以供参考)。简而言之,利用引物oCal-15(SEQ ID NO:23)和oCal-6(SEQ ID NO:24),自质粒CalFad2-2(描述于PCT专利公布WO 01/12800)扩增出DNA片段(ca1a24-4;SEQ ID NO:22)。用BgIII和BamHI消化DNA片段ca1a24-4(SEQ ID NO:22),并克隆至pKS123的BamHI位点以构建出pKR53B(SEQ ID NO:25)。将包含Bcon/NotI/Phas3框的pKR53B XbaI/SbfI片段,克隆至包含细菌潮霉素磷酸转移酶基因的pKR72(ATCC登录号PTA-6019;SEQ ID NO:26)XbaI/SbfI片段,以构建出pKR85(SEQ ID NO:27)。pKR72的特征如下所述。作为起始质粒的pKR72(ATCC登录号PTA-6019;SEQ ID NO:26),是此前已描述于PCT专利公布WO 02/008269(该专利的内容特此引入本文以供参考)的pKS123的一种衍生质粒,其包含两翼分别与T7启动子和转录终止子相接的潮霉素B磷酸转移酶(HPT)基因(Gritz,L.and Davies,J.,Gene 25:179-188(1983))(T7启动子/hpt/T7终止子框),以及一个用于在细菌(例如E.coli)中选择和复制的细菌复制起点(ori)。此外,pKR72还包含两翼分别与35S启动子(Odell等人,Nature 313:810-812(1985))和NOS 3′转录终止子(Depicker等人,J.Mol.Appl.Genet.1:561-570(1982))相接(35S/hpt/NOS3′框),从而可用于在植物,例如大豆中进行选择的潮霉素B磷酸转移酶基因。pKR72还包含一个两翼分别与β-大豆伴球蛋白(Beachy等人,EMBO J.4:3047-3053(1985))的α’亚单位和云扁豆蛋白(Doyle等人,J.Biol.Chem.261:9228-9238(1986))的3′转录终止区域相接的NotI限制性位点,从而允许克隆至所述NotI位点的基因在大豆种子中强烈的组织特异性表达。
利用引物oKR85-1(SEQ ID NO:28)和oKR85-2(SEQ ID NO:29)从质粒pKR85(SEQ ID NO:27)中扩增出Bcon/NotI/Phas3框,并按照生产商的说明书将得到的DNA片段克隆至(Stratgene),以构建出pPCR85(SEQ ID NO:30)。
将包含Bcon/NotI/Phas3框的pPCR85EcoRI/BglII片段,克隆至包含拟南芥二元载体主链的pZS199质粒(PCT专利公布WO93/11245;以及公布于1993年6月10日的美国专利5,952,544;此二专利的公开内容特此引入本文以供参考)EcoRI/BamHI片段,以构建出pKR91(SEQ ID NO:31)。
通过AscI消化将上述Bcon/NotI/Phas3框从pKR91上切下,制备出包含惟一AscI克隆位点的重连后的二元载体,称之为pKR92(SEQ ID NO:32)。
pKR92 YL DGAT2的构建:
表达质粒KS362的构建描述于实施例5。
将包含与β-大豆伴球蛋白启动子和云扁豆蛋白终止子融合的YL DGAT2基因,以及与Kti启动子和终止子序列融合的DsRed基因的一个表达框,作为6.4kb的AscI片段自KS362上切下。将上述DNA与经AscI线性化的去磷酸化pKR92载体DNA连接,以构建出pKR92 YL DGAT2(SEQ ID NO:33)。
pKR92 YL DGAT1/YL DGAT2的构建:
表达质粒KS364的构建描述于实施例5。将YL DGAT1(SEQ IDNO:1)和YL DGAT2(SEQ ID NO:9)基因分别与大豆球蛋白1和β-大豆伴球蛋白启动子以及相同的云扁豆蛋白终止子序列融合所构成的一个表达框,作为7kb的AscI片段自KS364上切下。将上述DNA与经AscI线性化的去磷酸化pKR92载体DNA连接,以构建出pKR92YL DGAT1/YL DGAT2(SEQ ID NO:34)。
转基因拟南芥品系的获得与分析:
通过电穿孔将pKR92YL DGAT2和pKR92YL DGAT1/YLDGAT2质粒DNA导入根癌农杆菌NTL4(Luo等人,Molecular Plant-Microbe Interactions 14(1):98-103(2001))。简言之,将1μg质粒DNA与100μL电转感受态细胞在冰上混合。将上述细胞悬液转入一个100μL电穿孔杯(间隙宽度为1mm),并用参数调至1kV、400Ω和25μF的BIORAD电穿孔仪进行电穿孔。将细胞转入1mLLB培养基,并在30℃孵育2h。将细胞铺板于包含50μg/mL卡那霉素的LB培养基上。将平板在30℃培养60h。从转染的农杆菌细胞单克隆接种出重组的农杆菌培养物(500mL LB,50μg/mL卡那霉素),并在30℃培养60h。离心(5000xg,10min)收集细胞,并重悬于包含0.05%(V/V)Silwet的1L 5%(W/V)蔗糖中。在metromix 360土壤混合物上以每100cm230植株的密度将拟南芥盆栽培养4周(22℃,持续光照,100μE m-2s-1)。将拟南芥植株反复浸入携带上述二元载体的农杆菌悬液,并置于黑暗、高湿度环境24h。植株在如前所述的标准植物生长条件下培养四至五周,然后收集植物材料并置于纸袋中在环境温度下干燥一周。用0.425mm筛眼的黄铜筛收集种子。
将干净的拟南芥种子(2g,相当于约100,000粒种子)依次用下列溶液清洗消毒:45mL 80%酒精,0.01%triton X-100,45mL 30%(V/V)溶于水的家用漂白剂,0.01%triton X-100,最后用无菌水反复冲洗。将含20,000粒种子的等份转入包含150mL无菌的植物生长培养基的方板(20x 20cm)中,所述培养基包含固化于10g/L琼脂中的0.5x MS盐、1.0%(W/V)蔗糖、0.05MES/KOH(pH 5.8)、200μg/mL特美汀,以及50μg/mL卡那霉素。通过将悬于水中的种子悬液与等体积融化熔化的植物生长培养基混匀,将上述种子均匀分散于上述培养基中。平板在标准生长条件下孵育十天。将卡那霉素抗性幼苗转入不含选择剂的植物生长培养基中,经8至10周发育至成熟(22℃,固定光照,100-200μE m-2s-1)。植株种植于包含36个插孔的培养盆中。在每一培养盆中,将至少6株非转化的野生型对照组植株与约三十株T2植株相邻种植。从植株个体收集种子,并通过NMR测量种子的油脂含量。
基于NMR的种子油脂含量分析:
种子油脂含量用Maran Ultra NMR分析仪(Resonance Instruments Ltd,Whitney,Oxfordshire,UK)测定。将样品(单粒的大豆种子或每批重量为5至200mg之间的拟南芥种子)置入事先称重并用区别的条形码标识符标记过的2mL聚丙烯管(Corning Inc,Corning NY,USA;Part no.430917)。然后将样品置入96孔载样皿并用Adept Cobra 600SCARA机器人系统按下列步骤操作。
1.拾起聚丙烯管(机器人臂配有一个真空捡拾装置)
2.读取条形码
3.用防静电装置处理聚丙烯管(以保证拟南芥种子不会粘附在管壁上)
4.称量聚丙烯管重量(包括样品),精确度至0.0001g
5.NMR读数;测量为将22.95MHz信号照射样品1msec后的质子自旋回波强度(收集对每份样品进行的32次NMR扫描数据)
6.将管放回架上
7.对下一管重复以上过程
条形码、聚丙烯管重量和NMR读数用一台与上述系统相连的计算机进行记录。通过用包含样品的聚丙烯管重量减去聚丙烯管自身重量,得到样品的重量。
大豆种子的油脂含量计算方法如下:
通过精确称量Corning管(见上)中大豆油脂样品(重量范围自0.0050至0.0700g,间隔约0.0050g,称量至0.0001g精确度)重量并进行NMR分析,测定出校正参数。建立起油脂含量(%种子基准重量;假定一粒标准种子的重量为0.1500g)对NMR值的校正曲线。
用NMR测量出的种子油脂含量和用传统的分析化学方法测得的绝对油脂含量之间的相关性测定如下。将五十粒经挑选以涵盖一定范围油脂含量的大豆种子,置于强制通风的烘箱内40℃烘干48h。如上所述,将单粒种子分别进行NMR分析,然后用GenoGrinder研磨仪(SPEX Centriprep(Metuchen,N.J.,U.S.A.);振动频率为1500次/分钟,时间1分钟)研磨为细粉。称取70至100mg之间的等分试样(精确至0.0001g),置入配有内衬的旋盖的13x 100mm玻璃管中;。将每粒大豆的残余粉末置于强制通风的烘箱内105℃烘干18h,通过烘干前后的重量差异测定出水分含量向上述含有粉末的玻璃管中加入庚烷(3mL),振荡混匀后将样品置入直立圆筒搅拌器,室温萃取1h。将萃取物1500x g离心10min,将上清轻轻倒入一个干净管子,沉淀用1mL庚烷再萃取两次(每次1h)。合并三次萃取的上清并加入50μL内标物(三十七烷酸;10mg/mL甲苯),然后在氮气流下室温蒸发至干燥状态;分别在5mL庚烷中包含0、0.0050、0.0100、0.0150、0.0200和0.0300g大豆油脂的标准物也以相同方式制备。向上述干燥后的团块加入1mL 5%硫酸(v∶v.,溶于无水甲醇),然后将混合物80℃加热30min并不时振荡混匀,从而使脂肪转变为脂肪酸甲基酯(FAMEs)。令上述样品冷却至室温,然后依次加入1mL 25%氯化钠水溶液和0.8mL庚烷。振荡混匀后令各相分离,将上层有机相转入一个样品瓶并进行GC分析。
将NMR测定的油脂含量与GC测定的油脂含量进行比较,得到油脂介于9.66和26.27%之间(GC值;%种子基准重量)、斜率为1.0225、R2为0.9744的线性关系;所基于的种子水分含量为平均2.6+/-0.8%。
拟南芥种子的油脂含量计算方法如下:
mg油脂=(NMR信号-2.1112)/37.514
%油脂(%基准重量)=(mg油脂/1000)/样本重量)*100
在引入此方程式计算之前,先按下述方法萃取拟南芥种子油脂。用研钵和研杵将约5g成熟的拟南芥种子(哥伦比亚生态型)研磨为细粉。将上述细粉置入一个33×94mm的纸壳筒(Ahlstrom#7100-3394;Ahlstrom,Mount Holly Springs,PA,USA),在索氏抽提器中用石油醚(BP 39.9至51.7℃)经约40个萃取循环萃取出油脂。使萃取物冷却,然后通过在旋转式蒸发器中真空除去溶剂而回收得到原油。通过精确称取11份部分纯化的拟南芥油脂标准样品(样品分别包含3.6、6.3、7.9、9.6、12.8、16.3、20.3、28.2、32.1、39.9和60mg部分纯化的拟南芥油脂;称量精确度为0.0001g)至2mL聚丙烯管(Corning Inc,Corning NY,USA;Part no.430917)并对其进行NMR分析,测定出校正参数。建立起油脂含量(%种子基准重量)对NMR值的校正曲线。
用于培植pKR92 YL DGAT2 T2的种子共计种植293个独立的事件,同时种植75个野生型对照。YL DGAT2转基因植株的油脂含量范围为27.9至47.3%。平均油脂含量为43.7%。野生型对照的油脂含量范围为39.5至49.6%。野生型对照的平均油脂含量为44.8%。
用于培植pKR92 YL DGAT1/YL DGAT2 T2的种子共计种植295个独立的植株,同时种植77个野生型对照。YL DGAT1/YLDGAT2转基因植株的油脂含量范围为34.5至47.6%。平均油脂含量为44%。野生型对照的油脂含量范围为41.3至46.6%。野生型对照的平均油脂含量为45%。总而言之,上述结果表明,YL DGAT基因的种子特异性表达并未提高拟南芥种子的油脂含量。
单独表达YL DGAT2或同时还表达YL DGAT1的拟南芥种子的 脂肪酸谱分析:
利用对FAME进行的GC分析,研究YL DGAT的表达是否会改变拟南芥种子的脂肪酸谱。将约100粒F2种子分配至96孔八联管的各孔中。向每一联排管中加入50μL三甲基氢氧化硫(TMSH)和0.5mL己烷,并在室温下摇动孵育30min以进行酯交换反应。分离脂肪酸甲基酯(1μL注射自己烷层)并用配套了Omegawax 320熔融石英毛细管柱(Supelco Inc.,Catalog No.24152)的Hewlett-Packard6890气相色谱仪定量。炉温程序设定为220℃保持2.6min,然后以20℃/min速率升温至240℃并维持2.4min。载气通过Whatman氢气发生器供应。将保留时间与市售标准物的甲基酯的保留时间进行比较。结果总结于表5。
表5
结果清楚地表明,YL DAGT2在拟南芥种子中的表达导致了种子脂质中油酸的增加,并且共表达YL DGAT1和YL DGAT2所得出的结果还要更为明显,这提供了YL DGAT1和YL DGAT2活性蛋白可以在转基因种子中得到合成的第一个指征。
实施例5
解脂耶氏酵母DGAT基因在大豆体细胞胚中的表达
用于构建在种子中特异性过表达YL DGAT基因的质粒的启动子和终止子序列列于表6和表7中。
表6
种子特异性启动子
表7
转录终止子
转录终止子 | 生物 | 参照序列 |
云扁豆蛋白3’ | 菜豆 | WO 2004/071467 |
库尼兹胰蛋白酶抑制剂3’ | 大豆 | WO 2004/071467 |
构建一种用于在大豆球蛋白Gy1启动子控制下表达YL DGAT1基因的质粒构造(KS349)(图3)。
大豆的大豆球蛋白Gy1启动子的分离按下述过程进行。基于NCBI数据库中的大豆的大豆球蛋白Gy1基因序列(GenBank登录号X15121;SEQ ID NO:35),设计了两条5′端带有BamHI或NcoI位点的寡核苷酸,用以扩增大豆的大豆球蛋白Gy1启动子(SEQ ID NO:36)。上述两条寡核苷酸的序列如下:
SEQ ID NO:37(oGy1-1):CGCGGATCCTAGCCTAAGTACGTACTC AAAATGCCA
SEQ ID NO:38(oGy1-2):GAATTCCCATGGGGTGATGACTGATGA GTGTTTAAGGAC
质粒pKS349由若干不同的中间载体经多个步骤构建而成。用BamHI和NcoI消化上述扩增后的大豆的大豆球蛋白Gy1启动子片段并纯化,然后克隆至p24K-G4G-@SalI(PCT专利公布WO98/59062)的BamHI和NcoI位点,以构建出pZBL114(SEQ ID NO:39)。用包含一种大豆GY1基因(SEQ ID NO:40)和合成的大麦高赖氨酸8(BHL8)基因(US 6800726 B1)(SEQ ID NO:41;)的融合产物的NcoI/KpnI片段,替换包含GUS的NcoI/KpnI片段,以构建出pZBL133(SEQ ID NO:43)。大豆GY1基因和BHL8基因的融合产物的DNA序列如SEQ ID NO:42所示。将云扁豆蛋白终止子从pZBL133(XbaI/插入)上切除,并代之以pKS123(PCT专利公布WO02/08269)(平末端)中的云扁豆蛋白终止子,以构建出pKS238(SEQID NO:44)。按下述步骤用天然的GY1序列替换上述GY1-BHL8融合物。用KpnI/BglI消化pKS238,将残余的载体条带(5.1kb)与天然GY1基因(SEQ ID NO:40)的一个DNA片段(BglI/KpnI)连接,以构建出pKS240(SEQ ID NO:45)。将包含Gy1/GM-GY1/Phas3′的BamHI/SalI片段自pKS240上切下,并连接至pKS120(SEQ ID NO:46)的BamHI/SalI位点,以构建出pKS242(SEQ ID NO:47)。质粒pKS120上已被移除了包含Bcon/NotI/Phas3′框的HindIII片段,除此以外与(上述)pKS 123相同。用包含来自pYDA1的YL-DGAT1的NcoI/NotI片段替换包含GM-GY1的NcoI/NotI片段,以构建出pKS349(SEQ ID NO:48)。
一种用于在β-大豆伴球蛋白启动子控制下表达YL DGAT2基因 的质粒(KS362)的构建(图3):
质粒pKS362由若干不同的中间载体经多个步骤构建而成。将包含来自pKS121(PCT专利公布WO 02/00904)的Kti/NotI/Kti3′的AscI框平末端连接至pBluescript II SK+(Stratagene)上的NotI(插入)位点,以构建出pKS121/BS。将来自pDsRed-表达载体(Clontech)的NcoI/NotI片段平末端连接至pKS121/BS的NotI(插入)位点,以构建出包含pDS-RED的KS121/BS(SEQ ID NO:49)。将包含Kti/DsRed/Kti3′的BamHI框(SEQ ID NO:50)连接至pKS123(PCT专利公布WO 02/08269)的BamHI位点,以构建出pKS332(SEQ ID NO:51),其中所述Kti/DsRed/Kti3′位于包含pDS-RED的KS121/BS中。用引物通过PCR将NotI位点引入YL-DGAT2基因的两端,从而合成出用于表达YL-DGAT2的基因(参见实施例1)。所得到的PCR产物用NotI限制酶消化,然后连接至pKS332的NotI位点,以构建出pKS362(SEQ ID NO:52)。
对照组质粒(KS352)的构建(图2):
根据所克隆的大豆P34启动子(WO 2004/071467)(SEQ ID NO:53)序列,设计两条5′端带有BamHI或NotI位点的寡核苷酸,用以重扩增上述P34启动子。上述两条寡核苷酸的序列如下:
SEQ ID NO:54(oP34-1):
CGCGGATCCAACTAAAAAAAGCTCTCAAATTACATTTTGAG
SEQ ID NO:55(oP34-2):
GAATTCGCGGCCGCAACTTGGTGGAAGAATTTTATGATTTGAAA
用BamHI和NotI消化重扩增的P34启动子片段并纯化,然后克隆至质粒pZBL115(SEQ ID NO:56)的BamHI和NotI位点,以构建出pJS89(SEQ ID NO:57)。用包含来自pRB322的复制起点、由T7启动子和T7终止子驱动的细菌HPT潮霉素抗性基因,以及一个35S启动子-HPT-Nos3′基因的pZBL115质粒,作为潮霉素抗性植株的一种选择标记。将高山被孢霉Δ-6脱氢酶基因(美国5,968,809)(SEQID NO:58)按有义方向克隆至pJS89(SEQ ID NO:57)的NotI位点,以构建出植物表达框和pJS93(SEQ ID NO:59)。
用SalI NotI双酶切将P34启动子从pJS93(SEQ ID NO:59)上切下,并连接至经SalI/NotI线性化的pKS127载体(US Pat.App.No.11/476,510)(SEQ ID NO:60),以构建出pKS343(SEQ ID NO:61)。将包含Kti/DsRed/Kti3′的BamHI框末端填平并连接至pKS343的HindIII(插入)位点,以构建出pKS352(SEQ ID NO:62),其中所述Kti/DsRed/Kti3′位于包含pDS-RED的KS121/BS中。
一种用于共表达YL DGAT1和YL DGAT2的质粒(KS364)的构 建:
通过将来自pKS362(SEQ ID NO:52)的包含Bcongl PRO/YL-DGAT2/Phas TER的3.3kb HindIII框,连接至在pKS349(SEQ IDNO:48)中位于Gy1启动子下游的惟一HindIII位点,构建出质粒pKS364(SEQ ID NO:63)。
转基因体细胞胚的生成:
如下所述,将大豆组织与KS349和KS362的混合物共轰击,以实现在YL DGAT1和YL DGAT2在大豆体细胞胚中的共表达。简言之,用限制酶PstI、XhoI消化KS349的DNA,以灭活选择性标记基因框(CaMV35S PRO/HPT/CaMV NOS TER)。将所述DNA与经Sall线性化的KS362质粒DNA按10∶1的比例混合,并用于如下文所述的大豆转化过程。或者如下文所述,将用于生成大豆体细胞胚的大豆组织,与同时包含YL DGAT1和YL DGAT2功能性表达框的完整的KS364质粒DNA共轰击。对于YL DGAT1的单独表达,则用未经切割的KS349质粒DNA来进行胚胎组织的基因枪轰击。类似地,对于YL DGAT2的单独表达,则用未经切割的KS362质粒DNA来进行胚胎组织的基因枪轰击。此外,仅包含选择性标记的质粒DNA(KS352)也被以相同方式用于大豆组织的转化。
培养条件:
大豆胚胎生成悬浮培养物(cv.Jack)培养于35mL液体SB 196培养基(参见下文),培养条件为150rpm摇床培养、26℃和按16∶8h昼/夜光周期以60-85μE/m2/s光强用白色冷光荧光灯照射。每7天至两周,通过将约35mg的组织接种至35mL新鲜的液体SB196,对培养物进行传代培养(优选的传代培养间隔时间为7天)。
用大豆表达质粒对大豆胚胎生成悬浮培养物进行的转化,通过的是基因枪轰击的方法(Klein等人,Nature 327:70(1987)),所有转化均用一台DuPont Biolistic PDS1000/HE设备(氦气型)进行。
大豆胚胎生成悬浮培养物的诱导:
每月诱导大豆培养物两次,每次诱导之间相隔5至7天。将种植45至55天后的存活大豆植株的带有未成熟种子的豆英摘下,将其去壳后置入一个灭菌后的Magenta盒子中。将大豆种子置于加入了1滴象牙皂的5%次氯酸钠溶液(即95mL灭菌蒸馏水加上5mL次氯酸钠和1滴象牙皂,混匀)中摇动消毒15min。用2瓶1升瓶子的无菌蒸馏水清洗种子,将小于4mm的种子分别置于显微镜载片上。将上述种子的较小一端切开,将子叶从种皮中挤出。子叶被转入含SB199培养基的平版(每板25至30个子叶)培养2周,然后转入SB 1培养2至4周。用纤维带包裹平板。此次之后,将次级胚切下,并置于SB196液体培养基7天。
用于轰击的DNA的准备:
采用包含所感兴趣的基因和选择性标记基因的完整质粒或质粒DNA片段进行轰击。
将一份包含1mg金颗粒的50μg灭菌水等份加入5μL的1μg/μL DNA溶液(完整质粒或如前所述制备的DNA片段)、50μL的2.5M CaCl2和20μL的0.1M亚精胺中。将混合物用调至4档的旋涡振荡器脉冲振荡5次,然后用台式微量离心机离心5sec。用150μL 100%酒精洗一次,然后在85μL 100%酒精中超声处理使沉淀块悬浮。向上述Biolistic PDS 1000/HE设备的每个飞盘中装入5μLDNA悬液。每一轰击反应(即每个盘)的5μL等份中包含约0.058mg金颗粒。
组织制备和用DNA轰击:
将约100至150mg 7天的胚胎悬浮培养物置于一个空的60×15mm无菌皮氏培养皿中,并将所述培养皿置入一个空的150×25mm皮氏培养皿。组织在每个盘轰击一次,膜破裂压力设为650PSI,小室抽真空至27至28英寸汞柱。组织置于距维持/终止屏约2.5英寸处。
转化胚的选择:
用潮霉素作为选择性标记,进行转化胚的选择。具体而言,轰击后的组织被置入新鲜的SB196培养基,如前所述进行培养。轰击后6至8天,将上述SB196替换为包含30mg/L潮霉素的新鲜SB196。选择性培养基每周更新一次。选择后四至六周,观察到绿色的转化组织从坏死的未转化胚胎发生块中长出。将绿色组织分离,接种至多孔板,以产生新的、克隆繁殖的转化的胚胎生成悬浮培养物。
胚胎的成熟:
转化的胚胎生成块在白色冷荧光灯(Phillips cool white EconowattF40/CW/RS/EW)和农用灯泡(Phillips F40Agro)(40瓦)以16∶8h光周期和90至120μE/m2s光强照射下,用SB196于26℃培养一至三周。然后将胚胎块移入一种固体琼脂培养基SB 166培养1周。然后用SB103培养基传代培养3周。或者,将胚胎块移入盛于250mL锥形瓶中的35mL SB228(ShaM)液体培养基,培养2至3周。组织于SB228中的培养条件为,130rpm摇床培养、26℃和按16∶8h昼/夜光周期以60至85μE/m2/s光强用白色冷光荧光灯照射。在此期间,将胚胎个体自培养块移出,并如上文所述筛选其脂肪酸组成的变化。
培养基配方:
SB 196-FN Lite液体增殖培养基(每升)
MS FeEDTA-100x储备液1 10mL
MS硫酸盐-100x储备液2 10mL
FN Lite卤化物-100x储备液3 10mL
FN Lite P,B,Mo-100x储备液4 10mL
B5维生素(1mL/L) 1.0mL
2,4-D(10mg/L终浓度) 1.0mL
KNO3 2.83gm
(NH4)2SO4 0.463gm
天冬酰胺 1.0gm
蔗糖(1%) 10gm
pH 5.8
FN Lite储备液
储备液编号 1000mL 500mL
1 MS Fe EDTA 100x储备液
Na2EDTA* 3.724g 1.862g
FeSO4-7H2O 2.784g 1.392g
*首先添加,搅拌溶解于深色瓶中
2 MS硫酸盐100x储备液
MgSO4-7H2O 37.0g 18.5g
MnSO4-H2O 1.69g 0.845g
ZnSO4-7H2O 0.86g 0.43g
CuSO4-5H2O 0.0025g 0.00125g
3 FN Lite卤化物100x储备液
CaCl2-2H2O 30.0g 15.0g
KI 0.083g 0.0715g
CoCl2-6H2O 0.0025g 0.00125g
4 FN Lite P,B,Mo 100x储备液
KH2PO4 18.5g 9.25g
H3BO3 0.62g 0.31g
Na2MoO4-2H2O 0.025g 0.0125g
SB1固体培养基(每升)
1个包装的MS盐(Gibco/BRL-Cat.No.11117-066)
1mL B5维生素1000X储备液
31.5g葡萄糖
2mL 2,4-D(20mg/L终浓度)
pH 5.7
8g TC琼脂
SB199固体培养基(每升)
1个包装的MS盐(Gibco/BRL-Cat.No.11117-066)
1mL B5维生素1000X储备液
30g蔗糖
4mL 2,4-D(40mg/L终浓度)
pH 7.0
2gm固化剂
SB 166体培养基(每升)
1个包装的MS盐(Gibco/BRL-Cat.No.11117-066)
1mL B5维生素1000X储备液
60g麦芽糖
750mg MgCl2六水合物
5g活性炭
pH 5.7
2g固化剂
SB 103固体培养基(每升)
1个包装的MS盐(Gibco/BRL-Cat.No.11117-066)
1mL B5维生素1000X储备液
60g麦芽糖
750mg MgCl2六水合物
pH 5.7
2g固化剂
SB 71-4固体培养基(每升)
1瓶Gamborg′s B5盐w/蔗糖(Gibco/BRL-Cat.No.21153-036)
pH 5.7
5g TC琼脂
2,4-D储备液
Phytotech Cat.No.D 295预配液-浓度1mg/mL
B5维生素储备液(每100mL)
分装保存于-20℃
10g肌肌醇
100mg尼克酸
100mg吡哆醇HCl
1g硫胺
若溶解不够迅速,可将溶液用热搅拌器加以微热。
SB 228-大豆组织分化和成熟培养基(SHaM)(每升)
DDI H2O 600mL
用于ShaM的FN-Lite Macro盐10X 100mL
MS Micro盐1000x 1mL
MS FeEDTA 100x 10mL
CaCl 100x 6.82mL
B5维生素1000x 1mL
L-甲硫氨酸 0.149g
蔗糖 30g
山梨醇 30g
调整体积至900mL
pH 5.8
高压灭菌
向冷却后的培养基(≤30C)加入:
*谷氨酰胺(Final conc.30mM)4%110mL
*注意:加入谷氨酰胺后,终体积将变为1010mL。
因谷氨酰胺降解相对迅速,可能最好在即将使用前加入培养基。培养基在谷氨酰胺加入后2周过期;不含谷氨酰胺的基本培养基可存放更长时间。
用于SHAM的FN-lite Macro 10X-储备液#1(每升)
(NH4)2SO4(硫酸铵) 4.63g
KNO3(硝酸钾) 28.3g
MgSO4*7H2O(七水硫酸镁) 3.7g
KH2PO4(磷酸二氢钾) 1.85g
加水至终体积
高压灭菌
MS Micro 1000X-储备液#2(每升)
H3BO3(硼酸) 6.2g
MnSO4*H2O(一水硫酸锰) 16.9g
ZnSO4*7H2O(七水硫酸锌) 8.6g
Na2MoO4*2H2O(二水钼酸钠) 0.25g
CuSO4*5H2O(无水硫酸铜)_ 0.025g
CoCl2*6H2O(六水氯化钴) 0.025g
KI(碘化钾)_ 0.8300g
加水至终体积
高压灭菌
FeEDTA 100X-储备液#3(per liter)
Na2EDTA*(EDTA钠盐)_ 3.73g
FeSO4*7H2O(七水硫酸铁) 2.78g
*在加入铁离子前EDTA必须完全溶解。
加水至终体积
此溶液对光敏感。盛装的瓶子应以铝箔包裹以避光。
高压灭菌
Ca 100X-储备液#4(每升)
CaCl2*2H2O(二水氯化钙) 44g
加水至终体积
高压灭菌
B5维生素1000X-储备液#5(每升)
硫胺素*HCl 10g
尼克酸_ 1g
吡哆醇*HCl 1g
肌肌醇 100g
加水至终体积
冷冻保存
4%谷氨酰胺-储备液#6(每升)
DDI水加热至30℃ 900mL
L-谷氨酰胺_ 40g
边搅拌边逐渐加入并施以微热。
温度勿超过35℃。
加水至终体积
过滤除菌
冷冻保存*
*注意:在31℃水浴中热解冻储备液以令结晶完全溶解。
油脂分析:
在液体成熟培养基SB228(SHaM)中培养两周后收集体细胞胚。在用KS352、KS349/KS362,以及KS362和KS364进行的转化终制造了约30个植株系。一个特定植株系中产生的所有胚胎均批量收集,并处理如下。将胚胎置于干冰上或-80℃冰箱中冷冻两小时,然后冷冻干燥48h。
用一个genogrinder瓶(1/2″X2″聚碳酸酯)和一粒钢珠(SPEXCentriprep(Metuchen,N.J.,U.S.A.)将干燥后的胚胎研磨为细粉。研磨时间为30sec,振动频率为1450次/分钟。对于每一事件,称取约10mg的组织一式三份置入Eppendorf管中。上述组织在室温下用200μL庚烷持续摇动萃取2h。离心令庚烷萃取物澄清,取25μL萃取物用于制备脂肪酸甲基酯衍生物如下。将1mL的25%甲醇钠储备液加入24mL HPLC级甲醇。甲醇钠保存于惰性气体中。
将5μL的17:0TAG(Nu-Chek Prep,Elysian,MN,USA)储备液(10mg/mL)同25μL的庚烷组织萃取物一起加入至一个玻璃培养管,并加入500μL的1%甲醇钠。样品在50℃水浴中反应15min。令样品冷却至室温,加入1mL 1M NaCl,然后短暂混匀。FAME被萃取至1mL的庚烷中,通过GC分析对4μL样品进行定量。
通过求油酸含量(%总FAME)对总FAME含量(%DW)的函数,进行数据分析。表8显示,所生成的携带对照组载体(KS352)的体细胞胚,其油酸含量几乎没有波动,油脂含量表现出一些波动但很可能是由于再生过程中引入的生物学变异所致。例如,在收获时胚胎所处的发育阶段就经常表现出变异。在用对照组质粒制备的胚胎中,在油酸含量和油脂含量之间没有观察到相关性(R2=0.1142)(表8)。在用在种子特异性强启动子控制下分别单独表达YL DGAT1和YL DGAT2基因的质粒KS349和KS362,或同时表达YL DGAT1和DGAT2基因的质粒(KS349/KS362,KS364)制备的胚胎中,油酸含量和总的酯化脂肪酸含量均表现出大幅波动。此外,如图4和5所示,在分别单独用KS349和KS362,或组合使用KS349和KS362,或用包含YL DGAT1和YL DAT2基因表达框的转化质粒KS364制备的体细胞胚中,均观察到油酸含量和总酯化脂肪酸含量之间存在强相关(R2≥0.59)。
表8
大豆体细胞胚的酯化脂肪酸和油酸含量
总之,上述数据显示,与拟南芥种子相似,在大豆体细胞胚中YLDGAT基因的表达与总酯化脂肪酸组分中油酸的增加相关。然而,与拟南芥中的情况相反的是,在大豆体细胞胚中增加的油酸含量与酯化脂肪酸总量的累积密切相关。换句话说,在大豆体细胞胚中,YLDGAT2的单独表达,或YL DGAT1和YL DAGT2的共表达均导致脂肪酸生物合成和向总酯化脂肪酸组分的整合的增加。上述发现总体上有力地说明,YL DGAT基因的表达提供了在大豆种子中实现总油脂含量提高的一种有效策略。
实施例6
解脂耶氏酵母DGAT基因在大豆种子中的表达
仅包含CaMV 35S PRO/HPT/NOS TER和Kti PRO/DsRed/Kti TER表达框的对照组质粒(KS332)(图2)的构建描述于实施例5中。其序列如SEQ ID NO:51所示。
通过基因枪轰击的方法(Klein等人,Nature(London)327:70-73(1987);美国专利4,945,050),利用KS332、KS362以及如实施例3中所述制备的10∶1的KS349和KS362的混合物,用一台BIORADBiolistic PDS1000/He设备制备出转基因大豆品系。大豆植株的转化和再生利用了下列储备液和培养基:
储备液:
100X硫酸盐储备液:
37.0g MgSO4.7H2O,1.69g MnSO4.H2O,0.86g ZnSO4.7H2O,0.0025gCuSO4.5H2O
100X卤化物储备液:
30.0g CaCl2.2H2O,0.083g KI,0.0025g CoCl2.6H2O
100X P,B,Mo储备液:
18.5g KH2PO4,0.62g H3BO3,0.025g Na2MoO4.2H2O
100X Fe EDTA储备液:
3.724g Na2EDTA,2.784g FeSO4.7H2O
2,4-D储备液:
10mg/mL维生素B51000X储备液:10.0g肌肌醇,0.10g尼克酸,0.10g吡哆醇HCl,1g硫胺。
培养基(每升):
SB196:上述储备液各10mL,B5维生素储备液1mL,0.463g(NH4)2SO4,2.83g KNO3,1mL 2,4-D储备液,1g天门冬酰胺,10g蔗糖,pH 5.7。
SB103:
1个包装Murashige&Skoog盐混合物,1mL B5维生素储备液,750mg MgCl2六水合物,60g麦芽糖,2g固化剂,pH 5.7。
SB166:
向SB103添加5g/L的活性炭而成。
SB71-4:
Gamborg′s B5盐,1mL B5维生素储备液,30g蔗糖,5g TC琼脂,pH 5.7。
将大豆胚胎生成悬浮培养物培养于35mL液体培养基(SB196),以制备用于转化的组织,培养条件为摇床(150rpm)培养、28℃,并按16h昼/8h夜的周期提供荧光。通过将约35mg组织接种至35mL的新鲜液体培养基,培养物每两周传代培养一次。
在基因枪轰击过程中,可以使用纯化的1)完整质粒DNA;或2)仅包含所感兴趣的重组DNA表达框的DNA片段。对于每十七次轰击转化,每种DNA质粒制备包含每碱基对1至90皮克(pg)的质粒DNA的85μL悬液。如下所述,两种重组DNA质粒均共沉淀至金颗粒上。将悬液中的DNA加入50μL 20至60mg/mL 0.6μm的金颗粒悬液,然后与50μL CaCl2(2.5M)和20μL亚精胺(0.1M)合并。混合物振荡混匀5sec,用微离心机离心5sec,去除上清。用150μL 100%酒精清洗包被了DNA的金颗粒一次,再次振荡并用微离心机离心,然后重悬于85μL无水酒精。然后将5μL DNA包被的金颗粒上样至每一大载物盘。
将约150至250mg已培养两周的悬浮培养物置入一个空的60mmX 15mm皮氏培养皿,并用移液器将残留的液体自上述组织吸走。将上述组织置于距维持屏约3.5英寸处,每一盘组织轰击一次。膜破裂压力设为650PSI,小室抽真空至-28英寸汞柱。对三个盘进行了轰击,轰击之后将来自每一盘的组织分为两份,放回含液体培养基的瓶中,并如上文所述进行培养。
轰击后7天,将上述液体培养基替换为含30至50mg/L潮霉素的新鲜SB196培养基。上述选择性培养基随后每周或每两周更新一次。轰击后七周,观察到鲜绿色的转化组织从坏死或萎黄的未转化胚胎生成块中长出。将绿色组织分离,接种至6孔培养皿的各个孔中,以产生新的、克隆繁殖的转化的胚胎生成悬浮培养物。由此,将每一新的品系处理为一个单独培养孔中的独立转化事件。然后,可以通过传代培养将上述悬液作为集中处于未成熟发育阶段的胚胎进行悬浮培养,或者也可以通过各个体细胞胚的熟化和萌发,再生为整株的植物。
在各细胞培养孔中培养两周后,将转化的胚胎生成块自液体培养基中移出,并置于不含激素或抗生素的固体培养基(SB166)上培养一周。胚胎于26℃在荧光和白炽光以16h昼/8h夜周期混合照射下培养。一周后,将培养物转入SB103培养基,并在相同的生长条件下再培养3周。
如此培养四周后,体细胞胚变得适宜萌发,此时将其从成熟培养基中移出,并置于空的皮氏培养皿中干燥一至五天。然后将干燥的胚胎种植于SB71-4培养基中,胚胎可在上述相同的光照和温度条件下于其中萌发。萌发的胚胎被转入无菌土壤中生长至成熟,以产生种子。
以每次金颗粒制备使用15pg/bp质粒DNA的浓度,用完整的KS362质粒DNA共计产生了29个带有种子的转基因品系。对于每一事件,就DS标记基因的存在对20粒种子进行了评分。简言之,在一台采用UV光源的体视显微镜(Leica MZ Fluo III)下对种子进行了观察。使用了具有下列特性的一组用于观察与DsRed表达相关的荧光的滤光镜:激发波长λ=540至580nm/发射波长λ≥570nm。在所获得的种子数量少于20粒的情况下,则将所有种子以上述方式进行评分。然后如实施例3所述通过NMR对大豆种子油脂含量进行了测量。用KS362产生的十九个事件中包含了DsRed阳性的种子。其中,11个事件表现出在DsRed阳性的转基因分离子的油脂含量和DsRed阴性的无转基因分离子的油脂含量之间存在可检测的差异。结果总结于表9。
表9
用KS362产生的T1大豆种子的油脂含量
用KS332的DNA共计产生了10个带有种子的转基因品系。如上所述,对于每一事件,就DS标记基因的存在对20粒种子进行了评分。在所获得的种子数量少于20粒的情况下,则将所有种子以上述方式进行评分。然后如实施例4所述通过NMR对大豆种子油脂含量进行了测量。用KS332产生的七个事件中包含了DsRed阳性的种子。结果总结于表10。
表10
用KS332产生的T1大豆种子的油脂含量
与用KS362产生的种子相反,用KS332产生的转基因种子中,没有能与DsRed标记在T1分离子中的存在相关联的稳定的油脂含量升高。
对用KS362产生的四个事件进行了DsRed状态的分析和所有获得的T1种子的油脂NMR测量。结果总结于表11。
表11
用KS362产生的T1大豆种子的油脂含量
总之,上述各表中的数据和图7显示,种子特异性的YL DAGT2表达会导致大豆种子成熟期油脂生物合成的增加,从而提供了用于提高大豆中油脂积累的一种有效的代谢工程工具。
用按10∶1比例混合的KS349和KS362 DNA共轰击(参见实施例4)共计产生了16个带有种子的转基因品系。上述DNA混合物以每次金颗粒制备使用15pg/bp质粒DNA的终浓度用于大豆的转化。简言之,在轰击之前,先用PstI和XhoI消化KS349 DNA,以令该质粒上的选择性标记基因失活。KS362的DNA用SalI线性化。有理由假定,由于上述对DNA的预处理,所有转化的限制性标记基因均来自于以较低的DNA浓度接受轰击的上述质粒(KS362)。在荧光立体式显微镜下对种子进行的初步检查显示,此次转化只有极少的事件包含DsRed标记基因阳性的T1种子。这种结果可能是由于DsRed表达框与用于大豆转化的KS362的SaII限制性片段的末端位置接近。这可能导致了不包含功能性的DsRed表达框的KS362DNA片段的整合。出于此原因,通过检测种子的脂肪酸组成,对不含转基因来源的YL DGAT的T1种子进行了筛选。对于每一事件,用GC对20粒种子进行了分析。50粒未转化的大豆种子也以相同方式进行了处理。用剃须刀片避开胚轴将大豆种子切开,得到种子切片。将约2mg的种子切片置入一个包含50μL三甲基氢氧化硫和0.5mL己烷的小瓶。上述切片一边摇动一边在室温下孵育30min。将5μL的上述己烷层注入配套了Omegawax 320熔融石英毛细管柱(Supelco Cat.No.24152)的Hewlett-Packard 6890气相色谱仪。炉内条件如下:维持220℃起始温度2.7分钟,用1分钟升温至240℃并维持于240℃。总运行时间为6min将保留时间与市售标准物(Nu-Chek Prep,Inc.Cat.No.U-99-A)进行比较。通过在0.5mL H2SO4甲醇(2.5%)中对各标准物直接进行转酯化反应,测定出脂肪酸含量。加入等体积的水后,将脂肪酸甲基酯从上述甲醇溶液中萃取至己烷。
鉴定出包含的T1种子具有≥25%油酸含量的十个事件。由于在未转化的大豆种子中并未观察到这样的油酸含量(参见图6和表12),并且在拟南芥种子和大豆体细胞中,均已发现油酸含量的增加与YL DGAT2的单独表达和YL DGAT1与YL DGAT2的共表达相联系,相信油酸以≥25%水平的存在提供了鉴定YL DGAT阳性T1种子的有效方法。在通过GC分析进行YL DGAT基因分型之后,如前所述(参见实施例3)通过NMR对种子进行了油脂测量。求油酸含量对总油脂含量的函数,结果在10个T1种子油酸≥25%的事件中,有七个表现为油脂和油酸含量之间具有R2≥0.3的相关性。这些事件的性质详细描述于表12中。
表12
用KS349/KS362产生的T1大豆种子的油脂含量
在与用于产生T1种子的条件相似的条件下培育Jack野生型种子,并通过GC和NMR进行分析。观察到油酸含量和油脂含量分别在7.6至20.5%和18.6至25.2%范围内波动。在未转化的大豆种子中没有观察到油酸含量和油脂含量之间存在相关性。
将用KS349/KS362产生的四个事件的所有获得的T1种子用GC和NMR进行分析。结果总结于表13。
表13
用KS349/KS362产生的T1大豆种子的油脂含量
此结果以及前述各表和图6和7中的数据有力支持了YL DGAT1和YL DGAT2基因的共表达,如同YL DGAT2基因的单独表达一样,提供了在大豆种子中实现总油脂含量提高的一种有效策略这一结论。此外,应当注意到在所筛选的少数转基因事件中,有大量的事件可以被鉴定为在无转基因分离子和转基因分离子之间具有≥2%分数的油脂差异。
实施例7
解脂耶氏酵母DGAT基因在玉米中的表达
基于本申请的实施例4、5和6中所公开的结果,YL DGAT1和YL DGAT2基因可以在玉米的种子胚胎中得到表达,以提高该组织中的油脂含量。如下所述,可以通过用表达框转化玉米实现此结果,其中所述表达框由胚胎偏好的启动子,例如玉米16kDa油体蛋白基因(Lee,K.and Huang,A.H.,Plant Mol.Biol.26:1981-1987(1984))的启动子和玉米胚胎早期高丰度启动子(EAP1)、来自解脂耶氏酵母并在其5′端与上述启动子可操作地连接的DGAT1和DGAT2的开放阅读框以及终止子(US 2006272058A1)组成。
用本领域所知的方法和技术,构建了由来自玉米16kDa油体蛋白基因的启动子(OLE PRO)、YL DGAT2基因的编码序列(SEQ ID NO:9)和来自根癌农杆菌胭脂碱合酶(NOS)基因的多聚腺苷酸信号序列/终止子组成的一个表达框。第二个表达框包括处于玉米胚胎高丰度蛋白(EAP1)启动子和终止子转录控制下的YL DGAT1基因,以及插入至上述启动子和编码序列之间以增强表达的玉米ADH1内含子1。将上述两个表达框以及一个编码选择性标记的基因,连接于一个适用于农杆菌介导的玉米转化的二元载体中。
一种基于农杆菌的方法可用于玉米的转化(见下)。将所得到的二元载体引入农杆菌LBA4404(PHP10523)细胞,优选通过电穿孔。通过体内重组,在所引入的二元载体和寄宿于农杆菌细胞的vir质粒(PHP10523)之间形成了一种共整合质粒。所得到的农杆菌细胞被用于转化玉米。
农杆菌介导的玉米的转化:
将新鲜分离的授粉后约十天(DAP)的未成熟玉米胚胎,与上述农杆菌共同孵育。优选的用于转化的基因型为高度可转化的基因型Hi-II(Armstrong,Maize Gen.Coop.Newsletter,65:92-93(1991))。也可以使用通过将Hi-II与一种自交原种杂交得到的F1杂种。未成熟的胚胎在用农杆菌处理后,可培养于包含毒性水平的除草剂的培养基上。只有获得了除草剂抗性基因以及与之连接的基因的细胞,才能生长于选择性培养基上。如此选择后的转基因事件可以得到增殖并再生为整株植物、制造种子并将转基因传递给后代。
农杆菌的制备:
经过改造的根癌农杆菌LBA4404可被构建为包含用于种子偏好地表达YL DGAT1和YL DGAT2基因的质粒,如美国专利5,591,616中所公开(该专利的内容特此引入本文以供参考)。通过将上述细菌在基本AB培养基上孵育并允许其于28℃孵育约三天,可制备出用于种子偏好地表达YL DGAT1和YL DGAT2基因的质粒转化的一块单菌落原始平板,以将改造后的构建物用于植物转化。基本AB培养基的组成和制备已描述于PCT专利公布WO 02/009040中(该专利的内容特此引入本文以供参考)。然后可通过将转化的农杆菌划线接种至包含50μg/mL奇放线菌素的YP培养基(0.5%(w/v)酵母提取物,1%(w/v)蛋白胨,0.5%(w/v)氯化钠,1.5%(w/v)琼脂),制备出工作平板。
此后,可以在进行玉米转化的前一天制备用于植物转染和共培养的转化的农杆菌。将50mg/mL奇放线菌素、100mM乙酰丁香酮和来自一至二天工作平板的约1/8环量的农杆菌加入至一个包含30mL基本A培养基(如PCT专利公布WO 02/009040中所述进行制备)的烧瓶。接下来将上述农杆菌于28℃200rpm摇床培养约14h。在对数生长中期,可使用标准的微生物学技术收集上述农杆菌,并以3至5X 108CFU/mL密度重悬于包含100mM乙酰丁香酮的561Q培养基中。561Q培养基的组成和制备描述于PCT专利公布WO02/009040。
未成熟胚芽制备:
在对玉米植株进行人工授粉后九至十天,发育中的未成熟胚胎为不透明的,长约1至1.5mm。这样的长度是用PHP18749转化的农杆菌进行转染的最佳大小。可将去壳的穗在50%的市售漂白剂和一滴Tween-20中灭菌30分钟,然后用灭菌水清洗两遍。此时可将未成熟的胚胎从颖果上无菌地剥离,并置入2mL由包含100μM乙酰丁香酮的561Q培养基组成的灭菌的维持溶液。
农杆菌对胚胎的感染和共培养:
将维持溶液自离体的未成熟胚胎缓缓倒走,代之以转化的农杆菌。在温和混匀并孵育约5分钟后,可将农杆菌自上述未成熟胚胎缓缓倒走。然后将未成熟胚胎移至一块562P培养基平板,所述平板的组成已描述于PCT专利公布WO 02/009040。将上述未成熟胚胎按盾片表面向上的方式置于上述培养基上,于20℃暗培养三天。此步骤之后可继之以在包含100μg/mL羧苄青霉素的562P培养基上于28℃暗培养三天,如美国专利5,981,840中所述。
转基因事件的选择:
将培养后的未成熟胚胎转入5630培养基,该培养基可按PCT专利公布WO 02/009040中所述进行制备。所述培养基包含用于选择转基因植物细胞的双丙氨膦,与大麦HGGT表达框相连的BAR基因可赋予转基因植物细胞抗性。间隔十至十四天后,将胚胎转入5630培养基。在5630培养基上培养六至八周后,可观察到推定的转基因胚胎生成组织的活跃生长。
T
0
植株的再生:
将转基因的胚胎生成组织转入288W培养基,并于28℃暗培养至体细胞胚成熟,或约十至十八天。将具有明显的盾片和胚芽鞘的各成熟的体细胞胚转入272胚萌发培养基,并于28℃进行光培养。当芽和根出现之后,将植株个体于土壤中盆栽,并利用典型的园艺方法进行耐寒锻炼。
288W培养基包含下列成分:950mL去离子水;4.3g MS盐(Gibco);0.1g肌肌醇;5mL MS维生素储备液(Gibco);1mL玉米素(5mg/mL溶液);60g蔗糖;8g琼脂(Sigma A-7049,Purified);2mL吲哚乙酸(0.5mg/mL溶液*);1mL 0.1mM ABA*;3mL双丙氨膦(1mg/mL solution*);以及2mL羧苄青霉素(50mg/mL溶液)。上述溶液的pH调节至pH 5.6将上述溶液高压灭菌。标以星号(*)的成分在培养基冷却至60℃之后加入。
272培养基包含下列成分:950mL去离子水;4.3g MS盐(Gibco);0.1g肌肌醇;5mL MS维生素储备液(Gibco);40g蔗糖;以及1.5g固化剂。将此溶液调节为pH 5.6然后高压灭菌。
实施例8
种仁油含量分析
核磁共振(NMR)分析:
种子于4℃在蒸馏水中浸泡12至24小时。将胚胎切下并保存于一块48孔板中。用Virtis 24x48冻干器将样品过夜冻干。NMR(Process Control Technologies-PCT(Ft.Collins,CO))按照生产商的说明书进行设置。用一系列包含精确量的玉米油(Mazola)的5mm NMR管对NMR进行校准。校准标准物为3、6、9、12、15、18、21、27、33和40mg。
实施例9
YL DGAT1和YL DGAT2基因的合成
设计编码YL DGAT1和YL DGAT2的核苷酸序列,以令其在大豆种子中的表达最优化,所用的方法与下列文献中所描述的相似:Wu,G等人,Nucleic Acids Research(2007),35:D76-D79;Villalobos,A.等人,BMC Bioinformatics(2006),7No pp.given;Wu,G.等人蛋白质Expression and Purification(2006),47:441-445;Richardson,S.M.等人Genome Research(2006),16:550-556;Jayaraj,S.等人Nucleic Acids Research(2005)33:3011-3016。DNA分子用DNA 2.0(Menlo Park,CA,USA)进行合成。表达最优化的YL DGAT1和YL DGAT2DNA序列分别如SEQ ID NO:64和SEQ ID NO:66所示。大豆最优化的YLDGAT酶的氨基酸序列如SEQ ID NO:65(YL DGAT1)和SEQ ID NO:67(YL DGAT2)所示,其分别与SEQ ID NO:1和SEQ ID NO:9的翻译产物序列一致。
实施例10
表达YL DGAT基因的大豆体细胞胚的脂肪酸组成
利用KS352、KS349、KS362和KS364质粒构造制备了转基因体细胞胚。上述DNA构造的构建和转化过程详细描述于实施例5。通过对脂肪酸甲基酯的GC分析测定出脂肪酸组成,其中所述脂肪酸甲基酯通过对庚烷萃取物的甲醇钠衍生作用而生成。结果总结于表14。表14将用缺少YL DGAT基因的一种对照组质粒产生的100个事件的脂肪酸组成,与用分别包含YL DGAT1(KS349)和YL DGAT2(KS362)或同时包含此二基因(KS364)的质粒产生的事件的脂肪酸组成进行了比较。对于用包含YL DGAT的DNA构造产生的事件,列出了所有油酸大于30%的事件的平均脂肪酸组成。
表14
用KS 352、349、362、349和362以及364制备的大豆体细胞
胚的脂肪酸组成
表14显示,单独表达YL DGAT1或YL DGAT2基因以及共表达此二基因均改变了大豆体细胞胚的脂肪酸谱。最显著的改变是油酸的升高和亚麻酸的降低,所测试的全部DNA构造均被观察到伴随有此现象。并且,YL DGAT基因的表达还导致了棕榈酸和亚油酸的降低和硬脂酸的升高。
实施例11
表达YL DGAT基因的大豆种子的脂肪酸组成
如实施例6中所述,利用KS362质粒DNA产生了事件AFS4822.1.13.1。转基因T1种子与来自相同T1植物的无转基因分离子种子相比,表现出油脂含量提高了24.6%。这一现象有力地支持了该事件中表达了YL DGAT2这一结论。令含有或不含YL DGAT2转基因的AFS4822.1.13.1的T1种子萌发,并在生长室中培养三个月。令事件AFS4703.1.6的DS-red阳性的T1种子萌发,并栽培于上述YL DGAT2事件旁边。AFS4703.1.6系利用KS332而产生,KS332为一种包含DS-red标记基因但不含YL DGAT2的质粒载体(参见实施例5)。
有几个T1分离子被鉴定为仅产生DS-red阳性的T2种子,表明这些品系相应的转基因为纯合的。从无转基因分离子后代收集的T2种子为DS-red阴性的,这证实了这些品系很可能不含功能性的转基因。
对于每一选择,将六粒种子切开,通过TMSH衍生作用并继之以气相色谱,分析了种子切片的脂肪酸组成,如实施例6中所述。表15A将AFS4822.1.13.1的DS-red阳性分离子的六粒种子切片的平均脂肪酸组成,与来自一无转基因分离子植株的种子切片的平均脂肪酸组成和仅包含DS-red标记基因的事件AFS4703.1.6的种子切片的平均脂肪酸组成进行了比较。结果证明YL DGAT2的表达改变了大豆种子的脂肪酸谱。最显著的改变是油酸的升高和亚麻酸的降低。并且,YL DGAT基因的表达还导致了棕榈酸和亚油酸的降低和硬脂酸的升高。
表15A
利用KS362产生的T2大豆种子的脂肪酸组成
表15B
通过共转化KS349和KS362产生的T2大豆种子的脂肪酸组成
事件AFS4818.1.9和AFS4818.1.3系利用来自质粒KS349(YLDGAT1)和KS362(YL DGAT2)的DNA片段的混合物而产生,如实施例5中所述。此二事件的转基因T1种子与来自相同T1植物的无转基因分离子种子相比,分别表现出油脂含量提高了22.9%和16.2%。尽管上述现象有力地支持了两个事件均表达转基因来源的YL DGAT这一结论,但并不清楚此二事件究竟包含用于转化的DNA混合物中存在的全部两种DGAT基因的完整拷贝,还是仅包含其中一种。对事件AFS4818.1.2、AFS4818.1.3、AFS4818.1.5、AFS48182.6和AFS4818.1.9油脂和油酸含量升高的T1种子(参加实施例6)进行了种植。分离了DNA,用EcoRI和HindIII两种限制酶消化并用标准操作方法转至尼龙膜。制备两份印迹,并分别与对应于YL DGAT1基因的一种1.21kb限制性片段(通过用NcoI/EcoRI消化KS349产生)和完整的YL DGAT2基因(通过用NotI消化KS 362产生)的探针杂交。基于KS 349(SEQ ID NO:48)和KS 362(SEQ ID NO:52)的序列,大小为≥1.908和≥3.335kb的限制性片段的强杂交信号,可分别指示YL DGAT1和YL DGAT2基因完整拷贝向大豆基因组中的插入。与此相符的是,当使用一种YL DGAT1探针时,所有事件均表现出≥1.908kb的强杂交条带(图8A)。而当使用来自未经改造的大豆的DNA时,并未观察到杂交信号。(泳道11和12,图8A和B)。这表明所有受测的事件均含有存在于KS349上的完整YLDGAT1表达框的至少一个拷贝的插入。不过,当将YL DGAT2的DNA用于杂交实验时,事件4818.1.9仅表现出一条非常弱的高分子量杂交条带,而受测的其他事件全部表现出≥3.335kb的强杂交条带(泳道9,图8B)。接下来将全部五个事件的基因组DNA用BstXI消化,转至尼龙膜并用如前所述制备的完整YL DGAT2DNA进行探针杂交(图9)。0.584kb(内部片段)的强杂交条带和另外两条分别为≥0.2kb和≥0.77kb的片段,可指示YL DGAT2表达框的一个完整拷贝的插入。除4818.1.9外的全部事件均表现出指示存在YLDGAT2的完整插入的杂交模式。推断4818.1.9仅包含YL DGAT1的一个功能性表达单位。
将事件4818.1.9和4818.1.3的T1植株培育至成熟并收集种子,其中所述T1植株来源于事件4818.1.9和4818.1.3的油脂和油酸含量升高了的种子。通过对来自T2种子的切片进行TMSH衍生和GC分析,测定了T2种子的脂肪酸组成。来自4818.1.9和4818.1.3的T2植株的转基因和无转基因分离子种子的脂肪酸组成比较于表中(表15B)。其证明了YL DGAT1的表达如同YL DGAT1和YLDGAT2的共表达一样,改变了大豆种子的脂肪酸谱。最显著的改变是油酸的升高和亚麻酸的降低。并且,YL DGAT基因的表达还导致了棕榈酸和亚油酸的降低和硬脂酸的升高。
实施例12
转基因事件的分析:生长室
本实施例描述了对来自携带纯合或杂合的转基因的T2植株的大豆进行的油脂含量测量,其中所述转基因包括分别的YL DGAT1或YL DGAT2,以及YL DGAT1和YL DGAT2。T2植株生长于受控环境(生长室)中。
利用NMR对来自生长于植物生长室的植株的T2大豆种子进行了油脂分析。种子分别收集自单个植株。从杂合植株的种子选择表现出DS red标记的分离,其中所述杂合植株来自用解脂耶氏酵母的DGAT2基因进行的转化。来自相同植株的DS red阳性种子(带有DGAT2转基因)和无转基因分离子种子的油脂含量显示于表16中。在所述表中,比较了包含DGAT2转基因的种子的油脂含量和来自相同植株的非转基因的、无转基因分离子种子的油脂含量。
表16
来自用于分离解脂耶氏酵母DGAT2转基因的转基因大豆T2植
株的转基因种子和无转基因分离子种子的油脂含量
通过共转化KS349和KS362而产生的事件的T2种子选择,系如上所述(实施例6)通过对种子切片的GC分析进行筛选。种子分别收集自单个植株。令来自杂合植株的种子选择分离,以分离出油酸含量升高(>22%总脂肪酸)的种子选择,其中所述杂合植株来自用解脂耶氏酵母DGAT基因进行的转化。实施例11描述了事件4818.1.9仅包含来自解脂耶氏酵母的DGAT1基因的一个完整表达框。来自相同植株的油酸含量升高的种子(带有DGAT1转基因)和无转基因分离子种子的油脂含量显示于表17中。在该表中,比较了包含DGAT 1转基因的种子的油脂含量和来自相同植株的非转基因的、无转基因分离子植株种子的油脂含量。
表17
来自用于分离解脂耶氏酵母DGAT1转基因的转基因大豆T1植
株的转基因种子和无转基因分离子种子的油脂含量
实施例11中描述了通过共转化KS349和KS362产生的其他事件包含解脂耶氏酵母两种DGAT基因的一个完整表达框。来自相同植株的油酸含量升高的种子(带有两种DGAT转基因)和无转基因分离子种子的油脂含量显示于表18中。在该表中,比较了包含两种DGAT转基因的种子的油脂含量和来自相同植株的非转基因的、无转基因分离子植株的油脂含量。
表18
来自用于分离耶氏酵母DGAT1和DGAT2转基因的转基因大豆
T1植株的转基因种子和无转基因分离子种子的油脂含量
KS362来源的耶氏酵母DGAT2基因表达框纯合的T2种子选择,其DS-red标记不再分离。通过NMR对48粒种子(或者当所获得的种子少于48粒时对获得的全部种子)进行了油脂含量测量。对于每一事件,将DS-red阴性的T1种子种植于栽培DGAT转基因纯合的T1植株所用的同一生长室中,并从长出的植株收集了无转基因分离子的T2种子。来自无转基因分离子选择的种子和来自DGAT转基因纯合品系的种子的油脂含量显示于表19中
表19
来自耶氏酵DGAT2转基因纯合的转基因大豆T1植株的无转
基因分离子种子和转基因种子的油脂含量
KS349来源的耶氏酵母DGAT1和KS362来源的DGAT2表达框纯合的T2种子选择,其与耶氏酵母DGAT基因的表达相联系的油酸升高(油酸≥22%)这一表型不再分离。通过NMR对48粒种子(或者当所获得的种子少于48粒时对获得的全部种子)进行了油脂含量测量。对于每一事件,将未表现出油酸升高的T1无转基因分离子种子种植于栽培DGAT转基因纯合的T1植株所用的同一生长室中,并从长出的植株收集了无转基因分离子的T2种子。来自无转基因分离子选择的种子和来自DGAT转基因纯合品系的种子的油脂含量显示于表20中。
表20
来自耶氏酵母DGAT1和DGAT2转基因纯合的转基因大豆T1
植株转基因种子和无转基因分离子种子的油脂含量
总之,从生长室获得的结果显示,大豆种子中与过表达耶氏酵母两种DGAT基因之一或共表达耶氏酵母两种DGAT基因相联系的油脂增加这一性状有极好的遗传力。与单独表达一种耶氏酵母DGAT基因相联系的油脂增加(与无转基因分离子种子相比)至少为10.9%,最高达25.1%。与共表达两种耶氏酵母DGAT基因相联系的油脂增加(与无转基因分离子种子相比)至少为14.4%,最高达24.2%。
实施例13A
转基因事件的分析:大田
本实施例描述了对来自携带纯合或杂合的转基因的T2植株的大豆进行的油脂含量测量,其中所述转基因包括分别的YL DGAT1或YL DGAT2,以及YL DGAT1和YL DGAT2。T2植株生长于非受控环境(大田)中。
利用YL DGAT2(包含于KS362中)产生的转基因事件的DS red阳性T1种子,和相应的DS red阴性的无转基因分离子种子,于2007年夏季种植于爱荷华州的一块大田中。通过利用YL DGAT1和YL DGAT2共转化而产生且油酸含量升高的T1种子,和相应的具有正常油酸水平的无转基因分离子种子,也以相似方式进行种植。自单个植株分别收集T2种子,并对其进行NMR分析以测量油脂含量。表21显示了来自KS362转基因纯合的事件的48粒均为DS red阳性的种子的油脂含量,和来自培育于相同环境中的相同事件的DS-red阴性的无转基因分离子种子的油脂含量。
表21
来自耶氏酵母DGAT2转基因纯合的大田生长转基因大豆T1植
株的无转基因分离子种子和转基因种子的油脂含量
表22显示了来自YL DGAT1和YL DGAT2纯合的分离子的48粒种子的油脂含量。所有种子均收集自表现出与YL DGAT表达相联系的油酸含量升高的纯合T2种子选择。在表22中,比较了上述品系的油脂含量,和相同事件的油酸水平未变的无转基因分离子种子的油脂含量,其中所述无转基因分离子种子来自与前者种植于相同环境中的植株。
表22
来自耶氏酵母DGAT1和DGAT2转基因纯合的大田生长转基因
大豆T1植株的无转基因分离子种子和转基因种子的油脂含量
实施例11描述了事件4818.1.9仅包含来自耶氏酵母的DGAT1基因的一个完整表达框。来自油酸含量升高的T1种子的上述事件的三株T1植株,与来自油酸含量未变的T1种子的无转基因分离子植株一起栽培于上述爱荷华州大田内。全部三个转基因分离子的T2种子仍然表现出油酸含量升高这一表型的分离,这表明亲本品系依然为DGAT1转基因杂合的。利用GC分析将所有转基因阳性的种子从这些品系中鉴定出来,并通过NMR对其进行油脂分析。在表23中,将这些种子的油脂含量与来自生长于相同环境中的T1植株的无转基因分离子种子的油脂含量进行了比较。
表23
来自耶氏酵母DGAT1转基因杂合的大田生长转基因大豆T1植
株的无转基因分离子种子和转基因种子的油脂含量
总之,在大田环境下获得的结果显示,大豆种子中与过表达耶氏酵母两种DGAT基因之一或共表达耶氏酵母两种DGAT基因相联系的油脂增加这一性状有极好的遗传力。与单独表达一种耶氏酵母DGAT基因相联系的油脂增加(与无转基因分离子种子相比)至少为11%,最高达14%。与共表达耶氏酵母DGAT基因相联系的油脂增加(与无转基因分离子种子相比)至少为19%,最高达20%。
实施例13B
大豆种子的组成分析
本实施例描述了对大豆种子的组成参数,如蛋白质含量和可溶性碳水化合物含量的测量,其中所述大豆种子来自单独表达一种YLDGAT基因(YL DGAT2)或同时表达两种YL DGAT基因的转基因事件。
对大豆种子与YL DGAT基因的表达相联系的组成变化进行了测量。出于此目的,测量了蛋白质、可溶性碳水化合物和淀粉的浓度如下。
非结构性碳水化合物和蛋白质分析。
将大豆干种子用GenoGrinder研磨仪研磨为细粉,称取子样品(精确度为0.1mg)并置入13x 100mm玻璃管,所述玻璃管配有内衬的密封旋盖。所测试的每一样品设三个重复。通过称量子样品在置于强制通风的烘箱内105℃烘干18h之前和之后的重量,计算出组织干重。
通过向每一玻璃管加入2mL庚烷进行脂质的萃取。将上述玻璃进行振荡混匀,并置入充满60℃热水的超声波浴缸(VWR ScientificModel 750D)。将样品满功率(约360W)超声处理15min,然后离心(5min×1700g)。将上清转入干净的13×100mm玻璃管,用庚烷(第二次萃取2mL,第三次萃取1mL)将沉淀再萃取两次,合并每次萃取的上清。进行脂质萃取之后,向沉淀加入1mL丙酮,振荡混匀以令材料完全分散,然后用Speedvac真空离心浓缩仪进行干燥。
非结构性碳水化合物萃取和分析。
向上述干燥后的沉淀加入2mL 80%酒精。将样品彻底地振荡混匀,直至植物材料完全分散于溶剂中,然后于60℃超声处理15min。5min×1700g离心之后,将上清轻轻倒入干净的13×100mm玻璃管。用80%酒精再进行两次萃取,合并每次萃取的上清。将萃取后的沉淀悬浮于丙酮并干燥(如上所述)。每次萃取时在用Speedvac干燥之前,加入β-苯基吡喃葡萄糖苷(100μL 0.5000+/-0.0010g/100mL储备液)作为内部标准。将萃取物保存于干燥器留待进一步分析。
将上述丙酮干燥后的粉末悬浮于0.9mL包含100U热稳定的α-淀粉酶(来自地衣芽孢杆菌;Sigma A-4551)的MOPS(50mM 3-N(吗啉代)丙烷-磺酸,5mM CaCl2,pH7.0)缓冲液。将样品置于加热块(90℃)中75min,每15min振荡混匀一次。令样品冷却至室温,向每一样品加入0.6mL包含5U淀粉葡萄糖苷酶(Roche 110 202 367001)的醋酸盐(285mM,pH 4.5)缓冲液。将样品在配有往复式摇动器的水浴器内于55℃孵育15至18h,掺入可溶性马铃薯淀粉(SigmaS-2630)作为标准物,以确保淀粉消化完全。
消化完成后萃取释放出的碳水化合物,然后进行分析。向每一管中加入无水酒精(6mL),将样品振荡混匀之后于60℃超声处理15min。将样品离心(5min×1700g),然后将上清轻轻倒入干净的13×100mm玻璃管。用3mL 80%酒精将沉淀再萃取两次,合并所得到的上清。每一样品在用Speedvac干燥之前先加入内部标准(100μL β-苯基吡喃葡萄糖苷,如上所述)。
样品的制备和分析
令来自上述可溶性和淀粉萃取的干燥后的样品,溶解于包含30mg/mL盐酸羟胺(Sigma-Aldrich 159417)的无水吡啶(Sigma-AldrichP57506)中。将样品置于定轨摇床(300rpm)过夜,然后加热1h(75℃),加热过程中每15min剧烈振荡混匀一次。冷却至室温后,加入1mL六甲基二硅氮烷(Sigma-Aldrich H-4875)和100μL三氟乙酸(Sigma-Aldrich T-6508)。将样品振荡混匀,令沉淀物沉降,然后将上清转移至GC样品瓶。
在一台配有DB-17MS毛细管柱(15m×0.32mm×0.25μm膜)的Agilent 6890气象色谱仪上对样品进行分析。入口和检测器温度均为275℃。注射(2μL,20∶1分流)后,将柱温以3℃/min的速率从初始温度(150℃)升高至180℃,然后再以25℃/min的速率升高至320℃终点温度。终点温度维持10min。载气为51cm/sec线速度的H2。通过火焰电离进行检测。数据分析利用Agilent ChemStation软件进行。每种糖相对于内部标准进行定量,并记录每种单独的碳水化合物的检测器响应(从与每组样品一起受测的标准物计算)。最终的碳水化合物浓度以组织干重作为基准表示。
蛋白质分析
通过在一台Thermo Finnigan Flash 1112EA燃烧分析器上进行燃烧分析,估计出蛋白质含量。用一台Mettler-Toledo MX5微量天平以0.001mg精确度称量4至8mg的样品用于分析。将通过分析器测定出的%N乘以6.25,计算出蛋白质含量。最终的蛋白质含量以占组织干重基准的%表示。
表24
来自两株分别为无转基因分离子或YL DGAT1 YL DGAT2转基因纯合的T1植株的大豆种子的组成分析。所述植株生长于相同的生长室环境中。数值单位为g/kg DW,除非另有说明。
表25
来自一株YL DGAT2转基因杂合的T1植株的大豆种子的组成分析。所述植株生长于生长室中。基于作为可见标记的DS red的表达,对YL DGAT转基因种子和无转基因分离子种子进行了选择。如上所述,将八粒DS red阳性的和DS red阴性的种子合并并进行了分析。数值单位为g/kg DW,除非另有说明。
表26
来自YL DGAT2转基因杂合或YL DGAT1 YL DGAT2转基因杂合的T1植株的大豆种子的组成分析。所述植株生长于大田中。基于作为可见标记的DS red的表达(4822.2.10),或基于GC分析测定为油酸含量升高(4818.1.2),对YL DGAT转基因种子和无转基因分离子种子进行了选择。如上所述,将八粒转基因阳性的和转基因阴性的种子合并并进行了分析。数值单位为g/kg DW,除非另有说明。
表24至26说明,一种或两种YL DGAT基因在不同环境(生长室,大田)中的表达,与种子组成的一种一致改变相联系,其中所述改变的特征在于可溶性碳水化合物的减少,也就是蔗糖的减少和水苏糖的更小程度的减少。最重要的是,当油脂积累通过YL DGAT基因的表达增加时,并未观察到蛋白质含量的减少。
实施例14
发育中的种子和体细胞胚中DGAT活性的测量
本实施例描述了单独包含耶氏酵母DGAT2或同时包含耶氏酵母DGAT2和DGAT1的大豆表达载体的构建、上述基因在大豆种子或体细胞胚中的表达以及在这些组织内的DGAT酶活性。
包含YL DGAT2的pKR1234的构建
将KS362(SEQ ID NO:52)包含YL DGAT2的NotI片段,克隆至pKR72(SEQ ID NO:26;实施例4)的NotI片段,以构建出pKR1234(SEQ ID NO:68)。
包含YL DGAT1和DGAT2的pKR1236的构建
利用引物oSGly-2(SEQ ID NO:69)和oSGly-3(SEQ ID NO:70),通过PCR将大豆球蛋白Gy1启动子从pZBL119(描述于PCT专利公布WO 2004/071467,该专利的内容特此引入本文以供参考)上扩增出来。按生产商的说明书,将所获得的PCR片段亚克隆至中间克隆载体pCR-Script AMP SK(+)(Stratagene),以构建出质粒pPSgly32(SEQ ID NO:71)。
将质粒pSGly32(SEQ ID NO:71)包含Gy1启动子的PstI/NotI片段,克隆至质粒pKR142(描述于PCT专利公布WO 2004/071467)的PstI/NotI片段,以构建出pKR264(SEQ ID NO:72),其中所述质粒pKR142包含豆球蛋白A23′转录终止区域、一个氨苄青霉素抗性基因和细菌复制起点。如此,载体pKR264包含了一个两侧分别为大豆球蛋白Gy1基因启动子和豆球蛋白A23′转录终止区域的NotI位点(Gy1/NotI/legA2框)。
将质粒KS349(SEQ ID NO:48)包含耶氏酵母DGAT1的NcoI/XbaI片段,克隆至质粒pKR908(US Pat.Pub US20080095915)的NcoI/XbaI位点,以构建出pKR1212(SEQ ID NO:73),其中所述质粒pKR908包含两侧为NotI位点的NcoI/XbaI位点。
将pKR1212(SEQ ID NO:73)包含耶氏酵母DGAT1基因的NotI片段,克隆至pKR264(SEQ ID NO:72)的NotI片段,以构建出pKR1235(SEQ ID NO:74)。
将pKR1235(74)包含耶氏酵母DGAT1的BsiWI片段,克隆至pKR1234(SEQ ID NO:68)的BsiWI位点,以构建出pKR1236(SEQ IDNO:75)。
对来自发育中T2种子的微粒体提取物进行的DGAT分析
如本文(实施例6)所述,用KS362转化了大豆胚胎生成悬浮培养物(cv.Jack),其中所述KS362如本文所述包含耶氏酵母DGAT2,并且种植了来自大豆植株AFS4822.1.13.1(种子称为7GR11-58)和AFS4822.2.10.1(种子称为7GR11-66)的T1种子,植物的栽培如实施例12中所述。在上述两个事件中,均发现转基因阳性种子与同一事件的无转基因分离子种子相比,油脂浓度有提高。
类似地,如本文(实施例6)所述,用KS362(包含YL DGAT2)和KS349(包含YL DGAT1)共转化了大豆胚胎生成悬浮培养物(cv.Jack)。来自事件AFS4818.1.2.1的转基因阳性T1种子被命名为7GR11-2。在该事件中,发现转基因阳性种子与同一事件(实施例6)的无转基因分离子种子相比,油脂浓度有提高。来自AFS4818.1.2.1和AFS4818.1.3.17的转基因阴性的无转基因分离子T1种子分别被命名为7GR11-7和7GR11-15。种植了这些种子,植物的栽培如实施例12中所述。
从经选择的开花后30天(DAF)的植株收集约1g T2种子,在液氮中急冻,然后保存于-80℃留待处理。用研钵和研杵将1g冷冻的种子组织在液氮中磨碎,然后加入3mL植物匀浆缓冲液(300mM蔗糖;1mM EDTA;10mM Tris.HCl,pH 8.0;1mM DTT;0.1%交联聚乙烯基吡咯烷酮),并用polytron匀浆器将组织进一步匀浆1分钟。用3层粗滤布真空过滤,继而以1层mira cloth神奇滤布过滤,收集滤渣。将得到的滤液于1,500×g离心15min两次,然后将上清于100,000x g离心60min。用移液器轻轻吹打令所得到的沉淀重悬于约0.5至1mL微粒体缓冲液(100mM磷酸钾,pH 7.2)中,并在一个涂有特氟隆的2mL尺寸杜恩斯匀浆器中进一步重悬。用Bradford试剂(Sigma-Aldrich)测定蛋白质浓度,将微粒体在液氮中急冻,保存于-80℃留待分析。
DGAT分析于25℃在植物分析缓冲液(500mM三(羟甲基)甲基甘氨酸,pH 7.8;28mM氯化钠;0.06%CHAPS)中进行,反应体系还包含20μM 1-14C-标记的油酰-辅酶A(50mCi/mmol,Perkin Elmer)、1.5mM二油酰基甘油(Sigma-Aldrich)和20μg的微粒体蛋白质,总反应体积为100μL,反应时间为5min。每一反应开始之前,先将其余的反应组分加好,最后加入微粒体膜。通过加入1mL热异丙醇(75℃)并于75℃加热10min终止反应。待反应混合液冷却至室温,加入1.5mL己烷并将样品混匀。加入1.25mL 500mM硫酸钠,低速离心分相,将上相转入另一玻璃管。然后在氮气中干燥上相。将来自每一反应的脂质重溶于75μL己烷,并加入1μL大豆油。将脂质加到一块Partisil K6 Silica Gel 60A TLC平板(Whatman,250μL厚,20cm×20cm),通过用80∶20∶1(v/v/v)的己烷∶二乙醚∶乙酸处理,使甘油三酯于其他脂质分开。通过在一个缸内用碘蒸气轻微染色,使甘油三酯显色并将其标记。将平板自碘缸中移出,待颜色褪去后,刮掉甘油三酯,通过液体闪烁计数测定放射性,以dpm/min表示。利用下列公式,将总活性测定为每分钟每mg蛋白质掺入甘油三酯中的放射性同位素标记的油酸的量:([dpm]/[2200000dpm/uCi]/[50uCi/umol]/[5min.]/[0.02mg蛋白质]×[1000nmol/umol])。所述每一样品的DGAT活性显示于表27中。
表27
经选择的发育中T2种子的DGAT活性。
来自大豆体细胞胚的微粒体萃取物的DGAT分析
用包含耶氏酵母DGAT2的pKR1234(SEQ ID NO:68)转化大豆胚胎生成悬浮培养物(cv.Jack),实验编号为MSE2181;用包含耶氏酵母DGAT2和DGAT1的pKR1236(SEQ ID NO:75)转化大豆胚胎生成悬浮培养物(cv.Jack),实验编号为MSE2182。对事件进行选择,体细胞胚在ShaM中的成熟如实施例5所述。
在ShaM中成熟2周后,将来自每一事件的约1g组织在液氮中冷冻,并用研钵和研杵磨碎,如处理大豆发育中种子的方法中所述。将少量研磨后的组织(约100mg)冻干过夜,剩余组织保存于-80℃。
利用GC方法和17:0内部标准物,对来自每一事件的约10mg冻干的组织测定油脂浓度,完全如实施例5中所述,油脂浓度和油酸含量(%总FAME)显示于表28中。微粒体蛋白质的制备、蛋白质浓度的测定和对经选择的事件的DGAT分析的进行完全如上述处理种子组织的方法中所述,其中所述经选择的事件已被测定为具有一定范围的油脂浓度。DGAT分析的结果也显示于表28中。
表28
用pKR1234或pKR1236转化的大豆体细胞胚的酯化脂肪酸、
油酸含量和DGAT活性
与具有野生型油脂水平的事件相比,用pKR1234转化并且具有最高的一些油脂浓度的事件的DGAT活性升高达到了9倍。与具有野生型油脂水平的事件相比,用pKR1236转化并且具有最高的一些油脂浓度的事件的DGAT活性升高达到了15.8倍。
此外还用包含耶氏酵母DGAT2和DGAT1的KS364(SEQ ID NO:63)转化了大豆胚胎生成悬浮培养物(cv.Jack)(实验编号MSE2134),并分析了各个事件的脂肪酸谱和油脂浓度,如实施例5中所述。基于上述数据,选择了两个事件进行DGAT分析,一个事件(事件54)具有高油酸(占总脂肪酸的32.83%)和油脂浓度(12.5%DCWt),另一个事件(事件33)具有野生型的油酸(占总脂肪酸的15.52%)和油脂浓度(8.2%DCWt)。将来自每一事件的转化的胚胎生成悬浮培养物培养于SB 196培养液中,并令胚胎在ShaM中成熟,如实施例5中所述。
在ShaM中成熟2周后,将来自每一事件的约1g组织在液氮中冷冻,制备微粒体蛋白质,对每一事件进行DGAT分析,完全如上文所述,结果显示于表29中。同样也提取了总脂质,并如下文所述测定了油酸和油脂浓度,结果报告于表29中。
表29
用KS364转化的大豆体细胞胚的酯化脂肪酸、油酸含量和
DGAT活性
与具有野生型油脂水平的事件(事件33)相比,具有高油脂浓度的事件(事件54)的DGAT活性升高达到10.7倍。
实施例15
表达DGAT基因的转基因种子和体细胞胚的脂质成分分析
将用来自一个具有野生型油脂和油酸浓度的事件(事件33)和来自一个具有具有高油脂和油酸浓度的事件(事件54)的KS364转化的大豆体细胞胚,冻干48小时,并用genogrinder研磨仪将组织磨碎,完全如实施例5中所述。
总脂质萃取
利用在Bligh,E.G.和Dyer,W.J.方法(Can.J.Biochem.Physiol.37:911-917(1959))基础上作了一些改进的方法,从每一事件提取了总脂质。简言之,将来自每一事件的约100mg研磨后的组织加入至一个16mm×125mm尺寸的配有内衬特氟隆的旋盖的试管中。加入甲醇∶氯仿/2∶1混合物(6mL),温和搅拌萃取样品1小时,然后加入2mL氯仿,并继续搅拌30min。然后,加入3.6mL水,剧烈振荡试管,用临床离心机离心分相。将下层有机相小心转移至第二个玻璃试管,用2mL氯仿将上层水相再萃取一次。再次离心,并将下层有机相与第一次的有机相合并。将样品于50℃在氮气流中干燥,通过称重估算出总脂质,将脂质溶于氯仿∶甲醇/6∶1至浓度为约10mg/mL。利用本文(实施例4)所述的硫酸/甲醇方法,对约50μg的每一样品进行FAME分析,结果显示于表30中。
中性和极性脂质的分离
依次用5mL甲醇、5mL甲醇∶氯仿/1∶1和5mL氯仿平衡Sep-pak氨基-丙基固相萃取柱(Waters;6cc柱,WAT054560)。向每一柱中加入溶于氯仿∶甲醇/6∶1的约5mg总脂质,然后加入5x 1mL的氯仿等份以洗脱中性脂质,收集并合并所有流分,于50℃在氮气流中干燥。然后依次用5×1mL等分试样的甲醇∶氯仿/1∶1等份和5×1mL的甲醇等份将极性脂质从每一柱上洗脱,合并所有流分并在氮气中干燥。将中性脂质溶解于约1mL的CHCl3∶MeOH/6∶1中,将极性脂质溶解于约200μL的CHCl3∶MeOH/6∶1中。利用本文(实施例4)所述的硫酸/甲醇方法,对约50μg的中性脂质进行FAME分析,结果显示于表30中。
通过TLC对TAG、PC和PE的分离
将约100μL中性脂质萃取物上样至一块Partisil K6 Silica Gel 60A TLC平板(Whatman,250μL厚,20cm x 20cm)距底部2cm处。类似地,将约200μL极性脂质流分上样至相同的TLC平板上。TAG、PC和PE的标准溶液(10mg/mL,溶于氯仿∶甲醇/6∶1)也同样上样至平板。用CHCl3∶MeOH∶AcOH/65∶35∶8混合液处理TLC平板,直到溶剂前沿达至平板的约一半处。将TLC平板置于空气中干燥10min,然后将其完全暴露于70∶30∶1(v/v/v)的己烷∶二乙醚∶乙酸中。通过在碘缸内轻微染色,使标准物显色,并将TAG、PC和PE所对应的条带自TLC平板上挖去。利用本文(实施例4)所述的硫酸/甲醇方法,直接衍生包含各种脂质的硅胶,结果显示于表30中。
TAG的脂肪酸位置分析
利用猪胰脂肪酶移除TAG sn1和sn3位上的酰基,继而通过所得到的单酰甘油(MAG)的酯交换反应,测定了TAG sn2位的脂肪酸谱。将约5mg的中性脂质萃取物悬浮于2mL 1M Tris.HCl,pH 8.0,置于一个带旋盖的玻璃试管中,并加入0.2mL 2.2%氯化钙和0.5mL0.05%胆盐。将脂质于37℃孵育5min,直接加入5mg猪胰脂肪酶,将悬液于37℃在摇动中孵育20min。孵育后,加入1mL乙醇并继而加入1mL 6M HCl,以终止反应。混匀之后加入2.5mL二乙醚,离心分相,小心转移上层有机相。重复二乙醚萃取一次,将两次的上层二乙醚相合并。用无水硫酸钠干燥,然后于50℃在氮气流中令二乙醚蒸发,将所得到的脂质溶于200μL氯仿∶甲醇/6∶1中。将上述脂质与三酰甘油(TAG)、二酰甘油(DAG)、单酰甘油(MAG)和游离脂肪酸(FFA)标准物一起上样至一块Partisil K6TLC平板,并将平板如本文所述进行处理。然后,通过轻微的碘染色令标准物显色,将MAG条带切下,并用如本文中所述的硫酸/甲醇方法衍化。来自上述MAG条带的FAME脂肪酸谱结果,代表了TAG sn2位(即甘油的C2上的酰基)的脂肪酸谱,所述结果与sn1和sn3位的计算值一起显示于表30中。表30中,在TAG sn1和sn3位上,每种脂肪酸(即16:0、18:0、18:1、18:2和18:3)占总脂肪酸的%通过下列公式计算:=([TAGx]-[sn2x]/3)*3/2;其中x指代所感兴趣的脂肪酸。
表30
TAG中各种脂质的脂肪酸组成和位置分布
在TAG中观察到的同YL DGAT的表达相联系的脂肪酸谱的变化,在极性脂质中也同样有观察到。
实施例16
氨基酸序列发生改变的耶氏酵母DGAT突变体
本实施例描述了突变体形式的耶氏酵母DGAT2的构建,突变基因向酵母表达载体的克隆,以及对微粒体蛋白组分的DGAT活性的分析。.
包含突变型耶氏酵母DGAT2的酿酒酵母表达载体的构建
按照生产商的说明书,利用PhusionTM高保真DNA聚合酶(Cat.No.F553S,Finnzymes Oy,Finland)以及寡核苷酸引物oYDG2-1(SEQ ID NO:76)和oYDG2-2(SEQ ID NO:77),将耶氏酵母DGAT2自pKR1234(SEQ ID NO:68;实施例14)中扩增出来。按照生产商的说明书,利用ZeroPCR克隆试剂盒(Invitrogen Corporation),将得到的DNA片段克隆至克隆载体,从而得到pKR1254(SEQ ID NO:78)。
按照生产商的说明书,利用定点突变试剂盒(Cat.No.200518,Stratagene,La Jolla,CA)以及寡核苷酸Y1D2_Y326F-5(SEQ ID NO:79)和Y1D2_Y326F-3(SEQ ID NO:80)使耶氏酵母DGAT2序列上的一个密码子发生改变,所述密码子编码耶氏酵母DGAT2序列所对应的氨基酸序列(SEQ ID NO:10)上的氨基酸Y326。经过大量测序,选择了一个克隆用于进一步研究,该克隆编码的一种氨基酸序列除Y326变为了F326外,与耶氏酵母DGAT2(SEQ IDNO:10)完全一致。该克隆被命名为pKR1254_Y326F(SEQ ID NO:81)。编码序列(Y1DGAT2_Y326F)改变后的核苷酸序列如SEQ IDNO:82所示,对应的氨基酸序列如SEQ ID NO:83所示。
按照生产商的说明书,利用定点突变试剂盒(Cat.No.200518,Stratagene,La Jolla,CA)以及寡核苷酸Y1D2_Y326L-5(SEQ ID NO:84)和Y1D2_Y326L-3(SEQ ID NO:85)使耶氏酵母DGAT2序列上的一个密码子发生改变,所述密码子编码耶氏酵母DGAT2序列所对应的氨基酸序列(SEQ ID NO:10)上的氨基酸Y326。经过大量测序,选择了一个克隆用于进一步研究,该克隆编码的一种氨基酸序列除Y326变为了L326外,与耶氏酵母DGAT2(SEQ IDNO:10)完全一致。该克隆被命名为pKR1254_Y326L(SEQ ID NO:86)。编码序列(Y1DGAT2 Y326L)改变后的核苷酸序列如SEQ IDNO:87所示,对应的氨基酸序列如SEQ ID NO:88所示。
按照生产商的说明书,利用定点突变试剂盒(Cat.No.200518,Stratagene,La Jolla,CA)以及寡核苷酸Y1D2_R327K-5(SEQ ID NO:89)和Y1D2_R327K-3(SEQ ID NO:90)使耶氏酵母DGAT2序列上的一个密码子发生改变,所述密码子编码耶氏酵母DGAT2序列所对应的氨基酸序列(SEQ ID NO:10)上的氨基酸R327。经过大量测序,选择了一个克隆用于进一步研究,该克隆编码的一种氨基酸序列除R327变为了K327外,与耶氏酵母DGAT2(SEQ IDNO:10)完全一致。该克隆被命名为pKR1254_R327K(SEQ ID NO:91)。编码序列(Y1DGAT2_Y326L)改变后的核苷酸序列如SEQ ID NO:92所示,对应的氨基酸序列如SEQ ID NO:93所示。
将pKR1254、pKR1254_Y326F、pKR1254_Y326L和pKR1254_R327K各自包含一个野生型或突变型Y1DGAT2基因的NotI片段,分别克隆至pY75(SEQ ID NO:3;实施例1)的NotI位点,从而分别获得pY191(SEQ ID NO:94)、pY192(SEQ ID NO:95)、pY193(SEQ ID NO:96)和pY194(SEQ ID NO:97)。
对突变型耶氏酵母DGAT2的DGAT活性的分析
自Open Biosystems(http://www.openbiosystems.com/)获得了一种敲除了内源性DGAT2基因并具有下列基因型(BY4741,MATa his3Δ1leu2Δ0met15Δ0 ura3Δ0)的酿酒酵母突变型菌株(DGA1)。用pY191、pY192、pY193或pY194转化该菌株并如本文所述分离转化子。将每一转化的三个独立的转化子在2mL添加了CSM-leu的DOBA培养液中于30℃孵育16h,获得培养物。将细胞(1mL)转入50mL上述DOBA培养液中并于30℃再培养16h。离心令细胞沉淀,在液氮中冷冻,然后保存于-80℃留待使用。
将沉淀重悬于2mL酵母匀浆缓冲液(20mM Tris.HCl,pH 8.0;10mM MgCl2;1mM EDTA;5%glycerol;1mM DTT;0.3M(NH4)2SO4),并将悬液加入至包含约1mL 0.5mm玻璃珠的2mL旋盖管中。通过旋涡振荡除去气穴,此时悬液充至管的上部,盖上管盖,用珠磨器5000rpm研磨三次以破碎细胞,每次1min,每次研磨后置冰上5min。1,500x g 4℃离心酵母匀浆物15min,然后将得到的上清液100,000xg离心60min。用移液器轻轻吹打令所得到的沉淀重悬于约0.2至1mL微粒体缓冲液(100mM磷酸钾,pH 7.2)中,并在一个涂有特氟隆的2mL尺寸杜恩斯匀浆器中进一步重悬。用Bradford试剂(Sigma-Aldrich)测定蛋白质浓度,将微粒体在液氮中急冻,保存于-80℃留待分析。
DGAT分析于25℃在植物分析缓冲液(50mM磷酸钾(pH 7.2))中进行,反应体系还包含20μM 1-14C-标记的油酰-辅酶A(50mCi/mmol,Perkin Elmer)和20μg的微粒体蛋白,总反应体积为100μL,反应时间为1min。每一反应开始之前,先将其余的反应组分加好,最后加入微粒体膜。除了所用的公式针对1min的反应时间作了相应改变以外,分析的终止和TAG中放射性的测定完全如对植物的DGAT分析中所述。assay time(i.e.[dpm]/[2200000dpm/uCi]/[50uCi/umol]/[5min.]/[0.02mg protein]×[1000nmol/umol]).所述每一样品的DGAT活性以及平均值显示于表31中。
表31
用pY191、pY192、pY193或pY194转化的DGA1的DGAT活 性。
从表31来看,Y326F和Y326L氨基酸变化似乎对在酵母中检测的耶氏酵母DGAT2活性影响轻微,选择将这两个突变用于在大豆体细胞胚中表达。
构建包含突变型耶氏酵母DGAT2的大豆表达载体
将pKR1254(SEQ ID NO:78)、pKR1254_Y326F(SEQ ID NO:81)或pKR1254_Y326L(SEQ ID NO:86)各自包含野生型或突变型耶氏酵母DGAT2基因的NotI片段,分别克隆至pKR72(SEQ ID NO:26;实施例4)的NotI片段,从而分别获得pKR1256(SEQ ID NO:98)、pKR1277(SEQ ID NO:99)或pKR1278(SEQ ID NO:100)。
测定表达突变型耶氏酵母DGAT2的大豆体细胞胚的油脂浓度
用包含野生型耶氏酵母DGAT2的pKR1256(SEQ ID NO:98)转化大豆胚胎生成悬浮培养物(cv.Jack),实验编号为MSE2228;或用包含耶氏酵母DGAT2_Y326F的pKR1277(SEQ ID NO:99)转化大豆胚胎生成悬浮培养物(cv.Jack),实验编号为MSE2229;或用包含耶氏酵母DGAT2_Y326L的pKR1278(SEQ ID NO:100)转化大豆胚胎生成悬浮培养物(cv.Jack),实验编号为MSE2230。对事件进行选择,体细胞胚在ShaM中的成熟如实施例5所述。对每一事件,利用NMR测定油脂浓度,并通过GC测定脂肪酸谱,完全如本文中所述,油脂浓度和脂肪酸含量(%总FAME)的结果显示于表32中。
表32
用pKR1256、pKR1277或pKR1278转化的大豆体细胞胚的油脂
浓度
在大豆体细胞胚中,携带Y326F突变的YL DGAT2蛋白变异体所导致的油脂浓度升高和油脂的脂肪酸谱改变的程度,至少也同野生型耶氏酵母DGAT2所导致的程度一样。
实施例17
针对表达进行过优化的DAGT基因
针对在大豆植物中的表达而优化过的编码YL DGAT1和YLDGAT2基因的序列如SEQ ID NO:64和SEQ ID NO:66所示。上述序列的设计描述于实施例9中。包括两翼为NotI限制性位点的上述DNA序列的DNA分子通过DNA 2.0(California,USA)进行合成。用NotI消化含有上述合成的基因的质粒DNA。将包含DGAT基因的NotI限制性片段,与经NotI线性化的去磷酸化的KS332DNA连接,所述KS332描述于实施例5中。在所得到的DNA构造中,耶氏酵母DGAT1或耶氏酵母DGAT2基因针对表达而优化过的变异体的表达,处于β-大豆伴球蛋白启动子的控制下,将所述DNA构造分别命名为KS392和KS393。它们的序列如SEQ ID NO:101和SEQ IDNO:102所示。此外还构建了质粒KS391。为了构建KS391,首先用NotI和NcoI消化KS349的DNA。将所得到的DGAT1限制性片段的两端与经NotI线性化的KS332DNA连接,使前者完全插入至质粒中。得到的质粒构造称为KS391。在该质粒构造中,天然的YLDGAT1序列处于β-大豆伴球蛋白启动子的控制下。KS391的序列如SEQ ID NO:103所示。如上所述(实施例5),用质粒KS391、KS392、KS 362和KS393的DNA进行基因枪轰击,进而再生出转基因大豆体细胞胚。利用NMR测量了体细胞胚的油脂含量。如上所述(实施例4),将短暂冻干的胚胎组织置于genogrnder瓶中粉碎。将20至200mg组织粉末转入NMR管。如实施例4中所述,从NMR信号计算出上述体细胞胚组织粉末的油脂含量。
表33
用pKS392和pKS391转化的大豆体细胞胚的油脂含量
ID 质粒构造 %油脂
1 2196.1.08 KS392 15.7
2 2196.3.03 KS392 13.9
3 2196.1.05 KS392 13.7
4 2196.1.15 KS392 13.4
5 2196.1.02 KS392 12.2
6 2196.3.07 KS392 12.1
7 2196.3.04 KS392 12.1
8 2196.3.05 KS392 11.6
9 2196.1.06 KS392 11.4
10 2196.1.07 KS392 11.4
11 2196.1.03 KS392 10.4
12 2196.1.14 KS392 10.2
13 2196.1.12 KS392 9.4
14 2196.3.08 KS392 9.1
15 2196.3.09 KS392 8.1
16 2196.4.02 KS392 8.1
17 2196.1.09 KS392 7.8
18 2196.2.01 KS392 7.6
19 2196.5.01 KS392 7.4
20 2196.3.01 KS392 7.4
21 2196.1.04 KS392 7.3
22 2196.2.02 KS392 7.3
23 2196.3.02 KS392 6.9
24 2196.1.10 KS392 6.5
25 2196.4.03 KS392 6.5
26 2196.1.11 KS392 6.0
27 2196.4.01 KS392 5.7
28 2196.3.06 KS392 5.4
29 2196.5.02 KS392 5.3
30 2196.1.13 KS392 3.5
平均%油脂9.1
1 2195.3.15 KS391 12.8
2 2195.5.02 KS391 11.5
3 2195.3.01 KS391 11.2
4 2195.3.03 KS391 11.0
5 2195.3.02 KS391 10.6
6 2195.4.06 KS391 10.4
7 2195.3.08 KS391 10.2
8 2195.2.01 KS391 10.1
9 2195.3.04 KS391 9.3
10 2195.3.05 KS391 9.2
11 2195.5.01 KS391 9.1
12 2195.2.04 KS391 8.8
13 2195.3.14 KS391 8.6
14 2195.3.10 KS391 7.8
15 2195.4.01 KS391 7.7
16 2195.5.03 KS391 6.5
17 2195.4.02 KS391 6.4
18 2195.3.09 KS391 6.3
19 2195.4.07 KS391 6.2
20 2195.3.06 KS391 6.1
21 2195.3.13 KS391 6.0
22 2195.5.05 KS391 5.9
23 2195.4.03 KS391 5.6
24 2195.5.04 KS391 5.5
25 2196.1.01 KS391 5.3
26 2195.4.04 KS391 5.3
27 2195.3.11 KS391 5.2
28 2195.2.03 KS391 5.0
29 2195.4.05 KS391 4.9
30 2195.3.07 KS391 4.4
31 2195.2.02 KS391 4.3
平均%油脂 7.6
表33比较了用KS392和KS391分别产生的30个和31个事件的油脂含量。用KS392产生的所有事件的平均油脂含量为9.1%,而用KS391产生的所有事件的平均油脂含量为7.6%。此外,用KS392转化的事件中最高的油脂含量为15.7%,而用KS391转化的最高为12.8%。申请人已证明,与天然的YL DGAT1基因相比,YL DGAT1针对表达所作的优化导致了发育中大豆胚胎油脂含量的升高。
表34
用pKS393和pKS362转化的大豆体细胞胚的油脂含量
样品# ID 质粒构造 %油脂
1 2207.5.05 KS393 12.3
2 2207.5.08 KS393 12.2
3 2207.5.06 KS393 11.7
4 2207.5.03 KS393 10.8
5 2207.5.01 KS393 10.6
6 2207.5.04 KS393 10.3
7 2207.4.04 KS393 10.3
8 2207.3.09 KS393 9.5
9 2207.5.07 KS393 9.3
10 2207.4.01 KS393 8.8
11 2207.4.02 KS393 8.7
12 2207.3.06 KS393 8.0
13 2207.3.04 KS393 7.9
14 2207.4.06 KS393 7.8
15 2207.4.03 KS393 7.7
16 2207.4.08 KS393 7.4
17 2207.3.14 KS393 7.1
18 2207.4.05 KS393 7.0
19 2207.3.12 KS393 7.0
20 2207.3.01 KS393 6.9
21 2207.5.02 KS393 6.8
22 2207.3.02 KS393 6.7
23 2207.3.07 KS393 6.7
24 2207.3.03 KS393 6.6
25 2207.3.13 KS393 6.5
26 2207.3.11 KS393 6.3
27 2207.3.05 KS393 5.8
28 2207.3.10 KS393 5.8
29 2207.4.07 KS393 5.5
30 2207.3.08 KS393 5.4
31 2207.4.09 KS393 5.4
平均%油脂 8.0
1 2208.2.08 KS362 11.6
2 2208.5.04 KS362 11.5
3 2208.2.04 KS362 11.5
4 2208.2.10 KS362 11.2
5 2208.2.09 KS362 10.3
6 2208.2.02 KS362 10.2
7 2208.5.02 KS362 10.0
8 2208.3.10 KS362 9.8
9 2208.3.12 KS362 9.8
10 2208.3.06 KS362 9.5
11 2208.3.04 KS362 8.8
12 2208.5.05 KS362 8.6
13 2208.2.07 KS362 8.1
14 2208.2.03 KS362 8.0
15 2208.3.03 KS362 8.0
16 2208.5.01 KS362 8.0
17 2208.2.06 KS362 7.8
18 2208.2.11 KS362 7.4
19 2208.5.07 KS362 7.3
20 2208.3.14 KS362 7.2
21 2208.3.02 KS362 7.0
22 2208.3.08 KS362 6.4
23 2208.3.07 KS362 6.1
24 2208.2.12 KS362 6.1
25 2208.3.09 KS362 6.0
26 2208.3.11 KS362 6.0
27 2208.2.01 KS362 5.8
28 2208.2.05 KS362 5.7
29 2208.3.01 KS362 5.4
30 2208.5.03 KS362 5.1
31 2208.3.13 KS362 4.7
32 2208.5.06 KS362 4.1
33 2208.3.05 KS362 3.2
平均%油脂 7.8
表34比较了用KS393和KS362分别产生的31个和33个事件的油脂含量。用KS393产生的所有事件的平均油脂含量为8.0%,而用KS393产生的所有事件的平均油脂含量为7.8%。此外,用KS393转化的事件中最高的油脂含量为12.3%,而用KS362转化的最高为11.6%。申请人已证明,与天然的YL DGAT2基因相比,YL DGAT2针对表达所作的优化导致了发育中大豆胚胎油脂含量的极少量的升高。
实施例18
YL DGAT1和一个FAD2/TE2下调构造在大豆体细胞胚中的共
表达
本实施例描述了包含耶氏酵母DGAT1(YL DGAT1)的大豆表达载体pKR1274,以及包含一种大豆脂肪酸去饱和酶2(GM FAD2)/硫酯酶2(GM TE2)下调构造的大豆表达载体pKR1267或pKR1269的构建。pKR1267和pKR1269中的GM FAD2-TE2下调区域是相同的,并且都受KTi3启动子驱动,不过pKR1267仅包含KTi3终止子,而pKR1269既包含KTi3终止子又包含大豆白蛋白终止子。
包含YL DGAT1的pKR1274的构建
作为起始质粒的pKR85(SEQ ID NO:27),此前已描述于实施例4中,其包含两翼分别与T7启动子和转录终止子相接(构成T7启动子/hpt/T7终止子框)的潮霉素B磷酸转移酶(HPT)基因(Gritz L.and Davies,J.,Gene 25:179-188(1983)),以及一个用于在细菌(例如E.coli)中选择和复制的细菌复制起点(ori)。此外,pKR72还包含两翼分别与35S启动子(Odell等人,Nature 313:810-812(1985))和NOS 3′转录终止子(Depicker等人,J.Mol.Appl.Genet.1:561-570(1982))相接(35S/hpt/NOS3′框),从而可用于在植物,例如大豆中进行选择的潮霉素B磷酸转移酶基因。质粒pKR85(SEQ ID NO:27)还包含一个两翼分别与β-大豆伴球蛋白(Beachy等人,EMBO J.4:3047-3053(1985))的α亚单位和云扁豆蛋白(Doyle等人,J.Biol.Chem.261:9228-9238(1986))的3′转录终止区域相接的NotI限制性位点,称为Bcon/NotI/Phas3′框。
通过HindIII消化,将上述Bcon/NotI/Phas3′框从pKR85(SEQ ID NO:27)上切下,并将得到的片段重新连接以构建pKR278(SEQ ID NO:104)。
将包含YL DGAT1的pKR1235(SEQ ID NO:74,实施例14)BsiWI片段,克隆至pKR278(SEQ ID NO:104)的BsiWI位点,以构建出pKR1274(SEQ ID NO:105),其中所述pKR278此前已描述于发表于US20080095915中的BB 1574中(该专利的内容特此引入本文以供参考)。
包含GM FAD2-TE2下调框的pKR1267的构建
按照生产商的说明书,利用PhusionTM高保真DNA聚合酶(Cat.No.F553S,Finnzymes Oy,Finland)以及寡核苷酸引物GmTE2_5-1(SEQID NO:108)和GmTE2_3-1(SEQ ID NO:109),将GM TE2(SEQ ID NO:106)的5′端自pTC4(SEQ ID NO:107)上扩增出来,其中所述pTC4此前已描述于WO1996006936A1中(该专利的内容特此引入本文以供参考)。按照生产商的说明书,利用PhusionTM高保真DNA聚合酶(Cat.No.F553S,Finnzymes Oy,Finland)以及寡核苷酸引物GmTE2_5-2(SEQ ID NO:110)和GmTE2_3-2(SEQ ID NO:111),将GM TE2(SEQ ID NO:106)的3′端自pTC4(SEQ ID NO:107)上扩增出来。合并所得到的两种PCR产物,并按照生产商的说明书,利用PhusionTM高保真DNA聚合酶(Cat.No.F553S,Finnzymes Oy,Finland)以及GmTE2_5-1(SEQ ID NO:108)和GmTE2_3-2(SEQ IDNO:111)进行扩增。按照生产商的说明书,利用ZeroPCR克隆试剂盒(Invitrogen Corporation),将所得到的DNA片段克隆至克隆载体,以构建出pKR1258(SEQ ID NO:112)。
按照生产商的说明书,利用PhusionTM高保真DNA聚合酶(Cat.No.F553S,Finnzymes Oy,Finland)以及寡核苷酸引物GmFAD2-1_5-1(SEQ ID NO:115)和GmFAD2-1_3-1(SEQ ID NO:116),将GM FAD2(SEQ ID NO:113)的5′端自pBS43(SEQ ID NO:114)上扩增出来,其中所述pBS43此前已描述于WO1997047731A2中(该专利的内容特此引入本文以供参考)。按照生产商的说明书,利用PhusionTM高保真DNA聚合酶(Cat.No.F553S,Finnzymes Oy,Finland)以及寡核苷酸引物GmFAD2-1_5-2(SEQ ID NO:117)和GmFAD2-1_3-2(SEQ IDNO:118),将GM FAD2-1(SEQ ID NO:113)的3′端自pBS43(SEQID NO:114)上扩增出来。合并所得到的两种PCR产物,并按照生产商的说明书,利用PhusionTM高保真DNA聚合酶(Cat.No.F553S,Finnzymes Oy,Finland)以及GmFAD2-1_5-1(SEQ ID NO:115)和GmFAD2-1_3-2(SEQ ID NO:118)进行扩增。按照生产商的说明书,利用ZeroPCR克隆试剂盒(Invitrogen Corporation),将所得到的DNA片段克隆至克隆载体,以构建出PCRblunt-Fad2-1(SEQ ID NO:119)。
将包含GM TE2的pKR1258(SEQ ID NO:112)MluI片段,克隆至包含GM FAD2-1的PCRblunt-Fad2-1(SEQ ID NO:119)MluI片段,以构建出pKR1259(SEQ ID NO:120)。
将包含GM FAD2/TE2片段5′端的pKR1259(SEQ ID NO:120)EcoRI片段,克隆至KR1259(SEQ ID NO:122)的MfeI位点,以构建出pKR1261(SEQ ID NO:121),并形成两翼为NotI位点的GMFAD2-TE2-TE2loop-TE2-FAD2发夹结构。
将包含GM FAD2-TE2-TE2loop-TE2-FAD2的pKR1261(SEQ IDNO:121)NotI片段,克隆至pKR123R(SEQ ID NO:122)的NotI位点,以构建出pKR1266(SEQ ID NO:123),其中所述pKR123R此前已描述于WO2004071467A2中(该专利的内容特此引入本文以供参考)。
将包含GM FAD2-TE2-TE2loop-TE2-FAD2的pKR1266(SEQ IDNO:123)BsiWI/PstI片段,克隆至pKR278(SEQ ID NO:104)的BsiWI/SbfI片段,以构建出pKR1267(SEQ ID NO:124)。
包含GM FAD2-TE2下调框的pKR1269的构建
将包含GM FAD2-TE2-TE2loop-TE2-FAD2的pKR1261(SEQ IDNO:121)NotI片段,克隆至pKR457(SEQ ID NO:125)的NotI位点,以构建出pKR1264(SEQ ID NO:126),其中所述pKR457此前已描述于PCT专利公布WO 2005/047479中(该专利的内容特此引入本文以供参考)。
将包含GM FAD2-TE2-TE2loop-TE2-FAD2的pKR 1264(SEQ IDNO:126)PstI片段,克隆至pKR277(SEQ ID NO:127)的SbfI片段,以构建出pKR1269(SEQ ID NO:128),其中所述pKR277此前已描述于PCT专利公布WO 2004/071467中。
GM FAD2-TE2-TE2loop-TE2-FAD2下调构造的单独表达或与
YL DGAT2的共表达
单独用pKR1267(SEQ ID NO:124)转化大豆胚胎生成悬浮培养物(cv.Jack),实验编号为MSE2213;或用pKR1274(SEQ ID NO:105)和pKR1269(SEQ ID NO:128)的BsiWI片段转化大豆胚胎生成悬浮培养物(cv.Jack),实验编号为MSE2210。对事件进行选择,体细胞胚在ShaM中的成熟如实施例5所述。如实施例4中所述,测定MSE2213和MSE2210中的油脂浓度和脂肪酸谱,每一实验的结果分别显示于表35和表36中。
表35
来自MSE2213的事件的油脂浓度和脂肪酸谱。
MSE2213(GM FAD2-TE2-TE2loop-TE2-FAD2)
事件 16:0 18:0 18:1 18:2 18:3 %油脂
2213-16 5.8 2.6 72.7 10.5 8.5 16.8
2213-26 12.0 5.7 24.8 47.2 10.4 16.5
2213-30 11.7 5.3 18.5 53.1 11.4 15.4
2213-23 9.0 3.2 52.2 24.7 11.0 15.3
2213-29 13.1 3.7 13.3 57.1 12.8 15.0
2213-7 6.4 3.1 66.3 15.6 8.6 14.9
2213-31 11.7 3.0 26.9 46.5 11.9 14.5
2213-13 6.7 3.3 64.5 16.6 8.9 14.4
2213-5 3.5 3.0 78.0 7.5 8.1 13.8
2213-9 7.6 3.4 57.6 20.3 11.0 13.2
2213-20 7.8 4.1 56.7 20.8 10.6 13.1
2213-4 4.1 2.7 72.1 11.1 10.0 12.7
2213-17 7.7 5.2 57.0 19.7 10.4 12.5
2213-3 7.4 3.6 64.4 14.4 10.2 12.5
2213-24 12.9 7.3 29.4 39.0 11.4 12.1
2213-1 13.8 7.1 22.1 44.6 12.4 11.9
2213-6 6.7 2.2 57.2 22.2 11.7 11.8
2213-11 9.2 5.2 58.2 16.4 11.0 10.9
2213-2 7.8 4.3 45.3 31.0 11.7 10.6
2213-14 7.3 4.6 63.0 15.0 10.2 10.5
2213-27 8.5 6.0 48.9 25.0 11.6 10.1
2213-19 8.0 4.0 53.7 21.4 12.8 9.9
2213-21 11.1 5.6 28.3 40.6 14.4 9.8
2213-18 7.4 4.1 57.2 17.7 13.5 9.4
2213-12 6.4 4.3 63.2 15.5 10.6 9.4
2213-10 13.8 7.0 24.0 42.1 13.2 9.4
2213-28 8.8 4.1 54.1 19.1 13.8 9.1
2213-25 6.9 3.5 53.9 21.9 13.8 9.0
2213-15 5.2 3.4 61.7 17.6 12.1 8.0
2213-8 14.4 6.6 20.0 42.9 16.1 7.6
2213-22 14.3 6.3 21.3 41.9 16.2 7.2
平均值 8.9 4.4 47.9 27.1 11.6 11.9
表36
来自MSE2210的事件的油脂浓度和脂肪酸谱
MSE2210(YL DGAT1 & GM FAD2-TE2-TE2loop-TE2-FAD2)
事件 16:0 18:0 18:1 18:2 18:3 %油脂
2210-2 3.8 2.7 74.7 13.1 5.8 22.1
2210-23 10.3 6.8 32.0 44.4 6.5 20.0
2210-29 7.5 5.5 63.9 17.1 6.0 20.0
2210-19 3.9 3.5 79.1 8.1 5.4 19.7
2210-10 7.3 3.8 66.6 15.6 6.7 19.6
2210-12 5.7 3.6 67.6 17.6 5.6 19.6
2210-14 4.5 2.4 67.9 17.8 7.4 19.3
2210-25 5.1 3.5 71.8 13.1 6.4 18.4
2210-5 9.0 4.1 43.1 35.9 7.9 17.8
2210-24 13.7 4.3 20.5 52.5 9.1 17.5
2210-13 10.2 5.5 36.8 40.4 7.0 17.4
2210-11 10.9 7.8 30.6 43.2 7.5 16.6
2210-1 4.1 3.0 75.6 9.6 7.7 15.9
2210-6 2.7 1.7 83.9 5.3 6.4 15.5
2210-16 8.4 3.7 48.5 31.3 8.1 15.3
2210-26 7.1 4.9 55.2 24.7 8.1 14.4
2210-7 4.3 3.2 62.9 20.8 8.7 14.3
2210-20 6.8 4.3 65.2 15.7 7.9 13.8
2210-27 10.9 7.4 42.4 30.0 9.3 13.8
2210-30 5.0 2.4 65.1 17.1 10.4 13.7
2210-3 7.1 4.8 52.9 27.4 7.9 13.6
2210-4 6.0 3.7 66.0 15.9 8.5 13.3
2210-22 3.2 3.3 77.7 8.1 7.8 13.2
2210-8 9.1 4.9 49.7 27.3 9.0 13.1
2210-21 6.0 3.5 67.8 14.9 7.8 12.8
2210-18 12.7 5.2 20.2 49.7 12.1 12.5
2210-31 4.4 2.9 73.1 10.1 9.5 11.7
2210-9 4.0 3.1 74.1 9.3 9.6 11.2
2210-15 3.5 2.4 72.5 11.0 10.6 11.1
2210-28 14.1 5.7 20.2 45.5 14.5 10.1
2217-29 12.0 7.5 28.0 38.8 13.8 8.7
平均值 7.2 4.2 56.6 23.6 8.4 15.4
对表35和36中的结果进行的比较证明,YL DGAT1的表达和GM FAD2-1与GM TE2的下调联合导致的脂肪酸谱改变程度,超过了仅有对FAD2-1和TE2基因的抑制时所观察到的改变程度。
序列表
<110>E.I.DU PONT DE NEMOURS AND COMPANY
<120>用于在大豆中提高种子贮藏油脂的生成和改变脂肪酸谱的来自解脂耶氏酵母的DGAT基因
<130>BB1623
<140>60/939872
<141>2007-05-24
<140>61/013406
<141>2007-12-13
<160>128
<170>PatentIn版本3.5
<210>1
<211>1581
<212>DNA
<213>解脂耶氏酵母
<400>1
atggaggtcc gacgacgaaa gatagacgtg ctcaaggccc agaaaaacgg ctacgaatcg 60
ggcccaccat ctcgacaatc gtcgcagccc tcctcaagag catcgtccag aacccgcaac 120
aaacactcct cgtccaccct gtcgctcagc ggactgacca tgaaagtcca gaagaaacct 180
gcgggacccc cggcgaactc caaaacgcca ttcctacaca tcaagcccgt gcacacgtgc 240
tgctccacat caatgctttc gcgcgattat gacggctcca accccagctt caagggcttc 300
aaaaacatcg gcatgatcat tctcattgtg ggaaatctac ggctcgcatt cgaaaactac 360
ctcaaatacg gcatttccaa cccgttcttc gaccccaaaa ttactccttc cgagtggcag 420
ctctcaggct tgctcatagt cgtggcctac gcacatatcc tcatggccta cgctattgag 480
agcgctgcca agctgctgtt cctctctagc aaacaccact acatggccgt ggggcttctg 540
cataccatga acactttgtc gtccatctcg ttgctgtcct acgtcgtcta ctactacctg 600
cccaaccccg tggcaggcac aatagtcgag tttgtggccg ttattctgtc tctcaaactc 660
gcctcatacg ccctcactaa ctcggatctc cgaaaagccg caattcatgc ccagaagctc 720
gacaagacgc aagacgataa cgaaaaggaa tccacctcgt cttcctcttc ttcagatgac 780
gcagagactt tggcagacat tgacgtcatt cctgcatact acgcacagct gccctacccc 840
cagaatgtga cgctgtcgaa cctgctgtac ttctggtttg ctcccacact ggtctaccag 900
cccgtgtacc ccaagacgga gcgtattcga cccaagcacg tgatccgaaa cctgtttgag 960
ctcgtctctc tgtgcatgct tattcagttt ctcatcttcc agtacgccta ccccatcatg 1020
cagtcgtgtc tggctctgtt cttccagccc aagctcgatt atgccaacat ctccgagcgc 1080
ctcatgaagt tggcctccgt gtctatgatg gtctggctca ttggattcta cgctttcttc 1140
cagaacggtc tcaatcttat tgccgagctc acctgttttg gaaacagaac cttctaccag 1200
cagtggtgga attcccgctc cattggccag tactggactc tatggaacaa gccagtcaac 1260
cagtacttta gacaccacgt ctacgtgcct cttctcgctc ggggcatgtc gcggttcaat 1320
gcgtcggtgg tggttttctt tttctccgcc gtcatccatg aactgcttgt cggcatcccc 1380
actcacaaca tcatcggagc cgccttcttc ggcatgatgt cgcaggtgcc tctgatcatg 1440
gctactgaga accttcagca tattaactcc tctctgggcc ccttccttgg caactgtgca 1500
ttctggttca cctttttcct gggacaaccc acttgtgcat tcctttacta tttggcctac 1560
aactacaagc agaaccagta g 1581
<210>2
<211>8325
<212>DNA
<213>人工序列
<220>
<223>质粒pYDA1
<400>2
aaacggtagg ttagtgcttg gtatatgagt tgtaggcatg acaatttgga aaggggtgga 60
ctttgggaat attgtgggat ttcaatacct tagtttgtac agggtaattg ttacaaatga 120
tacaaagaac tgtatttctt ttcatttgtt ttaattggtt gtatatcaag tccgttagac 180
gagctcagtg ccttggcttt tggcactgta tttcattttt agaggtacac tacattcagt 240
gaggtatggt aaggttgagg gcataatgaa ggcaccttgt actgacagtc acagacctct 300
caccgagaat tttatgagat atactcgggt tcattttagg ctcatcgatc aggagagacc 360
gggttggcgg cgtatttgtg tcccaaaaaa cagccccaat tgccccaatt gaccccaaat 420
tgacccagta gcgggcccaa ccccggcgag agcccccttc accccacata tcaaacctcc 480
cccggttccc acacttgccg ttaagggcgt agggtactgc agtctggaat ctacgcttgt 540
tcagactttg tactagtttc tttgtctggc catccgggta acccatgccg gacgcaaaat 600
agactactga aaattttttt gctttgtggt tgggacttta gccaagggta taaaagacca 660
ccgtccccga attacctttc ctcttctttt ctctctctcc ttgtcaactc acacccgaaa 720
tcgttaagca tttccttctg agtataagaa tcattcacca tggaggtccg acgacgaaag 780
atagacgtgc tcaaggccca gaaaaacggc tacgaatcgg gcccaccatc tcgacaatcg 840
tcgcagccct cctcaagagc atcgtccaga acccgcaaca aacactcctc gtccaccctg 900
tcgctcagcg gactgaccat gaaagtccag aagaaacctg cgggaccccc ggcgaactcc 960
aaaacgccat tcctacacat caagcccgtg cacacgtgct gctccacatc aatgctttcg 1020
cgcgattatg acggctccaa ccccagcttc aagggcttca aaaacatcgg catgatcatt 1080
ctcattgtgg gaaatctacg gctcgcattc gaaaactacc tcaaatacgg catttccaac 1140
ccgttcttcg accccaaaat tactccttcc gagtggcagc tctcaggctt gctcatagtc 1200
gtggcctacg cacatatcct catggcctac gctattgaga gcgctgccaa gctgctgttc 1260
ctctctagca aacaccacta catggccgtg gggcttctgc ataccatgaa cactttgtcg 1320
tccatctcgt tgctgtccta cgtcgtctac tactacctgc ccaaccccgt ggcaggcaca 1380
atagtcgagt ttgtggccgt tattctgtct ctcaaactcg cctcatacgc cctcactaac 1440
tcggatctcc gaaaagccgc aattcatgcc cagaagctcg acaagacgca agacgataac 1500
gaaaaggaat ccacctcgtc ttcctcttct tcagatgacg cagagacttt ggcagacatt 1560
gacgtcattc ctgcatacta cgcacagctg ccctaccccc agaatgtgac gctgtcgaac 1620
ctgctgtact tctggtttgc tcccacactg gtctaccagc ccgtgtaccc caagacggag 1680
cgtattcgac ccaagcacgt gatccgaaac ctgtttgagc tcgtctctct gtgcatgctt 1740
attcagtttc tcatcttcca gtacgcctac cccatcatgc agtcgtgtct ggctctgttc 1800
ttccagccca agctcgatta tgccaacatc tccgagcgcc tcatgaagtt ggcctccgtg 1860
tctatgatgg tctggctcat tggattctac gctttcttcc agaacggtct caatcttatt 1920
gccgagctca cctgttttgg aaacagaacc ttctaccagc agtggtggaa ttcccgctcc 1980
attggccagt actggactct atggaacaag ccagtcaacc agtactttag acaccacgtc 2040
tacgtgcctc ttctcgctcg gggcatgtcg cggttcaatg cgtcggtggt ggttttcttt 2100
ttctccgccg tcatccatga actgcttgtc ggcatcccca ctcacaacat catcggagcc 2160
gccttcttcg gcatgatgtc gcaggtgcct ctgatcatgg ctactgagaa ccttcagcat 2220
attaactcct ctctgggccc cttccttggc aactgtgcat tctggttcac ctttttcctg 2280
ggacaaccca cttgtgcatt cctttactat ttggcctaca actacaagca gaaccagtag 2340
cggccgcaag tgtggatggg gaagtgagtg cccggttctg tgtgcacaat tggcaatcca 2400
agatggatgg attcaacaca gggatatagc gagctacgtg gtggtgcgag gatatagcaa 2460
cggatattta tgtttgacac ttgagaatgt acgatacaag cactgtccaa gtacaatact 2520
aaacatactg tacatactca tactcgtacc cgggcaacgg tttcacttga gtgcagtggc 2580
tagtgctctt actcgtacag tgtgcaatac tgcgtatcat agtctttgat gtatatcgta 2640
ttcattcatg ttagttgcgt acgggcgtcg ttgcttgtgt gatttttgag gacccatccc 2700
tttggtatat aagtatactc tggggttaag gttgcccgtg tagtctaggt tatagttttc 2760
atgtgaaata ccgagagccg agggagaata aacgggggta tttggacttg tttttttcgc 2820
ggaaaagcgt cgaatcaacc ctgcgggcct tgcaccatgt ccacgacgtg tttctcgccc 2880
caattcgccc cttgcacgtc aaaattaggc ctccatctag acccctccat aacatgtgac 2940
tgtggggaaa agtataaggg aaaccatgca accatagacg acgtgaaaga cggggaggaa 3000
ccaatggagg ccaaagaaat ggggtagcaa cagtccagga gacagacaag gagacaagga 3060
gagggcgccc gaaagatcgg aaaaacaaac atgtccaatt ggggcagtga cggaaacgac 3120
acggacactt cagtacaatg gaccgaccat ctccaagcca gggttattcc ggtatcacct 3180
tggccgtaac ctcccgctgg tacctgatat tgtacacgtt cacattcaat atactttcag 3240
ctacaataag agaggctgtt tgtcgggcat gtgtgtccgt cgtatggggt gatgtccgag 3300
ggcgaaattc gctacaagct taactctggc gcttgtccag tatgaataga caagtcaaga 3360
ccagtggtgc catgattgac agggaggtac aagacttcga tactcgagca ttactcggac 3420
ttgtggcgat tgaacagacg ggcgatcgct tctcccccgt attgccggcg cgccagctgc 3480
attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 3540
cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 3600
caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 3660
caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 3720
ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 3780
cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 3840
ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 3900
tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 3960
gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 4020
ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 4080
ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 4140
gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 4200
aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 4260
tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 4320
ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 4380
tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 4440
aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 4500
tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 4560
ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 4620
gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 4680
gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 4740
taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 4800
tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 4860
ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 4920
tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 4980
ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 5040
tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 5100
ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 5160
aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 5220
actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 5280
aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 5340
tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 5400
aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 5460
ctgatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg 5520
taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta 5580
accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt 5640
tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca 5700
aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa 5760
gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat 5820
ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag 5880
gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg 5940
ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg 6000
ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 6060
tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 6120
ggccagtgaa ttgtaatacg actcactata gggcgaattg ggcccgacgt cgcatgcgct 6180
gatgacactt tggtctgaaa gagatgcatt ttgaatccca aacttgcagt gcccaagtga 6240
catacatctc cgcgttttgg aaaatgttca gaaacagttg attgtgttgg aatggggaat 6300
ggggaatgga aaaatgactc aagtatcaat tccaaaaact tctctggctg gcagtaccta 6360
ctgtccatac tactgcattt tctccagtca ggccactcta tactcgacga cacagtagta 6420
aaacccagat aatttcgaca taaacaagaa aacagaccca ataatattta tatatagtca 6480
gccgtttgtc cagttcagac tgtaatagcc gaaaaaaaat ccaaagtttc tattctagga 6540
aaatatattc caatattttt aattcttaat ctcatttatt ttattctagc gaaatacatt 6600
tcagctactt gagacatgtg atacccacaa atcggattcg gactcggttg ttcagaagag 6660
catatggcat tcgtgctcgc ttgttcacgt attcttcctg ttccatctct tggccgacaa 6720
tcacacaaaa atggggtttt ttttttaatt ctaatgattc attacagcaa aattgagata 6780
tagcagacca cgtattccat aatcaccaag gaagttcttg ggcgtcttaa ttaagtcata 6840
cacaagtcag ctttcttcga gcctcatata agtataagta gttcaacgta ttagcactgt 6900
acccagcatc tccgtatcga gaaacacaac aacatgcccc attggacaga tcatgcggat 6960
acacaggttg tgcagtatca tacatactcg atcagacagg tcgtctgacc atcatacaag 7020
ctgaacaagc gctccatact tgcacgctct ctatatacac agttaaatta catatccata 7080
gtctaacctc taacagttaa tcttctggta agcctcccag ccagccttct ggtatcgctt 7140
ggcctcctca ataggatctc ggttctggcc gtacagacct cggccgacaa ttatgatatc 7200
cgttccggta gacatgacat cctcaacagt tcggtactgc tgtccgagag cgtctccctt 7260
gtcgtcaaga cccaccccgg gggtcagaat aagccagtcc tcagagtcgc ccttaggtcg 7320
gttctgggca atgaagccaa ccacaaactc ggggtcggat cgggcaagct caatggtctg 7380
cttggagtac tcgccagtgg ccagagagcc cttgcaagac agctcggcca gcatgagcag 7440
acctctggcc agcttctcgt tgggagaggg gactaggaac tccttgtact gggagttctc 7500
gtagtcagag acgtcctcct tcttctgttc agagacagtt tcctcggcac cagctcgcag 7560
gccagcaatg attccggttc cgggtacacc gtgggcgttg gtgatatcgg accactcggc 7620
gattcggtga caccggtact ggtgcttgac agtgttgcca atatctgcga actttctgtc 7680
ctcgaacagg aagaaaccgt gcttaagagc aagttccttg agggggagca cagtgccggc 7740
gtaggtgaag tcgtcaatga tgtcgatatg ggttttgatc atgcacacat aaggtccgac 7800
cttatcggca agctcaatga gctccttggt ggtggtaaca tccagagaag cacacaggtt 7860
ggttttcttg gctgccacga gcttgagcac tcgagcggca aaggcggact tgtggacgtt 7920
agctcgagct tcgtaggagg gcattttggt ggtgaagagg agactgaaat aaatttagtc 7980
tgcagaactt tttatcggaa ccttatctgg ggcagtgaag tatatgttat ggtaatagtt 8040
acgagttagt tgaacttata gatagactgg actatacggc tatcggtcca aattagaaag 8100
aacgtcaatg gctctctggg cgtcgccttt gccgacaaaa atgtgatcat gatgaaagcc 8160
agcaatgacg ttgcagctga tattgttgtc ggccaaccgc gccgaaaacg cagctgtcag 8220
acccacagcc tccaacgaag aatgtatcgt caaagtgatc caagcacact catagttgga 8280
gtcgtactcc aaaggcggca atgacgagtc agacagatac tcgtc 8325
<210>3
<211>7518
<212>DNA
<213>人工序列
<220>
<223>质粒pY75
<400>3
ggccgcaggg cggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg 60
acctcgaggg ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca 120
ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 180
cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 240
ccttcccaac agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca 300
ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 360
gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 420
caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 480
cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 540
tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 600
acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 660
gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 720
ttaacgttta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 780
accgcatatc gacggtcgag gagaacttct agtatatcca catacctaat attattgcct 840
tattaaaaat ggaatcccaa caattacatc aaaatccaca ttctcttcaa aatcaattgt 900
cctgtacttc cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct 960
atttctcaac aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat 1020
atcttgaccg cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc 1080
tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac 1140
agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata 1200
cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gaggccggaa ccggcttttc 1260
atatagaata gagaagcgtt catgactaaa tgcttgcatc acaatacttg aagttgacaa 1320
tattatttaa ggacctattg ttttttccaa taggtggtta gcaatcgtct tactttctaa 1380
cttttcttac cttttacatt tcagcaatat atatatatat ttcaaggata taccattcta 1440
atgtctgccc ctatgtctgc ccctaagaag atcgtcgttt tgccaggtga ccacgttggt 1500
caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt tcgttccaat 1560
gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc tacaggtgtc 1620
ccacttccag atgaggcgct ggaagcctcc aagaaggttg atgccgtttt gttaggtgct 1680
gtggctggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt actaaaaatc 1740
cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc cgactctctt 1800
ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt tgttgtcaga 1860
gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga tggtgtcgct 1920
tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat ggccgctttc 1980
atggccctac aacatgagcc accattgcct atttggtcct tggataaagc taatcttttg 2040
gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga attccctaca 2100
ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa gaacccaacc 2160
cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc cgatgaagcc 2220
tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc tttgccagac 2280
aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga tttgccaaag 2340
aataaggttg accctatcgc cactatcttg tctgctgcaa tgatgttgaa attgtcattg 2400
aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt ggatgcaggt 2460
atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga tgctgtcgcc 2520
gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata 2580
aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca 2640
tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag 2700
gatagtaaag gaatacaggt aagcaaattg atactaatgg ctcaacgtga taaggaaaaa 2760
gaattgcact ttaacattaa tattgacaag gaggagggca ccacacaaaa agttaggtgt 2820
aacagaaaat catgaaacta cgattcctaa tttgatattg gaggattttc tctaaaaaaa 2880
aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa 2940
taccttcttg aagcatttcc cataatggtg aaagttccct caagaatttt actctgtcag 3000
aaacggcctt acgacgtagt cgatatggtg cactctcagt acaatctgct ctgatgccgc 3060
atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 3120
gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 3180
gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt 3240
ataggttaat gtcatgataa taatggtttc ttagtatgat ccaatatcaa aggaaatgat 3300
agcattgaag gatgagacta atccaattga ggagtggcag catatagaac agctaaaggg 3360
tagtgctgaa ggaagcatac gataccccgc atggaatggg ataatatcac aggaggtact 3420
agactacctt tcatcctaca taaatagacg catataagta cgcatttaag cataaacacg 3480
cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac 3540
gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg 3600
aaacgctttg aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcaga 3660
gcgcttttga aaaccaaaag cgctctgaag acgcactttc aaaaaaccaa aaacgcaccg 3720
gactgtaacg agctactaaa atattgcgaa taccgcttcc acaaacattg ctcaaaagta 3780
tctctttgct atatatctct gtgctatatc cctatataac ctacccatcc acctttcgct 3840
ccttgaactt gcatctaaac tcgacctcta cattttttat gtttatctct agtattactc 3900
tttagacaaa aaaattgtag taagaactat tcatagagtg aatcgaaaac aatacgaaaa 3960
tgtaaacatt tcctatacgt agtatataga gacaaaatag aagaaaccgt tcataatttt 4020
ctgaccaatg aagaatcatc aacgctatca ctttctgttc acaaagtatg cgcaatccac 4080
atcggtatag aatataatcg gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc 4140
tagtaatcag taaacgcggg aagtggagtc aggctttttt tatggaagag aaaatagaca 4200
ccaaagtagc cttcttctaa ccttaacgga cctacagtgc aaaaagttat caagagactg 4260
cattatagag cgcacaaagg agaaaaaaag taatctaaga tgctttgtta gaaaaatagc 4320
gctctcggga tgcatttttg tagaacaaaa aagaagtata gattctttgt tggtaaaata 4380
gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa 444O
ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa gcacagattc ttcgttggta 4500
aaatagcgct ttcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg 4560
aaaaattagc gctctcgcgt tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt 4620
caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 4680
attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 4740
aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 4800
tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 4860
agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 4920
gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 4980
cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 5040
agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 5100
taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 5160
tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 5220
taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 5280
acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 5340
ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 5400
cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 5460
agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 5520
tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 5580
agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 5640
tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 5700
ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 5760
tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 5820
aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 5880
tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 5940
agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 6000
taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 6060
caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6120
agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 6180
aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 6240
gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 6300
tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 6360
gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 6420
ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 6480
ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 6540
aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 6600
aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 6660
atgtgagtta cctcactcat taggcacccc aggctttaca ctttatgctt ccggctccta 6720
tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 6780
acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggga 6840
tttcgaaact aagttcttgg tgttttaaaa ctaaaaaaaa gactaactat aaaagtagaa 6900
tttaagaagt ttaagaaata gatttacaga attacaatca atacctaccg tctttatata 6960
cttattagtc aagtagggga ataatttcag ggaactggtt tcaacctttt ttttcagctt 7020
tttccaaatc agagagagca gaaggtaata gaaggtgtaa gaaaatgaga tagatacatg 7080
cgtgggtcaa ttgccttgtg tcatcattta ctccaggcag gttgcatcac tccattgagg 7140
ttgtgcccgt tttttgcctg tttgtgcccc tgttctctgt agttgcgcta agagaatgga 7200
cctatgaact gatggttggt gaagaaaaca atattttggt gctgggattc tttttttttc 7260
tggatgccag cttaaaaagc gggctccatt atatttagtg gatgccagga ataaactgtt 7320
cacccagaca cctacgatgt tatatattct gtgtaacccg ccccctattt tgggcatgta 7380
cgggttacag cagaattaaa aggctaattt tttgactaaa taaagttagg aaaatcacta 7440
ctattaatta tttacgtatt ctttgaaatg gcagtattga taatgataaa ctcgaaatca 7500
ctagtggatc cgcccagc 7518
<210>4
<211>9109
<212>DNA
<213>人工序列
<220>
<223>质粒pY75 YLDGAT1
<400>4
catggaggtc cgacgacgaa agatagacgt gctcaaggcc cagaaaaacg gctacgaatc 60
gggcccacca tctcgacaat cgtcgcagcc ctcctcaaga gcatcgtcca gaacccgcaa 120
caaacactcc tcgtccaccc tgtcgctcag cggactgacc atgaaagtcc agaagaaacc 180
tgcgggaccc ccggcgaact ccaaaacgcc attcctacac atcaagcccg tgcacacgtg 240
ctgctccaca tcaatgcttt cgcgcgatta tgacggctcc aaccccagct tcaagggctt 300
caaaaacatc ggcatgatca ttctcattgt gggaaatcta cggctcgcat tcgaaaacta 360
cctcaaatac ggcatttcca acccgttctt cgaccccaaa attactcctt ccgagtggca 420
gctctcaggc ttgctcatag tcgtggccta cgcacatatc ctcatggcct acgctattga 480
gagcgctgcc aagctgctgt tcctctctag caaacaccac tacatggccg tggggcttct 540
gcataccatg aacactttgt cgtccatctc gttgctgtcc tacgtcgtct actactacct 600
gcccaacccc gtggcaggca caatagtcga gtttgtggcc gttattctgt ctctcaaact 660
cgcctcatac gccctcacta actcggatct ccgaaaagcc gcaattcatg cccagaagct 720
cgacaagacg caagacgata acgaaaagga atccacctcg tcttcctctt cttcagatga 780
cgcagagact ttggcagaca ttgacgtcat tcctgcatac tacgcacagc tgccctaccc 840
ccagaatgtg acgctgtcga acctgctgta cttctggttt gctcccacac tggtctacca 900
gcccgtgtac cccaagacgg agcgtattcg acccaagcac gtgatccgaa acctgtttga 960
gctcgtctct ctgtgcatgc ttattcagtt tctcatcttc cagtacgcct accccatcat 1020
gcagtcgtgt ctggctctgt tcttccagcc caagctcgat tatgccaaca tctccgagcg 1080
cctcatgaag ttggcctccg tgtctatgat ggtctggctc attggattct acgctttctt 1140
ccagaacggt ctcaatctta ttgccgagct cacctgtttt ggaaacagaa ccttctacca 1200
gcagtggtgg aattcccgct ccattggcca gtactggact ctatggaaca agccagtcaa 1260
ccagtacttt agacaccacg tctacgtgcc tcttctcgct cggggcatgt cgcggttcaa 1320
tgcgtcggtg gtggttttct ttttctccgc cgtcatccat gaactgcttg tcggcatccc 1380
cactcacaac atcatcggag ccgccttctt cggcatgatg tcgcaggtgc ctctgatcat 1440
ggctactgag aaccttcagc atattaactc ctctctgggc cccttccttg gcaactgtgc 1500
attctggttc acctttttcc tgggacaacc cacttgtgca ttcctttact atttggccta 1560
caactacaag cagaaccagt agcggccggc cgcagggcgg atcccccggg ctgcaggaat 1620
tcgatatcaa gcttatcgat accgtcgacc tcgagggggg gcccggtacc caattcgccc 1680
tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa 1740
aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt 1800
aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa 1860
tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc 1920
gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt 1980
ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc 2040
cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt 2100
agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt 2160
aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt 2220
gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa 2280
aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcctgatg cggtattttc 2340
tccttacgca tctgtgcggt atttcacacc gcatatcgac ggtcgaggag aacttctagt 2400
atatccacat acctaatatt attgccttat taaaaatgga atcccaacaa ttacatcaaa 2460
atccacattc tcttcaaaat caattgtcct gtacttcctt gttcatgtgt gttcaaaaac 2520
gttatattta taggataatt atactctatt tctcaacaag taattggttg tttggccgag 2580
cggtctaagg cgcctgattc aagaaatatc ttgaccgcag ttaactgtgg gaatactcag 2640
gtatcgtaag atgcaagagt tcgaatctct tagcaaccat tatttttttc ctcaacataa 2700
cgagaacaca caggggcgct atcgcacaga atcaaattcg atgactggaa attttttgtt 2760
aatttcagag gtcgcctgac gcatatacct ttttcaactg aaaaattggg agaaaaagga 2820
aaggtgagag gccggaaccg gcttttcata tagaatagag aagcgttcat gactaaatgc 2880
ttgcatcaca atacttgaag ttgacaatat tatttaagga cctattgttt tttccaatag 2940
gtggttagca atcgtcttac tttctaactt ttcttacctt ttacatttca gcaatatata 3000
tatatatttc aaggatatac cattctaatg tctgccccta tgtctgcccc taagaagatc 3060
gtcgttttgc caggtgacca cgttggtcaa gaaatcacag ccgaagccat taaggttctt 3120
aaagctattt ctgatgttcg ttccaatgtc aagttcgatt tcgaaaatca tttaattggt 3180
ggtgctgcta tcgatgctac aggtgtccca cttccagatg aggcgctgga agcctccaag 3240
aaggttgatg ccgttttgtt aggtgctgtg gctggtccta aatggggtac cggtagtgtt 3300
agacctgaac aaggtttact aaaaatccgt aaagaacttc aattgtacgc caacttaaga 3360
ccatgtaact ttgcatccga ctctctttta gacttatctc caatcaagcc acaatttgct 3420
aaaggtactg acttcgttgt tgtcagagaa ttagtgggag gtatttactt tggtaagaga 3480
aaggaagacg atggtgatgg tgtcgcttgg gatagtgaac aatacaccgt tccagaagtg 3540
caaagaatca caagaatggc cgctttcatg gccctacaac atgagccacc attgcctatt 3600
tggtccttgg ataaagctaa tcttttggcc tcttcaagat tatggagaaa aactgtggag 3660
gaaaccatca agaacgaatt ccctacattg aaggttcaac atcaattgat tgattctgcc 3720
gccatgatcc tagttaagaa cccaacccac ctaaatggta ttataatcac cagcaacatg 3780
tttggtgata tcatctccga tgaagcctcc gttatcccag gttccttggg tttgttgcca 3840
tctgcgtcct tggcctcttt gccagacaag aacaccgcat ttggtttgta cgaaccatgc 3900
cacggttctg ctccagattt gccaaagaat aaggttgacc ctatcgccac tatcttgtct 3960
gctgcaatga tgttgaaatt gtcattgaac ttgcctgaag aaggtaaggc cattgaagat 4020
gcagttaaaa aggttttgga tgcaggtatc agaactggtg atttaggtgg ttccaacagt 4080
accaccgaag tcggtgatgc tgtcgccgaa gaagttaaga aaatccttgc ttaaaaagat 4140
tctctttttt tatgatattt gtacataaac tttataaatg aaattcataa tagaaacgac 4200
acgaaattac aaaatggaat atgttcatag ggtagacgaa actatatacg caatctacat 4260
acatttatca agaaggagaa aaaggaggat agtaaaggaa tacaggtaag caaattgata 4320
ctaatggctc aacgtgataa ggaaaaagaa ttgcacttta acattaatat tgacaaggag 4380
gagggcacca cacaaaaagt taggtgtaac agaaaatcat gaaactacga ttcctaattt 4440
gatattggag gattttctct aaaaaaaaaa aaatacaaca aataaaaaac actcaatgac 4500
ctgaccattt gatggagttt aagtcaatac cttcttgaag catttcccat aatggtgaaa 4560
gttccctcaa gaattttact ctgtcagaaa cggccttacg acgtagtcga tatggtgcac 4620
tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc 4680
cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac 4740
cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg 4800
aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta 4860
gtatgatcca atatcaaagg aaatgatagc attgaaggat gagactaatc caattgagga 4920
gtggcagcat atagaacagc taaagggtag tgctgaagga agcatacgat accccgcatg 4980
gaatgggata atatcacagg aggtactaga ctacctttca tcctacataa atagacgcat 5040
ataagtacgc atttaagcat aaacacgcac tatgccgttc ttctcatgta tatatatata 5100
caggcaacac gcagatatag gtgcgacgtg aacagtgagc tgtatgtgcg cagctcgcgt 5160
tgcattttcg gaagcgctcg ttttcggaaa cgctttgaag ttcctattcc gaagttccta 5220
ttctctagaa agtataggaa cttcagagcg cttttgaaaa ccaaaagcgc tctgaagacg 5280
cactttcaaa aaaccaaaaa cgcaccggac tgtaacgagc tactaaaata ttgcgaatac 5340
cgcttccaca aacattgctc aaaagtatct ctttgctata tatctctgtg ctatatccct 5400
atataaccta cccatccacc tttcgctcct tgaacttgca tctaaactcg acctctacat 5460
tttttatgtt tatctctagt attactcttt agacaaaaaa attgtagtaa gaactattca 5520
tagagtgaat cgaaaacaat acgaaaatgt aaacatttcc tatacgtagt atatagagac 5580
aaaatagaag aaaccgttca taattttctg accaatgaag aatcatcaac gctatcactt 5640
tctgttcaca aagtatgcgc aatccacatc ggtatagaat ataatcgggg atgcctttat 5700
cttgaaaaaa tgcacccgca gcttcgctag taatcagtaa acgcgggaag tggagtcagg 5760
ctttttttat ggaagagaaa atagacacca aagtagcctt cttctaacct taacggacct 5820
acagtgcaaa aagttatcaa gagactgcat tatagagcgc acaaaggaga aaaaaagtaa 5880
tctaagatgc tttgttagaa aaatagcgct ctcgggatgc atttttgtag aacaaaaaag 5940
aagtatagat tctttgttgg taaaatagcg ctctcgcgtt gcatttctgt tctgtaaaaa 6000
tgcagctcag attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttttacaa 6060
aaatgaagca cagattcttc gttggtaaaa tagcgctttc gcgttgcatt tctgttctgt 6120
aaaaatgcag ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttc 6180
tacaaaatga agcacagatg cttcgttcag gtggcacttt tcggggaaat gtgcgcggaa 6240
cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 6300
cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 6360
tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 6420
tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 6480
atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 6640
gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 6600
aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 6660
aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 6720
gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 6780
cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 6840
atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt 6900
tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 6960
ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 7020
ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 7080
ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 7140
tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 7200
tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 7260
aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 7320
tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 7380
tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 7440
gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 7500
agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 7560
tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 7620
ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 7680
cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 7740
tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 7800
acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 7860
gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 7920
ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 7980
tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 8040
attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 8100
cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc 8160
ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga 8220
aagcgggcag tgagcgcaac gcaattaatg tgagttacct cactcattag gcaccccagg 8280
ctttacactt tatgcttccg gctcctatgt tgtgtggaat tgtgagcgga taacaatttc 8340
acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg 8400
aacaaaagct ggagctccac cgcgggattt cgaaactaag ttcttggtgt tttaaaacta 8460
aaaaaaagac taactataaa agtagaattt aagaagttta agaaatagat ttacagaatt 8520
acaatcaata cctaccgtct ttatatactt attagtcaag taggggaata atttcaggga 8580
actggtttca accttttttt tcagcttttt ccaaatcaga gagagcagaa ggtaatagaa 8640
ggtgtaagaa aatgagatag atacatgcgt gggtcaattg ccttgtgtca tcatttactc 8700
caggcaggtt gcatcactcc attgaggttg tgcccgtttt ttgcctgttt gtgcccctgt 8760
tctctgtagt tgcgctaaga gaatggacct atgaactgat ggttggtgaa gaaaacaata 8820
ttttggtgct gggattcttt ttttttctgg atgccagctt aaaaagcggg ctccattata 8880
tttagtggat gccaggaata aactgttcac ccagacacct acgatgttat atattctgtg 8940
taacccgccc cctattttgg gcatgtacgg gttacagcag aattaaaagg ctaatttttt 9000
gactaaataa agttaggaaa atcactacta ttaattattt acgtattctt tgaaatggca 9060
gtattgataa tgataaactc gaaatcacta gtggatccgc ccagcggcc 9109
<210>5
<211>6849
<212>DNA
<213>人工序列
<220>
<223>质粒pRS425
<400>5
tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60
cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120
ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180
accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc 240
accattatgg gaaatgcttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca 300
ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat 360
taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc 420
ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc 480
aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt 540
agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg 600
tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct 660
ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg 720
ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct 780
tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac 840
aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat 900
ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc 960
aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg 1020
ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca 1080
gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc 1140
acagtttttc tccataatct tgaagaggcc aaaagattag ctttatccaa ggaccaaata 1200
ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact 1260
tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc 1320
ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca 1380
aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt 1440
aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca 1500
ctaccggtac cccatttagg accagccaca gcacctaaca aaacggcatc aaccttcttg 1560
gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca 1620
attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga 1680
accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc 1740
ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata 1800
tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat 1860
tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat 1920
ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct 1980
ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca 2040
aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat 2100
gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga 2160
gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg 2220
gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt 2280
ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg 2340
atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta 2400
gaagttctcc tcgaccgtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa 2460
ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt 2520
taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa 2580
gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag 2640
aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt 2700
gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac 2760
cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag 2820
gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg 2880
cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg 2940
ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc 3000
cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc 3060
cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc 3120
gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaattc 3180
ctgcagcccg ggggatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt 3240
ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc 3300
tgtgtgaaat tgttatccgc tcacaattcc acacaacata ggagccggaa gcataaagtg 3360
taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc gctcactgcc 3420
cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 3480
gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 3540
ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 3600
agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 3660
ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 3720
caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 3780
gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 3840
cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 3900
tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 3960
gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 4020
cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 4080
tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 4140
tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 4200
caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 4260
aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 4320
cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 4380
ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 4440
tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 4500
atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 4560
tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 4620
aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 4680
catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 4740
gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 4800
ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 4860
aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 4920
atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 4980
cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 5040
gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 5100
agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 5160
gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 5220
caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 5280
ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 5340
tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 5400
aggggttccg cgcacatttc cccgaaaagt gccacctgaa cgaagcatct gtgcttcatt 5460
ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca 5520
tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt 5580
catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag 5640
ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta 5700
tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc 5760
ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc 5820
actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa 5880
aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt 5940
tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga 6000
acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct 6060
attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca 6120
ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat 6180
aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt 6240
tatataggga tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg 6300
aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga 6360
aagtgcgtct tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta 6420
gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa 6480
atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata tctgcgtgtt 6540
gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct taaatgcgta 6600
cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt gatattatcc 6660
cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt tctatatgct 6720
gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt gatattggat 6780
catactaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc 6840
cctttcgtc 6849
<210>6
<211>7494
<212>DNA
<213>人工序列
<220>
<223>质粒pGPD425
<400>6
ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat 60
ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacataggag 120
ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc acattaattg 180
cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 240
tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 300
ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 360
taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 420
agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 480
cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 540
tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 600
tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 660
gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 720
acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 780
acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 840
cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 900
gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 960
gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 1020
agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 1080
ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 1140
ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 1200
atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 1260
tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 1320
gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 1380
ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 1440
caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 1500
cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 1560
cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 1620
cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 1680
agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 1740
tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 1800
agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 1860
atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 1920
ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 1980
cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 2040
caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 2100
attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 2160
agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgaacgaa 2220
gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca 2280
aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa 2340
cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc 2400
aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt 2460
accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt 2520
tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct 2580
cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta 2640
ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag 2700
ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat 2760
tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt 2820
atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg 2880
tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa 2940
tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa 3000
aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt 3060
tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc 3120
gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa 3180
gttcctatac tttctagaga ataggaactt cggaatagga acttcaaagc gtttccgaaa 3240
acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac agctcactgt tcacgtcgca 3300
cctatatctg cgtgttgcct gtatatatat atacatgaga agaacggcat agtgcgtgtt 3360
tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg atgaaaggta gtctagtacc 3420
tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc actacccttt 3480
agctgttcta tatgctgcca ctcctcaatt ggattagtct catccttcaa tgctatcatt 3540
tcctttgata ttggatcata ctaagaaacc attattatca tgacattaac ctataaaaat 3600
aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga 3660
cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa 3720
gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca 3780
tcagagcaga ttgtactgag agtgcaccat atcgactacg tcgtaaggcc gtttctgaca 3840
gagtaaaatt cttgagggaa ctttcaccat tatgggaaat gcttcaagaa ggtattgact 3900
taaactccat caaatggtca ggtcattgag tgttttttat ttgttgtatt tttttttttt 3960
tagagaaaat cctccaatat caaattagga atcgtagttt catgattttc tgttacacct 4020
aactttttgt gtggtgccct cctccttgtc aatattaatg ttaaagtgca attctttttc 4080
cttatcacgt tgagccatta gtatcaattt gcttacctgt attcctttac tatcctcctt 4140
tttctccttc ttgataaatg tatgtagatt gcgtatatag tttcgtctac cctatgaaca 4200
tattccattt tgtaatttcg tgtcgtttct attatgaatt tcatttataa agtttatgta 4260
caaatatcat aaaaaaagag aatcttttta agcaaggatt ttcttaactt cttcggcgac 4320
agcatcaccg acttcggtgg tactgttgga accacctaaa tcaccagttc tgatacctgc 4380
atccaaaacc tttttaactg catcttcaat ggccttacct tcttcaggca agttcaatga 4440
caatttcaac atcattgcag cagacaagat agtggcgata gggtcaacct tattctttgg 4500
caaatctgga gcagaaccgt ggcatggttc gtacaaacca aatgcggtgt tcttgtctgg 4560
caaagaggcc aaggacgcag atggcaacaa acccaaggaa cctgggataa cggaggcttc 4620
atcggagatg atatcaccaa acatgttgct ggtgattata ataccattta ggtgggttgg 4680
gttcttaact aggatcatgg cggcagaatc aatcaattga tgttgaacct tcaatgtagg 4740
gaattcgttc ttgatggttt cctccacagt ttttctccat aatcttgaag aggccaaaag 4800
attagcttta tccaaggacc aaataggcaa tggtggctca tgttgtaggg ccatgaaagc 4860
ggccattctt gtgattcttt gcacttctgg aacggtgtat tgttcactat cccaagcgac 4920
accatcacca tcgtcttcct ttctcttacc aaagtaaata cctcccacta attctctgac 4980
aacaacgaag tcagtacctt tagcaaattg tggcttgatt ggagataagt ctaaaagaga 5040
gtcggatgca aagttacatg gtcttaagtt ggcgtacaat tgaagttctt tacggatttt 5100
tagtaaacct tgttcaggtc taacactacc ggtaccccat ttaggaccag ccacagcacc 5160
taacaaaacg gcatcaacct tcttggaggc ttccagcgcc tcatctggaa gtgggacacc 5220
tgtagcatcg atagcagcac caccaattaa atgattttcg aaatcgaact tgacattgga 5280
acgaacatca gaaatagctt taagaacctt aatggcttcg gctgtgattt cttgaccaac 5340
gtggtcacct ggcaaaacga cgatcttctt aggggcagac ataggggcag acattagaat 5400
ggtatatcct tgaaatatat atatatattg ctgaaatgta aaaggtaaga aaagttagaa 5460
agtaagacga ttgctaacca cctattggaa aaaacaatag gtccttaaat aatattgtca 5520
acttcaagta ttgtgatgca agcatttagt catgaacgct tctctattct atatgaaaag 5580
ccggttccgg cctctcacct ttcctttttc tcccaatttt tcagttgaaa aaggtatatg 5640
cgtcaggcga cctctgaaat taacaaaaaa tttccagtca tcgaatttga ttctgtgcga 5700
tagcgcccct gtgtgttctc gttatgttga ggaaaaaaat aatggttgct aagagattcg 5760
aactcttgca tcttacgata cctgagtatt cccacagtta actgcggtca agatatttct 5820
tgaatcaggc gccttagacc gctcggccaa acaaccaatt acttgttgag aaatagagta 5880
taattatcct ataaatataa cgtttttgaa cacacatgaa caaggaagta caggacaatt 5940
gattttgaag agaatgtgga ttttgatgta attgttggga ttccattttt aataaggcaa 6000
taatattagg tatgtggata tactagaagt tctcctcgac cgtcgatatg cggtgtgaaa 6060
taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaaacg ttaatatttt 6120
gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat 6180
cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt 6240
ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt 6300
ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag 6360
gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg 6420
aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc 6480
gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc 6540
gctacagggc gcgtcgcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 6600
ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 6660
aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg 6720
cgcgtaatac gactcactat agggcgaatt gggtaccggg ccccccctcg aggtcgacgg 6780
tatcgataag cttgatatcg aattcctgca gcccggggga tccactagtg atttcgagtt 6840
tatcattatc aatactgcca tttcaaagaa tacgtaaata attaatagta gtgattttcc 6900
taactttatt tagtcaaaaa attagccttt taattctgct gtaacccgta catgcccaaa 6960
atagggggcg ggttacacag aatatataac atcgtaggtg tctgggtgaa cagtttattc 7020
ctggcatcca ctaaatataa tggagcccgc tttttaagct ggcatccaga aaaaaaaaga 7080
atcccagcac caaaatattg ttttcttcac caaccatcag ttcataggtc cattctctta 7140
gcgcaactac agagaacagg ggcacaaaca ggcaaaaaac gggcacaacc tcaatggagt 7200
gatgcaacct gcctggagta aatgatgaca caaggcaatt gacccacgca tgtatctatc 7260
tcattttctt acaccttcta ttaccttctg ctctctctga tttggaaaaa gctgaaaaaa 7320
aaggttgaaa ccagttccct gaaattattc ccctacttga ctaataagta tataaagacg 7380
gtaggtattg attgtaattc tgtaaatcta tttcttaaac ttcttaaatt ctacttttat 7440
agttagtctt ttttttagtt ttaaaacacc aagaacttag tttcgaaatc ccgc 7494
<210>7
<211>36
<212>DNA
<213>人工序列
<220>
<223>质粒oYLDGAT2-1
<400>7
gcggccgcat gactatcgac tcacaatact acaagt 36
<210>8
<211>36
<212>DNA
<213>人工序列
<220>
<223>质粒oYLDGAT2-2
<400>8
gcggccgctt actcaatcat tcggaactct ggggct 36
<210>9
<211>1545
<212>DNA
<213>解脂耶氏酵母
<400>9
atgactatcg actcacaata ctacaagtcg cgagacaaaa acgacacggc acccaaaatc 60
gcgggaatcc gatatgcccc gctatcgaca ccattactca accgatgtga gaccttctct 120
ctggtctggc acattttcag cattcccact ttcctcacaa ttttcatgct atgctgcgca 180
attccactgc tctggccatt tgtgattgcg tatgtagtgt acgctgttaa agacgactcc 240
ccgtccaacg gaggagtggt caagcgatac tcgcctattt caagaaactt cttcatctgg 300
aagctctttg gccgctactt ccccataact ctgcacaaga cggtggatct ggagcccacg 360
cacacatact accctctgga cgtccaggag tatcacctga ttgctgagag atactggccg 420
cagaacaagt acctccgagc aatcatcacc accatcgagt actttctgcc cgccttcatg 480
aaacggtctc tttctatcaa cgagcaggag cagcctgccg agcgagatcc tctcctgtct 540
cccgtttctc ccagctctcc gggttctcaa cctgacaagt ggattaacca cgacagcaga 600
tatagccgtg gagaatcatc tggctccaac ggccacgcct cgggctccga acttaacggc 660
aacggcaaca acggcaccac taaccgacga cctttgtcgt ccgcctctgc tggctccact 720
gcatctgatt ccacgcttct taacgggtcc ctcaactcct acgccaacca gatcattggc 780
gaaaacgacc cacagctgtc gcccacaaaa ctcaagccca ctggcagaaa atacatcttc 840
ggctaccacc cccacggcat tatcggcatg ggagcctttg gtggaattgc caccgaggga 900
gctggatggt ccaagctctt tccgggcatc cctgtttctc ttatgactct caccaacaac 960
ttccgagtgc ctctctacag agagtacctc atgagtctgg gagtcgcttc tgtctccaag 1020
aagtcctgca aggccctcct caagcgaaac cagtctatct gcattgtcgt tggtggagca 1080
caggaaagtc ttctggccag acccggtgtc atggacctgg tgctactcaa gcgaaagggt 1140
tttgttcgac ttggtatgga ggtcggaaat gtcgcccttg ttcccatcat ggcctttggt 1200
gagaacgacc tctatgacca ggttagcaac gacaagtcgt ccaagctgta ccgattccag 1260
cagtttgtca agaacttcct tggattcacc cttcctttga tgcatgcccg aggcgtcttc 1320
aactacgatg tcggtcttgt cccctacagg cgacccgtca acattgtggt tggttccccc 1380
attgacttgc cttatctccc acaccccacc gacgaagaag tgtccgaata ccacgaccga 1440
tacatcgccg agctgcagcg aatctacaac gagcacaagg atgaatattt catcgattgg 1500
accgaggagg gcaaaggagc cccagagttc cgaatgattg agtaa 1545
<210>10
<211>514
<212>PRT
<213>解脂耶氏酵母
<400>10
Met Thr Ile Asp Ser Gln Tyr Tyr Lys Set Arg Asp Lys Asn Asp Thr
1 5 10 15
Ala Pro Lys Ile Ala Gly Ile Arg Tyr Ala Pro Leu Ser Thr Pro Leu
20 25 30
Leu Asn Arg Cys Glu Thr Phe Ser Leu Val Trp His Ile Phe Ser Ile
35 40 45
Pro Thr Phe Leu Thr Ile Phe Met Leu Cys Cys Ala Ile Pro Leu Leu
50 55 60
Trp Pro Phe Val Ile Ala Tyr Val Val Tyr Ala Val Lys Asp Asp Ser
65 70 75 80
Pro Ser Asn Gly Gly Val Val Lys Arg Tyr Ser Pro Ile Ser Arg Asn
85 90 95
Phe Phe Ile Trp Lys Leu Phe Gly Arg Tyr Phe Pro Ile Thr Leu His
100 105 110
Lys Thr Val Asp Leu Glu Pro Thr His Thr Tyr Tyr Pro Leu Asp Val
115 120 125
Gln Glu Tyr His Leu Ile Ala Glu Arg Tyr Trp Pro Gln Asn Lys Tyr
130 135 140
Leu Arg Ala Ile Ile Thr Thr Ile Glu Tyr Phe Leu Pro Ala Phe Met
145 150 155 160
Lys Arg Set Leu Ser Ile Asn Glu Gln Glu Gln Pro Ala Glu Arg Asp
165 170 175
Pro Leu Leu Ser Pro Val Ser Pro Ser Ser Pro Gly Ser Gln Pro Asp
180 185 190
Lys Trp Ile Asn His Asp Ser Arg Tyr Ser Arg Gly Glu Ser Ser Gly
195 200 205
Ser Asn Gly His Ala Ser Gly Ser Glu Leu Asn Gly Asn Gly Asn Asn
210 215 220
Gly Thr Thr Asn Arg Arg Pro Leu Ser Ser Ala Ser Ala Gly Ser Thr
225 230 235 240
Ala Ser Asp Ser Thr Leu Leu Asn Gly Ser Leu Asn Ser Tyr Ala Asn
245 250 255
Gln Ile Ile Gly Glu Asn Asp Pro Gln Leu Ser Pro Thr Lys Leu Lys
260 265 270
Pro Thr Gly Arg Lys Tyr Ile Phe Gly Tyr His Pro His Gly Ile Ile
275 280 285
Gly Met Gly Ala Phe Gly Gly Ile Ala Thr Glu Gly Ala Gly Trp Ser
290 295 300
Lys Leu Phe Pro Gly Ile Pro Val Ser Leu Met Thr Leu Thr Asn Asn
305 310 315 320
Phe Arg Val Pro Leu Tyr Arg Glu Tyr Leu Met Ser Leu Gly Val Ala
325 330 335
Ser Val Ser Lys Lys Ser Cys Lys Ala Leu Leu Lys Arg Asn Gln Ser
340 345 350
Ile Cys Ile Val Val Gly Gly Ala Gln Glu Ser Leu Leu Ala Arg Pro
355 360 365
Gly Val Met Asp Leu Val Leu Leu Lys Arg Lys Gly Phe Val Arg Leu
370 375 380
Gly Met Glu Val Gly Asn Val Ala Leu Val Pro Ile Met Ala Phe Gly
385 390 395 400
Glu Asn Asp Leu Tyr Asp Gln Val Ser Asn Asp Lys Ser Ser Lys Leu
405 410 415
Tyr Arg Phe Gln Gln Phe Val Lys Asn Phe Leu Gly Phe Thr Leu Pro
420 425 430
Leu Met His Ala Arg Gly Val Phe Asn Tyr Asp Val Gly Leu Val Pro
435 440 445
Tyr Arg Arg Pro Val Asn Ile Val Val Gly Ser Pro Ile Asp Leu Pro
450 455 460
Tyr Leu Pro His Pro Thr Asp Glu Glu Val Ser Glu Tyr His Asp Arg
465 470 475 480
Tyr Ile Ala Glu Leu Gln Arg Ile Tyr Asn Glu His Lys Asp Glu Tyr
485 490 495
Phe Ile Asp Trp Thr Glu Glu Gly Lys Gly Ala Pro Glu Phe Arg Met
500 505 510
Ile Glu
<210>11
<211>9070
<212>DNA
<213>人工序列
<220>
<223>质粒pY75 YLDGAT2
<400>11
ggccgcaggg cggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg 60
acctcgaggg ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca 120
ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 180
cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 240
ccttcccaac agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca 300
ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 360
gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 420
caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 480
cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 540
tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 600
acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 660
gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 720
ttaacgttta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 780
accgcatatc gacggtcgag gagaacttct agtatatcca catacctaat attattgcct 840
tattaaaaat ggaatcccaa caattacatc aaaatccaca ttctcttcaa aatcaattgt 900
cctgtacttc cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct 960
atttctcaac aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat 1020
atcttgaccg cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc 1080
tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac 1140
agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata 1200
cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gaggccggaa ccggcttttc 1260
atatagaata gagaagcgtt catgactaaa tgcttgcatc acaatacttg aagttgacaa 1320
tattatttaa ggacctattg ttttttccaa taggtggtta gcaatcgtct tactttctaa 1380
cttttcttac cttttacatt tcagcaatat atatatatat ttcaaggata taccattcta 1440
atgtctgccc ctatgtctgc ccctaagaag atcgtcgttt tgccaggtga ccacgttggt 1500
caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt tcgttccaat 1560
gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc tacaggtgtc 1620
ccacttccag atgaggcgct ggaagcctcc aagaaggttg atgccgtttt gttaggtgct 1680
gtggctggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt actaaaaatc 1740
cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc cgactctctt 1800
ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt tgttgtcaga 1860
gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga tggtgtcgct 1920
tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat ggccgctttc 1980
atggccctac aacatgagcc accattgcct atttggtcct tggataaagc taatcttttg 2040
gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga attccctaca 2100
ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa gaacccaacc 2160
cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc cgatgaagcc 2220
tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc tttgccagac 2280
aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga tttgccaaag 2340
aataaggttg accctatcgc cactatcttg tctgctgcaa tgatgttgaa attgtcattg 2400
aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt ggatgcaggt 2460
atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga tgctgtcgcc 2520
gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata 2580
aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca 2640
tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag 2700
gatagtaaag gaatacaggt aagcaaattg atactaatgg ctcaacgtga taaggaaaaa 2760
gaattgcact ttaacattaa tattgacaag gaggagggca ccacacaaaa agttaggtgt 2820
aacagaaaat catgaaacta cgattcctaa tttgatattg gaggattttc tctaaaaaaa 2880
aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa 2940
taccttcttg aagcatttcc cataatggtg aaagttccct caagaatttt actctgtcag 3000
aaacggcctt acgacgtagt cgatatggtg cactctcagt acaatctgct ctgatgccgc 3060
atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 3120
gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 3180
gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt 3240
ataggttaat gtcatgataa taatggtttc ttagtatgat ccaatatcaa aggaaatgat 3300
agcattgaag gatgagacta atccaattga ggagtggcag catatagaac agctaaaggg 3360
tagtgctgaa ggaagcatac gataccccgc atggaatggg ataatatcac aggaggtact 3420
agactacctt tcatcctaca taaatagacg catataagta cgcatttaag cataaacacg 3480
cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac 3540
gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg 3600
aaacgctttg aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcaga 3660
gcgcttttga aaaccaaaag cgctctgaag acgcactttc aaaaaaccaa aaacgcaccg 3720
gactgtaacg agctactaaa atattgcgaa taccgcttcc acaaacattg ctcaaaagta 3780
tctctttgct atatatctct gtgctatatc cctatataac ctacccatcc acctttcgct 3840
ccttgaactt gcatctaaac tcgacctcta cattttttat gtttatctct agtattactc 3900
tttagacaaa aaaattgtag taagaactat tcatagagtg aatcgaaaac aatacgaaaa 3960
tgtaaacatt tcctatacgt agtatataga gacaaaatag aagaaaccgt tcataatttt 4020
ctgaccaatg aagaatcatc aacgctatca ctttctgttc acaaagtatg cgcaatccac 4080
atcggtatag aatataatcg gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc 4140
tagtaatcag taaacgcggg aagtggagtc aggctttttt tatggaagag aaaatagaca 4200
ccaaagtagc cttcttctaa ccttaacgga cctacagtgc aaaaagttat caagagactg 4260
cattatagag cgcacaaagg agaaaaaaag taatctaaga tgctttgtta gaaaaatagc 4320
gctctcggga tgcatttttg tagaacaaaa aagaagtata gattctttgt tggtaaaata 4380
gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa 4440
ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa gcacagattc ttcgttggta 4500
aaatagcgct ttcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg 4560
aaaaattagc gctctcgcgt tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt 4620
caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 4680
attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 4740
aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 4800
tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 4860
agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 4920
gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 4980
cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 5040
agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 5100
taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 5160
tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 5220
taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 5280
acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 5340
ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 5400
cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 5460
agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 5520
tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 5580
agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 5640
tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 5700
ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 5760
tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 5820
aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 5880
tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 5940
agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 6000
taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 6060
caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6120
agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 6180
aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 6240
gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 6300
tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 6360
gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 6420
ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 6480
ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 6540
aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 6600
aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 6660
atgtgagtta cctcactcat taggcacccc aggctttaca ctttatgctt ccggctccta 6720
tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 6780
acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggga 6840
tttcgaaact aagttcttgg tgttttaaaa ctaaaaaaaa gactaactat aaaagtagaa 6900
tttaagaagt ttaagaaata gatttacaga attacaatca atacctaccg tctttatata 6960
cttattagtc aagtagggga ataatttcag ggaactggtt tcaacctttt ttttcagctt 7020
tttccaaatc agagagagca gaaggtaata gaaggtgtaa gaaaatgaga tagatacatg 7080
cgtgggtcaa ttgccttgtg tcatcattta ctccaggcag gttgcatcac tccattgagg 7140
ttgtgcccgt tttttgcctg tttgtgcccc tgttctctgt agttgcgcta agagaatgga 7200
cctatgaact gatggttggt gaagaaaaca atattttggt gctgggattc tttttttttc 7260
tggatgccag cttaaaaagc gggctccatt atatttagtg gatgccagga ataaactgtt 7320
cacccagaca cctacgatgt tatatattct gtgtaacccg ccccctattt tgggcatgta 7380
cgggttacag cagaattaaa aggctaattt tttgactaaa taaagttagg aaaatcacta 7440
ctattaatta tttacgtatt ctttgaaatg gcagtattga taatgataaa ctcgaaatca 7500
ctagtggatc cgcccagcgg ccgcatgact atcgactcac aatactacaa gtcgcgagac 7560
aaaaacgaca cggcacccaa aatcgcggga atccgatatg ccccgctatc gacaccatta 7620
ctcaaccgat gtgagacctt ctctctggtc tggcacattt tcagcattcc cactttcctc 7680
acaattttca tgctatgctg cgcaattcca ctgctctggc catttgtgat tgcgtatgta 7740
gtgtacgctg ttaaagacga ctccccgtcc aacggaggag tggtcaagcg atactcgcct 7800
atttcaagaa acttcttcat ctggaagctc tttggccgct acttccccat aactctgcac 7860
aagacggtgg atctggagcc cacgcacaca tactaccctc tggacgtcca ggagtatcac 7920
ctgattgctg agagatactg gccgcagaac aagtacctcc gagcaatcat ctccaccatc 7980
gagtactttc tgcccgcctt catgaaacgg tctctttcta tcaacgagca ggagcagcct 8040
gccgagcgag atcctctcct gtctcccgtt tctcccagct ctccgggttc tcaacctgac 8100
aagtggatta accacgacag cagatatagc cgtggagaat catctggctc caacggccac 8160
gcctcgggct ccgaacttaa cggcaacggc aacaatggca ccactaaccg acgacctttg 8220
tcgtccgcct ctgctggctc cactgcatct gattccacgc ttcttaacgg gtccctcaac 8280
tcctacgcca accagatcat tggcgaaaac gacccacagc tgtcgcccac aaaactcaag 8340
cccactggca gaaaatacat cttcggctac cacccccacg gcattatcgg catgggagcc 8400
tttggtggaa ttgccaccga gggagctgga tggtccaagc tctttccggg catccctgtt 8460
tctcttatga ctctcaccaa caacttccga gtgcctctct acagagagta cctcatgagt 8520
ctgggagtcg cttctgtctc caagaagtcc tgcaaggccc tcctcaagcg aaaccagtct 8580
atctgcattg tcgttggtgg agcacaggaa agtcttctgg ccagacccgg tgtcatggac 8640
ctggtgctac tcaagcgaaa gggttttgtt cgacttggta tggaggtcgg aaatgtcgcc 8700
cttgttccca tcatggcctt tggtgagaac gacctctatg accaggttag caacgacaag 8760
tcgtccaagc tgtaccgatt ccagcagttt gtcaagaact tccttggatt cacccttcct 8820
ttgatgcatg cccgaggcgt cttcaactac gatgtcggtc ttgtccccta caggcgaccc 8880
gtcaacattg tggttggttc ccccattgac ttgccttatc tcccacaccc caccgacgaa 8940
gaagtgtccg aataccacga ccgatacatc gccgagctgc agcgaatcta caacgagcac 9000
aaggatgaat atttcatcga ttggaccgag gagggcaaag gagcccagag ttccgaatga 9060
ttgagtaagc 9070
<210>12
<211>32
<212>DNA
<213>人工序列
<220>
<223>引物YDGAT1-F
<400>12
gatcatccat ggaagtccga cgacgaaaaa tc 32
<210>13
<211>36
<212>DNA
<213>人工序列
<220>
<223>引物YDGAT1-R
<400>13
gatcaagcgg ccgcctactg gttctgcttg tagttg 36
<210>14
<211>35
<212>DNA
<213>人工序列
<220>
<223>引物YDGAT2-F
<400>14
gatcatccat ggctatcgac tcacaatact acaag 35
<210>15
<211>37
<212>DNA
<213>人工序列
<220>
<223>引物YDGAT2-R
<400>15
gatcatgcgg ccgcttactc aatcattcgg aactctg 37
<210>16
<211>1603
<212>DNA
<213>解脂耶氏酵母
<400>16
gatcatccat ggaagtccga cgacgaaaaa tcgacgtgct caaggcccag aaaaacggct 60
acgaatcggg cccaccatct cgacaatcgt cgcagccctc ctcaagagca tcgtccagaa 120
cccgcaacaa acactcctcg tccaccctgt cgctcagcgg actgaccatg aaagtccaga 180
agaaacctgc gggacccccg gcgaactcca aaacgccatt cctacacatc aagcccgtgc 240
acacgtgctg ctccacatca atgctttcgc gcgattatga cggctccaac cccagcttca 300
agggcttcaa aaacatcggc atgatcattc tcattgtggg aaatctacgg ctcgcattcg 360
aaaactacct caaatacggc atttccaacc cgttcttcga ccccaaaatt actccttccg 420
agtggcagct ctcaggcttg ctcatagtcg tggcctacgc acatatcctc atggcctacg 480
ctattgagag cgctgccaag ctgctgttcc tctctagcaa acaccactac atggccgtgg 540
ggcttctgca taccatgaac actttgtcgt ccatctcgtt gctgtcctac gtcgtctact 600
actacctgcc caaccccgtg gcaggcacaa tagtcgagtt tgtggccgtt attctgtctc 660
tcaaactcgc ctcatacgcc ctcactaact cggatctccg aaaagccgca attcatgccc 720
agaagctcga caagacgcaa gacgataacg aaaaggaatc cacctcgtct tcctcttctt 780
cagatgacgc agagactttg gcagacattg acgtcattcc tgcatactac gcacagctgc 840
cctaccccca gaatgtgacg ctgtcgaacc tgctgtactt ctggtttgct cccacactgg 900
tctaccagcc cgtgtacccc aagacggagc gtattcgacc caagcacgtg atccgaaacc 960
tgtttgagct cgtctctctg tgcatgctta ttcagtttct catcttccag tacgcctacc 1020
ccatcatgca gtcgtgtctg gctctgttct tccagcccaa gctcgattat gccaacatct 1080
ccgagcgcct catgaagttg gcctccgtgt ctatgatggt ctggctcatt ggattctacg 1140
ctttcttcca gaacggtctc aatcttattg ccgagctcac ctgttttgga aacagaacct 1200
tctaccagca gtggtggaat tcccgctcca ttggccagta ctggactcta tggaacaagc 1260
cagtcaacca gtactttaga caccacgtct acgtgcctct tctcgctcgg ggcatgtcgc 1320
ggttcaatgc gtcggtggtg gttttctttt tctccgccgt catccatgaa ctgcttgtcg 1380
gcatccccac tcacaacatc atcggagccg ccttcttcgg catgatgtcg caggtgcctc 1440
tgatcatggc tactgagaac cttcagcata ttaactcctc tctgggcccc ttccttggca 1500
actgtgcatt ctggttcacc tttttcctgg gacaacccac ttgtgcattc ctttattatc 1560
tggcttacaa ctacaagcag aaccagtagg cggccgcttg atc 1603
<210>17
<211>1567
<212>DNA
<213>解脂耶氏酵母
<400>17
gatcatccat ggctatcgac tcacaatact acaagtcgcg agacaaaaac gacacggcac 60
ccaaaatcgc gggaatccga tatgccccgc tatcgacacc attactcaac cgatgtgaga 120
ccttctctct ggtctggcac attttcagca ttcccacttt cctcacaatt ttcatgctat 180
gctgcgcaat tccactgctc tggccatttg tgattgcgta tgtagtgtac gctgttaaag 240
acgactcccc gtccaacgga ggagtggtca agcgatactc gcctatttca agaaacttct 300
tcatctggaa gctctttggc cgctacttcc ccataactct gcacaagacg gtggatctgg 360
agcccacgca cacatactac cctctggacg tccaggagta tcacctgatt gctgagagat 420
actggccgca gaacaagtac ctccgagcaa tcatctccac catcgagtac tttctgcccg 480
ccttcatgaa acggtctctt tctatcaacg agcaggagca gcctgccgag cgagatcctc 540
tcctgtctcc cgtttctccc agctctccgg gttctcaacc tgacaagtgg attaaccacg 600
acagcagata tagccgtgga gaatcatctg gctccaacgg ccacgcctcg ggctccgaac 660
ttaacggcaa cggcaacaat ggcaccacta accgacgacc tttgtcgtcc gcctctgctg 720
gctccactgc atctgattcc acgcttctta acgggtccct caactcctac gccaaccaga 780
tcattggcga aaacgaccca cagctgtcgc ccacaaaact caagcccact ggcagaaaat 840
acatcttcgg ctaccacccc cacggcatta tcggcatggg agcctttggt ggaattgcca 900
ccgagggagc tggatggtcc aagctctttc cgggcatccc tgtttctctt atgactctca 960
ccaacaactt ccgagtgcct ctctacagag agtacctcat gagtctggga gtcgcttctg 1020
tctccaagaa gtcctgcaag gccctcctca agcgaaacca gtctatctgc attgtcgttg 1080
gtggagcaca ggaaagtctt ctggccagac ccggtgtcat ggacctggtg ctactcaagc 1140
gaaagggttt tgttcgactt ggtatggagg tcggaaatgt cgcccttgtt cccatcatgg 1200
cctttggtga gaacgacctc tatgaccagg ttagcaacga caagtcgtcc aagctgtacc 1260
gattccagca gtttgtcaag aacttccttg gattcaccct tcctttgatg catgcccgag 1320
gcgtcttcaa ctacgatgtc ggtcttgtcc cctacaggcg acccgtcaac attgtggttg 1380
gttcccccat tgacttgcct tatctcccac accccaccga cgaagaagtg tccgaatacc 1440
acgaccgata catcgccgag ctgcagcgaa tctacaacga gcacaaggat gaatatttca 1500
tcgattggac cgaggagggc aaaggagccc cagagttccg aatgattgag taagcggccg 1560
catgatc 1567
<210>18
<211>6991
<212>DNA
<213>人工序列
<220>
<223>质粒pFBAIN-MOD-1
<400>18
gcggccgcaa gtgtggatgg ggaagtgagt gcccggttct gtgtgcacaa ttggcaatcc 60
aagatggatg gattcaacac agggatatag cgagctacgt ggtggtgcga ggatatagca 120
acggatattt atgtttgaca cttgagaatg tacgatacaa gcactgtcca agtacaatac 180
taaacatact gtacatactc atactcgtac ccgggcaacg gtttcacttg agtgcagtgg 240
ctagtgctct tactcgtaca gtgtgcaata ctgcgtatca tagtctttga tgtatatcgt 300
attcattcat gttagttgcg tacgagccgg aagcataaag tgtaaagcct ggggtgccta 360
atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 420
cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 480
tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 540
agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 600
aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 660
gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 720
tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 780
cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 840
ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 900
cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 960
atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 1020
agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 1080
gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 1140
gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 1200
tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 1260
agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 1320
gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 1380
aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 1440
aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 1500
ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 1560
gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 1620
aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 1680
ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 1740
tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 1800
ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 1860
cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 1920
agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 1980
gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 2040
gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 2100
acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 2160
acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 2220
agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 2280
aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 2340
gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 2400
tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 2460
ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 2520
cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 2580
ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 2640
tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 2700
gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 2760
ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 2820
gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat 2880
tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta 2940
cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt 3000
tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc 3060
gaattgggta ccgggccccc cctcgaggtc gatggtgtcg ataagcttga tatcgaattc 3120
atgtcacaca aaccgatctt cgcctcaagg aaacctaatt ctacatccga gagactgccg 3180
agatccagtc tacactgatt aattttcggg ccaataattt aaaaaaatcg tgttatataa 3240
tattatatgt attatatata tacatcatga tgatactgac agtcatgtcc cattgctaaa 3300
tagacagact ccatctgccg cctccaactg atgttctcaa tatttaaggg gtcatctcgc 3360
attgtttaat aataaacaga ctccatctac cgcctccaaa tgatgttctc aaaatatatt 3420
gtatgaactt atttttatta cttagtatta ttagacaact tacttgcttt atgaaaaaca 3480
cttcctattt aggaaacaat ttataatggc agttcgttca tttaacaatt tatgtagaat 3540
aaatgttata aatgcgtatg ggaaatctta aatatggata gcataaatga tatctgcatt 3600
gcctaattcg aaatcaacag caacgaaaaa aatcccttgt acaacataaa tagtcatcga 3660
gaaatatcaa ctatcaaaga acagctattc acacgttact attgagatta ttattggacg 3720
agaatcacac actcaactgt ctttctctct tctagaaata caggtacaag tatgtactat 3780
tctcattgtt catacttcta gtcatttcat cccacatatt ccttggattt ctctccaatg 3840
aatgacattc tatcttgcaa attcaacaat tataataaga tataccaaag tagcggtata 3900
gtggcaatca aaaagcttct ctggtgtgct tctcgtattt atttttattc taatgatcca 3960
ttaaaggtat atatttattt cttgttatat aatccttttg tttattacat gggctggata 4020
cataaaggta ttttgattta attttttgct taaattcaat cccccctcgt tcagtgtcaa 4080
ctgtaatggt aggaaattac catacttttg aagaagcaaa aaaaatgaaa gaaaaaaaaa 4140
atcgtatttc caggttagac gttccgcaga atctagaatg cggtatgcgg tacattgttc 4200
ttcgaacgta aaagttgcgc tccctgagat attgtacatt tttgctttta caagtacaag 4260
tacatcgtac aactatgtac tactgttgat gcatccacaa cagtttgttt tgtttttttt 4320
tgtttttttt ttttctaatg attcattacc gctatgtata cctacttgta cttgtagtaa 4380
gccgggttat tggcgttcaa ttaatcatag acttatgaat ctgcacggtg tgcgctgcga 4440
gttactttta gcttatgcat gctacttggg tgtaatattg ggatctgttc ggaaatcaac 4500
ggatgctcaa tcgatttcga cagtaattaa ttaagtcata cacaagtcag ctttcttcga 4560
gcctcatata agtataagta gttcaacgta ttagcactgt acccagcatc tccgtatcga 4620
gaaacacaac aacatgcccc attggacaga tcatgcggat acacaggttg tgcagtatca 4680
tacatactcg atcagacagg tcgtctgacc atcatacaag ctgaacaagc gctccatact 4740
tgcacgctct ctatatacac agttaaatta catatccata gtctaacctc taacagttaa 4800
tcttctggta agcctcccag ccagccttct ggtatcgctt ggcctcctca ataggatctc 4860
ggttctggcc gtacagacct cggccgacaa ttatgatatc cgttccggta gacatgacat 4920
cctcaacagt tcggtactgc tgtccgagag cgtctccctt gtcgtcaaga cccaccccgg 4980
gggtcagaat aagccagtcc tcagagtcgc ccttaggtcg gttctgggca atgaagccaa 5040
ccacaaactc ggggtcggat cgggcaagct caatggtctg cttggagtac tcgccagtgg 5100
ccagagagcc cttgcaagac agctcggcca gcatgagcag acctctggcc agcttctcgt 5160
tgggagaggg gactaggaac tccttgtact gggagttctc gtagtcagag acgtcctcct 5220
tcttctgttc agagacagtt tcctcggcac cagctcgcag gccagcaatg attccggttc 5280
cgggtacacc gtgggcgttg gtgatatcgg accactcggc gattcggtga caccggtact 5340
ggtgcttgac agtgttgcca atatctgcga actttctgtc ctcgaacagg aagaaaccgt 5400
gcttaagagc aagttccttg agggggagca cagtgccggc gtaggtgaag tcgtcaatga 5460
tgtcgatatg ggttttgatc atgcacacat aaggtccgac cttatcggca agctcaatga 5520
gctccttggt ggtggtaaca tccagagaag cacacaggtt ggttttcttg gctgccacga 5580
gcttgagcac tcgagcggca aaggcggact tgtggacgtt agctcgagct tcgtaggagg 5640
gcattttggt ggtgaagagg agactgaaat aaatttagtc tgcagaactt tttatcggaa 5700
ccttatctgg ggcagtgaag tatatgttat ggtaatagtt acgagttagt tgaacttata 5760
gatagactgg actatacggc tatcggtcca aattagaaag aacgtcaatg gctctctggg 5820
cgtcgccttt gccgacaaaa atgtgatcat gatgaaagcc agcaatgacg ttgcagctga 5880
tattgttgtc ggccaaccgc gccgaaaacg cagctgtcag acccacagcc tccaacgaag 5940
aatgtatcgt caaagtgatc caagcacact catagttgga gtcgtactcc aaaggcggca 6000
atgacgagtc agacagatac tcgtcgaaaa cagtgtacgc agatctacta tagaggaaca 6060
tttaaattgc cccggagaag acggccaggc cgcctagatg acaaattcaa caactcacag 6120
ctgactttct gccattgcca ctaggggggg gcctttttat atggccaagc caagctctcc 6180
acgtcggttg ggctgcaccc aacaataaat gggtagggtt gcaccaacaa agggatggga 6240
tggggggtag aagatacgag gataacgggg ctcaatggca caaataagaa cgaatactgc 6300
cattaagact cgtgatccag cgactgacac cattgcatca tctaagggcc tcaaaactac 6360
ctcggaactg ctgcgctgat ctggacacca cagaggttcc gagcacttta ggttgcacca 6420
aatgtcccac caggtgcagg cagaaaacgc tggaacagcg tgtacagttt gtcttaacaa 6480
aaagtgaggg cgctgaggtc gagcagggtg gtgtgacttg ttatagcctt tagagctgcg 6540
aaagcgcgta tggatttggc tcatcaggcc agattgaggg tctgtggaca catgtcatgt 6600
tagtgtactt caatcgcccc ctggatatag ccccgacaat aggccgtggc ctcatttttt 6660
tgccttccgc acatttccat tgctcggtac ccacaccttg cttctcctgc acttgccaac 6720
cttaatactg gtttacattg accaacatct tacaagcggg gggcttgtct agggtatata 6780
taaacagtgg ctctcccaat cggttgccag tctctttttt cctttctttc cccacagatt 6840
cgaaatctaa actacacatc acagaattcc gagccgtgag tatccacgac aagatcagtg 6900
tcgagacgac gcgttttgtg taatgacaca atccgaaagt cgctagcaac acacactctc 6960
tacacaaact aacccagctc tggtaccatg g 6991
<210>19
<211>8568
<212>DNA
<213>人工序列
<220>
<223>质粒pFBAIN-YLDGAT1
<400>19
ggccgcaagt gtggatgggg aagtgagtgc ccggttctgt gtgcacaatt ggcaatccaa 60
gatggatgga ttcaacacag ggatatagcg agctacgtgg tggtgcgagg atatagcaac 120
ggatatttat gtttgacact tgagaatgta cgatacaagc actgtccaag tacaatacta 180
aacatactgt acatactcat actcgtaccc gggcaacggt ttcacttgag tgcagtggct 240
agtgctctta ctcgtacagt gtgcaatact gcgtatcata gtctttgatg tatatcgtat 300
tcattcatgt tagttgcgta cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 360
gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 420
tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 480
ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 540
cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 600
gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 660
tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 720
agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 780
tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 840
cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 900
ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 960
ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 1020
ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 1080
ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 1140
cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 1200
gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 1260
atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 1320
ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 1380
gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 1440
tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 1500
ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 1560
taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 1620
gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 1680
gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 1740
ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 1800
aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 1860
gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 1920
cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 1980
actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 2040
caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 2100
gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 2160
ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 2220
caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 2280
tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 2340
gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 2400
cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 2460
ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 2520
tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 2580
ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 2640
atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 2700
ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 2760
tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 2820
tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc 2880
gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg 2940
ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc 3000
ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga 3060
attgggtacc gggccccccc tcgaggtcga tggtgtcgat aagcttgata tcgaattcat 3120
gtcacacaaa ccgatcttcg cctcaaggaa acctaattct acatccgaga gactgccgag 3180
atccagtcta cactgattaa ttttcgggcc aataatttaa aaaaatcgtg ttatataata 3240
ttatatgtat tatatatata catcatgatg atactgacag tcatgtccca ttgctaaata 3300
gacagactcc atctgccgcc tccaactgat gttctcaata tttaaggggt catctcgcat 3360
tgtttaataa taaacagact ccatctaccg cctccaaatg atgttctcaa aatatattgt 3420
atgaacttat ttttattact tagtattatt agacaactta cttgctttat gaaaaacact 3480
tcctatttag gaaacaattt ataatggcag ttcgttcatt taacaattta tgtagaataa 3540
atgttataaa tgcgtatggg aaatcttaaa tatggatagc ataaatgata tctgcattgc 3600
ctaattcgaa atcaacagca acgaaaaaaa tcccttgtac aacataaata gtcatcgaga 3660
aatatcaact atcaaagaac agctattcac acgttactat tgagattatt attggacgag 3720
aatcacacac tcaactgtct ttctctcttc tagaaataca ggtacaagta tgtactattc 3780
tcattgttca tacttctagt catttcatcc cacatattcc ttggatttct ctccaatgaa 3840
tgacattcta tcttgcaaat tcaacaatta taataagata taccaaagta gcggtatagt 3900
ggcaatcaaa aagcttctct ggtgtgcttc tcgtatttat ttttattcta atgatccatt 3960
aaaggtatat atttatttct tgttatataa tccttttgtt tattacatgg gctggataca 4020
taaaggtatt ttgatttaat tttttgctta aattcaatcc cccctcgttc agtgtcaact 4080
gtaatggtag gaaattacca tacttttgaa gaagcaaaaa aaatgaaaga aaaaaaaaat 4140
cgtatttcca ggttagacgt tccgcagaat ctagaatgcg gtatgcggta cattgttctt 4200
cgaacgtaaa agttgcgctc cctgagatat tgtacatttt tgcttttaca agtacaagta 4260
catcgtacaa ctatgtacta ctgttgatgc atccacaaca gtttgttttg tttttttttg 4320
tttttttttt ttctaatgat tcattaccgc tatgtatacc tacttgtact tgtagtaagc 4380
cgggttattg gcgttcaatt aatcatagac ttatgaatct gcacggtgtg cgctgcgagt 4440
tacttttagc ttatgcatgc tacttgggtg taatattggg atctgttcgg aaatcaacgg 4500
atgctcaatc gatttcgaca gtaattaatt aagtcataca caagtcagct ttcttcgagc 4560
ctcatataag tataagtagt tcaacgtatt agcactgtac ccagcatctc cgtatcgaga 4620
aacacaacaa catgccccat tggacagatc atgcggatac acaggttgtg cagtatcata 4680
catactcgat cagacaggtc gtctgaccat catacaagct gaacaagcgc tccatacttg 4740
cacgctctct atatacacag ttaaattaca tatccatagt ctaacctcta acagttaatc 4800
ttctggtaag cctcccagcc agccttctgg tatcgcttgg cctcctcaat aggatctcgg 4860
ttctggccgt acagacctcg gccgacaatt atgatatccg ttccggtaga catgacatcc 4920
tcaacagttc ggtactgctg tccgagagcg tctcccttgt cgtcaagacc caccccgggg 4980
gtcagaataa gccagtcctc agagtcgccc ttaggtcggt tctgggcaat gaagccaacc 5040
acaaactcgg ggtcggatcg ggcaagctca atggtctgct tggagtactc gccagtggcc 5100
agagagccct tgcaagacag ctcggccagc atgagcagac ctctggccag cttctcgttg 5160
ggagagggga ctaggaactc cttgtactgg gagttctcgt agtcagagac gtcctccttc 5220
ttctgttcag agacagtttc ctcggcacca gctcgcaggc cagcaatgat tccggttccg 5280
ggtacaccgt gggcgttggt gatatcggac cactcggcga ttcggtgaca ccggtactgg 5340
tgcttgacag tgttgccaat atctgcgaac tttctgtcct cgaacaggaa gaaaccgtgc 5400
ttaagagcaa gttccttgag ggggagcaca gtgccggcgt aggtgaagtc gtcaatgatg 5460
tcgatatggg ttttgatcat gcacacataa ggtccgacct tatcggcaag ctcaatgagc 5520
tccttggtgg tggtaacatc cagagaagca cacaggttgg ttttcttggc tgccacgagc 5580
ttgagcactc gagcggcaaa ggcggacttg tggacgttag ctcgagcttc gtaggagggc 5640
attttggtgg tgaagaggag actgaaataa atttagtctg cagaactttt tatcggaacc 5700
ttatctgggg cagtgaagta tatgttatgg taatagttac gagttagttg aacttataga 5760
tagactggac tatacggcta tcggtccaaa ttagaaagaa cgtcaatggc tctctgggcg 5820
tcgcctttgc cgacaaaaat gtgatcatga tgaaagccag caatgacgtt gcagctgata 5880
ttgttgtcgg ccaaccgcgc cgaaaacgca gctgtcagac ccacagcctc caacgaagaa 5940
tgtatcgtca aagtgatcca agcacactca tagttggagt cgtactccaa aggcggcaat 6000
gacgagtcag acagatactc gtcgaaaaca gtgtacgcag atctactata gaggaacatt 6060
taaattgccc cggagaagac ggccaggccg cctagatgac aaattcaaca actcacagct 6120
gactttctgc cattgccact aggggggggc ctttttatat ggccaagcca agctctccac 6180
gtcggttggg ctgcacccaa caataaatgg gtagggttgc accaacaaag ggatgggatg 6240
gggggtagaa gatacgagga taacggggct caatggcaca aataagaacg aatactgcca 6300
ttaagactcg tgatccagcg actgacacca ttgcatcatc taagggcctc aaaactacct 6360
cggaactgct gcgctgatct ggacaccaca gaggttccga gcactttagg ttgcaccaaa 6420
tgtcccacca ggtgcaggca gaaaacgctg gaacagcgtg tacagtttgt cttaacaaaa 6480
agtgagggcg ctgaggtcga gcagggtggt gtgacttgtt atagccttta gagctgcgaa 6540
agcgcgtatg gatttggctc atcaggccag attgagggtc tgtggacaca tgtcatgtta 6600
gtgtacttca atcgccccct ggatatagcc ccgacaatag gccgtggcct catttttttg 6660
ccttccgcac atttccattg ctcggtaccc acaccttgct tctcctgcac ttgccaacct 6720
taatactggt ttacattgac caacatctta caagcggggg gcttgtctag ggtatatata 6780
aacagtggct ctcccaatcg gttgccagtc tcttttttcc tttctttccc cacagattcg 6840
aaatctaaac tacacatcac agaattccga gccgtgagta tccacgacaa gatcagtgtc 6900
gagacgacgc gttttgtgta atgacacaat ccgaaagtcg ctagcaacac acactctcta 6960
cacaaactaa cccagctctg gtaccatgga agtccgacga cgaaaaatcg acgtgctcaa 7020
ggcccagaaa aacggctacg aatcgggccc accatctcga caatcgtcgc agccctcctc 7080
aagagcatcg tccagaaccc gcaacaaaca ctcctcgtcc accctgtcgc tcagcggact 7140
gaccatgaaa gtccagaaga aacctgcggg acccccggcg aactccaaaa cgccattcct 7200
acacatcaag cccgtgcaca cgtgctgctc cacatcaatg ctttcgcgcg attatgacgg 7260
ctccaacccc agcttcaagg gcttcaaaaa catcggcatg atcattctca ttgtgggaaa 7320
tctacggctc gcattcgaaa actacctcaa atacggcatt tccaacccgt tcttcgaccc 7380
caaaattact ccttccgagt ggcagctctc aggcttgctc atagtcgtgg cctacgcaca 7440
tatcctcatg gcctacgcta ttgagagcgc tgccaagctg ctgttcctct ctagcaaaca 7500
ccactacatg gccgtggggc ttctgcatac catgaacact ttgtcgtcca tctcgttgct 7560
gtcctacgtc gtctactact acctgcccaa ccccgtggca ggcacaatag tcgagtttgt 7620
ggccgttatt ctgtctctca aactcgcctc atacgccctc actaactcgg atctccgaaa 7680
agccgcaatt catgcccaga agctcgacaa gacgcaagac gataacgaaa aggaatccac 7740
ctcgtcttcc tcttcttcag atgacgcaga gactttggca gacattgacg tcattcctgc 7800
atactacgca cagctgccct acccccagaa tgtgacgctg tcgaacctgc tgtacttctg 7860
gtttgctccc acactggtct accagcccgt gtaccccaag acggagcgta ttcgacccaa 7920
gcacgtgatc cgaaacctgt ttgagctcgt ctctctgtgc atgcttattc agtttctcat 7980
cttccagtac gcctacccca tcatgcagtc gtgtctggct ctgttcttcc agcccaagct 8040
cgattatgcc aacatctccg agcgcctcat gaagttggcc tccgtgtcta tgatggtctg 8100
gctcattgga ttctacgctt tcttccagaa cggtctcaat cttattgccg agctcacctg 8160
ttttggaaac agaaccttct accagcagtg gtggaattcc cgctccattg gccagtactg 8220
gactctatgg aacaagccag tcaaccagta ctttagacac cacgtctacg tgcctcttct 8280
cgctcggggc atgtcgcggt tcaatgcgtc ggtggtggtt ttctttttct ccgccgtcat 8340
ccatgaactg cttgtcggca tccccactca caacatcatc ggagccgcct tcttcggcat 8400
gatgtcgcag gtgcctctga tcatggctac tgagaacctt cagcatatta actcctctct 8460
gggccccttc cttggcaact gtgcattctg gttcaccttt ttcctgggac aacccacttg 8520
tgcattcctt tattatctgg cttacaacta caagcagaac cagtaggc 8568
<210>20
<211>8532
<212>DNA
<213>人工序列
<220>
<223>质粒pFBAIN-YLDGAT2
<400>20
ggccgcaagt gtggatgggg aagtgagtgc ccggttctgt gtgcacaatt ggcaatccaa 60
gatggatgga ttcaacacag ggatatagcg agctacgtgg tggtgcgagg atatagcaac 120
ggatatttat gtttgacact tgagaatgta cgatacaagc actgtccaag tacaatacta 180
aacatactgt acatactcat actcgtaccc gggcaacggt ttcacttgag tgcagtggct 240
agtgctctta ctcgtacagt gtgcaatact gcgtatcata gtctttgatg tatatcgtat 300
tcattcatgt tagttgcgta cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 360
gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 420
tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 480
ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 540
cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 600
gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 660
tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 720
agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 780
tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 840
cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 900
ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 960
ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 1020
ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 1080
ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 1140
cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 1200
gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 1260
atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 1320
ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 1380
gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 1440
tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 1500
ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 1560
taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 1620
gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 1680
gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 1740
ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 1800
aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 1860
gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 1920
cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 1980
actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 2040
caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 2100
gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 2160
ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 2220
caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 2280
tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 2340
gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 2400
cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 2460
ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 2520
tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 2580
ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 2640
atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 2700
ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 2760
tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 2820
tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc 2880
gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg 2940
ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc 3000
ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga 3060
attgggtacc gggccccccc tcgaggtcga tggtgtcgat aagcttgata tcgaattcat 3120
gtcacacaaa ccgatcttcg cctcaaggaa acctaattct acatccgaga gactgccgag 3180
atccagtcta cactgattaa ttttcgggcc aataatttaa aaaaatcgtg ttatataata 3240
ttatatgtat tatatatata catcatgatg atactgacag tcatgtccca ttgctaaata 3300
gacagactcc atctgccgcc tccaactgat gttctcaata tttaaggggt catctcgcat 3360
tgtttaataa taaacagact ccatctaccg cctccaaatg atgttctcaa aatatattgt 3420
atgaacttat ttttattact tagtattatt agacaactta cttgctttat gaaaaacact 3480
tcctatttag gaaacaattt ataatggcag ttcgttcatt taacaattta tgtagaataa 3540
atgttataaa tgcgtatggg aaatcttaaa tatggatagc ataaatgata tctgcattgc 3600
ctaattcgaa atcaacagca acgaaaaaaa tcccttgtac aacataaata gtcatcgaga 3660
aatatcaact atcaaagaac agctattcac acgttactat tgagattatt attggacgag 3720
aatcacacac tcaactgtct ttctctcttc tagaaataca ggtacaagta tgtactattc 3780
tcattgttca tacttctagt catttcatcc cacatattcc ttggatttct ctccaatgaa 3840
tgacattcta tcttgcaaat tcaacaatta taataagata taccaaagta gcggtatagt 3900
ggcaatcaaa aagcttctct ggtgtgcttc tcgtatttat ttttattcta atgatccatt 3960
aaaggtatat atttatttct tgttatataa tccttttgtt tattacatgg gctggataca 4020
taaaggtatt ttgatttaat tttttgctta aattcaatcc cccctcgttc agtgtcaact 4080
gtaatggtag gaaattacca tacttttgaa gaagcaaaaa aaatgaaaga aaaaaaaaat 4140
cgtatttcca ggttagacgt tccgcagaat ctagaatgcg gtatgcggta cattgttctt 4200
cgaacgtaaa agttgcgctc cctgagatat tgtacatttt tgcttttaca agtacaagta 4260
catcgtacaa ctatgtacta ctgttgatgc atccacaaca gtttgttttg tttttttttg 4320
tttttttttt ttctaatgat tcattaccgc tatgtatacc tacttgtact tgtagtaagc 4380
cgggttattg gcgttcaatt aatcatagac ttatgaatct gcacggtgtg cgctgcgagt 4440
tacttttagc ttatgcatgc tacttgggtg taatattggg atctgttcgg aaatcaacgg 4500
atgctcaatc gatttcgaca gtaattaatt aagtcataca caagtcagct ttcttcgagc 4560
ctcatataag tataagtagt tcaacgtatt agcactgtac ccagcatctc cgtatcgaga 4620
aacacaacaa catgccccat tggacagatc atgcggatac acaggttgtg cagtatcata 4680
catactcgat cagacaggtc gtctgaccat catacaagct gaacaagcgc tccatacttg 4740
cacgctctct atatacacag ttaaattaca tatccatagt ctaacctcta acagttaatc 4800
ttctggtaag cctcccagcc agccttctgg tatcgcttgg cctcctcaat aggatctcgg 4860
ttctggccgt acagacctcg gccgacaatt atgatatccg ttccggtaga catgacatcc 4920
tcaacagttc ggtactgctg tccgagagcg tctcccttgt cgtcaagacc caccccgggg 4980
gtcagaataa gccagtcctc agagtcgccc ttaggtcggt tctgggcaat gaagccaacc 5040
acaaactcgg ggtcggatcg ggcaagctca atggtctgct tggagtactc gccagtggcc 5100
agagagccct tgcaagacag ctcggccagc atgagcagac ctctggccag cttctcgttg 5160
ggagagggga ctaggaactc cttgtactgg gagttctcgt agtcagagac gtcctccttc 5220
ttctgttcag agacagtttc ctcggcacca gctcgcaggc cagcaatgat tccggttccg 5280
ggtacaccgt gggcgttggt gatatcggac cactcggcga ttcggtgaca ccggtactgg 5340
tgcttgacag tgttgccaat atctgcgaac tttctgtcct cgaacaggaa gaaaccgtgc 5400
ttaagagcaa gttccttgag ggggagcaca gtgccggcgt aggtgaagtc gtcaatgatg 5460
tcgatatggg ttttgatcat gcacacataa ggtccgacct tatcggcaag ctcaatgagc 5520
tccttggtgg tggtaacatc cagagaagca cacaggttgg ttttcttggc tgccacgagc 5580
ttgagcactc gagcggcaaa ggcggacttg tggacgttag ctcgagcttc gtaggagggc 5640
attttggtgg tgaagaggag actgaaataa atttagtctg cagaactttt tatcggaacc 5700
ttatctgggg cagtgaagta tatgttatgg taatagttac gagttagttg aacttataga 5760
tagactggac tatacggcta tcggtccaaa ttagaaagaa cgtcaatggc tctctgggcg 5820
tcgcctttgc cgacaaaaat gtgatcatga tgaaagccag caatgacgtt gcagctgata 5880
ttgttgtcgg ccaaccgcgc cgaaaacgca gctgtcagac ccacagcctc caacgaagaa 5940
tgtatcgtca aagtgatcca agcacactca tagttggagt cgtactccaa aggcggcaat 6000
gacgagtcag acagatactc gtcgaaaaca gtgtacgcag atctactata gaggaacatt 6060
taaattgccc cggagaagac ggccaggccg cctagatgac aaattcaaca actcacagct 6120
gactttctgc cattgccact aggggggggc ctttttatat ggccaagcca agctctccac 6180
gtcggttggg ctgcacccaa caataaatgg gtagggttgc accaacaaag ggatgggatg 6240
gggggtagaa gatacgagga taacggggct caatggcaca aataagaacg aatactgcca 6300
ttaagactcg tgatccagcg actgacacca ttgcatcatc taagggcctc aaaactacct 6360
cggaactgct gcgctgatct ggacaccaca gaggttccga gcactttagg ttgcaccaaa 6420
tgtcccacca ggtgcaggca gaaaacgctg gaacagcgtg tacagtttgt cttaacaaaa 6480
agtgagggcg ctgaggtcga gcagggtggt gtgacttgtt atagccttta gagctgcgaa 6540
agcgcgtatg gatttggctc atcaggccag attgagggtc tgtggacaca tgtcatgtta 6600
gtgtacttca atcgccccct ggatatagcc ccgacaatag gccgtggcct catttttttg 6660
ccttccgcac atttccattg ctcggtaccc acaccttgct tctcctgcac ttgccaacct 6720
taatactggt ttacattgac caacatctta caagcggggg gcttgtctag ggtatatata 6780
aacagtggct ctcccaatcg gttgccagtc tcttttttcc tttctttccc cacagattcg 6840
aaatctaaac tacacatcac agaattccga gccgtgagta tccacgacaa gatcagtgtc 6900
gagacgacgc gttttgtgta atgacacaat ccgaaagtcg ctagcaacac acactctcta 6960
cacaaactaa cccagctctg gtaccatggc tatcgactca caatactaca agtcgcgaga 7020
caaaaacgac acggcaccca aaatcgcggg aatccgatat gccccgctat cgacaccatt 7080
actcaaccga tgtgagacct tctctctggt ctggcacatt ttcagcattc ccactttcct 7140
cacaattttc atgctatgct gcgcaattcc actgctctgg ccatttgtga ttgcgtatgt 7200
agtgtacgct gttaaagacg actccccgtc caacggagga gtggtcaagc gatactcgcc 7260
tatttcaaga aacttcttca tctggaagct ctttggccgc tacttcccca taactctgca 7320
caagacggtg gatctggagc ccacgcacac atactaccct ctggacgtcc aggagtatca 7380
cctgattgct gagagatact ggccgcagaa caagtacctc cgagcaatca tctccaccat 7440
cgagtacttt ctgcccgcct tcatgaaacg gtctctttct atcaacgagc aggagcagcc 7500
tgccgagcga gatcctctcc tgtctcccgt ttctcccagc tctccgggtt ctcaacctga 7560
caagtggatt aaccacgaca gcagatatag ccgtggagaa tcatctggct ccaacggcca 7620
cgcctcgggc tccgaactta acggcaacgg caacaatggc accactaacc gacgaccttt 7680
gtcgtccgcc tctgctggct ccactgcatc tgattccacg cttcttaacg ggtccctcaa 7740
ctcctacgcc aaccagatca ttggcgaaaa cgacccacag ctgtcgccca caaaactcaa 7800
gcccactggc agaaaataca tcttcggcta ccacccccac ggcattatcg gcatgggagc 7860
ctttggtgga attgccaccg agggagctgg atggtccaag ctctttccgg gcatccctgt 7920
ttctcttatg actctcacca acaacttccg agtgcctctc tacagagagt acctcatgag 7980
tctgggagtc gcttctgtct ccaagaagtc ctgcaaggcc ctcctcaagc gaaaccagtc 8040
tatctgcatt gtcgttggtg gagcacagga aagtcttctg gccagacccg gtgtcatgga 8100
cctggtgcta ctcaagcgaa agggttttgt tcgacttggt atggaggtcg gaaatgtcgc 8160
ccttgttccc atcatggcct ttggtgagaa cgacctctat gaccaggtta gcaacgacaa 8220
gtcgtccaag ctgtaccgat tccagcagtt tgtcaagaac ttccttggat tcacccttcc 8280
tttgatgcat gcccgaggcg tcttcaacta cgatgtcggt cttgtcccct acaggcgacc 8340
cgtcaacatt gtggttggtt cccccattga cttgccttat ctcccacacc ccaccgacga 8400
agaagtgtcc gaataccacg accgatacat cgccgagctg cagcgaatct acaacgagca 8460
caaggatgaa tatttcatcg attggaccga ggagggcaaa ggagccccag agttccgaat 8520
gattgagtaa gc 8532
<210>21
<211>7049
<212>DNA
<213>人工序列
<220>
<223>质粒pKS123
<400>21
agcttggatc cgtcgacggc gcgcccgatc atccggatat agttcctcct ttcagcaaaa 60
aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc tcagcggtgg 120
cagcagccaa ctcagcttcc tttcgggctt tgttagcagc cggatcgatc caagctgtac 180
ctcactattc ctttgccctc ggacgagtgc tggggcgtcg gtttccacta tcggcgagta 240
cttctacaca gccatcggtc cagacggccg cgcttctgcg ggcgatttgt gtacgcccga 300
cagtcccggc tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat 360
cgaaattgcc gtcaaccaag ctctgataga gttggtcaag accaatgcgg agcatatacg 420
cccggagccg cggcgatcct gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct 480
gctccataca agccaaccac ggcctccaga agaagatgtt ggcgacctcg tattgggaat 540
ccccgaacat cgcctcgctc cagtcaatga ccgctgttat gcggccattg tccgtcagga 600
cattgttgga gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag tcctcggccc 660
aaagcatcag ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg tccatcacag 720
tttgccagtg atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt 780
attgaccgat tccttgcggt ccgaatgggc cgaacccgct cgtctggcta agatcggccg 840
cagcgatcgc atccatagcc tccgcgaccg gctgcagaac agcgggcagt tcggtttcag 900
gcaggtcttg caacgtgaca ccctgtgcac ggcgggagat gcaataggtc aggctctcgc 960
tgaattcccc aatgtcaagc acttccggaa tcgggagcgc ggccgatgca aagtgccgat 1020
aaacataacg atctttgtag aaaccatcgg cgcagctatt tacccgcagg acatatccac 1080
gccctcctac atcgaagctg aaagcacgag attcttcgcc ctccgagagc tgcatcaggt 1140
cggagacgct gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg gtgagttcag 1200
gcttttccat gggtatatct ccttcttaaa gttaaacaaa attatttcta gagggaaacc 1260
gttgtggtct ccctatagtg agtcgtatta atttcgcggg atcgagatcg atccaattcc 1320
aatcccacaa aaatctgagc ttaacagcac agttgctcct ctcagagcag aatcgggtat 1380
tcaacaccct catatcaact actacgttgt gtataacggt ccacatgccg gtatatacga 1440
tgactggggt tgtacaaagg cggcaacaaa cggcgttccc ggagttgcac acaagaaatt 1500
tgccactatt acagaggcaa gagcagcagc tgacgcgtac acaacaagtc agcaaacaga 1560
caggttgaac ttcatcccca aaggagaagc tcaactcaag cccaagagct ttgctaaggc 1620
cctaacaagc ccaccaaagc aaaaagccca ctggctcacg ctaggaacca aaaggcccag 1680
cagtgatcca gccccaaaag agatctcctt tgccccggag attacaatgg acgatttcct 1740
ctatctttac gatctaggaa ggaagttcga aggtgaaggt gacgacacta tgttcaccac 1800
tgataatgag aaggttagcc tcttcaattt cagaaagaat gctgacccac agatggttag 1860
agaggcctac gcagcaggtc tcatcaagac gatctacccg agtaacaatc tccaggagat 1920
caaatacctt cccaagaagg ttaaagatgc agtcaaaaga ttcaggacta attgcatcaa 1980
gaacacagag aaagacatat ttctcaagat cagaagtact attccagtat ggacgattca 2040
aggcttgctt cataaaccaa ggcaagtaat agagattgga gtctctaaaa aggtagttcc 2100
tactgaatct aaggccatgc atggagtcta agattcaaat cgaggatcta acagaactcg 2160
ccgtgaagac tggcgaacag ttcatacaga gtcttttacg actcaatgac aagaagaaaa 2220
tcttcgtcaa catggtggag cacgacactc tggtctactc caaaaatgtc aaagatacag 2280
tctcagaaga ccaaagggct attgagactt ttcaacaaag gataatttcg ggaaacctcc 2340
tcggattcca ttgcccagct atctgtcact tcatcgaaag gacagtagaa aaggaaggtg 2400
gctcctacaa atgccatcat tgcgataaag gaaaggctat cattcaagat gcctctgccg 2460
acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa gaagacgttc 2520
caaccacgtc ttcaaagcaa gtggattgat gtgacatctc cactgacgta agggatgacg 2580
cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca tttcatttgg 2640
agaggacacg ctcgagctca tttctctatt acttcagcca taacaaaaga actcttttct 2700
cttcttatta aaccatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga 2760
tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg 2820
ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg 2880
gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg 2940
aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac 3000
agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg 3060
cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat 3120
tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg 3180
atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc 3240
aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc 3300
acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg 3360
actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga 3420
ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc 3480
ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc 3540
agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa 3600
tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg 3660
tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc 3720
gtccgagggc aaaggaatag tgaggtacct aaagaaggag tgcgtcgaag cagatcgttc 3780
aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat 3840
catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt 3900
atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga 3960
aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact 4020
agatcgatgt cgaatctgat caacctgcat taatgaatcg gccaacgcgc ggggagaggc 4080
ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 4140
cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 4200
ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 4260
aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 4320
cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 4380
cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 4440
gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt 4500
tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 4560
cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 4620
ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 4680
gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 4740
gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 4800
accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 4860
ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 4920
tcacgttaag ggattttggt catgacatta acctataaaa ataggcgtat cacgaggccc 4980
tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag 5040
acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca 5100
gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg 5160
agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata cataacctta 5220
tgtatcatac acatacgatt taggtgacac tatagaacgg cgcgccaagc ttttgatcca 5280
tgcccttcat ttgccgctta ttaattaatt tggtaacagt ccgtactaat cagttactta 5340
tccttccccc atcataatta atcttggtag tctcgaatgc cacaacactg actagtctct 5400
tggatcataa gaaaaagcca aggaacaaaa gaagacaaaa cacaatgaga gtatcctttg 5460
catagcaatg tctaagttca taaaattcaa acaaaaacgc aatcacacac agtggacatc 5520
acttatccac tagctgatca ggatcgccgc gtcaagaaaa aaaaactgga ccccaaaagc 5580
catgcacaac aacacgtact cacaaaggtg tcaatcgagc agcccaaaac attcaccaac 5640
tcaacccatc atgagccctc acatttgttg tttctaaccc aacctcaaac tcgtattctc 5700
ttccgccacc tcatttttgt ttatttcaac acccgtcaaa ctgcatgcca ccccgtggcc 5760
aaatgtccat gcatgttaac aagacctatg actataaata gctgcaatct cggcccaggt 5820
tttcatcatc aagaaccagt tcaatatcct agtacaccgt attaaagaat ttaagatata 5880
ctgcggccgc aagtatgaac taaaatgcat gtaggtgtaa gagctcatgg agagcatgga 5940
atattgtatc cgaccatgta acagtataat aactgagctc catctcactt cttctatgaa 6000
taaacaaagg atgttatgat atattaacac tctatctatg caccttattg ttctatgata 6060
aatttcctct tattattata aatcatctga atcgtgacgg cttatggaat gcttcaaata 6120
gtacaaaaac aaatgtgtac tataagactt tctaaacaat tctaacctta gcattgtgaa 6180
cgagacataa gtgttaagaa gacataacaa ttataatgga agaagtttgt ctccatttat 6240
atattatata ttacccactt atgtattata ttaggatgtt aaggagacat aacaattata 6300
aagagagaag tttgtatcca tttatatatt atatactacc catttatata ttatacttat 6360
ccacttattt aatgtcttta taaggtttga tccatgatat ttctaatatt ttagttgata 6420
tgtatatgaa agggtactat ttgaactctc ttactctgta taaaggttgg atcatcctta 6480
aagtgggtct atttaatttt attgcttctt acagataaaa aaaaaattat gagttggttt 6540
gataaaatat tgaaggattt aaaataataa taaataacat ataatatatg tatataaatt 6600
tattataata taacatttat ctataaaaaa gtaaatattg tcataaatct atacaatcgt 6660
ttagccttgc tggacgaatc tcaattattt aaacgagagt aaacatattt gactttttgg 6720
ttatttaaca aattattatt taacactata tgaaattttt ttttttatca gcaaagaata 6780
aaattaaatt aagaaggaca atggtgtccc aatccttata caaccaactt ccacaagaaa 6840
gtcaagtcag agacaacaaa aaaacaagca aaggaaattt tttaatttga gttgtcttgt 6900
ttgctgcata atttatgcag taaaacacta cacataaccc ttttagcagt agagcaatgg 6960
ttgaccgtgt gcttagcttc ttttatttta tttttttatc agcaaagaat aaataaaata 7020
aaatgagaca cttcagggat gtttcaaca 7049
<210>22
<211>1098
<212>DNA
<213>人工序列
<220>
<223>cal a24-4
<400>22
aggatcctgc aggcagccaa gaaggttttg gagcgagttc caatctcaaa accgccattc 60
gaatacaatg atctgaagaa agcagtacca ccacattgtt tttcacgacc actttcccga 120
tccttgtatt tcctctttca cgacattatt gtaacatgta tccttttcta cgtagcatca 180
aactacattc atatgctccc tcgtttcctt tcctgcatcg tatggcctgt ttactggatc 240
tcccaaggag tttttctcgg cagattgtgg atgatcggcc acgaatgcgg tcatcatagc 300
ttcagtaatt accgttgggt cgacgataca gtcggttttc taatccatac ggccaccctc 360
actccctatt tttccttcaa atatagccac cgtaatcacc atgcacacac caattccatg 420
gaatacgacg aggttcatat cccgaaacgc aaatcagaag ctctctactt tgaatttctg 480
ggcaacaacc caatcggctt aatgatcacc atgctatgta aactgacttt cggatatgca 540
gcttacatta tgttcaatta cacaggtaag aagcacaaat ctgggggctt agcgagccac 600
ttctacccac aaagccctct ctttaacgac agcgaacgta accatgtttt gttctctgac 660
atcgggattt gcatcgtctt gtacgcgtgt taccgtattg tgacggtcac aggggcaatg 720
ccggcatttt atgtgtacgg tattccttgg gttataatga gtgctattct ctttgcagca 780
acttatttac aacacactca tccttcaatc cctcattatg atacaacgga gtggaactgg 840
cttagagggg ctttatcgac aattgataga gatttagggt tcttcaacat gaacaaaaca 900
cattatcatg ttatccacca tttgtttcct gtcattccgg aataccatgc acaagaggca 960
accgaggcca tcaagcccat cttaggtcaa tattacaagt atgatggtac tccgtttcta 1020
aaggccttgt ggagagaaat gaaggagtgt atttatgtag aatccgatga aggtcagaag 1080
aaacctgcag gagatctt 1098
<210>23
<211>30
<212>DNA
<213>人工序列
<220>
<223>引物oCal-15
<400>23
aggatcctgc aggcagccaa gaaggttttg 30
<210>24
<211>29
<212>DNA
<213>人工序列
<220>
<223>引物oCal-6
<400>24
aagatctcct gcaggtttct tctgacctc 29
<210>25
<211>8138
<212>DNA
<213>人工序列
<220>
<223>质粒pKR53B
<400>25
gatccgtcga cggcgcgccc gatcatccgg atatagttcc tcctttcagc aaaaaacccc 60
tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag 120
ccaactcagc ttcctttcgg gctttgttag cagccggatc gatccaagct gtacctcact 180
attcctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta 240
cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc 300
cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat 360
tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga 420
gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca 480
tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga 540
acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt 600
tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca 660
tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc 720
agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac 780
cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga 840
tcgcatccat agcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt 900
cttgcaacgt gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaatt 960
ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat 1020
aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc 1080
ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga 1140
cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt 1200
ccatgggtat atctccttct taaagttaaa caaaattatt tctagaggga aaccgttgtg 1260
gtctccctat agtgagtcgt attaatttcg cgggatcgag atcgatccaa ttccaatccc 1320
acaaaaatct gagcttaaca gcacagttgc tcctctcaga gcagaatcgg gtattcaaca 1380
ccctcatatc aactactacg ttgtgtataa cggtccacat gccggtatat acgatgactg 1440
gggttgtaca aaggcggcaa caaacggcgt tcccggagtt gcacacaaga aatttgccac 1500
tattacagag gcaagagcag cagctgacgc gtacacaaca agtcagcaaa cagacaggtt 1560
gaacttcatc cccaaaggag aagctcaact caagcccaag agctttgcta aggccctaac 1620
aagcccacca aagcaaaaag cccactggct cacgctagga accaaaaggc ccagcagtga 1680
tccagcccca aaagagatct cctttgcccc ggagattaca atggacgatt tcctctatct 1740
ttacgatcta ggaaggaagt tcgaaggtga aggtgacgac actatgttca ccactgataa 1800
tgagaaggtt agcctcttca atttcagaaa gaatgctgac ccacagatgg ttagagaggc 1860
ctacgcagca ggtctcatca agacgatcta cccgagtaac aatctccagg agatcaaata 1920
ccttcccaag aaggttaaag atgcagtcaa aagattcagg actaattgca tcaagaacac 1980
agagaaagac atatttctca agatcagaag tactattcca gtatggacga ttcaaggctt 2040
gcttcataaa ccaaggcaag taatagagat tggagtctct aaaaaggtag ttcctactga 2100
atctaaggcc atgcatggag tctaagattc aaatcgagga tctaacagaa ctcgccgtga 2160
agactggcga acagttcata cagagtcttt tacgactcaa tgacaagaag aaaatcttcg 2220
tcaacatggt ggagcacgac actctggtct actccaaaaa tgtcaaagat acagtctcag 2280
aagaccaaag ggctattgag acttttcaac aaaggataat ttcgggaaac ctcctcggat 2340
tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct 2400
acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatgcctct gccgacagtg 2460
gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca 2520
cgtcttcaaa gcaagtggat tgatgtgaca tctccactga cgtaagggat gacgcacaat 2580
cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga 2640
cacgctcgag ctcatttctc tattacttca gccataacaa aagaactctt ttctcttctt 2700
attaaaccat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa 2760
agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca 2820
gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct 2880
acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc 2940
ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg 3000
tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg 3060
ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac 3120
cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc 3180
atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc 3240
tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg 3300
atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga 3360
gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt 3420
ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag 3480
gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct 3540
tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc 3600
gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga 3660
ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga 3720
gggcaaagga atagtgaggt acctaaagaa ggagtgcgtc gaagcagatc gttcaaacat 3780
ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata 3840
atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat 3900
gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa 3960
aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg 4020
atgtcgaatc gatcaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4080
gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4140
ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4200
acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4260
cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4320
caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4380
gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4440
tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt 4500
aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 4560
ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 4620
cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 4680
tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 4740
tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 4800
ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 4860
aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 4920
aagggatttt ggtcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 4980
tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 5040
cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 5100
ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 5160
accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca 5220
tacacatacg atttaggtga cactatagaa cggcgcgcca agcttttgat ccatgccctt 5280
catttgccgc ttattaatta atttggtaac agtccgtact aatcagttac ttatccttcc 5340
cccatcataa ttaatcttgg tagtctcgaa tgccacaaca ctgactagtc tcttggatca 5400
taagaaaaag ccaaggaaca aaagaagaca aaacacaatg agagtatcct ttgcatagca 5460
atgtctaagt tcataaaatt caaacaaaaa cgcaatcaca cacagtggac atcacttatc 5520
cactagctga tcaggatcgc cgcgtcaaga aaaaaaaact ggaccccaaa agccatgcac 5580
aacaacacgt actcacaaag gtgtcaatcg agcagcccaa aacattcacc aactcaaccc 5640
atcatgagcc ctcacatttg ttgtttctaa cccaacctca aactcgtatt ctcttccgcc 5700
acctcatttt tgtttatttc aacacccgtc aaactgcatg ccaccccgtg gccaaatgtc 5760
catgcatgtt aacaagacct atgactataa atagctgcaa tctcggccca ggttttcatc 5820
atcaagaacc agttcaatat cctagtacac cgtattaaag aatttaagat atactgcggc 5880
cgcaagtatg aactaaaatg catgtaggtg taagagctca tggagagcat ggaatattgt 5940
atccgaccat gtaacagtat aataactgag ctccatctca cttcttctat gaataaacaa 6000
aggatgttat gatatattaa cactctatct atgcacctta ttgttctatg ataaatttcc 6060
tcttattatt ataaatcatc tgaatcgtga cggcttatgg aatgcttcaa atagtacaaa 6120
aacaaatgtg tactataaga ctttctaaac aattctaacc ttagcattgt gaacgagaca 6180
taagtgttaa gaagacataa caattataat ggaagaagtt tgtctccatt tatatattat 6240
atattaccca cttatgtatt atattaggat gttaaggaga cataacaatt ataaagagag 6300
aagtttgtat ccatttatat attatatact acccatttat atattatact tatccactta 6360
tttaatgtct ttataaggtt tgatccatga tatttctaat attttagttg atatgtatat 6420
gaaagggtac tatttgaact ctcttactct gtataaaggt tggatcatcc ttaaagtggg 6480
tctatttaat tttattgctt cttacagata aaaaaaaaat tatgagttgg tttgataaaa 6540
tattgaagga tttaaaataa taataaataa catataatat atgtatataa atttattata 6600
atataacatt tatctataaa aaagtaaata ttgtcataaa tctatacaat cgtttagcct 6660
tgctggacga atctcaatta tttaaacgag agtaaacata tttgactttt tggttattta 6720
acaaattatt atttaacact atatgaaatt ttttttttta tcagcaaaga ataaaattaa 6780
attaagaagg acaatggtgt cccaatcctt atacaaccaa cttccacaag aaagtcaagt 6840
cagagacaac aaaaaaacaa gcaaaggaaa ttttttaatt tgagttgtct tgtttgctgc 6900
ataatttatg cagtaaaaca ctacacataa cccttttagc agtagagcaa tggttgaccg 6960
tgtgcttagc ttcttttatt ttattttttt atcagcaaag aataaataaa ataaaatgag 7020
acacttcagg gatgtttcaa caagcttgga tctcctgcag gtttcttctg accttcatcg 7080
gattctacat aaatacactc cttcatttct ctccacaagg cctttagaaa cggagtacca 7140
tcatacttgt aatattgacc taagatgggc ttgatggcct cggttgcctc ttgtgcatgg 7200
tattccggaa tgacaggaaa caaatggtgg ataacatgat aatgtgtttt gttcatgttg 7260
aagaacccta aatctctatc aattgtcgat aaagcccctc taagccagtt ccactccgtt 7320
gtatcataat gagggattga aggatgagtg tgttgtaaat aagttgctgc aaagagaata 7380
gcactcatta taacccaagg aataccgtac acataaaatg ccggcattgc ccctgtgacc 7440
gtcacaatac ggtaacacgc gtacaagacg atgcaaatcc cgatgtcaga gaacaaaaca 7500
tggttacgtt cgctgtcgtt aaagagaggg ctttgtgggt agaagtggct cgctaagccc 7560
ccagatttgt gcttcttacc tgtgtaattg aacataatgt aagctgcata tccgaaagtc 7620
agtttacata gcatggtgat cattaagccg attgggttgt tgcccagaaa ttcaaagtag 7680
agagcttctg atttgcgttt cgggatatga acctcgtcgt attccatgga attggtgtgt 7740
gcatggtgat tacggtggct atatttgaag gaaaaatagg gagtgagggt ggccgtatgg 7800
attagaaaac cgactgtatc gtcgacccaa cggtaattac tgaagctatg atgaccgcat 7860
tcgtggccga tcatccacaa tctgccgaga aaaactcctt gggagatcca gtaaacaggc 7920
catacgatgc aggaaaggaa acgagggagc atatgaatgt agtttgatgc tacgtagaaa 7980
aggatacatg ttacaataat gtcgtgaaag aggaaataca aggatcggga aagtggtcgt 8040
gaaaaacaat gtggtggtac tgctttcttc agatcattgt attcgaatgg cggttttgag 8100
attggaactc gctccaaaac cttcttggct gcctgcag 8138
<210>26
<211>7085
<212>DNA
<213>人工序列
<220>
<223>质粒pKR72
<400>26
gtacggatcc gtcgacggcg cgcccgatca tccggatata gttcctcctt tcagcaaaaa 60
acccctcaag acccgtttag aggccccaag gggttatgct agttattgct cagcggtggc 120
agcagccaac tcagcttcct ttcgggcttt gttagcagcc ggatcgatcc aagctgtacc 180
tcactattcc tttgccctcg gacgagtgct ggggcgtcgg tttccactat cggcgagtac 240
ttctacacag ccatcggtcc agacggccgc gcttctgcgg gcgatttgtg tacgcccgac 300
agtcccggct ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc 360
gaaattgccg tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga gcatatacgc 420
ccggagccgc ggcgatcctg caagctccgg atgcctccgc tcgaagtagc gcgtctgctg 480
ctccatacaa gccaaccacg gcctccagaa gaagatgttg gcgacctcgt attgggaatc 540
cccgaacatc gcctcgctcc agtcaatgac cgctgttatg cggccattgt ccgtcaggac 600
attgttggag ccgaaatccg cgtgcacgag gtgccggact tcggggcagt cctcggccca 660
aagcatcagc tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt 720
ttgccagtga tacacatggg gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta 780
ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa gatcggccgc 840
agcgatcgca tccatagcct ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg 900
caggtcttgc aacgtgacac cctgtgcacg gcgggagatg caataggtca ggctctcgct 960
gaattcccca atgtcaagca cttccggaat cgggagcgcg gccgatgcaa agtgccgata 1020
aacataacga tctttgtaga aaccatcggc gcagctattt acccgcagga catatccacg 1080
ccctcctaca tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc 1140
ggagacgctg tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg 1200
cttttccatg ggtatatctc cttcttaaag ttaaacaaaa ttatttctag agggaaaccg 1260
ttgtggtctc cctatagtga gtcgtattaa tttcgcggga tcgagatcga tccaattcca 1320
atcccacaaa aatctgagct taacagcaca gttgctcctc tcagagcaga atcgggtatt 1380
caacaccctc atatcaacta ctacgttgtg tataacggtc cacatgccgg tatatacgat 1440
gactggggtt gtacaaaggc ggcaacaaac ggcgttcccg gagttgcaca caagaaattt 1500
gccactatta cagaggcaag agcagcagct gacgcgtaca caacaagtca gcaaacagac 1560
aggttgaact tcatccccaa aggagaagct caactcaagc ccaagagctt tgctaaggcc 1620
ctaacaagcc caccaaagca aaaagcccac tggctcacgc taggaaccaa aaggcccagc 1680
agtgatccag ccccaaaaga gatctccttt gccccggaga ttacaatgga cgatttcctc 1740
tatctttacg atctaggaag gaagttcgaa ggtgaaggtg acgacactat gttcaccact 1800
gataatgaga aggttagcct cttcaatttc agaaagaatg ctgacccaca gatggttaga 1860
gaggcctacg cagcaggtct catcaagacg atctacccga gtaacaatct ccaggagatc 1920
aaataccttc ccaagaaggt taaagatgca gtcaaaagat tcaggactaa ttgcatcaag 1980
aacacagaga aagacatatt tctcaagatc agaagtacta ttccagtatg gacgattcaa 2040
ggcttgcttc ataaaccaag gcaagtaata gagattggag tctctaaaaa ggtagttcct 2100
actgaatcta aggccatgca tggagtctaa gattcaaatc gaggatctaa cagaactcgc 2160
cgtgaagact ggcgaacagt tcatacagag tcttttacga ctcaatgaca agaagaaaat 2220
cttcgtcaac atggtggagc acgacactct ggtctactcc aaaaatgtca aagatacagt 2280
ctcagaagac caaagggcta ttgagacttt tcaacaaagg ataatttcgg gaaacctcct 2340
cggattccat tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaaggtgg 2400
ctcctacaaa tgccatcatt gcgataaagg aaaggctatc attcaagatg cctctgccga 2460
cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc 2520
aaccacgtct tcaaagcaag tggattgatg tgacatctcc actgacgtaa gggatgacgc 2580
acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga 2640
gaggacacgc tcgagctcat ttctctatta cttcagccat aacaaaagaa ctcttttctc 2700
ttcttattaa accatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat 2760
cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc 2820
tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg 2880
tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga 2940
agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca 3000
gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc 3060
ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt 3120
cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga 3180
tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca 3240
ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca 3300
cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga 3360
ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag 3420
gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct 3480
tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca 3540
gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat 3600
cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt 3660
ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg 3720
tccgagggca aaggaatagt gaggtaccta aagaaggagt gcgtcgaagc agatcgttca 3780
aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc 3840
atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta 3900
tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa 3960
aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta 4020
gatcgatgtc gaatcgatca acctgcatta atgaatcggc caacgcgcgg ggagaggcgg 4080
tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 4140
gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4200
ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4260
ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 4320
acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 4380
tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 4440
ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc 4500
ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 4560
ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 4620
actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4680
gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 4740
tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 4800
caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4860
atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4920
acgttaaggg attttggtca tgacattaac ctataaaaat aggcgtatca cgaggccctt 4980
tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 5040
ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 5100
gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag 5160
agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg 5220
tatcatacac atacgattta ggtgacacta tagaacggcg cgccaagctt gttgaaacat 5280
ccctgaagtg tctcatttta ttttatttat tctttgctga taaaaaaata aaataaaaga 5340
agctaagcac acggtcaacc attgctctac tgctaaaagg gttatgtgta gtgttttact 5400
gcataaatta tgcagcaaac aagacaactc aaattaaaaa atttcctttg cttgtttttt 5460
tgttgtctct gacttgactt tcttgtggaa gttggttgta taaggattgg gacaccattg 5520
tccttcttaa tttaatttta ttctttgctg ataaaaaaaa aaatttcata tagtgttaaa 5580
taataatttg ttaaataacc aaaaagtcaa atatgtttac tctcgtttaa ataattgaga 5640
ttcgtccagc aaggctaaac gattgtatag atttatgaca atatttactt ttttatagat 5700
aaatgttata ttataataaa tttatataca tatattatat gttatttatt attattttaa 5760
atccttcaat attttatcaa accaactcat aatttttttt ttatctgtaa gaagcaataa 5820
aattaaatag acccacttta aggatgatcc aacctttata cagagtaaga gagttcaaat 5880
agtacccttt catatacata tcaactaaaa tattagaaat atcatggatc aaaccttata 5940
aagacattaa ataagtggat aagtataata tataaatggg tagtatataa tatataaatg 6000
gatacaaact tctctcttta taattgttat gtctccttaa catcctaata taatacataa 6060
gtgggtaata tataatatat aaatggagac aaacttcttc cattataatt gttatgtctt 6120
cttaacactt atgtctcgtt cacaatgcta aggttagaat tgtttagaaa gtcttatagt 6180
acacatttgt ttttgtacta tttgaagcat tccataagcc gtcacgattc agatgattta 6240
taataataag aggaaattta tcatagaaca ataaggtgca tagatagagt gttaatatat 6300
cataacatcc tttgtttatt catagaagaa gtgagatgga gctcagttat tatactgtta 6360
catggtcgga tacaatattc catgctctcc atgagctctt acacctacat gcattttagt 6420
tcatacttgc ggccgcagta tatcttaaat tctttaatac ggtgtactag gatattgaac 6480
tggttcttga tgatgaaaac ctgggccgag attgcagcta tttatagtca taggtcttgt 6540
taacatgcat ggacatttgg ccacggggtg gcatgcagtt tgacgggtgt tgaaataaac 6600
aaaaatgagg tggcggaaga gaatacgagt ttgaggttgg gttagaaaca acaaatgtga 6660
gggctcatga tgggttgagt tggtgaatgt tttgggctgc tcgattgaca cctttgtgag 6720
tacgtgttgt tgtgcatggc ttttggggtc cagttttttt ttcttgacgc ggcgatcctg 6780
atcagctagt ggataagtga tgtccactgt gtgtgattgc gtttttgttt gaattttatg 6840
aacttagaca ttgctatgca aaggatactc tcattgtgtt ttgtcttctt ttgttccttg 6900
gctttttctt atgatccaag agactagtca gtgttgtggc attcgagact accaagatta 6960
attatgatgg gggaaggata agtaactgat tagtacggac tgttaccaaa ttaattaata 7020
agcggcaaat gaagggcatg gatcaaaagc ttggatctcc tgcaggatct ggccggccgg 7080
atctc 7085
<210>27
<211>7085
<212>DNA
<213>人工序列
<220>
<223>质粒pKR85
<400>27
cgcgccaagc ttttgatcca tgcccttcat ttgccgctta ttaattaatt tggtaacagt 60
ccgtactaat cagttactta tccttccccc atcataatta atcttggtag tctcgaatgc 120
cacaacactg actagtctct tggatcataa gaaaaagcca aggaacaaaa gaagacaaaa 180
cacaatgaga gtatcctttg catagcaatg tctaagttca taaaattcaa acaaaaacgc 240
aatcacacac agtggacatc acttatccac tagctgatca ggatcgccgc gtcaagaaaa 300
aaaaactgga ccccaaaagc catgcacaac aacacgtact cacaaaggtg tcaatcgagc 360
agcccaaaac attcaccaac tcaacccatc atgagccctc acatttgttg tttctaaccc 420
aacctcaaac tcgtattctc ttccgccacc tcatttttgt ttatttcaac acccgtcaaa 480
ctgcatgcca ccccgtggcc aaatgtccat gcatgttaac aagacctatg actataaata 540
gctgcaatct cggcccaggt tttcatcatc aagaaccagt tcaatatcct agtacaccgt 600
attaaagaat ttaagatata ctgcggccgc aagtatgaac taaaatgcat gtaggtgtaa 660
gagctcatgg agagcatgga atattgtatc cgaccatgta acagtataat aactgagctc 720
catctcactt cttctatgaa taaacaaagg atgttatgat atattaacac tctatctatg 780
caccttattg ttctatgata aatttcctct tattattata aatcatctga atcgtgacgg 840
cttatggaat gcttcaaata gtacaaaaac aaatgtgtac tataagactt tctaaacaat 900
tctaacctta gcattgtgaa cgagacataa gtgttaagaa gacataacaa ttataatgga 960
agaagtttgt ctccatttat atattatata ttacccactt atgtattata ttaggatgtt 1020
aaggagacat aacaattata aagagagaag tttgtatcca tttatatatt atatactacc 1080
catttatata ttatacttat ccacttattt aatgtcttta taaggtttga tccatgatat 1140
ttctaatatt ttagttgata tgtatatgaa agggtactat ttgaactctc ttactctgta 1200
taaaggttgg atcatcctta aagtgggtct atttaatttt attgcttctt acagataaaa 1260
aaaaaattat gagttggttt gataaaatat tgaaggattt aaaataataa taaataacat 1320
ataatatatg tatataaatt tattataata taacatttat ctataaaaaa gtaaatattg 1380
tcataaatct atacaatcgt ttagccttgc tggacgaatc tcaattattt aaacgagagt 1440
aaacatattt gactttttgg ttatttaaca aattattatt taacactata tgaaattttt 1500
ttttttatca gcaaagaata aaattaaatt aagaaggaca atggtgtccc aatccttata 1560
caaccaactt ccacaagaaa gtcaagtcag agacaacaaa aaaacaagca aaggaaattt 1620
tttaatttga gttgtcttgt ttgctgcata atttatgcag taaaacacta cacataaccc 1680
ttttagcagt agagcaatgg ttgaccgtgt gcttagcttc ttttatttta tttttttatc 1740
agcaaagaat aaataaaata aaatgagaca cttcagggat gtttcaacaa gcttggatct 1800
cctgcaggat ctggccggcc ggatctcgta cggatccgtc gacggcgcgc ccgatcatcc 1860
ggatatagtt cctcctttca gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg 1920
ttatgctagt tattgctcag cggtggcagc agccaactca gcttcctttc gggctttgtt 1980
agcagccgga tcgatccaag ctgtacctca ctattccttt gccctcggac gagtgctggg 2040
gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga cggccgcgct 2100
tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga ttgcgtcgca 2160
tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct gatagagttg 2220
gtcaagacca atgcggagca tatacgcccg gagccgcggc gatcctgcaa gctccggatg 2280
cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc tccagaagaa 2340
gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt caatgaccgc 2400
tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt gcacgaggtg 2460
ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct gcgcgacgga 2520
cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat cagcaatcgc 2580
gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga atgggccgaa 2640
cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atagcctccg cgaccggctg 2700
cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg 2760
ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg 2820
gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac catcggcgca 2880
gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag cacgagattc 2940
ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga tcagaaactt 3000
ctcgacagac gtcgcggtga gttcaggctt ttccatgggt atatctcctt cttaaagtta 3060
aacaaaatta tttctagagg gaaaccgttg tggtctccct atagtgagtc gtattaattt 3120
cgcgggatcg agatcgatcc aattccaatc ccacaaaaat ctgagcttaa cagcacagtt 3180
gctcctctca gagcagaatc gggtattcaa caccctcata tcaactacta cgttgtgtat 3240
aacggtccac atgccggtat atacgatgac tggggttgta caaaggcggc aacaaacggc 3300
gttcccggag ttgcacacaa gaaatttgcc actattacag aggcaagagc agcagctgac 3360
gcgtacacaa caagtcagca aacagacagg ttgaacttca tccccaaagg agaagctcaa 3420
ctcaagccca agagctttgc taaggcccta acaagcccac caaagcaaaa agcccactgg 3480
ctcacgctag gaaccaaaag gcccagcagt gatccagccc caaaagagat ctcctttgcc 3540
ccggagatta caatggacga tttcctctat ctttacgatc taggaaggaa gttcgaaggt 3600
gaaggtgacg acactatgtt caccactgat aatgagaagg ttagcctctt caatttcaga 3660
aagaatgctg acccacagat ggttagagag gcctacgcag caggtctcat caagacgatc 3720
tacccgagta acaatctcca ggagatcaaa taccttccca agaaggttaa agatgcagtc 3780
aaaagattca ggactaattg catcaagaac acagagaaag acatatttct caagatcaga 3840
agtactattc cagtatggac gattcaaggc ttgcttcata aaccaaggca agtaatagag 3900
attggagtct ctaaaaaggt agttcctact gaatctaagg ccatgcatgg agtctaagat 3960
tcaaatcgag gatctaacag aactcgccgt gaagactggc gaacagttca tacagagtct 4020
tttacgactc aatgacaaga agaaaatctt cgtcaacatg gtggagcacg acactctggt 4080
ctactccaaa aatgtcaaag atacagtctc agaagaccaa agggctattg agacttttca 4140
acaaaggata atttcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat 4200
cgaaaggaca gtagaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa 4260
ggctatcatt caagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag 4320
gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga 4380
catctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc 4440
tatataagga agttcatttc atttggagag gacacgctcg agctcatttc tctattactt 4500
cagccataac aaaagaactc ttttctcttc ttattaaacc atgaaaaagc ctgaactcac 4560
cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca 4620
gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt 4680
cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt 4740
tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct 4800
gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga 4860
actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct 4920
tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg 4980
gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga 5040
cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga 5100
ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga 5160
caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata 5220
cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg 5280
ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct 5340
ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc 5400
ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac 5460
acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga 5520
tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagtgag gtacctaaag 5580
aaggagtgcg tcgaagcaga tcgttcaaac atttggcaat aaagtttctt aagattgaat 5640
cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta 5700
ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg 5760
caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta 5820
tcgcgcgcgg tgtcatctat gttactagat cgatgtcgaa tcgatcaacc tgcattaatg 5880
aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 5940
cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 6000
ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 6060
ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 6120
cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 6180
actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 6240
cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 6300
atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 6360
gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 6420
caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 6480
agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 6540
tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 6600
tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 6660
gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 6720
gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga cattaaccta 6780
taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 6840
cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 6900
cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 6960
tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg 7020
cggctacaat taatacataa ccttatgtat catacacata cgatttaggt gacactatag 7080
aacgg 7085
<210>28
<211>22
<212>DNA
<213>人工序列
<220>
<223>引物oKR85-1
<400>28
actcgaggcg cgccgtcgac gg 22
<210>29
<211>18
<212>DNA
<213>人工序列
<220>
<223>引物oKR85-2
<400>29
aagatctggc gcgccaag 18
<210>30
<211>4827
<212>DNA
<213>人工序列
<220>
<223>质粒pPCR85
<400>30
actcgaggcg cgccgtcgac ggatccgtac gagatccggc cggccagatc ctgcaggaga 60
tccaagcttg ttgaaacatc cctgaagtgt ctcattttat tttatttatt ctttgctgat 120
aaaaaaataa aataaaagaa gctaagcaca cggtcaacca ttgctctact gctaaaaggg 180
ttatgtgtag tgttttactg cataaattat gcagcaaaca agacaactca aattaaaaaa 240
tttcctttgc ttgttttttt gttgtctctg acttgacttt cttgtggaag ttggttgtat 300
aaggattggg acaccattgt ccttcttaat ttaattttat tctttgctga taaaaaaaaa 360
aatttcatat agtgttaaat aataatttgt taaataacca aaaagtcaaa tatgtttact 420
ctcgtttaaa taattgagat tcgtccagca aggctaaacg attgtataga tttatgacaa 480
tatttacttt tttatagata aatgttatat tataataaat ttatatacat atattatatg 540
ttatttatta ttattttaaa tccttcaata ttttatcaaa ccaactcata attttttttt 600
tatctgtaag aagcaataaa attaaataga cccactttaa ggatgatcca acctttatac 660
agagtaagag agttcaaata gtaccctttc atatacatat caactaaaat attagaaata 720
tcatggatca aaccttataa agacattaaa taagtggata agtataatat ataaatgggt 780
agtatataat atataaatgg atacaaactt ctctctttat aattgttatg tctccttaac 840
atcctaatat aatacataag tgggtaatat ataatatata aatggagaca aacttcttcc 900
attataattg ttatgtcttc ttaacactta tgtctcgttc acaatgctaa ggttagaatt 960
gtttagaaag tcttatagta cacatttgtt tttgtactat ttgaagcatt ccataagccg 1020
tcacgattca gatgatttat aataataaga ggaaatttat catagaacaa taaggtgcat 1080
agatagagtg ttaatatatc ataacatcct ttgtttattc atagaagaag tgagatggag 1140
ctcagttatt atactgttac atggtcggat acaatattcc atgctctcca tgagctctta 1200
cacctacatg cattttagtt catacttgcg gccgcagtat atcttaaatt ctttaatacg 1260
gtgtactagg atattgaact ggttcttgat gatgaaaacc tgggccgaga ttgcagctat 1320
ttatagtcat aggtcttgtt aacatgcatg gacatttggc cacggggtgg catgcagttt 1380
gacgggtgtt gaaataaaca aaaatgaggt ggcggaagag aatacgagtt tgaggttggg 1440
ttagaaacaa caaatgtgag ggctcatgat gggttgagtt ggtgaatgtt ttgggctgct 1500
cgattgacac ctttgtgagt acgtgttgtt gtgcatggct tttggggtcc agtttttttt 1560
tcttgacgcg gcgatcctga tcagctagtg gataagtgat gtccactgtg tgtgattgcg 1620
tttttgtttg aattttatga acttagacat tgctatgcaa aggatactct cattgtgttt 1680
tgtcttcttt tgttccttgg ctttttctta tgatccaaga gactagtcag tgttgtggca 1740
ttcgagacta ccaagattaa ttatgatggg ggaaggataa gtaactgatt agtacggact 1800
gttaccaaat taattaataa gcggcaaatg aagggcatgg atcaaaagct tggcgcgcca 1860
gatcttgggc tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga 1920
gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 1980
ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 2040
taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 2100
aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 2160
attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 2220
cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 2280
gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 2340
ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 2400
agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 2460
tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 2520
ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 2580
gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 2640
ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 2700
gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 2760
aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 2820
aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 2880
ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 2940
gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 3000
gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 3060
tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 3120
ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 3180
ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 3240
atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 3300
ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 3360
tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 3420
attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 3480
tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 3540
ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 3600
gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 3660
gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 3720
gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 3780
aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 3840
taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 3900
tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 3960
tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 4020
atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 4080
tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa 4140
atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata 4200
aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac 4260
tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc 4320
cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa 4380
atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg 4440
cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg 4500
tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc 4560
attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 4620
tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 4680
tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat 4740
agggcgaatt gggtaccggg ccccccctcg aggtcgacgg tatcgataag cttgatatcg 4800
aattcctgca gcccggggga tccgccc 4827
<210>31
<211>15114
<212>DNA
<213>人工序列
<220>
<223>质粒pKR91
<400>31
gatctggcgc gccaagcttt tgatccatgc ccttcatttg ccgcttatta attaatttgg 60
taacagtccg tactaatcag ttacttatcc ttcccccatc ataattaatc ttggtagtct 120
cgaatgccac aacactgact agtctcttgg atcataagaa aaagccaagg aacaaaagaa 180
gacaaaacac aatgagagta tcctttgcat agcaatgtct aagttcataa aattcaaaca 240
aaaacgcaat cacacacagt ggacatcact tatccactag ctgatcagga tcgccgcgtc 300
aagaaaaaaa aactggaccc caaaagccat gcacaacaac acgtactcac aaaggtgtca 360
atcgagcagc ccaaaacatt caccaactca acccatcatg agccctcaca tttgttgttt 420
ctaacccaac ctcaaactcg tattctcttc cgccacctca tttttgttta tttcaacacc 480
cgtcaaactg catgccaccc cgtggccaaa tgtccatgca tgttaacaag acctatgact 540
ataaatagct gcaatctcgg cccaggtttt catcatcaag aaccagttca atatcctagt 600
acaccgtatt aaagaattta agatatactg cggccgcaag tatgaactaa aatgcatgta 660
ggtgtaagag ctcatggaga gcatggaata ttgtatccga ccatgtaaca gtataataac 720
tgagctccat ctcacttctt ctatgaataa acaaaggatg ttatgatata ttaacactct 780
atctatgcac cttattgttc tatgataaat ttcctcttat tattataaat catctgaatc 840
gtgacggctt atggaatgct tcaaatagta caaaaacaaa tgtgtactat aagactttct 900
aaacaattct aaccttagca ttgtgaacga gacataagtg ttaagaagac ataacaatta 960
taatggaaga agtttgtctc catttatata ttatatatta cccacttatg tattatatta 1020
ggatgttaag gagacataac aattataaag agagaagttt gtatccattt atatattata 1080
tactacccat ttatatatta tacttatcca cttatttaat gtctttataa ggtttgatcc 1140
atgatatttc taatatttta gttgatatgt atatgaaagg gtactatttg aactctctta 1200
ctctgtataa aggttggatc atccttaaag tgggtctatt taattttatt gcttcttaca 1260
gataaaaaaa aaattatgag ttggtttgat aaaatattga aggatttaaa ataataataa 1320
ataacatata atatatgtat ataaatttat tataatataa catttatcta taaaaaagta 1380
aatattgtca taaatctata caatcgttta gccttgctgg acgaatctca attatttaaa 1440
cgagagtaaa catatttgac tttttggtta tttaacaaat tattatttaa cactatatga 1500
aatttttttt tttatcagca aagaataaaa ttaaattaag aaggacaatg gtgtcccaat 1560
ccttatacaa ccaacttcca caagaaagtc aagtcagaga caacaaaaaa acaagcaaag 1620
gaaatttttt aatttgagtt gtcttgtttg ctgcataatt tatgcagtaa aacactacac 1680
ataacccttt tagcagtaga gcaatggttg accgtgtgct tagcttcttt tattttattt 1740
ttttatcagc aaagaataaa taaaataaaa tgagacactt cagggatgtt tcaacaagct 1800
tggatctcct gcaggatctg gccggccgga tctcgtacgg atccgtcgac ggcgcgcctc 1860
gagtgggcgg atcccccggg ctgcaggaat tcactggccg tcgttttaca acgtcgtgac 1920
tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc 1980
tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat 2040
ggcgaatgga tcgatccatc gcgatgtacc ttttgttagt cagcctctcg attgctcatc 2100
gtcattacac agtaccgaag tttgatcgat ctagtaacat agatgacacc gcgcgcgata 2160
atttatccta gtttgcgcgc tatattttgt tttctatcgc gtattaaatg tataattgcg 2220
ggactctaat cataaaaacc catctcataa ataacgtcat gcattacatg ttaattatta 2280
catgcttaac gtaattcaac agaaattata tgataatcat cgcaagaccg gcaacaggat 2340
tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc tgcttcgacg cactccttct 2400
ttactccacc atctcgtcct tattgaaaac gtgggtagca ccaaaacgaa tcaagtcgct 2460
ggaactgaag ttaccaatca cgctggatga tttgccagtt ggattaatct tgcctttccc 2520
cgcatgaata atattgatga atgcatgcgt gaggggtagt tcgatgttgg caatagctgc 2580
aattgccgcg acatcctcca acgagcataa ttcttcagaa aaatagcgat gttccatgtt 2640
gtcagggcat gcatgatgca cgttatgagg tgacggtgct aggcagtatt ccctcaaagt 2700
ttcatagtca gtatcatatt catcattgca ttcctgcaag agagaattga gacgcaatcc 2760
acacgctgcg gcaaccttcc ggcgttcgtg gtctatttgc tcttggacgt tgcaaacgta 2820
agtgttggat cgatccgggg tgggcgaaga actccagcat gagatccccg cgctggagga 2880
tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg 2940
tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca 3000
gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg 3060
agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc 3120
aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca 3180
gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc 3240
atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc 3300
ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc 3360
catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc 3420
cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg 3480
agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct 3540
tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca 3600
cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac 3660
aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat 3720
tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc 3780
gtgcaatcca tcttgttcaa tcatgcgaaa cgatccccgc aagcttggag actggtgatt 3840
tcagcgtgtc ctctccaaat gaaatgaact tccttatata gaggaagggt cttgcgaagg 3900
atagtgggat tgtgcgtcat cccttacgtc agtggagata tcacatcaat ccacttgctt 3960
tgaagacgtg gttggaacgt cttctttttc cacgatgctc ctcgtgggtg ggggtccatc 4020
tttgggacca ctgtcggcag aggcatcttc aacgatggcc tttcctttat cgcaatgatg 4080
gcatttgtag gagccacctt ccttttccac tatcttcaca ataaagtgac agatagctgg 4140
gcaatggaat ccgaggaggt ttccggatat taccctttgt tgaaaagtct caattgccct 4200
ttggtcttct gagactgtat ctttgatatt tttggagtag acaagcgtgt cgtgctccac 4260
catgttgacg aagattttct tcttgtcatt gagtcgtaag agactctgta tgaactgttc 4320
gccagtcttt acggcgagtt ctgttaggtc ctctatttga atctttgact ccatggcctt 4380
tgattcagtg ggaactacct ttttagagac tccaatctct attacttgcc ttggtttgtg 4440
aagcaagcct tgaatcgtcc atactggaat agtacttctg atcttgagaa atatatcttt 4500
ctctgtgttc ttgatgcagt tagtcctgaa tcttttgact gcatctttaa ccttcttggg 4560
aaggtatttg atctcctgga gattattgct cgggtagatc gtcttgatga gacctgctgc 4620
gtaagcctct ctaaccatct gtgggttagc attctttctg aaattgaaaa ggctaatctt 4680
ctcattatca gtggtgaaca tggtatcgtc accttctccg tcgaacttcc tgactagatc 4740
gtagagatag aggaagtcgt ccattgtgat ctctggggca aaggagatct gaattaattc 4800
gatatggtgg atttatcaca aatgggaccc gccgccgaca gaggtgtgat gttaggccag 4860
gactttgaaa atttgcgcaa ctatcgtata gtggccgaca aattgacgcc gagttgacag 4920
actgcctagc atttgagtga attatgtgag gtaatgggct acactgaatt ggtagctcaa 4980
actgtcagta tttatgtata tgagtgtata ttttcgcata atctcagacc aatctgaaga 5040
tgaaatgggt atctgggaat ggcgaaatca aggcatcgat cgtgaagttt ctcatctaag 5100
cccccatttg gacgtgaatg tagacacgtc gaaataaaga tttccgaatt agaataattt 5160
gtttattgct ttcgcctata aatacgacgg atcgtaattt gtcgttttat caaaatgtac 5220
tttcatttta taataacgct gcggacatct acatttttga attgaaaaaa aattggtaat 5280
tactctttct ttttctccat attgaccatc atactcattg ctgatccatg tagatttccc 5340
ggacatgaag ccatttacaa ttgaatatat cctgccgccg ctgccgcttt gcacccggtg 5400
gagcttgcat gttggtttct acgcagaact gagccggtta ggcagataat ttccattgag 5460
aactgagcca tgtgcacctt ccccccaaca cggtgagcga cggggcaacg gagtgatcca 5520
catgggactt ttaaacatca tccgtcggat ggcgttgcga gagaagcagt cgatccgtga 5580
gatcagccga cgcaccgggc aggcgcgcaa cacgatcgca aagtatttga acgcaggtac 5640
aatcgagccg acgttcacgc ggaacgacca agcaagctag ctttaatgcg gtagtttatc 5700
acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt 5760
catcctcggc accgtcaccc tggatgctgt aggcataggc ttggttatgc cggtactgcc 5820
gggcctcttg cgggatatcg tccattccga cagcatcgcc agtcactatg gcgtgctgct 5880
agcgctatat gcgttgatgc aatttctatg cgcacccgtt ctcggagcac tgtccgaccg 5940
ctttggccgc cgcccagtcc tgctcgcttc gctacttgga gccactatcg actacgcgat 6000
catggcgacc acacccgtcc tgtggtccaa cccctccgct gctatagtgc agtcggcttc 6060
tgacgttcag tgcagccgtc ttctgaaaac gacatgtcgc acaagtccta agttacgcga 6120
caggctgccg ccctgccctt ttcctggcgt tttcttgtcg cgtgttttag tcgcataaag 6180
tagaatactt gcgactagaa ccggagacat tacgccatga acaagagcgc cgccgctggc 6240
ctgctgggct atgcccgcgt cagcaccgac gaccaggact tgaccaacca acgggccgaa 6300
ctgcacgcgg ccggctgcac caagctgttt tccgagaaga tcaccggcac caggcgcgac 6360
cgcccggagc tggccaggat gcttgaccac ctacgccctg gcgacgttgt gacagtgacc 6420
aggctagacc gcctggcccg cagcacccgc gacctactgg acattgccga gcgcatccag 6480
gaggccggcg cgggcctgcg tagcctggca gagccgtggg ccgacaccac cacgccggcc 6540
ggccgcatgg tgttgaccgt gttcgccggc attgccgagt tcgagcgttc cctaatcatc 6600
gaccgcaccc ggagcgggcg cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc 6660
cctaccctca ccccggcaca gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc 6720
accgtgaaag aggcggctgc actgcttggc gtgcatcgct cgaccctgta ccgcgcactt 6780
gagcgcagcg aggaagtgac gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac 6840
gcattgaccg aggccgacgc cctggcggcc gccgagaatg aacgccaaga ggaacaagca 6900
tgaaaccgca ccaggacggc caggacgaac cgtttttcat taccgaagag atcgaggcgg 6960
agatgatcgc ggccgggtac gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc 7020
atgaaatcct ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg 7080
ctgaagaaac cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc 7140
gtcatgcggt cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aagggaacgc 7200
atgaagttat cgctgtactt aaccagaaag gcgggtcagg caagacgacc atcgcaaccc 7260
atctagcccg cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat tccgatcccc 7320
agggcagtgc ccgcgattgg gcggccgtgc gggaagatca accgctaacc gttgtcggca 7380
tcgaccgccc gacgattgac cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga 7440
tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg 7500
tgctgattcc ggtgcagcca agcccttacg acatatgggc caccgccgac ctggtggagc 7560
tggttaagca gcgcattgag gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc 7620
gggcgatcaa aggcacgcgc atcggcggtg aggttgccga ggcgctggcc gggtacgagc 7680
tgcccattct tgagtcccgt atcacgcagc gcgtgagcta cccaggcact gccgccgccg 7740
gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg 7800
ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag 7860
cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg caacgttggc 7920
cagcctggca gacacgccag ccatgaagcg ggtcaacttt cagttgccgg cggaggatca 7980
caccaagctg aagatgtacg cggtacgcca aggcaagacc attaccgagc tgctatctga 8040
atacatcgcg cagctaccag agtaaatgag caaatgaata aatgagtaga tgaattttag 8100
cggctaaagg aggcggcatg gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc 8160
ccatgtgtgg aggaacgggc ggttggccag gcgtaagcgg ctgggttgtc tgccggccct 8220
gcaatggcac tggaaccccc aagcccgagg aatcggcgtg agcggtcgca aaccatccgg 8280
cccggtacaa atcggcgcgg cgctgggtga tgacctggtg gagaagttga aggccgcgca 8340
ggccgcccag cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc 8400
cgctgatcga atccgcaaag aatcccggca accgccggca gccggtgcgc cgtcgattag 8460
gaagccgccc aagggcgacg agcaaccaga ttttttcgtt ccgatgctct atgacgtggg 8520
cacccgcgat agtcgcagca tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg 8580
acgagctggc gaggtgatcc gctacgagct tccagacggg cacgtagagg tttccgcagg 8640
gccggccggc atggccagtg tgtgggatta cgacctggta ctgatggcgg tttcccatct 8700
aaccgaatcc atgaaccgat accgggaagg gaagggagac aagcccggcc gcgtgttccg 8760
tccacacgtt gcggacgtac tcaagttctg ccggcgagcc gatggcggaa agcagaaaga 8820
cgacctggta gaaacctgca ttcggttaaa caccacgcac gttgccatgc agcgtacgaa 8880
gaaggccaag aacggccgcc tggtgacggt atccgagggt gaagccttga ttagccgcta 8940
caagatcgta aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc tagctgattg 9000
gatgtaccgc gagatcacag aaggcaagaa cccggacgtg ctgacggttc accccgatta 9060
ctttttgatc gatcccggca tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg 9120
caaggcagaa gccagatggt tgttcaagac gatctacgaa cgcagtggca gcgccggaga 9180
gttcaagaag ttctgtttca ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta 9240
cgatttgaag gaggaggcgg ggcaggctgg cccgatccta gtcatgcgct accgcaacct 9300
gatcgagggc gaagcatccg ccggttccta atgtacggag cagatgctag ggcaaattgc 9360
cctagcaggg gaaaaaggtc gaaaaggtct ctttcctgtg gatagcacgt acattgggaa 9420
cccaaagccg tacattggga accggaaccc gtacattggg aacccaaagc cgtacattgg 9480
gaaccggtca cacatgtaag tgactgatat aaaagagaaa aaaggcgatt tttccgccta 9540
aaactcttta aaacttatta aaactcttaa aacccgcctg gcctgtgcat aactgtctgg 9600
ccagcgcaca gccgaagagc tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg 9660
ccccgccgct tcgcgtcggc ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg 9720
gccaggcaat ctaccagggc gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc 9780
acatcaaggc accctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 9840
agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 9900
agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg 9960
atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca 10020
ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc 10080
ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 10140
agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 10200
catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 10260
tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 10320
gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 10380
ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 10440
cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 10500
caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 10560
ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 10620
taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 10680
taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 10740
cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 10800
tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 10860
gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 10920
catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 10980
atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 11040
ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 11100
gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 11160
agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 11220
gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 11280
agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 11340
catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 11400
aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 11460
gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 11520
taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 11580
caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg 11640
ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aagacctgca 11700
gggggggggg ggcgctgagg tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc 11760
tgaatcgccc catcatccag ccagaaagtg agggagccac ggttgatgag agctttgttg 11820
taggtggacc agttggtgat tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg 11880
ggaagatgcg tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccgc 11940
cgtcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt 12000
agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac 12060
catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata 12120
ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta 12180
ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg 12240
aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc 12300
cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg 12360
cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat 12420
gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt 12480
cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat 12540
caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta 12600
gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca 12660
actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat 12720
tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc 12780
tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt 12840
aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga 12900
gattttgaga cacaacgtgg ctttcccccc cccccctgca ggtcaattcg gtcgatatgg 12960
ctattacgaa gaaggctcgt gcgcggagtc ccgtgaactt tcccacgcaa caagtgaacc 13020
gcaccgggtt tgccggaggc catttcgtta aaatgcgcag ccatggctgc ttcgtccagc 13080
atggcgtaat actgatcctc gtcttcggct ggcggtatat tgccgatggg cttcaaaagc 13140
cgccgtggtt gaaccagtct atccattcca aggtagcgaa ctcgaccgct tcgaagctcc 13200
tccatggtcc acgccgatga atgacctcgg ccttgtaaag accgttgatc gcttctgcga 13260
gggcgttgtc gtgctgtcgc cgacgcttcc gatagatggc tcgatacctg cttctgccaa 13320
ccgctcggaa tagcgaaagg acacgtattg aacaccgcga tccgagtgat gcactaggcc 13380
gccatgagcg ggacgccgat catgatgagc ctcctcgagg gcatcgagga caaagcctgc 13440
atgtgctgtc cggctcgccc gccatccgac aatgcgacgg gcgaagacgt cgatcacgaa 13500
ggccacgtag acgaagccct cccaagtggc gacataagta cggacatgcg caaaggcttt 13560
cccggtttgt cgctgatggt gcaagagacg ctgaagcgcg atccgatgcg caggcatctg 13620
ttcgtcttcc gcggtcgtgg cggtggcctg atcaaggtca ctcgccgaag agctgcatga 13680
ttggctcgaa accgagcggg ggaaattgtc gcgcagttct cccgtcgccg aggcgataaa 13740
ttacatgctc aagcgatggg atggcattac gtcattcctc gatgacggcc cgatttgcct 13800
gacgaacaat gctgccgaac gaacgctcag aggctatgta ctcggcagga agtcatggct 13860
gtttgccgga tcggatcgtt gtgctgaacg tgcggcgttc atggcgacac tgatcatgag 13920
cgccaagctc aataacatcg atccgcaggc ctggcttgcc gacgtccgcg ccgaccttgc 13980
ggacgctccg atcagcaggc ttgagcaaca gctgccgtgg aactggacat ccaagacact 14040
gagtgctcag gcggcctgac ctgcggcctt caccggatac ttaccccatt atcgcagatt 14100
gcgatgaagc atcagcgtca ttcagcaatc ttgccaaagt atgcaggctc gcgagaatcg 14160
acgtgcgaaa ccggctggtt gcgccaaaga tccgcttgcg gagcggtcga acattcatgc 14220
tgggacttca agaggtcgag tagaggaaga accggaaagg ttgcaccgga aaatatgcgt 14280
tcctttggag agcgcctcat ggacgtgaac aaatcgcccg gaccaaggat gccacggata 14340
caaaagctcg cgaagctcgg tcccgtgggt gttctgtcgt ctcgttgtac aacgaaatcc 14400
attcccattc cgcgctcaag atggcttccc ctcggcagtt catcagggct aaatcaatct 14460
agccgacttg tccggtgaaa tgggctgcac tccaacagaa acaatcaaac aaacatacac 14520
agcgacttat tcacacgagc tcaaattaca acggtatata tcctgccagt cagcatcatc 14580
acaccaaaag ttaggcccga atagtttgaa attagaaagc tcgcaattga ggtctacagg 14640
ccaaattcgc tcttagccgt acaatattac tcaccggtgc gatgcccccc atcgtaggtg 14700
aaggtggaaa ttaatgatcc atcttgagac cacaggccca caacagctac cagtttcctc 14760
aagggtccac caaaaacgta agcgcttacg tacatggtcg ataagaaaag gcaatttgta 14820
gatgttaaca tccaacgtcg ctttcaggga tcgatccaat acgcaaaccg cctctccccg 14880
cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca 14940
gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact 15000
ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa 15060
acagctatga ccatgattac gccaagcttg catgcctgca ggtcgactct agag 15114
<210>32
<211>13268
<212>DNA
<213>人工序列
<220>
<223>质粒pKR92
<400>32
cgcgcctcga gtgggcggat cccccgggct gcaggaattc actggccgtc gttttacaac 60
gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt 120
tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca 180
gcctgaatgg cgaatggatc gatccatcgc gatgtacctt ttgttagtca gcctctcgat 240
tgctcatcgt cattacacag taccgaagtt tgatcgatct agtaacatag atgacaccgc 300
gcgcgataat ttatcctagt ttgcgcgcta tattttgttt tctatcgcgt attaaatgta 360
taattgcggg actctaatca taaaaaccca tctcataaat aacgtcatgc attacatgtt 420
aattattaca tgcttaacgt aattcaacag aaattatatg ataatcatcg caagaccggc 480
aacaggattc aatcttaaga aactttattg ccaaatgttt gaacgatctg cttcgacgca 540
ctccttcttt actccaccat ctcgtcctta ttgaaaacgt gggtagcacc aaaacgaatc 600
aagtcgctgg aactgaagtt accaatcacg ctggatgatt tgccagttgg attaatcttg 660
cctttccccg catgaataat attgatgaat gcatgcgtga ggggtagttc gatgttggca 720
atagctgcaa ttgccgcgac atcctccaac gagcataatt cttcagaaaa atagcgatgt 780
tccatgttgt cagggcatgc atgatgcacg ttatgaggtg acggtgctag gcagtattcc 840
ctcaaagttt catagtcagt atcatattca tcattgcatt cctgcaagag agaattgaga 900
cgcaatccac acgctgcggc aaccttccgg cgttcgtggt ctatttgctc ttggacgttg 960
caaacgtaag tgttggatcg atccggggtg ggcgaagaac tccagcatga gatccccgcg 1020
ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa 1080
ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc 1140
gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 1200
gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 1260
tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc 1320
cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 1380
gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg 1440
aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 1500
ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 1560
caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 1620
tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 1680
cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 1740
gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg 1800
gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 1860
cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 1920
gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atccccgcaa gcttggagac 1980
tggtgatttc agcgtgtcct ctccaaatga aatgaacttc cttatataga ggaagggtct 2040
tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatatc acatcaatcc 2100
acttgctttg aagacgtggt tggaacgtct tctttttcca cgatgctcct cgtgggtggg 2160
ggtccatctt tgggaccact gtcggcagag gcatcttcaa cgatggcctt tcctttatcg 2220
caatgatggc atttgtagga gccaccttcc ttttccacta tcttcacaat aaagtgacag 2280
atagctgggc aatggaatcc gaggaggttt ccggatatta ccctttgttg aaaagtctca 2340
attgcccttt ggtcttctga gactgtatct ttgatatttt tggagtagac aagcgtgtcg 2400
tgctccacca tgttgacgaa gattttcttc ttgtcattga gtcgtaagag actctgtatg 2460
aactgttcgc cagtctttac ggcgagttct gttaggtcct ctatttgaat ctttgactcc 2520
atggcctttg attcagtggg aactaccttt ttagagactc caatctctat tacttgcctt 2580
ggtttgtgaa gcaagccttg aatcgtccat actggaatag tacttctgat cttgagaaat 2640
atatctttct ctgtgttctt gatgcagtta gtcctgaatc ttttgactgc atctttaacc 2700
ttcttgggaa ggtatttgat ctcctggaga ttattgctcg ggtagatcgt cttgatgaga 2760
cctgctgcgt aagcctctct aaccatctgt gggttagcat tctttctgaa attgaaaagg 2820
ctaatcttct cattatcagt ggtgaacatg gtatcgtcac cttctccgtc gaacttcctg 2880
actagatcgt agagatagag gaagtcgtcc attgtgatct ctggggcaaa ggagtctgaa 2940
ttaattcgat atggtggatt tatcacaaat gggacccgcc gccgacagag gtgtgatgtt 3000
aggccaggac tttgaaaatt tgcgcaacta tcgtatagtg gccgacaaat tgacgccgag 3060
ttgacagact gcctagcatt tgagtgaatt atgtgaggta atgggctaca ctgaattggt 3120
agctcaaact gtcagtattt atgtatatga gtgtatattt tcgcataatc tcagaccaat 3180
ctgaagatga aatgggtatc tgggaatggc gaaatcaagg catcgatcgt gaagtttctc 3240
atctaagccc ccatttggac gtgaatgtag acacgtcgaa ataaagattt ccgaattaga 3300
ataatttgtt tattgctttc gcctataaat acgacggatc gtaatttgtc gttttatcaa 3360
aatgtacttt cattttataa taacgctgcg gacatctaca tttttgaatt gaaaaaaaat 3420
tggtaattac tctttctttt tctccatatt gaccatcata ctcattgctg atccatgtag 3480
atttcccgga catgaagcca tttacaattg aatatatcct gccgccgctg ccgctttgca 3540
cccggtggag cttgcatgtt ggtttctacg cagaactgag ccggttaggc agataatttc 3600
cattgagaac tgagccatgt gcaccttccc cccaacacgg tgagcgacgg ggcaacggag 3660
tgatccacat gggactttta aacatcatcc gtcggatggc gttgcgagag aagcagtcga 3720
tccgtgagat cagccgacgc accgggcagg cgcgcaacac gatcgcaaag tatttgaacg 3780
caggtacaat cgagccgacg ttcacgcgga acgaccaagc aagctagctt taatgcggta 3840
gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc 3900
tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg gttatgccgg 3960
tactgccggg cctcttgcgg gatatcgtcc attccgacag catcgccagt cactatggcg 4020
tgctgctagc gctatatgcg ttgatgcaat ttctatgcgc acccgttctc ggagcactgt 4080
ccgaccgctt tggccgccgc ccagtcctgc tcgcttcgct acttggagcc actatcgact 4140
acgcgatcat ggcgaccaca cccgtcctgt ggtccaaccc ctccgctgct atagtgcagt 4200
cggcttctga cgttcagtgc agccgtcttc tgaaaacgac atgtcgcaca agtcctaagt 4260
tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt gttttagtcg 4320
cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca agagcgccgc 4380
cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga ccaaccaacg 4440
ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca ccggcaccag 4500
gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg acgttgtgac 4560
agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca ttgccgagcg 4620
catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg acaccaccac 4680
gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg agcgttccct 4740
aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg tgaagtttgg 4800
cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga tcgaccagga 4860
aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga ccctgtaccg 4920
cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg gtgccttccg 4980
tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac gccaagagga 5040
acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac cgaagagatc 5100
gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacgt ctcaaccgtg 5160
cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg gccggccagc 5220
ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac 5280
agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca aatacgcaag 5340
ggaacgcatg aagttatcgc tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc 5400
gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt agtcgattcc 5460
gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt 5520
gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc 5580
gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat caaggcagcc 5640
gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac cgccgacctg 5700
gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc ggcctttgtc 5760
gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg 5820
tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc 5880
gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg cgaggtccag 5940
gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa gagaaaatga 6000
gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa 6060
cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag ttgccggcgg 6120
aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt accgagctgc 6180
tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat gagtagatga 6240
attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac cgacgccgtg 6300
gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg ggttgtctgc 6360
cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac 6420
catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag aagttgaagg 6480
ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt gaatcgtggc 6540
aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt 6600
cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg atgctctatg 6660
acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc 6720
gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac gtagaggttt 6780
ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg atggcggttt 6840
cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag cccggccgcg 6900
tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat ggcggaaagc 6960
agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt gccatgcagc 7020
gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa gccttgatta 7080
gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag atcgagctag 7140
ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg acggttcacc 7200
ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg gcacgccgcg 7260
ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc agtggcagcg 7320
ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc 7380
cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc atgcgctacc 7440
gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag atgctagggc 7500
aaattgccct agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat agcacgtaca 7560
ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac ccaaagccgt 7620
acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa ggcgattttt 7680
ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac 7740
tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct 7800
ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca aaaatggctg 7860
gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc 7920
ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga 7980
cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa 8040
gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca 8100
cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga 8160
gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca 8220
ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 8280
cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 8340
gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 8400
tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 8460
agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 8520
tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 8580
cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 8640
ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 8700
ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 8760
ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 8820
ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 8880
cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 8940
gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 9000
atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 9060
ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 9120
gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 9180
tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 9240
ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 9300
taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 9360
gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 9420
gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 9480
ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 9540
aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 9600
gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 9660
cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 9720
actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 9780
caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaag 9840
acctgcaggg gggggggggc gctgaggtct gcctcgtgaa gaaggtgttg ctgactcata 9900
ccaggcctga atcgccccat catccagcca gaaagtgagg gagccacggt tgatgagagc 9960
tttgttgtag gtggaccagt tggtgatttt gaacttttgc tttgccacgg aacggtctgc 10020
gttgtcggga agatgcgtga tctgatcctt caactcagca aaagttcgat ttattcaaca 10080
aagccgccgt cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat 10140
tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta 10200
tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag 10260
ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata 10320
caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg 10380
acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca 10440
ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt 10500
gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga 10560
atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca 10620
ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat 10680
gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc 10740
cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc 10800
agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc 10860
ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat 10920
cgcggcctcg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg 10980
tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa 11040
catcagagat tttgagacac aacgtggctt tccccccccc ccctgcaggt caattcggtc 11100
gatatggcta ttacgaagaa ggctcgtgcg cggagtcccg tgaactttcc cacgcaacaa 11160
gtgaaccgca ccgggtttgc cggaggccat ttcgttaaaa tgcgcagcca tggctgcttc 11220
gtccagcatg gcgtaatact gatcctcgtc ttcggctggc ggtatattgc cgatgggctt 11280
caaaagccgc cgtggttgaa ccagtctatc cattccaagg tagcgaactc gaccgcttcg 11340
aagctcctcc atggtccacg ccgatgaatg acctcggcct tgtaaagacc gttgatcgct 11400
tctgcgaggg cgttgtcgtg ctgtcgccga cgcttccgat agatggctcg atacctgctt 11460
ctgccaaccg ctcggaatag cgaaaggaca cgtattgaac accgcgatcc gagtgatgca 11520
ctaggccgcc atgagcggga cgccgatcat gatgagcctc ctcgagggca tcgaggacaa 11580
agcctgcatg tgctgtccgg ctcgcccgcc atccgacaat gcgacgggcg aagacgtcga 11640
tcacgaaggc cacgtagacg aagccctccc aagtggcgac ataagtacgg acatgcgcaa 11700
aggctttccc ggtttgtcgc tgatggtgca agagacgctg aagcgcgatc cgatgcgcag 11760
gcatctgttc gtcttccgcg gtcgtggcgg tggcctgatc aaggtcactc gccgaagagc 11820
tgcatgattg gctcgaaacc gagcggggga aattgtcgcg cagttctccc gtcgccgagg 11880
cgataaatta catgctcaag cgatgggatg gcattacgtc attcctcgat gacggcccga 11940
tttgcctgac gaacaatgct gccgaacgaa cgctcagagg ctatgtactc ggcaggaagt 12000
catggctgtt tgccggatcg gatcgttgtg ctgaacgtgc ggcgttcatg gcgacactga 12060
tcatgagcgc caagctcaat aacatcgatc cgcaggcctg gcttgccgac gtccgcgccg 12120
accttgcgga cgctccgatc agcaggcttg agcaacagct gccgtggaac tggacatcca 12180
agacactgag tgctcaggcg gcctgacctg cggccttcac cggatactta ccccattatc 12240
gcagattgcg atgaagcatc agcgtcattc agcaatcttg ccaaagtatg caggctcgcg 12300
agaatcgacg tgcgaaaccg gctggttgcg ccaaagatcc gcttgcggag cggtcgaaca 12360
ttcatgctgg gacttcaaga ggtcgagtag aggaagaacc ggaaaggttg caccggaaaa 12420
tatgcgttcc tttggagagc gcctcatgga cgtgaacaaa tcgcccggac caaggatgcc 12480
acggatacaa aagctcgcga agctcggtcc cgtgggtgtt ctgtcgtctc gttgtacaac 12540
gaaatccatt cccattccgc gctcaagatg gcttcccctc ggcagttcat cagggctaaa 12600
tcaatctagc cgacttgtcc ggtgaaatgg gctgcactcc aacagaaaca atcaaacaaa 12660
catacacagc gacttattca cacgagctca aattacaacg gtatatatcc tgccagtcag 12720
catcatcaca ccaaaagtta ggcccgaata gtttgaaatt agaaagctcg caattgaggt 12780
ctacaggcca aattcgctct tagccgtaca atattactca ccggtgcgat gccccccatc 12840
gtaggtgaag gtggaaatta atgatccatc ttgagaccac aggcccacaa cagctaccag 12900
tttcctcaag ggtccaccaa aaacgtaagc gcttacgtac atggtcgata agaaaaggca 12960
atttgtagat gttaacatcc aacgtcgctt tcagggatcg atccaatacg caaaccgcct 13020
ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa 13080
gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct 13140
ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac 13200
acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga 13260
ggatctgg 13268
<210>33
<211>19604
<212>DNA
<213>人工序列
<220>
<223>质粒pKR92 YLDGAT2
<220>
<221>misc_feature
<222>(4526)..(4526)
<223>n为a,c,g,或t
<400>33
cgcgccaagc ttttgatcca tgcccttcat ttgccgctta ttaattaatt tggtaacagt 60
ccgtactaat cagttactta tccttccccc atcataatta atcttggtag tctcgaatgc 120
cacaacactg actagtctct tggatcataa gaaaaagcca aggaacaaaa gaagacaaaa 180
cacaatgaga gtatcctttg catagcaatg tctaagttca taaaattcaa acaaaaacgc 240
aatcacacac agtggacatc acttatccac tagctgatca ggatcgccgc gtcaagaaaa 300
aaaaactgga ccccaaaagc catgcacaac aacacgtact cacaaaggtg tcaatcgagc 360
agcccaaaac attcaccaac tcaacccatc atgagccctc acatttgttg tttctaaccc 420
aacctcaaac tcgtattctc ttccgccacc tcatttttgt ttatttcaac acccgtcaaa 480
ctgcatgcca ccccgtggcc aaatgtccat gcatgttaac aagacctatg actataaata 540
gctgcaatct cggcccaggt tttcatcatc aagaaccagt tcaatatcct agtacaccgt 600
attaaagaat ttaagatata ctgcggccgc atgactatcg actcacaata ctacaagtcg 660
cgagacaaaa acgacacggc acccaaaatc gcgggaatcc gatatgcccc gctatcgaca 720
ccattactca accgatgtga gaccttctct ctggtctggc acattttcag cattcccact 780
ttcctcacaa ttttcatgct atgctgcgca attccactgc tctggccatt tgtgattgcg 840
tatgtagtgt acgctgttaa agacgactcc ccgtccaacg gaggagtggt caagcgatac 900
tcgcctattt caagaaactt cttcatctgg aagctctttg gccgctactt ccccataact 960
ctgcacaaga cggtggatct ggagcccacg cacacatact accctctgga cgtccaggag 1020
tatcacctga ttgctgagag atactggccg cagaacaagt acctccgagc aatcatctcc 1080
accatcgagt actttctgcc cgccttcatg aaacggtctc tttctatcaa cgagcaggag 1140
cagcctgccg agcgagatcc tctcctgtct cccgtttctc ccagctctcc gggttctcaa 1200
cctgacaagt ggattaacca cgacagcaga tatagccgtg gagaatcatc tggctccaac 1260
ggccacgcct cgggctccga acttaacggc aacggcaaca atggcaccac taaccgacga 1320
cctttgtcgt ccgcctctgc tggctccact gcatctgatt ccacgcttct taacgggtcc 1380
ctcaactcct acgccaacca gatcattggc gaaaacgacc cacagctgtc gcccacaaaa 1440
ctcaagccca ctggcagaaa atacatcttc ggctaccacc cccacggcat tatcggcatg 1500
ggagcctttg gtggaattgc caccgaggga gctggatggt ccaagctctt tccgggcatc 1560
cctgtttctc ttatgactct caccaacaac ttccgagtgc ctctctacag agagtacctc 1620
atgagtctgg gagtcgcttc tgtctccaag aagtcctgca aggccctcct caagcgaaac 1680
cagtctatct gcattgtcgt tggtggagca caggaaagtc ttctggccag acccggtgtc 1740
atggacctgg tgctactcaa gcgaaagggt tttgttcgac ttggtatgga ggtcggaaat 1800
gtcgcccttg ttcccatcat ggcctttggt gagaacgacc tctatgacca ggttagcaac 1860
gacaagtcgt ccaagctgta ccgattccag cagtttgtca agaacttcct tggattcacc 1920
cttcctttga tgcatgcccg aggcgtcttc aactacgatg tcggtcttgt cccctacagg 1980
cgacccgtca acattgtggt tggttccccc attgacttgc cttatctccc acaccccacc 2040
gacgaagaag tgtccgaata ccacgaccga tacatcgccg agctgcagcg aatctacaac 2100
gagcacaagg atgaatattt catcgattgg accgaggagg gcaaaggagc cccagagttc 2160
cgaatgattg agtaagcggc cgcaagtatg aactaaaatg catgtaggtg taagagctca 2220
tggagagcat ggaatattgt atccgaccat gtaacagtat aataactgag ctccatctca 2280
cttcttctat gaataaacaa aggatgttat gatatattaa cactctatct atgcacctta 2340
ttgttctatg ataaatttcc tcttattatt ataaatcatc tgaatcgtga cggcttatgg 2400
aatgcttcaa atagtacaaa aacaaatgtg tactataaga ctttctaaac aattctaacc 2460
ttagcattgt gaacgagaca taagtgttaa gaagacataa caattataat ggaagaagtt 2520
tgtctccatt tatatattat atattaccca cttatgtatt atattaggat gttaaggaga 2580
cataacaatt ataaagagag aagtttgtat ccatttatat attatatact acccatttat 2640
atattatact tatccactta tttaatgtct ttataaggtt tgatccatga tatttctaat 2700
attttagttg atatgtatat gaaagggtac tatttgaact ctcttactct gtataaaggt 2760
tggatcatcc ttaaagtggg tctatttaat tttattgctt cttacagata aaaaaaaaat 2820
tatgagttgg tttgataaaa tattgaagga tttaaaataa taataaataa catataatat 2880
atgtatataa atttattata atataacatt tatctataaa aaagtaaata ttgtcataaa 2940
tctatacaat cgtttagcct tgctggacga atctcaatta tttaaacgag agtaaacata 3000
tttgactttt tggttattta acaaattatt atttaacact atatgaaatt ttttttttta 3060
tcagcaaaga ataaaattaa attaagaagg acaatggtgt cccaatcctt atacaaccaa 3120
cttccacaag aaagtcaagt cagagacaac aaaaaaacaa gcaaaggaaa ttttttaatt 3180
tgagttgtct tgtttgctgc ataatttatg cagtaaaaca ctacacataa cccttttagc 3240
agtagagcaa tggttgaccg tgtgcttagc ttcttttatt ttattttttt atcagcaaag 3300
aataaataaa ataaaatgag acacttcagg gatgtttcaa caagcttgga tcctcgaaga 3360
gaagggttaa taacacactt ttttaacatt tttaacacaa attttagtta tttaaaaatt 3420
tattaaaaaa tttaaaataa gaagaggaac tctttaaata aatctaactt acaaaattta 3480
tgatttttaa taagttttca ccaataaaaa atgtcataaa aatatgttaa aaagtatatt 3540
atcaatattc tctttatgat aaataaaaag aaaaaaaaaa taaaagttaa gtgaaaatga 3600
gattgaagtg actttaggtg tgtataaata tatcaacccc gccaacaatt tatttaatcc 3660
aaatatattg aagtatatta ttccatagcc tttatttatt tatatattta ttatataaaa 3720
gctttatttg ttctaggttg ttcatgaaat atttttttgg ttttatctcc gttgtaagaa 3780
aatcatgtgc tttgtgtcgc cactcactat tgcagctttt tcatgcattg gtcagattga 3840
cggttgattg tatttttgtt ttttatggtt ttgtgttatg acttaagtct tcatctcttt 3900
atctcttcat caggtttgat ggttacctaa tatggtccat gggtacatgc atggttaaat 3960
taggtggcca actttgttgt gaacgataga atttttttta tattaagtaa actattttta 4020
tattatgaaa taataataaa aaaaatattt tatcattatt aacaaaatca tattagttaa 4080
tttgttaact ctataataaa agaaatactg taacattcac attacatggt aacatctttc 4140
caccctttca tttgtttttt gtttgatgac tttttttctt gtttaaattt atttcccttc 4200
ttttaaattt ggaatacatt atcatcatat ataaactaaa atactaaaaa caggattaca 4260
caaatgataa ataataacac aaatatttat aaatctagct gcaatatatt taaactagct 4320
atatcgatat tgtaaaataa aactagctgc attgatactg ataaaaaaat atcatgtgct 4380
ttctggactg atgatgcagt atacttttga cattgccttt attttatttt tcagaaaagc 4440
tttcttagtt ctgggttctt cattatttgt ttcccatctc cattgtgaat tgaatcattt 4500
gcttcgtgtc acaaatacaa tttagntagg tacatgcatt ggtcagattc acggtttatt 4560
atgtcatgac ttaagttcat ggtagtacat tacctgccac gcatgcatta tattggttag 4620
atttgatagg caaatttggt tgtcaacaat ataaatataa ataatgtttt tatattacga 4680
aataacagtg atcaaaacaa acagttttat ctttattaac aagattttgt ttttgtttga 4740
tgacgttttt taatgtttac gctttccccc ttcttttgaa tttagaacac tttatcatca 4800
taaaatcaaa tactaaaaaa attacatatt tcataaataa taacacaaat atttttaaaa 4860
aatctgaaat aataatgaac aatattacat attatcacga aaattcatta ataaaaatat 4920
tatataaata aaatgtaata gtagttatat gtaggaaaaa agtactgcac gcataatata 4980
tacaaaaaga ttaaaatgaa ctattataaa taataacact aaattaatgg tgaatcatat 5040
caaaataatg aaaaagtaaa taaaatttgt aattaacttc tatatgtatt acacacacaa 5100
ataataaata atagtaaaaa aaattatgat aaatatttac catctcataa gatatttaaa 5160
ataatgataa aaatatagat tattttttat gcaactagct agccaaaaag agaacacggg 5220
tatatataaa aagagtacct ttaaattcta ctgtacttcc tttattcctg acgtttttat 5280
atcaagtgga catacgtgaa gattttaatt atcagtctaa atatttcatt agcacttaat 5340
acttttctgt tttattccta tcctataagt agtcccgatt ctcccaacat tgcttattca 5400
cacaactaac taagaaagtc ttccatagcc ccccaagcgg cccatggcct cctccgagga 5460
cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga 5520
gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct 5580
gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcca 5640
gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca agaagctgtc 5700
cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac 5760
cgtgacccag gactcctccc tgcaggacgg ctccttcatc tacaaggtga agttcatcgg 5820
cgtgaacttc ccctccgacg gccccgtaat gcagaagaag actatgggct gggaggcctc 5880
caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccaca aggccctgaa 5940
gctgaaggac ggcggccact acctggtgga gttcaagtcc atctacatgg ccaagaagcc 6000
cgtgcagctg cccggctact actacgtgga ctccaagctg gacatcacct cccacaacga 6060
ggactacacc atcgtggagc agtacgagcg cgccgagggc cgccaccacc tgttcctgta 6120
gcggccggcc gcgacacaag tgtgagagta ctaaataaat gctttggttg tacgaaatca 6180
ttacactaaa taaaataatc aaagcttata tatgccttcc gctaaggccg aatgcaaaga 6240
aattggttct ttctcgttat cttttgccac ttttactagt acgtattaat tactacttaa 6300
tcatctttgt ttacggctca ttatatccgt cgacggcgcg cctcgagtgg gcggatcccc 6360
cgggctgcag gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg 6420
ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag 6480
aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggatcgatc 6540
catcgcgatg taccttttgt tagtcagcct ctcgattgct catcgtcatt acacagtacc 6600
gaagtttgat cgatctagta acatagatga caccgcgcgc gataatttat cctagtttgc 6660
gcgctatatt ttgttttcta tcgcgtatta aatgtataat tgcgggactc taatcataaa 6720
aacccatctc ataaataacg tcatgcatta catgttaatt attacatgct taacgtaatt 6780
caacagaaat tatatgataa tcatcgcaag accggcaaca ggattcaatc ttaagaaact 6840
ttattgccaa atgtttgaac gatctgcttc gacgcactcc ttctttactc caccatctcg 6900
tccttattga aaacgtgggt agcaccaaaa cgaatcaagt cgctggaact gaagttacca 6960
atcacgctgg atgatttgcc agttggatta atcttgcctt tccccgcatg aataatattg 7020
atgaatgcat gcgtgagggg tagttcgatg ttggcaatag ctgcaattgc cgcgacatcc 7080
tccaacgagc ataattcttc agaaaaatag cgatgttcca tgttgtcagg gcatgcatga 7140
tgcacgttat gaggtgacgg tgctaggcag tattccctca aagtttcata gtcagtatca 7200
tattcatcat tgcattcctg caagagagaa ttgagacgca atccacacgc tgcggcaacc 7260
ttccggcgtt cgtggtctat ttgctcttgg acgttgcaaa cgtaagtgtt ggatcgatcc 7320
ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc 7380
ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg 7440
tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag 7500
aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa 7560
agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc 7620
aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa 7680
aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga 7740
tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc 7800
tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct 7860
cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc 7920
agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac 7980
aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca 8040
acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc 8100
tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc 8160
ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag 8220
tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt 8280
tcaatcatgc gaaacgatcc ccgcaagctt ggagactggt gatttcagcg tgtcctctcc 8340
aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aaggatagtg ggattgtgcg 8400
tcatccctta cgtcagtgga gatatcacat caatccactt gctttgaaga cgtggttgga 8460
acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg accactgtcg 8520
gcagaggcat cttcaacgat ggcctttcct ttatcgcaat gatggcattt gtaggagcca 8580
ccttcctttt ccactatctt cacaataaag tgacagatag ctgggcaatg gaatccgagg 8640
aggtttccgg atattaccct ttgttgaaaa gtctcaattg ccctttggtc ttctgagact 8700
gtatctttga tatttttgga gtagacaagc gtgtcgtgct ccaccatgtt gacgaagatt 8760
ttcttcttgt cattgagtcg taagagactc tgtatgaact gttcgccagt ctttacggcg 8820
agttctgtta ggtcctctat ttgaatcttt gactccatgg cctttgattc agtgggaact 8880
acctttttag agactccaat ctctattact tgccttggtt tgtgaagcaa gccttgaatc 8940
gtccatactg gaatagtact tctgatcttg agaaatatat ctttctctgt gttcttgatg 9000
cagttagtcc tgaatctttt gactgcatct ttaaccttct tgggaaggta tttgatctcc 9060
tggagattat tgctcgggta gatcgtcttg atgagacctg ctgcgtaagc ctctctaacc 9120
atctgtgggt tagcattctt tctgaaattg aaaaggctaa tcttctcatt atcagtggtg 9180
aacatggtat cgtcaccttc tccgtcgaac ttcctgacta gatcgtagag atagaggaag 9240
tcgtccattg tgatctctgg ggcaaaggag tctgaattaa ttcgatatgg tggatttatc 9300
acaaatggga cccgccgccg acagaggtgt gatgttaggc caggactttg aaaatttgcg 9360
caactatcgt atagtggccg acaaattgac gccgagttga cagactgcct agcatttgag 9420
tgaattatgt gaggtaatgg gctacactga attggtagct caaactgtca gtatttatgt 9480
atatgagtgt atattttcgc ataatctcag accaatctga agatgaaatg ggtatctggg 9540
aatggcgaaa tcaaggcatc gatcgtgaag tttctcatct aagcccccat ttggacgtga 9600
atgtagacac gtcgaaataa agatttccga attagaataa tttgtttatt gctttcgcct 9660
ataaatacga cggatcgtaa tttgtcgttt tatcaaaatg tactttcatt ttataataac 9720
gctgcggaca tctacatttt tgaattgaaa aaaaattggt aattactctt tctttttctc 9780
catattgacc atcatactca ttgctgatcc atgtagattt cccggacatg aagccattta 9840
caattgaata tatcctgccg ccgctgccgc tttgcacccg gtggagcttg catgttggtt 9900
tctacgcaga actgagccgg ttaggcagat aatttccatt gagaactgag ccatgtgcac 9960
cttcccccca acacggtgag cgacggggca acggagtgat ccacatggga cttttaaaca 10020
tcatccgtcg gatggcgttg cgagagaagc agtcgatccg tgagatcagc cgacgcaccg 10080
ggcaggcgcg caacacgatc gcaaagtatt tgaacgcagg tacaatcgag ccgacgttca 10140
cgcggaacga ccaagcaagc tagctttaat gcggtagttt atcacagtta aattgctaac 10200
gcagtcaggc accgtgtatg aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca 10260
ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc ttgcgggata 10320
tcgtccattc cgacagcatc gccagtcact atggcgtgct gctagcgcta tatgcgttga 10380
tgcaatttct atgcgcaccc gttctcggag cactgtccga ccgctttggc cgccgcccag 10440
tcctgctcgc ttcgctactt ggagccacta tcgactacgc gatcatggcg accacacccg 10500
tcctgtggtc caacccctcc gctgctatag tgcagtcggc ttctgacgtt cagtgcagcc 10560
gtcttctgaa aacgacatgt cgcacaagtc ctaagttacg cgacaggctg ccgccctgcc 10620
cttttcctgg cgttttcttg tcgcgtgttt tagtcgcata aagtagaata cttgcgacta 10680
gaaccggaga cattacgcca tgaacaagag cgccgccgct ggcctgctgg gctatgcccg 10740
cgtcagcacc gacgaccagg acttgaccaa ccaacgggcc gaactgcacg cggccggctg 10800
caccaagctg ttttccgaga agatcaccgg caccaggcgc gaccgcccgg agctggccag 10860
gatgcttgac cacctacgcc ctggcgacgt tgtgacagtg accaggctag accgcctggc 10920
ccgcagcacc cgcgacctac tggacattgc cgagcgcatc caggaggccg gcgcgggcct 10980
gcgtagcctg gcagagccgt gggccgacac caccacgccg gccggccgca tggtgttgac 11040
cgtgttcgcc ggcattgccg agttcgagcg ttccctaatc atcgaccgca cccggagcgg 11100
gcgcgaggcc gccaaggccc gaggcgtgaa gtttggcccc cgccctaccc tcaccccggc 11160
acagatcgcg cacgcccgcg agctgatcga ccaggaaggc cgcaccgtga aagaggcggc 11220
tgcactgctt ggcgtgcatc gctcgaccct gtaccgcgca cttgagcgca gcgaggaagt 11280
gacgcccacc gaggccaggc ggcgcggtgc cttccgtgag gacgcattga ccgaggccga 11340
cgccctggcg gccgccgaga atgaacgcca agaggaacaa gcatgaaacc gcaccaggac 11400
ggccaggacg aaccgttttt cattaccgaa gagatcgagg cggagatgat cgcggccggg 11460
tacgtgttcg agccgcccgc gcacgtctca accgtgcggc tgcatgaaat cctggccggt 11520
ttgtctgatg ccaagctggc ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc 11580
cgccgtctaa aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg 11640
tatatgatgc gatgagtaaa taaacaaata cgcaagggaa cgcatgaagt tatcgctgta 11700
cttaaccaga aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg 11760
caactcgccg gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat 11820
tgggcggccg tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt 11880
gaccgcgacg tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag 11940
gcggcggact tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag 12000
ccaagccctt acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt 12060
gaggtcacgg atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg 12120
cgcatcggcg gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc 12180
cgtatcacgc agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa 12240
tcagaacccg agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca 12300
aaactcattt gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg 12360
ccggccgtcc gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc 12420
cagccatgaa gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt 12480
acgcggtacg ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac 12540
cagagtaaat gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc 12600
atggaaaatc aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg 12660
ggcggttggc caggcgtaag cggctgggtt gtctgccggc cctgcaatgg cactggaacc 12720
cccaagcccg aggaatcggc gtgagcggtc gcaaaccatc cggcccggta caaatcggcg 12780
cggcgctggg tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac 12840
gcatcgaggc agaagcacgc cccggtgaat cgtggcaagc ggccgctgat cgaatccgca 12900
aagaatcccg gcaaccgccg gcagccggtg cgccgtcgat taggaagccg cccaagggcg 12960
acgagcaacc agattttttc gttccgatgc tctatgacgt gggcacccgc gatagtcgca 13020
gcatcatgga cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga 13080
tccgctacga gcttccagac gggcacgtag aggtttccgc agggccggcc ggcatggcca 13140
gtgtgtggga ttacgacctg gtactgatgg cggtttccca tctaaccgaa tccatgaacc 13200
gataccggga agggaaggga gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg 13260
tactcaagtt ctgccggcga gccgatggcg gaaagcagaa agacgacctg gtagaaacct 13320
gcattcggtt aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc 13380
gcctggtgac ggtatccgag ggtgaagcct tgattagccg ctacaagatc gtaaagagcg 13440
aaaccgggcg gccggagtac atcgagatcg agctagctga ttggatgtac cgcgagatca 13500
cagaaggcaa gaacccggac gtgctgacgg ttcaccccga ttactttttg atcgatcccg 13560
gcatcggccg ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat 13620
ggttgttcaa gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt 13680
tcaccgtgcg caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg 13740
cggggcaggc tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat 13800
ccgccggttc ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag 13860
gtcgaaaagg tctctttcct gtggatagca cgtacattgg gaacccaaag ccgtacattg 13920
ggaaccggaa cccgtacatt gggaacccaa agccgtacat tgggaaccgg tcacacatgt 13980
aagtgactga tataaaagag aaaaaaggcg atttttccgc ctaaaactct ttaaaactta 14040
ttaaaactct taaaacccgc ctggcctgtg cataactgtc tggccagcgc acagccgaag 14100
agctgcaaaa agcgcctacc cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc 14160
ggcctatcgc ggccgctggc cgctcaaaaa tggctggcct acggccaggc aatctaccag 14220
ggcgcggaca agccgcgccg tcgccactcg accgccggcg cccacatcaa ggcaccctgc 14280
ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 14340
acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 14400
gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact 14460
ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa 14520
taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca 14580
ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 14640
taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 14700
agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 14760
cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 14820
tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 14880
tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 14940
gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 15000
acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 15060
acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 15120
cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 15180
gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 15240
gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 15300
agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 15360
ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 15420
ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 15480
atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 15540
tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 15600
gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 15660
ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 15720
caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 15780
cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 15840
cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 15900
cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 15960
agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 16020
tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 16080
agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac 16140
atagcagaac tttaaaagtg ctcatcattg gaaaagacct gcaggggggg gggggcgctg 16200
aggtctgcct cgtgaagaag gtgttgctga ctcataccag gcctgaatcg ccccatcatc 16260
cagccagaaa gtgagggagc cacggttgat gagagctttg ttgtaggtgg accagttggt 16320
gattttgaac ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg 16380
atccttcaac tcagcaaaag ttcgatttat tcaacaaagc cgccgtcccg tcaagtcagc 16440
gtaatgctct gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc 16500
atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc 16560
cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg 16620
tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca 16680
aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc 16740
aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca 16800
aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat 16860
acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac 16920
actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat 16980
gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa 17040
tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct 17100
gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc 17160
ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta 17220
tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc 17280
cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt 17340
gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg 17400
tggctttccc ccccccccct gcaggtcaat tcggtcgata tggctattac gaagaaggct 17460
cgtgcgcgga gtcccgtgaa ctttcccacg caacaagtga accgcaccgg gtttgccgga 17520
ggccatttcg ttaaaatgcg cagccatggc tgcttcgtcc agcatggcgt aatactgatc 17580
ctcgtcttcg gctggcggta tattgccgat gggcttcaaa agccgccgtg gttgaaccag 17640
tctatccatt ccaaggtagc gaactcgacc gcttcgaagc tcctccatgg tccacgccga 17700
tgaatgacct cggccttgta aagaccgttg atcgcttctg cgagggcgtt gtcgtgctgt 17760
cgccgacgct tccgatagat ggctcgatac ctgcttctgc caaccgctcg gaatagcgaa 17820
aggacacgta ttgaacaccg cgatccgagt gatgcactag gccgccatga gcgggacgcc 17880
gatcatgatg agcctcctcg agggcatcga ggacaaagcc tgcatgtgct gtccggctcg 17940
cccgccatcc gacaatgcga cgggcgaaga cgtcgatcac gaaggccacg tagacgaagc 18000
cctcccaagt ggcgacataa gtacggacat gcgcaaaggc tttcccggtt tgtcgctgat 18060
ggtgcaagag acgctgaagc gcgatccgat gcgcaggcat ctgttcgtct tccgcggtcg 18120
tggcggtggc ctgatcaagg tcactcgccg aagagctgca tgattggctc gaaaccgagc 18180
gggggaaatt gtcgcgcagt tctcccgtcg ccgaggcgat aaattacatg ctcaagcgat 18240
gggatggcat tacgtcattc ctcgatgacg gcccgatttg cctgacgaac aatgctgccg 18300
aacgaacgct cagaggctat gtactcggca ggaagtcatg gctgtttgcc ggatcggatc 18360
gttgtgctga acgtgcggcg ttcatggcga cactgatcat gagcgccaag ctcaataaca 18420
tcgatccgca ggcctggctt gccgacgtcc gcgccgacct tgcggacgct ccgatcagca 18480
ggcttgagca acagctgccg tggaactgga catccaagac actgagtgct caggcggcct 18540
gacctgcggc cttcaccgga tacttacccc attatcgcag attgcgatga agcatcagcg 18600
tcattcagca atcttgccaa agtatgcagg ctcgcgagaa tcgacgtgcg aaaccggctg 18660
gttgcgccaa agatccgctt gcggagcggt cgaacattca tgctgggact tcaagaggtc 18720
gagtagagga agaaccggaa aggttgcacc ggaaaatatg cgttcctttg gagagcgcct 18780
catggacgtg aacaaatcgc ccggaccaag gatgccacgg atacaaaagc tcgcgaagct 18840
cggtcccgtg ggtgttctgt cgtctcgttg tacaacgaaa tccattccca ttccgcgctc 18900
aagatggctt cccctcggca gttcatcagg gctaaatcaa tctagccgac ttgtccggtg 18960
aaatgggctg cactccaaca gaaacaatca aacaaacata cacagcgact tattcacacg 19020
agctcaaatt acaacggtat atatcctgcc agtcagcatc atcacaccaa aagttaggcc 19080
cgaatagttt gaaattagaa agctcgcaat tgaggtctac aggccaaatt cgctcttagc 19140
cgtacaatat tactcaccgg tgcgatgccc cccatcgtag gtgaaggtgg aaattaatga 19200
tccatcttga gaccacaggc ccacaacagc taccagtttc ctcaagggtc caccaaaaac 19260
gtaagcgctt acgtacatgg tcgataagaa aaggcaattt gtagatgtta acatccaacg 19320
tcgctttcag ggatcgatcc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat 19380
taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt 19440
aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt 19500
atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat 19560
tacgccaagc ttgcatgcct gcaggtcgac tctagaggat ctgg 19604
<210>34
<211>20082
<212>DNA
<213>人工序列
<220>
<223>质粒pKR92 YLDGAT1 YLDGAT2
<400>34
cgcgccaagc ttttgatcca tgcccttcat ttgccgctta ttaattaatt tggtaacagt 60
ccgtactaat cagttactta tccttccccc atcataatta atcttggtag tctcgaatgc 120
cacaacactg actagtctct tggatcataa gaaaaagcca aggaacaaaa gaagacaaaa 180
cacaatgaga gtatcctttg catagcaatg tctaagttca taaaattcaa acaaaaacgc 240
aatcacacac agtggacatc acttatccac tagctgatca ggatcgccgc gtcaagaaaa 300
aaaaactgga ccccaaaagc catgcacaac aacacgtact cacaaaggtg tcaatcgagc 360
agcccaaaac attcaccaac tcaacccatc atgagccctc acatttgttg tttctaaccc 420
aacctcaaac tcgtattctc ttccgccacc tcatttttgt ttatttcaac acccgtcaaa 480
ctgcatgcca ccccgtggcc aaatgtccat gcatgttaac aagacctatg actataaata 540
gctgcaatct cggcccaggt tttcatcatc aagaaccagt tcaatatcct agtacaccgt 600
attaaagaat ttaagatata ctgcggccgc atgactatcg actcacaata ctacaagtcg 660
cgagacaaaa acgacacggc acccaaaatc gcgggaatcc gatatgcccc gctatcgaca 720
ccattactca accgatgtga gaccttctct ctggtctggc acattttcag cattcccact 780
ttcctcacaa ttttcatgct atgctgcgca attccactgc tctggccatt tgtgattgcg 840
tatgtagtgt acgctgttaa agacgactcc ccgtccaacg gaggagtggt caagcgatac 900
tcgcctattt caagaaactt cttcatctgg aagctctttg gccgctactt ccccataact 960
ctgcacaaga cggtggatct ggagcccacg cacacatact accctctgga cgtccaggag 1020
tatcacctga ttgctgagag atactggccg cagaacaagt acctccgagc aatcatctcc 1080
accatcgagt actttctgcc cgccttcatg aaacggtctc tttctatcaa cgagcaggag 1140
cagcctgccg agcgagatcc tctcctgtct cccgtttctc ccagctctcc gggttctcaa 1200
cctgacaagt ggattaacca cgacagcaga tatagccgtg gagaatcatc tggctccaac 1260
ggccacgcct cgggctccga acttaacggc aacggcaaca atggcaccac taaccgacga 1320
cctttgtcgt ccgcctctgc tggctccact gcatctgatt ccacgcttct taacgggtcc 1380
ctcaactcct acgccaacca gatcattggc gaaaacgacc cacagctgtc gcccacaaaa 1440
ctcaagccca ctggcagaaa atacatcttc ggctaccacc cccacggcat tatcggcatg 1500
ggagcctttg gtggaattgc caccgaggga gctggatggt ccaagctctt tccgggcatc 1560
cctgtttctc ttatgactct caccaacaac ttccgagtgc ctctctacag agagtacctc 1620
atgagtctgg gagtcgcttc tgtctccaag aagtcctgca aggccctcct caagcgaaac 1680
cagtctatct gcattgtcgt tggtggagca caggaaagtc ttctggccag acccggtgtc 1740
atggacctgg tgctactcaa gcgaaagggt tttgttcgac ttggtatgga ggtcggaaat 1800
gtcgcccttg ttcccatcat ggcctttggt gagaacgacc tctatgacca ggttagcaac 1860
gacaagtcgt ccaagctgta ccgattccag cagtttgtca agaacttcct tggattcacc 1920
cttcctttga tgcatgcccg aggcgtcttc aactacgatg tcggtcttgt cccctacagg 1980
cgacccgtca acattgtggt tggttccccc attgacttgc cttatctccc acaccccacc 2040
gacgaagaag tgtccgaata ccacgaccga tacatcgccg agctgcagcg aatctacaac 2100
gagcacaagg atgaatattt catcgattgg accgaggagg gcaaaggagc cccagagttc 2160
cgaatgattg agtaagcggc cgcaagtatg aactaaaatg catgtaggtg taagagctca 2220
tggagagcat ggaatattgt atccgaccat gtaacagtat aataactgag ctccatctca 2280
cttcttctat gaataaacaa aggatgttat gatatattaa cactctatct atgcacctta 2340
ttgttctatg ataaatttcc tcttattatt ataaatcatc tgaatcgtga cggcttatgg 2400
aatgcttcaa atagtacaaa aacaaatgtg tactataaga ctttctaaac aattctaacc 2460
ttagcattgt gaacgagaca taagtgttaa gaagacataa caattataat ggaagaagtt 2520
tgtctccatt tatatattat atattaccca cttatgtatt atattaggat gttaaggaga 2580
cataacaatt ataaagagag aagtttgtat ccatttatat attatatact acccatttat 2640
atattatact tatccactta tttaatgtct ttataaggtt tgatccatga tatttctaat 2700
attttagttg atatgtatat gaaagggtac tatttgaact ctcttactct gtataaaggt 2760
tggatcatcc ttaaagtggg tctatttaat tttattgctt cttacagata aaaaaaaaat 2820
tatgagttgg tttgataaaa tattgaagga tttaaaataa taataaataa catataatat 2880
atgtatataa atttattata atataacatt tatctataaa aaagtaaata ttgtcataaa 2940
tctatacaat cgtttagcct tgctggacga atctcaatta tttaaacgag agtaaacata 3000
tttgactttt tggttattta acaaattatt atttaacact atatgaaatt ttttttttta 3060
tcagcaaaga ataaaattaa attaagaagg acaatggtgt cccaatcctt atacaaccaa 3120
cttccacaag aaagtcaagt cagagacaac aaaaaaacaa gcaaaggaaa ttttttaatt 3180
tgagttgtct tgtttgctgc ataatttatg cagtaaaaca ctacacataa cccttttagc 3240
agtagagcaa tggttgaccg tgtgcttagc ttcttttatt ttattttttt atcagcaaag 3300
aataaataaa ataaaatgag acacttcagg gatgtttcaa caagcttgga tcctagccta 3360
agtacgtact caaaatgcca acaaataaaa aaaaagttgc tttaataatg ccaaaacaaa 3420
ttaataaaac acttacaaca ccggattttt tttaattaaa atgtgccatt taggataaat 3480
agttaatatt tttaataatt atttaaaaag ccgtatctac taaaatgatt tttatttggt 3540
tgaaaatatt aatatgttta aatcaacaca atctatcaaa attaaactaa aaaaaaaata 3600
agtgtacgtg gttaacatta gtacagtaat ataagaggaa aatgagaaat taagaaattg 3660
aaagcgagtc taatttttaa attatgaacc tgcatatata aaaggaaaga aagaatccag 3720
gaagaaaaga aatgaaacca tgcatggtcc cctcgtcatc acgagtttct gccatttgca 3780
atagaaacac tgaaacacct ttctctttgt cacttaattg agatgccgaa gccacctcac 3840
accatgaact tcatgaggtg tagcacccaa ggcttccata gccatgcata ctgaagaatg 3900
tctcaagctc agcaccctac ttctgtgacg tgtccctcat tcaccttcct ctcttcccta 3960
taaataacca cgcctcaggt tctccgcttc acaactcaaa cattctctcc attggtcctt 4020
aaacactcat cagtcatcac catggaggtc cgacgacgaa agatagacgt gctcaaggcc 4080
cagaaaaacg gctacgaatc gggcccacca tctcgacaat cgtcgcagcc ctcctcaaga 4140
gcatcgtcca gaacccgcaa caaacactcc tcgtccaccc tgtcgctcag cggactgacc 4200
atgaaagtcc agaagaaacc tgcgggaccc ccggcgaact ccaaaacgcc attcctacac 4260
atcaagcccg tgcacacgtg ctgctccaca tcaatgcttt cgcgcgatta tgacggctcc 4320
aaccccagct tcaagggctt caaaaacatc ggcatgatca ttctcattgt gggaaatcta 4380
cggctcgcat tcgaaaacta cctcaaatac ggcatttcca acccgttctt cgaccccaaa 4440
attactcctt ccgagtggca gctctcaggc ttgctcatag tcgtggccta cgcacatatc 4500
ctcatggcct acgctattga gagcgctgcc aagctgctgt tcctctctag caaacaccac 4560
tacatggccg tggggcttct gcataccatg aacactttgt cgtccatctc gttgctgtcc 4620
tacgtcgtct actactacct gcccaacccc gtggcaggca caatagtcga gtttgtggcc 4680
gttattctgt ctctcaaact cgcctcatac gccctcacta actcggatct ccgaaaagcc 4740
gcaattcatg cccagaagct cgacaagacg caagacgata acgaaaagga atccacctcg 4800
tcttcctctt cttcagatga cgcagagact ttggcagaca ttgacgtcat tcctgcatac 4860
tacgcacagc tgccctaccc ccagaatgtg acgctgtcga acctgctgta cttctggttt 4920
gctcccacac tggtctacca gcccgtgtac cccaagacgg agcgtattcg acccaagcac 4980
gtgatccgaa acctgtttga gctcgtctct ctgtgcatgc ttattcagtt tctcatcttc 5040
cagtacgcct accccatcat gcagtcgtgt ctggctctgt tcttccagcc caagctcgat 5100
tatgccaaca tctccgagcg cctcatgaag ttggcctccg tgtctatgat ggtctggctc 5160
attggattct acgctttctt ccagaacggt ctcaatctta ttgccgagct cacctgtttt 5220
ggaaacagaa ccttctacca gcagtggtgg aattcccgct ccattggcca gtactggact 5280
ctatggaaca agccagtcaa ccagtacttt agacaccacg tctacgtgcc tcttctcgct 5340
cggggcatgt cgcggttcaa tgcgtcggtg gtggttttct ttttctccgc cgtcatccat 5400
gaactgcttg tcggcatccc cactcacaac atcatcggag ccgccttctt cggcatgatg 5460
tcgcaggtgc ctctgatcat ggctactgag aaccttcagc atattaactc ctctctgggc 5520
cccttccttg gcaactgtgc attctggttc acctttttcc tgggacaacc cacttgtgca 5580
ttcctttact atttggccta caactacaag cagaaccagt agcggccgca agtatgaact 5640
aaaatgcatg taggtgtaag agctcatgga gagcatggaa tattgtatcc gaccatgtaa 5700
cagtataata actgagctcc atctcacttc ttctatgaat aaacaaagga tgttatgata 5760
tattaacact ctatctatgc accttattgt tctatgataa atttcctctt attattataa 5820
atcatctgaa tcgtgacggc ttatggaatg cttcaaatag tacaaaaaca aatgtgtact 5880
ataagacttt ctaaacaatt ctaaccttag cattgtgaac gagacataag tgttaagaag 5940
acataacaat tataatggaa gaagtttgtc tccatttata tattatatat tacccactta 6000
tgtattatat taggatgtta aggagacata acaattataa agagagaagt ttgtatccat 6060
ttatatatta tatactaccc atttatatat tatacttatc cacttattta atgtctttat 6120
aaggtttgat ccatgatatt tctaatattt tagttgatat gtatatgaaa gggtactatt 6180
tgaactctct tactctgtat aaaggttgga tcatccttaa agtgggtcta tttaatttta 6240
ttgcttctta cagataaaaa aaaaattatg agttggtttg ataaaatatt gaaggattta 6300
aaataataat aaataacata taatatatgt atataaattt attataatat aacatttatc 6360
tataaaaaag taaatattgt cataaatcta tacaatcgtt tagccttgct ggacgaatct 6420
caattattta aacgagagta aacatatttg actttttggt tatttaacaa attattattt 6480
aacactatat gaaatttttt tttttatcag caaagaataa aattaaatta agaaggacaa 6540
tggtgtccca atccttatac aaccaacttc cacaagaaag tcaagtcaga gacaacaaaa 6600
aaacaagcaa aggaaatttt ttaatttgag ttgtcttgtt tgctgcataa tttatgcagt 6660
aaaacactac acataaccct tttagcagta gagcaatggt tgaccgtgtg cttagcttct 6720
tttattttat ttttttatca gcaaagaata aataaaataa aatgagacac ttcagggatg 6780
tttcaacaag ctctagactg gaattcgtcg acggcgcgcc tcgagtgggc ggatcccccg 6840
ggctgcagga attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt 6900
acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag 6960
gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gatcgatcca 7020
tcgcgatgta ccttttgtta gtcagcctct cgattgctca tcgtcattac acagtaccga 7080
agtttgatcg atctagtaac atagatgaca ccgcgcgcga taatttatcc tagtttgcgc 7140
gctatatttt gttttctatc gcgtattaaa tgtataattg cgggactcta atcataaaaa 7200
cccatctcat aaataacgtc atgcattaca tgttaattat tacatgctta acgtaattca 7260
acagaaatta tatgataatc atcgcaagac cggcaacagg attcaatctt aagaaacttt 7320
attgccaaat gtttgaacga tctgcttcga cgcactcctt ctttactcca ccatctcgtc 7380
cttattgaaa acgtgggtag caccaaaacg aatcaagtcg ctggaactga agttaccaat 7440
cacgctggat gatttgccag ttggattaat cttgcctttc cccgcatgaa taatattgat 7500
gaatgcatgc gtgaggggta gttcgatgtt ggcaatagct gcaattgccg cgacatcctc 7560
caacgagcat aattcttcag aaaaatagcg atgttccatg ttgtcagggc atgcatgatg 7620
cacgttatga ggtgacggtg ctaggcagta ttccctcaaa gtttcatagt cagtatcata 7680
ttcatcattg cattcctgca agagagaatt gagacgcaat ccacacgctg cggcaacctt 7740
ccggcgttcg tggtctattt gctcttggac gttgcaaacg taagtgttgg atcgatccgg 7800
ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc 7860
ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg 7920
atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa 7980
ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag 8040
cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa 8100
cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa 8160
gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc 8220
ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg 8280
atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg 8340
ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag 8400
ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag 8460
gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac 8520
gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc 8580
gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc 8640
ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc 8700
atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc 8760
aatcatgcga aacgatcccc gcaagcttgg agactggtga tttcagcgtg tcctctccaa 8820
atgaaatgaa cttccttata tagaggaagg gtcttgcgaa ggatagtggg attgtgcgtc 8880
atcccttacg tcagtggaga tatcacatca atccacttgc tttgaagacg tggttggaac 8940
gtcttctttt tccacgatgc tcctcgtggg tgggggtcca tctttgggac cactgtcggc 9000
agaggcatct tcaacgatgg cctttccttt atcgcaatga tggcatttgt aggagccacc 9060
ttccttttcc actatcttca caataaagtg acagatagct gggcaatgga atccgaggag 9120
gtttccggat attacccttt gttgaaaagt ctcaattgcc ctttggtctt ctgagactgt 9180
atctttgata tttttggagt agacaagcgt gtcgtgctcc accatgttga cgaagatttt 9240
cttcttgtca ttgagtcgta agagactctg tatgaactgt tcgccagtct ttacggcgag 9300
ttctgttagg tcctctattt gaatctttga ctccatggcc tttgattcag tgggaactac 9360
ctttttagag actccaatct ctattacttg ccttggtttg tgaagcaagc cttgaatcgt 9420
ccatactgga atagtacttc tgatcttgag aaatatatct ttctctgtgt tcttgatgca 9480
gttagtcctg aatcttttga ctgcatcttt aaccttcttg ggaaggtatt tgatctcctg 9540
gagattattg ctcgggtaga tcgtcttgat gagacctgct gcgtaagcct ctctaaccat 9600
ctgtgggtta gcattctttc tgaaattgaa aaggctaatc ttctcattat cagtggtgaa 9660
catggtatcg tcaccttctc cgtcgaactt cctgactaga tcgtagagat agaggaagtc 9720
gtccattgtg atctctgggg caaaggagtc tgaattaatt cgatatggtg gatttatcac 9780
aaatgggacc cgccgccgac agaggtgtga tgttaggcca ggactttgaa aatttgcgca 9840
actatcgtat agtggccgac aaattgacgc cgagttgaca gactgcctag catttgagtg 9900
aattatgtga ggtaatgggc tacactgaat tggtagctca aactgtcagt atttatgtat 9960
atgagtgtat attttcgcat aatctcagac caatctgaag atgaaatggg tatctgggaa 10020
tggcgaaatc aaggcatcga tcgtgaagtt tctcatctaa gcccccattt ggacgtgaat 10080
gtagacacgt cgaaataaag atttccgaat tagaataatt tgtttattgc tttcgcctat 10140
aaatacgacg gatcgtaatt tgtcgtttta tcaaaatgta ctttcatttt ataataacgc 10200
tgcggacatc tacatttttg aattgaaaaa aaattggtaa ttactctttc tttttctcca 10260
tattgaccat catactcatt gctgatccat gtagatttcc cggacatgaa gccatttaca 10320
attgaatata tcctgccgcc gctgccgctt tgcacccggt ggagcttgca tgttggtttc 10380
tacgcagaac tgagccggtt aggcagataa tttccattga gaactgagcc atgtgcacct 10440
tccccccaac acggtgagcg acggggcaac ggagtgatcc acatgggact tttaaacatc 10500
atccgtcgga tggcgttgcg agagaagcag tcgatccgtg agatcagccg acgcaccggg 10560
caggcgcgca acacgatcgc aaagtatttg aacgcaggta caatcgagcc gacgttcacg 10620
cggaacgacc aagcaagcta gctttaatgc ggtagtttat cacagttaaa ttgctaacgc 10680
agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg caccgtcacc 10740
ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt gcgggatatc 10800
gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata tgcgttgatg 10860
caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg ccgcccagtc 10920
ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac cacacccgtc 10980
ctgtggtcca acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagccgt 11040
cttctgaaaa cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct 11100
tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga 11160
accggagaca ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg 11220
tcagcaccga cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca 11280
ccaagctgtt ttccgagaag atcaccggca ccaggcgcga ccgcccggag ctggccagga 11340
tgcttgacca cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc 11400
gcagcacccg cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc 11460
gtagcctggc agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg 11520
tgttcgccgg cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc 11580
gcgaggccgc caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac 11640
agatcgcgca cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg 11700
cactgcttgg cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga 11760
cgcccaccga ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg 11820
ccctggcggc cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg 11880
ccaggacgaa ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta 11940
cgtgttcgag ccgcccgcgc acgtctcaac cgtgcggctg catgaaatcc tggccggttt 12000
gtctgatgcc aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg 12060
ccgtctaaaa aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta 12120
tatgatgcga tgagtaaata aacaaatacg caagggaacg catgaagtta tcgctgtact 12180
taaccagaaa ggcgggtcag gcaagacgac catcgcaacc catctagccc gcgccctgca 12240
actcgccggg gccgatgttc tgttagtcga ttccgatccc cagggcagtg cccgcgattg 12300
ggcggccgtg cgggaagatc aaccgctaac cgttgtcggc atcgaccgcc cgacgattga 12360
ccgcgacgtg aaggccatcg gccggcgcga cttcgtagtg atcgacggag cgccccaggc 12420
ggcggacttg gctgtgtccg cgatcaaggc agccgacttc gtgctgattc cggtgcagcc 12480
aagcccttac gacatatggg ccaccgccga cctggtggag ctggttaagc agcgcattga 12540
ggtcacggat ggaaggctac aagcggcctt tgtcgtgtcg cgggcgatca aaggcacgcg 12600
catcggcggt gaggttgccg aggcgctggc cgggtacgag ctgcccattc ttgagtcccg 12660
tatcacgcag cgcgtgagct acccaggcac tgccgccgcc ggcacaaccg ttcttgaatc 12720
agaacccgag ggcgacgctg cccgcgaggt ccaggcgctg gccgctgaaa ttaaatcaaa 12780
actcatttga gttaatgagg taaagagaaa atgagcaaaa gcacaaacac gctaagtgcc 12840
ggccgtccga gcgcacgcag cagcaaggct gcaacgttgg ccagcctggc agacacgcca 12900
gccatgaagc gggtcaactt tcagttgccg gcggaggatc acaccaagct gaagatgtac 12960
gcggtacgcc aaggcaagac cattaccgag ctgctatctg aatacatcgc gcagctacca 13020
gagtaaatga gcaaatgaat aaatgagtag atgaatttta gcggctaaag gaggcggcat 13080
ggaaaatcaa gaacaaccag gcaccgacgc cgtggaatgc cccatgtgtg gaggaacggg 13140
cggttggcca ggcgtaagcg gctgggttgt ctgccggccc tgcaatggca ctggaacccc 13200
caagcccgag gaatcggcgt gagcggtcgc aaaccatccg gcccggtaca aatcggcgcg 13260
gcgctgggtg atgacctggt ggagaagttg aaggccgcgc aggccgccca gcggcaacgc 13320
atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg ccgctgatcg aatccgcaaa 13380
gaatcccggc aaccgccggc agccggtgcg ccgtcgatta ggaagccgcc caagggcgac 13440
gagcaaccag attttttcgt tccgatgctc tatgacgtgg gcacccgcga tagtcgcagc 13500
atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc gacgagctgg cgaggtgatc 13560
cgctacgagc ttccagacgg gcacgtagag gtttccgcag ggccggccgg catggccagt 13620
gtgtgggatt acgacctggt actgatggcg gtttcccatc taaccgaatc catgaaccga 13680
taccgggaag ggaagggaga caagcccggc cgcgtgttcc gtccacacgt tgcggacgta 13740
ctcaagttct gccggcgagc cgatggcgga aagcagaaag acgacctggt agaaacctgc 13800
attcggttaa acaccacgca cgttgccatg cagcgtacga agaaggccaa gaacggccgc 13860
ctggtgacgg tatccgaggg tgaagccttg attagccgct acaagatcgt aaagagcgaa 13920
accgggcggc cggagtacat cgagatcgag ctagctgatt ggatgtaccg cgagatcaca 13980
gaaggcaaga acccggacgt gctgacggtt caccccgatt actttttgat cgatcccggc 14040
atcggccgtt ttctctaccg cctggcacgc cgcgccgcag gcaaggcaga agccagatgg 14100
ttgttcaaga cgatctacga acgcagtggc agcgccggag agttcaagaa gttctgtttc 14160
accgtgcgca agctgatcgg gtcaaatgac ctgccggagt acgatttgaa ggaggaggcg 14220
gggcaggctg gcccgatcct agtcatgcgc taccgcaacc tgatcgaggg cgaagcatcc 14280
gccggttcct aatgtacgga gcagatgcta gggcaaattg ccctagcagg ggaaaaaggt 14340
cgaaaaggtc tctttcctgt ggatagcacg tacattggga acccaaagcc gtacattggg 14400
aaccggaacc cgtacattgg gaacccaaag ccgtacattg ggaaccggtc acacatgtaa 14460
gtgactgata taaaagagaa aaaaggcgat ttttccgcct aaaactcttt aaaacttatt 14520
aaaactctta aaacccgcct ggcctgtgca taactgtctg gccagcgcac agccgaagag 14580
ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac gccccgccgc ttcgcgtcgg 14640
cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac ggccaggcaa tctaccaggg 14700
cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc cacatcaagg caccctgcct 14760
cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac 14820
agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt 14880
tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg 14940
cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata 15000
ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact 15060
gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 15120
atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 15180
caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 15240
cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 15300
taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 15360
ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 15420
tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 15480
gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 15540
ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 15600
aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 15660
aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 15720
agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 15780
cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 15840
gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 15900
atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 15960
gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 16020
tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 16080
gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 16140
ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 16200
actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 16260
ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 16320
tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 16380
cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 16440
ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 16500
ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 16560
tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat 16620
agcagaactt taaaagtgct catcattgga aaagacctgc aggggggggg gggcgctgag 16680
gtctgcctcg tgaagaaggt gttgctgact cataccaggc ctgaatcgcc ccatcatcca 16740
gccagaaagt gagggagcca cggttgatga gagctttgtt gtaggtggac cagttggtga 16800
ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat 16860
ccttcaactc agcaaaagtt cgatttattc aacaaagccg ccgtcccgtc aagtcagcgt 16920
aatgctctgc cagtgttaca accaattaac caattctgat tagaaaaact catcgagcat 16980
caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg 17040
tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta 17100
tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa 17160
aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa 17220
aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa 17280
atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac 17340
gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac 17400
tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc 17460
tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg 17520
cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt 17580
aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt 17640
cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata 17700
cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg 17760
ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt 17820
tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg 17880
gctttccccc ccccccctgc aggtcaattc ggtcgatatg gctattacga agaaggctcg 17940
tgcgcggagt cccgtgaact ttcccacgca acaagtgaac cgcaccgggt ttgccggagg 18000
ccatttcgtt aaaatgcgca gccatggctg cttcgtccag catggcgtaa tactgatcct 18060
cgtcttcggc tggcggtata ttgccgatgg gcttcaaaag ccgccgtggt tgaaccagtc 18120
tatccattcc aaggtagcga actcgaccgc ttcgaagctc ctccatggtc cacgccgatg 18180
aatgacctcg gccttgtaaa gaccgttgat cgcttctgcg agggcgttgt cgtgctgtcg 18240
ccgacgcttc cgatagatgg ctcgatacct gcttctgcca accgctcgga atagcgaaag 18300
gacacgtatt gaacaccgcg atccgagtga tgcactaggc cgccatgagc gggacgccga 18360
tcatgatgag cctcctcgag ggcatcgagg acaaagcctg catgtgctgt ccggctcgcc 18420
cgccatccga caatgcgacg ggcgaagacg tcgatcacga aggccacgta gacgaagccc 18480
tcccaagtgg cgacataagt acggacatgc gcaaaggctt tcccggtttg tcgctgatgg 18540
tgcaagagac gctgaagcgc gatccgatgc gcaggcatct gttcgtcttc cgcggtcgtg 18600
gcggtggcct gatcaaggtc actcgccgaa gagctgcatg attggctcga aaccgagcgg 18660
gggaaattgt cgcgcagttc tcccgtcgcc gaggcgataa attacatgct caagcgatgg 18720
gatggcatta cgtcattcct cgatgacggc ccgatttgcc tgacgaacaa tgctgccgaa 18780
cgaacgctca gaggctatgt actcggcagg aagtcatggc tgtttgccgg atcggatcgt 18840
tgtgctgaac gtgcggcgtt catggcgaca ctgatcatga gcgccaagct caataacatc 18900
gatccgcagg cctggcttgc cgacgtccgc gccgaccttg cggacgctcc gatcagcagg 18960
cttgagcaac agctgccgtg gaactggaca tccaagacac tgagtgctca ggcggcctga 19020
cctgcggcct tcaccggata cttaccccat tatcgcagat tgcgatgaag catcagcgtc 19080
attcagcaat cttgccaaag tatgcaggct cgcgagaatc gacgtgcgaa accggctggt 19140
tgcgccaaag atccgcttgc ggagcggtcg aacattcatg ctgggacttc aagaggtcga 19200
gtagaggaag aaccggaaag gttgcaccgg aaaatatgcg ttcctttgga gagcgcctca 19260
tggacgtgaa caaatcgccc ggaccaagga tgccacggat acaaaagctc gcgaagctcg 19320
gtcccgtggg tgttctgtcg tctcgttgta caacgaaatc cattcccatt ccgcgctcaa 19380
gatggcttcc cctcggcagt tcatcagggc taaatcaatc tagccgactt gtccggtgaa 19440
atgggctgca ctccaacaga aacaatcaaa caaacataca cagcgactta ttcacacgag 19500
ctcaaattac aacggtatat atcctgccag tcagcatcat cacaccaaaa gttaggcccg 19560
aatagtttga aattagaaag ctcgcaattg aggtctacag gccaaattcg ctcttagccg 19620
tacaatatta ctcaccggtg cgatgccccc catcgtaggt gaaggtggaa attaatgatc 19680
catcttgaga ccacaggccc acaacagcta ccagtttcct caagggtcca ccaaaaacgt 19740
aagcgcttac gtacatggtc gataagaaaa ggcaatttgt agatgttaac atccaacgtc 19800
gctttcaggg atcgatccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 19860
atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa 19920
tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat 19980
gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta 20040
cgccaagctt gcatgcctgc aggtcgactc tagaggatct gg 20082
<210>35
<211>3527
<212>DNA
<213>大豆
<400>35
tagcctaagt acgtactcaa aatgccaaca aataaaaaaa aagttgcttt aataatgcca 60
aaacaaatta ataaaacact tacaacaccg gatttttttt aattaaaatg tgccatttag 120
gataaatagt taatattttt aataattatt taaaaagccg tatctactaa aatgattttt 180
atttggttga aaatattaat atgtttaaat caacacaatc tatcaaaatt aaactaaaaa 240
aaaaataagt gtacgtggtt aacattagta cagtaatata agaggaaaat gagaaattaa 300
gaaattgaaa gcgagtctaa tttttaaatt atgaacctgc atatataaaa ggaaagaaag 360
aatccaggaa gaaaagaaat gaaaccatgc atggtcccct cgtcatcacg agtttctgcc 420
atttgcaata gaaacactga aacacctttc tctttgtcac ttaattgaga tgccgaagcc 480
acctcacacc atgaacttca tgaggtgtag cacccaaggc ttccatagcc atgcatactg 540
aagaatgtct caagctcagc accctacttc tgtgacgttg tccctcattc accttcctct 600
cttccctata aataaccacg cctcaggttc tccgcttcac aactcaaaca ttctcctcca 660
ttggtcctta aacactcatc agtcatcacc atggccaagc tagttttttc cctttgtttt 720
ctgcttttca gtggctgctg cttcgctttc agttccagag agcagcctca gcaaaacgag 780
tgccagatcc aaaaactcaa tgccctcaaa ccggataacc gtatagagtc agaaggaggg 840
ctcattgaga catggaaccc taacaacaag ccattccagt gtgccggtgt tgccctctct 900
cgctgcaccc tcaaccgcaa cgcccttcgt agaccttcct acaccaacgg tccccaggaa 960
atctacatcc aacaaggtcc atcttgtcca aacttcacat ataaatatat aatagactta 1020
aatatgttta agggtttgat aaatgaggga attttatttt agatttttaa taattttttt 1080
tgttttgagt ttttatatat taaaattttt gttttgattt cttccatatg acgtaacata 1140
atcatatcat tgataatgtt gggttcctaa tttttgtttg tttgttgttt tgtaatatga 1200
ataggtaagg gtatttttgg catgatatac ccgggttgtc ctagcacatt tgaagagcct 1260
caacaacctc aacaaagagg acaaagcagc agaccacaag accgtcacca gaagatctat 1320
aacttcagag agggtgattt gatcgcagtg cctactggtg ttgcatggtg gatgtacaac 1380
aatgaagaca ctcctgttgt tgccgtttct attattgaca ccaacagctt ggagaaccag 1440
ctcgaccaga tgcctagggt gagccacata gcaatattag atattataat tctttaaagg 1500
tttaaatatc attttagttc gtggagttgc actttctaat ttagtaccta tagattaaaa 1560
tatgccaatt gaatccttat agttgtgttt ttttatccaa tttggttctt gtcttgaaat 1620
aaatggacaa tattgtagct gataaaaaaa ggaaactgga ctacattgta acgttaagat 1680
tagaattctt aagttctaat actagctggt tacagattga caactatttg ttttgacaat 1740
tcttggcaga gattctatct tgctgggaac caagagcaag agtttctaaa atatcagcaa 1800
gagcaaggag gtcatcaaag ccagaaagga aagcatcagc aagaagaaga aaacgaagga 1860
ggcagcatat tgagtggctt caccctggaa ttcttggaac atgcattcag cgtggacaag 1920
cagatagcga aaaacctaca aggagagaac gaaggggaag acaagggagc cattgtgaca 1980
gtgaaaggag gtctgagcgt gataaaacca cccacggacg agcagcaaca aagaccccag 2040
gaagaggaag aagaagaaga ggatgagaag ccacagtgca agggtaaaga caaacactgc 2100
caacgccccc gaggaagcca aagcaaaagc agaagaaatg gcattgacga gaccatatgc 2160
accatgagac ttcgccacaa cattggccag acttcatcac ctgacatcta caaccctcaa 2220
gccggtagcg tcacaaccgc caccagcctt gacttcccag ccctctcgtg gctcagactc 2280
agtgctgagt ttggatctct ccgcaaggta cgtacatcat tcatcaaaga tcaacataca 2340
tttatacatt aaactaatat ttgttgccaa atatttatta attttattga taattaattt 2400
ttttagaaaa tttgtttgat cacttttaat ggagtctttc atcttaatta cattatttat 2460
acttagacta atgatttatt gattaataat aatcttagat acactataaa atgtgtgacg 2520
gagttatctt aacacttgca tggattctat cttttctgtc tttatatata gaaatagaga 2580
gaaaaaaaaa gaaaagattg atgaaaaaag caaaacaaaa aatagtatta ttataaaaat 2640
attggatgaa tttgttgtga ctcttgcatg cattgatgta cgatgcagaa tgcaatgttc 2700
gtgccacact acaacctgaa cgcgaacagc ataatatacg cattgaatgg acgggcattg 2760
atacaagtgg tgaattgcaa cggtgagaga gtgtttgatg gagagctgca agagggacgg 2820
gtgctgatcg tgccacaaaa ctttgtggtg gctgcaagat cacagagtga caacttcgag 2880
tatgtgtcat tcaagaccaa tgatacaccc atgatcggca ctcttgcagg ggcaaactca 2940
ttgttgaacg cattaccaga ggaagtgatt cagcacactt tcaacctaaa aagccagcag 3000
gccaggcaga taaagaacaa caaccctttc aagttcctgg ttccacctca ggagtctcag 3060
aagagagctg tggcttagag ccctttttgt atgtgctacc ccacttttgt ctttttggca 3120
atagtgctag caaccaataa ataataataa taataatgaa taagaaaaca aaggctttag 3180
cttgcctttt gttcactgta aaataataat gtaagtactc tctataatga gtcacgaaac 3240
ttttgcggga ataaaaggag aaattccaat gagttttctg tcaaatcttc ttttgtctct 3300
ctctctctct cttttttttt tctttcttct gagcttcttg caaaacaaaa ggcaaacaat 3360
aacgattggt ccaatgatag ttagcttgat cgatgatatc tttaggaagt gttggcagga 3420
caggacatga tgtagaagac taaaattgaa agtattgcag acccaatagt tgaagattaa 3480
ctttaagaat gaagacgtct tatcaggttc ttcatgactt ggagctc 3527
<210>36
<211>690
<212>DNA
<213>大豆
<400>36
tagcctaagt acgtactcaa aatgccaaca aataaaaaaa aagttgcttt aataatgcca 60
aaacaaatta ataaaacact tacaacaccg gatttttttt aattaaaatg tgccatttag 120
gataaatagt taatattttt aataattatt taaaaagccg tatctactaa aatgattttt 180
atttggttga aaatattaat atgtttaaat caacacaatc tatcaaaatt aaactaaaaa 240
aaaaataagt gtacgtggtt aacattagta cagtaatata agaggaaaat gagaaattaa 300
gaaattgaaa gcgagtctaa tttttaaatt atgaacctgc atatataaaa ggaaagaaag 360
aatccaggaa gaaaagaaat gaaaccatgc atggtcccct cgtcatcacg agtttctgcc 420
atttgcaata gaaacactga aacacctttc tctttgtcac ttaattgaga tgccgaagcc 480
acctcacacc atgaacttca tgaggtgtag cacccaaggc ttccatagcc atgcatactg 540
aagaatgtct caagctcagc accctacttc tgtgacgttg tccctcattc accttcctct 600
cttccctata aataaccacg cctcaggttc tccgcttcac aactcaaaca ttctcctcca 660
ttggtcctta aacactcatc agtcatcacc 690
<210>37
<211>36
<212>DNA
<213>人工序列
<220>
<223>引物oGY1-1
<400>37
cgcggatcct agcctaagta cgtactcaaa atgcca 36
<210>38
<211>39
<212>DNA
<213>人工序列
<220>
<223>引物oGY1-2
<400>38
gaattcccat ggggtgatga ctgatgagtg tttaaggac 39
<210>39
<211>6660
<212>DNA
<213>人工序列
<220>
<223>质粒pZBL114
<400>39
cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 60
taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 120
agctgacttg gtcagcggcc gctctagaac tagtggatcc tagcctaagt acgtactcaa 180
aatgccaaca aataaaaaaa aagttgcttt aataatgcca aaacaaatta ataaaacact 240
tacaacaccg gatttttttt aattaaaatg tgccatttag gataaatagt taatattttt 300
aataattatt taaaaagccg tatctactaa aatgattttt atttggttga aaatattaat 360
atgtttaaat caacacaatc tatcaaaatt aaactaaaaa aaaaataagt gtacgtggtt 420
aacattagta cagtaatata agaggaaaat gagaaattaa gaaattgaaa gcgagtctaa 480
tttttaaatt atgaacctgc atatataaaa ggaaagaaag aatccaggaa gaaaagaaat 540
gaaaccatgc atggtcccct cgtcatcacg agtttctgcc atttgcaata gaaacactga 600
aacacctttc tctttgtcac ttaattgaga tgccgaagcc acctcacacc atgaacttca 660
tgaggtgtag cacccaaggc ttccatagcc atgcatactg aagaatgtct caagctcagc 720
accctacttc tgtgacgtgt ccctcattca ccttcctctc ttccctataa ataaccacgc 780
ctcaggttct ccgcttcaca actcaaacat tctctccatt ggtccttaaa cactcatcag 840
tcatcaccat ggtacgtcct gtagaaaccc caacccgtga aatcaaaaaa ctcgacggcc 900
tgtgggcatt cagtctggat cgcgaaaact gtggaattga tcagcgttgg tgggaaagcg 960
cgttacaaga aagccgggca attgctgtgc caggcagttt taacgatcag ttcgccgatg 1020
cagatattcg taattatgcg ggcaacgtct ggtatcagcg cgaagtcttt ataccgaaag 1080
gttgggcagg ccagcgtatc gtgctgcgtt tcgatgcggt cactcattac ggcaaagtgt 1140
gggtcaataa tcaggaagtg atggagcatc agggcggcta tacgccattt gaagccgatg 1200
tcacgccgta tgttattgcc gggaaaagtg tacgtatcac cgtttgtgtg aacaacgaac 1260
tgaactggca gactatcccg ccgggaatgg tgattaccga cgaaaacggc aagaaaaagc 1320
agtcttactt ccatgatttc tttaactatg ccggaatcca tcgcagcgta atgctctaca 1380
ccacgccgaa cacctgggtg gacgatatca ccgtggtgac gcatgtcgcg caagactgta 1440
accacgcgtc tgttgactgg caggtggtgg ccaatggtga tgtcagcgtt gaactgcgtg 1500
atgcggatca acaggtggtt gcaactggac aaggcactag cgggactttg caagtggtga 1560
atccgcacct ctggcaaccg ggtgaaggtt atctctatga actgtgcgtc acagccaaaa 1620
gccagacaga gtgtgatatc tacccgcttc gcgtcggcat ccggtcagtg gcagtgaagg 1680
gccaacagtt cctgattaac cacaaaccgt tctactttac tggctttggt cgtcatgaag 1740
atgcggactt acgtggcaaa ggattcgata acgtgctgat ggtgcacgac cacgcattaa 1800
tggactggat tggggccaac tcctaccgta cctcgcatta cccttacgct gaagagatgc 1860
tcgactgggc agatgaacat ggcatcgtgg tgattgatga aactgctgct gtcggcttta 1920
acctctcttt aggcattggt ttcgaagcgg gcaacaagcc gaaagaactg tacagcgaag 1980
aggcagtcaa cggggaaact cagcaagcgc acttacaggc gattaaagag ctgatagcgc 2040
gtgacaaaaa ccacccaagc gtggtgatgt ggagtattgc caacgaaccg gatacccgtc 2100
cgcaagtgca cgggaatatt tcgccactgg cggaagcaac gcgtaaactc gacccgacgc 2160
gtccgatcac ctgcgtcaat gtaatgttct gcgacgctca caccgatacc atcagcgatc 2220
tctttgatgt gctgtgcctg aaccgttatt acggatggta tgtccaaagc ggcgatttgg 2280
aaacggcaga gaaggtactg gaaaaagaac ttctggcctg gcaggagaaa ctgcatcagc 2340
cgattatcat caccgaatac ggcgtggata cgttagccgg gctgcactca atgtacaccg 2400
acatgtggag tgaagagtat cagtgtgcat ggctggatat gtatcaccgc gtctttgatc 2460
gcgtcagcgc cgtcgtcggt gaacaggtat ggaatttcgc cgattttgcg acctcgcaag 2520
gcatattgcg cgttggcggt aacaagaaag ggatcttcac tcgcgaccgc aaaccgaagt 2580
cggcggcttt tctgctgcaa aaacgctgga ctggcatgaa cttcggtgaa aaaccgcagc 2640
agggaggcaa acaatgaatc aacaactctc ctggcgcacc atcgtcggct acagcctcgg 2700
tggggaattc cccgggggta cctctagaca tggccacgtg catgaagtat gaactaaaat 2760
gcatgtaggt gtaagagctc atggagagca tggaatattg tatccgacca tgtaacagta 2820
taataactga gctccatctc acttcttcta tgaataaaca aaggatgtta tgatatatta 2880
acactctatc tatgcacctt attgttctat gataaatttc ctcttattat tataaatcat 2940
ctgaatcgtg acggcttatg gaatgcttca aatagtacaa aaacaaatgt gtactataag 3000
actttctaaa caattctaac tttagcattg tgaacgagac ataagtgtta agaagacata 3060
acaattataa tggaagaagt ttgtctccat ttatatatta tatattaccc acttatgtat 3120
tatattagga tgttaaggag acataacaat tataaagaga gaagtttgta tccatttata 3180
tattatatac tacccattta tatattatac ttatccactt atttaatgtc tttataaggt 3240
ttgatccatg atatttctaa tattttagtt gatatgtata tgaaagggta ctatttgaac 3300
tctcttactc tgtataaagg ttggatcatc cttaaagtgg gtctatttaa ttttattgct 3360
tcttacagat aaaaaaaaaa ttatgagttg gtttgataaa atattgaagg atttaaaata 3420
ataataaata ataaataaca tataatatat gtatataaat ttattataat ataacattta 3480
tctataaaaa agtaaatatt gtcataaatc tatacaatcg tttagccttg ctggacgact 3540
ctcaattatt taaacgagag taaacatatt tgactttttg gttatttaac aaattattat 3600
ttaacactat atgaaatttt ttttttttat cagcaaagaa ataaaattaa attaagaagg 3660
acaatggtgt gtcccaatcc ttatacaacc aacttccaca agaaagtcaa gtcagagaca 3720
acaaaaaaac aagcaaagga aattttttaa tttgagttgt cttgtttgct gcataattta 3780
tgcagtaaaa cactacacat aaccctttta gcagtagagc aatggttgac cgtgtgctta 3840
gcttctttta ttttattttt ttatcagcaa agaataaata aaataaaatg agacacttca 3900
gggatgtttc aacaagcttt gctctagact ggaattcgtc gacgagctcc ctatagtgag 3960
tcgtattaga ggccgacttg gccaaattcg taatcatggt catagctgtt tcctgtgtga 4020
aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 4080
tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 4140
cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 4200
ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 4260
cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 4320
ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 4380
aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 4440
cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 4500
cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 4560
gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 4620
tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 4680
cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 4740
ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 4800
gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 4860
gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 4920
accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 4980
ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 5040
tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 5100
aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 5160
taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 5220
gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 5280
agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 5340
cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 5400
tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 5460
gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 5520
agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 5580
gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 5640
atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 5700
gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 5760
tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 5820
atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 5880
agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 5940
gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 6000
cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 6060
tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 6120
ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg 6180
gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 6240
cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 6300
aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 6360
cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 6420
ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 6480
aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 6540
ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacaaaa 6600
tattaacgtt tacaatttcc cattcgccat tcaggctgcg caactgttgg gaagggcgat 6660
<210>40
<211>1437
<212>DNA
<213>人工序列
<220>
<223>GM GY1 gene
<400>40
atgggttcca gagagcagcc tcagcaaaac gagtgccaga tccaaaaact caatgccctc 60
aaaccggata accgtataga gtcagaagga gggctcattg agacatggaa ccctaacaac 120
aagccattcc agtgtgccgg tgttgccctc tctcgctgca ccctcaaccg caacgccctt 180
cgtagacctt cctacaccaa cggtccccag gaaatctaca tccaacaagg taagggtatt 240
tttggcatga tatacccggg ttgtcctagc acatttgaag agcctcaaca acctcaacaa 300
agaggacaaa gcagcagacc acaagaccgt caccagaaga tctataactt cagagagggt 360
gatttgatcg cagtgcctac tggtgttgca tggtggatgt acaacaatga agacactcct 420
gttgttgccg tttctattat tgacaccaac agcttggaga accagctcga ccagatgcct 480
aggagattct atcttgctgg gaaccaagag caagagtttc taaaatatca gcaagagcaa 540
ggaggtcatc aaagccagaa aggaaagcat cagcaagaag aagaaaacga aggaggcagc 600
atattgagtg gcttcaccct ggaattcttg gaacatgcat tcagcgtgga caagcagata 660
gcgaaaaacc tacaaggaga gaacgaaggg gaagacaagg gagccattgt gacagtgaaa 720
ggaggtctga gcgtgttaaa accacccacg gacgagcagc aacaaagacc ccaggaagag 780
gaagaagaag aagaggatga gaagccacag tgcaagggta aagacaaaca ctgccaacgc 840
ccccgaggaa gccaaagcaa aagcagaaga aatggcattg acgagaccat atgcaccatg 900
agacttcgcc acaacattgg ccagacttca tcacctgaca tctacaaccc tcaagccggt 960
agcgtcacaa ccgccaccag ccttgacttc ccagccctct cgtggctcag actcagtgct 1020
gagtttggat ctctccgcaa gaatgcaatg ttcgtgccac actacaacct gaacgcgaac 1080
agcataatat acgcattgaa tggacgggca ttgatacaag tggtgaattg caacggtgag 1140
agagtgtttg atggagagct gcaagaggga cgggtgctga tcgtgccaca aaactttgtg 1200
gtggctgcaa gatcacagag tgacaacttc gagtatgtgt cattcaagac caatgataca 1260
cccatgatcg gcactcttgc aggggcaaac tcattgttga acgcattacc agaggaagtg 1320
attcagcaca ctttcaacct aaaaagccag caggccaggc agataaagaa caacaaccct 1380
ttcaagttcc tggttccacc tcaggagtct cagaagagag ctgtggctta gggtacc 1437
<210>41
<211>204
<212>DNA
<213>人工序列
<220>
<223>synthetic BHL8 gene
<400>41
atggctaaga ttaagtgcac gtggcctgag ctggtgggca agaccgtgga gaaagccaag 60
aagatgatca tgaaggacaa gccagaggcg aagatcatgg ttctgccagt tgggaccaaa 120
gtgaccggtg aatggaagat ggatcgcgtc cgcctctggg tcgataagaa ggacaagatc 180
gccaagactc cgaagtgcgg ctag 204
<210>42
<211>1701
<212>DNA
<213>人工序列
<220>
<223>GY1-BHL8 fusion product
<400>42
atggccaagc tagttttttc cctttgtttt ctgcttttca gtggctgctg cttcgctttc 60
agttccagag agcagcctca gcaaaacgag tgccagatcc aaaaactcaa tgccctcaaa 120
ccggataacc gtatagagtc agaaggaggg ctcattgaga catggaaccc taacaacaag 180
ccattccagt gtgccggtgt tgccctctct cgctgcaccc tcaaccgcaa cgcccttcgt 240
agaccttcct acaccaacgg tccccaggaa atctacatcc aacaaggtaa gggtattttt 300
ggcatgatat acccgggttg tcctagcaca tttgaagagc ctcagcaacc tcaacaaaga 360
ggacaaagca gcagaccaca agaccgtcac cagaagatct ataacttcag agagggtgat 420
ttgatcgcag tgcctactgg tgttgcatgg tggatgtaca acaatgaaga cactcctgtt 480
gttgccgttt ctattattga caccaacagc ttggagaacc agctcgacca gatgcctagg 540
agattctatc ttgctgggaa ccaagagcaa gagtttctaa aatatcagca agagcaagga 600
ggtcatcaaa gccagaaagg aaagcatcag caagaagaag aaaacgaagg aggcagcata 660
ttgagtggct tcaccctgga attcttggaa catgcattca gcgtggacaa gcagatagcg 720
aaaaacctac aaggagagaa cgaaggggaa gacaagggag ccattgtgac agtgaaagga 780
ggtctgagcg tgttaaaacc acccacggac gagcagcaac aaagacccca ggaagaggaa 840
gaagaagaag aggatgagaa gccacagtgc aagggtaaag acaaacactg ccaacgcccc 900
cgaggaagcc aaagcaaaag cagaagaaat ggcattgacg agaccatatg caccatgaga 960
cttcgccaca acattggcca gacttcatca cctgacatct acaaccctca agccggtagc 1020
gtcacaaccg ccaccagcct tgacttccca gccctctcgt ggctcagact cagtgctgag 1080
tttggatctc tccgcaagaa tgcaatgttc gtgccacact acaacctgaa cgcgaacagc 1140
ataatatacg cattgaatgg acgggcattg atacaagtgg tgaattgcaa cggtgagaga 1200
gtgtttgatg gagagctgca agagggacgg gtgctgatcg tgccacaaaa ctttgtggtg 1260
gctgcaagat cacagagtga caacttcgag tatgtgtcat tcaagaccaa tgatacaccc 1320
atgatcggca ctcttgcagg ggcaaactca ttgttgaacg cattaccaga ggaagtgatt 1380
cagcacactt tcaacctaaa aagccagcag gccaggcaga taaagaacaa caaccctttc 1440
aagttcctgg ttccacctca ggagtctcag aagagagctg tggctggtgg tggatctatg 1500
gctaagatta agtgcacgtg gcctgagctg gtgggcaaga ccgtggagaa agccaagaag 1560
atgatcatga aggacaagcc agaggcgaag atcatggttc tgccagttgg gaccaaagtg 1620
accggtgaat ggaagatgga tcgcgtccgc ctctgggtcg ataagaagga caagatcgcc 1680
aagactccga agtgcggcta g 1701
<210>43
<211>6493
<212>DNA
<213>人工序列
<220>
<223>质粒pZBL133
<400>43
cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 60
taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 120
agctgacttg gtcagcggcc gctctagaac tagtggatcc tagcctaagt acgtactcaa 180
aatgccaaca aataaaaaaa aagttgcttt aataatgcca aaacaaatta ataaaacact 240
tacaacaccg gatttttttt aattaaaatg tgccatttag gataaatagt taatattttt 300
aataattatt taaaaagccg tatctactaa aatgattttt atttggttga aaatattaat 360
atgtttaaat caacacaatc tatcaaaatt aaactaaaaa aaaaataagt gtacgtggtt 420
aacattagta cagtaatata agaggaaaat gagaaattaa gaaattgaaa gcgagtctaa 480
tttttaaatt atgaacctgc atatataaaa ggaaagaaag aatccaggaa gaaaagaaat 540
gaaaccatgc atggtcccct cgtcatcacg agtttctgcc atttgcaata gaaacactga 600
aacacctttc tctttgtcac ttaattgaga tgccgaagcc acctcacacc atgaacttca 660
tgaggtgtag cacccaaggc ttccatagcc atgcatactg aagaatgtct caagctcagc 720
accctacttc tgtgacgtgt ccctcattca ccttcctctc ttccctataa ataaccacgc 780
ctcaggttct ccgcttcaca actcaaacat tctctccatt ggtccttaaa cactcatcag 840
tcatcaccat ggccaagcta gttttttccc tttgttttct gcttttcagt ggctgctgct 900
tcgctttcag ttccagagag cagcctcagc aaaacgagtg ccagatccaa aaactcaatg 960
ccctcaaacc ggataaccgt atagagtcag aaggagggct cattgagaca tggaacccta 1020
acaacaagcc attccagtgt gccggtgttg ccctctctcg ctgcaccctc aaccgcaacg 1080
cccttcgtag accttcctac accaacggtc cccaggaaat ctacatccaa caaggtaagg 1140
gtatttttgg catgatatac ccgggttgtc ctagcacatt tgaagagcct cagcaacctc 1200
aacaaagagg acaaagcagc agaccacaag accgtcacca gaagatctat aacttcagag 1260
agggtgattt gatcgcagtg cctactggtg ttgcatggtg gatgtacaac aatgaagaca 1320
ctcctgttgt tgccgtttct attattgaca ccaacagctt ggagaaccag ctcgaccaga 1380
tgcctaggag attctatctt gctgggaacc aagagcaaga gtttctaaaa tatcagcaag 1440
agcaaggagg tcatcaaagc cagaaaggaa agcatcagca agaagaagaa aacgaaggag 1500
gcagcatatt gagtggcttc accctggaat tcttggaaca tgcattcagc gtggacaagc 1560
agatagcgaa aaacctacaa ggagagaacg aaggggaaga caagggagcc attgtgacag 1620
tgaaaggagg tctgagcgtg ttaaaaccac ccacggacga gcagcaacaa agaccccagg 1680
aagaggaaga agaagaagag gatgagaagc cacagtgcaa gggtaaagac aaacactgcc 1740
aacgcccccg aggaagccaa agcaaaagca gaagaaatgg cattgacgag accatatgca 1800
ccatgagact tcgccacaac attggccaga cttcatcacc tgacatctac aaccctcaag 1860
ccggtagcgt cacaaccgcc accagccttg acttcccagc cctctcgtgg ctcagactca 1920
gtgctgagtt tggatctctc cgcaagaatg caatgttcgt gccacactac aacctgaacg 1980
cgaacagcat aatatacgca ttgaatggac gggcattgat acaagtggtg aattgcaacg 2040
gtgagagagt gtttgatgga gagctgcaag agggacgggt gctgatcgtg ccacaaaact 2100
ttgtggtggc tgcaagatca cagagtgaca acttcgagta tgtgtcattc aagaccaatg 2160
atacacccat gatcggcact cttgcagggg caaactcatt gttgaacgca ttaccagagg 2220
aagtgattca gcacactttc aacctaaaaa gccagcaggc caggcagata aagaacaaca 2280
accctttcaa gttcctggtt ccacctcagg agtctcagaa gagagctgtg gctggtggtg 2340
gatctatggc taagattaag tgcacgtggc ctgagctggt gggcaagacc gtggagaaag 2400
ccaagaagat gatcatgaag gacaagccag aggcgaagat catggttctg ccagttggga 2460
ccaaagtgac cggtgaatgg aagatggatc gcgtccgcct ctgggtcgat aagaaggaca 2520
agatcgccaa gactccgaag tgcggctagg gtacctctag acatggccac gtgcatgaag 2580
tatgaactaa aatgcatgta ggtgtaagag ctcatggaga gcatggaata ttgtatccga 2640
ccatgtaaca gtataataac tgagctccat ctcacttctt ctatgaataa acaaaggatg 2700
ttatgatata ttaacactct atctatgcac cttattgttc tatgataaat ttcctcttat 2760
tattataaat catctgaatc gtgacggctt atggaatgct tcaaatagta caaaaacaaa 2820
tgtgtactat aagactttct aaacaattct aactttagca ttgtgaacga gacataagtg 2880
ttaagaagac ataacaatta taatggaaga agtttgtctc catttatata ttatatatta 2940
cccacttatg tattatatta ggatgttaag gagacataac aattataaag agagaagttt 3000
gtatccattt atatattata tactacccat ttatatatta tacttatcca cttatttaat 3060
gtctttataa ggtttgatcc atgatatttc taatatttta gttgatatgt atatgaaagg 3120
gtactatttg aactctctta ctctgtataa aggttggatc atccttaaag tgggtctatt 3180
taattttatt gcttcttaca gataaaaaaa aaattatgag ttggtttgat aaaatattga 3240
aggatttaaa ataataataa ataataaata acatataata tatgtatata aatttattat 3300
aatataacat ttatctataa aaaagtaaat attgtcataa atctatacaa tcgtttagcc 3360
ttgctggacg actctcaatt atttaaacga gagtaaacat atttgacttt ttggttattt 3420
aacaaattat tatttaacac tatatgaaat tttttttttt tatcagcaaa gaaataaaat 3480
taaattaaga aggacaatgg tgtgtcccaa tccttataca accaacttcc acaagaaagt 3540
caagtcagag acaacaaaaa aacaagcaaa ggaaattttt taatttgagt tgtcttgttt 3600
gctgcataat ttatgcagta aaacactaca cataaccctt ttagcagtag agcaatggtt 3660
gaccgtgtgc ttagcttctt ttattttatt tttttatcag caaagaataa ataaaataaa 3720
atgagacact tcagggatgt ttcaacaagc tttgctctag actggaattc gtcgacgagc 3780
tccctatagt gagtcgtatt agaggccgac ttggccaaat tcgtaatcat ggtcatagct 3840
gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 3900
aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 3960
actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 4020
cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 4080
gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 4140
atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 4200
caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 4260
gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 4320
ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 4380
cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 4440
taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 4500
cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 4560
acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 4620
aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 4680
atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 4740
atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 4800
gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 4860
gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 4920
ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 4980
ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 5040
tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 5100
accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 5160
atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 5220
cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 5280
tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 5340
tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 5400
gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 5460
agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 5520
aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 5580
gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 5640
tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 5700
gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 5760
tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 5820
aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 5880
catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 5940
acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg 6000
cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 6060
cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 6120
ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct 6180
cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 6240
ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 6300
tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 6360
ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa 6420
aatattaaca aaatattaac gtttacaatt tcccattcgc cattcaggct gcgcaactgt 6480
tgggaagggc gat 6493
<210>44
<211>6472
<212>DNA
<213>人工序列
<220>
<223>质粒pKS238
<400>44
ctagactgga attcgtcgac gagctcccta tagtgagtcg tattagaggc cgacttggcc 60
aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc 120
acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta 180
actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 240
gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 300
cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 360
tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 420
gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 480
ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 540
aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 600
tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 660
ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 720
gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 780
tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 840
caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 900
ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt 960
cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 1020
ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 1080
cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 1140
gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 1200
aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 1260
acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 1320
gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 1380
cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 1440
cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 1500
tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 1560
cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 1620
gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 1680
cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 1740
ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 1800
gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 1860
taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 1920
gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 1980
acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 2040
aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 2100
cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 2160
atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 2220
gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag 2280
cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt 2340
tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt 2400
ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg 2460
tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt 2520
taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt 2580
tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca 2640
aaaatttaac gcgaatttta acaaaatatt aacaaaatat taacgtttac aatttcccat 2700
tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta 2760
cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt 2820
tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgacttggtc agcggccgct 2880
ctagaactag tggatcctag cctaagtacg tactcaaaat gccaacaaat aaaaaaaaag 2940
ttgctttaat aatgccaaaa caaattaata aaacacttac aacaccggat tttttttaat 3000
taaaatgtgc catttaggat aaatagttaa tatttttaat aattatttaa aaagccgtat 3060
ctactaaaat gatttttatt tggttgaaaa tattaatatg tttaaatcaa cacaatctat 3120
caaaattaaa ctaaaaaaaa aataagtgta cgtggttaac attagtacag taatataaga 3180
ggaaaatgag aaattaagaa attgaaagcg agtctaattt ttaaattatg aacctgcata 3240
tataaaagga aagaaagaat ccaggaagaa aagaaatgaa accatgcatg gtcccctcgt 3300
catcacgagt ttctgccatt tgcaatagaa acactgaaac acctttctct ttgtcactta 3360
attgagatgc cgaagccacc tcacaccatg aacttcatga ggtgtagcac ccaaggcttc 3420
catagccatg catactgaag aatgtctcaa gctcagcacc ctacttctgt gacgtgtccc 3480
tcattcacct tcctctcttc cctataaata accacgcctc aggttctccg cttcacaact 3540
caaacattct ctccattggt ccttaaacac tcatcagtca tcaccatggc caagctagtt 3600
ttttcccttt gttttctgct tttcagtggc tgctgcttcg ctttcagttc cagagagcag 3660
cctcagcaaa acgagtgcca gatccaaaaa ctcaatgccc tcaaaccgga taaccgtata 3720
gagtcagaag gagggctcat tgagacatgg aaccctaaca acaagccatt ccagtgtgcc 3780
ggtgttgccc tctctcgctg caccctcaac cgcaacgccc ttcgtagacc ttcctacacc 3840
aacggtcccc aggaaatcta catccaacaa ggtaagggta tttttggcat gatatacccg 3900
ggttgtccta gcacatttga agagcctcag caacctcaac aaagaggaca aagcagcaga 3960
ccacaagacc gtcaccagaa gatctataac ttcagagagg gtgatttgat cgcagtgcct 4020
actggtgttg catggtggat gtacaacaat gaagacactc ctgttgttgc cgtttctatt 4080
attgacacca acagcttgga gaaccagctc gaccagatgc ctaggagatt ctatcttgct 4140
gggaaccaag agcaagagtt tctaaaatat cagcaagagc aaggaggtca tcaaagccag 4200
aaaggaaagc atcagcaaga agaagaaaac gaaggaggca gcatattgag tggcttcacc 4260
ctggaattct tggaacatgc attcagcgtg gacaagcaga tagcgaaaaa cctacaagga 4320
gagaacgaag gggaagacaa gggagccatt gtgacagtga aaggaggtct gagcgtgtta 4380
aaaccaccca cggacgagca gcaacaaaga ccccaggaag aggaagaaga agaagaggat 4440
gagaagccac agtgcaaggg taaagacaaa cactgccaac gcccccgagg aagccaaagc 4500
aaaagcagaa gaaatggcat tgacgagacc atatgcacca tgagacttcg ccacaacatt 4560
ggccagactt catcacctga catctacaac cctcaagccg gtagcgtcac aaccgccacc 4620
agccttgact tcccagccct ctcgtggctc agactcagtg ctgagtttgg atctctccgc 4680
aagaatgcaa tgttcgtgcc acactacaac ctgaacgcga acagcataat atacgcattg 4740
aatggacggg cattgataca agtggtgaat tgcaacggtg agagagtgtt tgatggagag 4800
ctgcaagagg gacgggtgct gatcgtgcca caaaactttg tggtggctgc aagatcacag 4860
agtgacaact tcgagtatgt gtcattcaag accaatgata cacccatgat cggcactctt 4920
gcaggggcaa actcattgtt gaacgcatta ccagaggaag tgattcagca cactttcaac 4980
ctaaaaagcc agcaggccag gcagataaag aacaacaacc ctttcaagtt cctggttcca 5040
cctcaggagt ctcagaagag agctgtggct ggtggtggat ctatggctaa gattaagtgc 5100
acgtggcctg agctggtggg caagaccgtg gagaaagcca agaagatgat catgaaggac 5160
aagccagagg cgaagatcat ggttctgcca gttgggacca aagtgaccgg tgaatggaag 5220
atggatcgcg tccgcctctg ggtcgataag aaggacaaga tcgccaagac tccgaagtgc 5280
ggctagggta cctctagggt ggcggccgca agtatgaact aaaatgcatg taggtgtaag 5340
agctcatgga gagcatggaa tattgtatcc gaccatgtaa cagtataata actgagctcc 5400
atctcacttc ttctatgaat aaacaaagga tgttatgata tattaacact ctatctatgc 5460
accttattgt tctatgataa atttcctctt attattataa atcatctgaa tcgtgacggc 5520
ttatggaatg cttcaaatag tacaaaaaca aatgtgtact ataagacttt ctaaacaatt 5580
ctaaccttag cattgtgaac gagacataag tgttaagaag acataacaat tataatggaa 5640
gaagtttgtc tccatttata tattatatat tacccactta tgtattatat taggatgtta 5700
aggagacata acaattataa agagagaagt ttgtatccat ttatatatta tatactaccc 5760
atttatatat tatacttatc cacttattta atgtctttat aaggtttgat ccatgatatt 5820
tctaatattt tagttgatat gtatatgaaa gggtactatt tgaactctct tactctgtat 5880
aaaggttgga tcatccttaa agtgggtcta tttaatttta ttgcttctta cagataaaaa 5940
aaaaattatg agttggtttg ataaaatatt gaaggattta aaataataat aaataacata 6000
taatatatgt atataaattt attataatat aacatttatc tataaaaaag taaatattgt 6060
cataaatcta tacaatcgtt tagccttgct ggacgaatct caattattta aacgagagta 6120
aacatatttg actttttggt tatttaacaa attattattt aacactatat gaaatttttt 6180
tttttatcag caaagaataa aattaaatta agaaggacaa tggtgtccca atccttatac 6240
aaccaacttc cacaagaaag tcaagtcaga gacaacaaaa aaacaagcaa aggaaatttt 6300
ttaatttgag ttgtcttgtt tgctgcataa tttatgcagt aaaacactac acataaccct 6360
tttagcagta gagcaatggt tgaccgtgtg cttagcttct tttattttat ttttttatca 6420
gcaaagaata aataaaataa aatgagacac ttcagggatg tttcaacaag ct 6472
<210>45
<211>6259
<212>DNA
<213>人工序列
<220>
<223>质粒pKS240
<400>45
ctctagggtg gcggccgcaa gtatgaacta aaatgcatgt aggtgtaaga gctcatggag 60
agcatggaat attgtatccg accatgtaac agtataataa ctgagctcca tctcacttct 120
tctatgaata aacaaaggat gttatgatat attaacactc tatctatgca ccttattgtt 180
ctatgataaa tttcctctta ttattataaa tcatctgaat cgtgacggct tatggaatgc 240
ttcaaatagt acaaaaacaa atgtgtacta taagactttc taaacaattc taaccttagc 300
attgtgaacg agacataagt gttaagaaga cataacaatt ataatggaag aagtttgtct 360
ccatttatat attatatatt acccacttat gtattatatt aggatgttaa ggagacataa 420
caattataaa gagagaagtt tgtatccatt tatatattat atactaccca tttatatatt 480
atacttatcc acttatttaa tgtctttata aggtttgatc catgatattt ctaatatttt 540
agttgatatg tatatgaaag ggtactattt gaactctctt actctgtata aaggttggat 600
catccttaaa gtgggtctat ttaattttat tgcttcttac agataaaaaa aaaattatga 660
gttggtttga taaaatattg aaggatttaa aataataata aataacatat aatatatgta 720
tataaattta ttataatata acatttatct ataaaaaagt aaatattgtc ataaatctat 780
acaatcgttt agccttgctg gacgaatctc aattatttaa acgagagtaa acatatttga 840
ctttttggtt atttaacaaa ttattattta acactatatg aaattttttt ttttatcagc 900
aaagaataaa attaaattaa gaaggacaat ggtgtcccaa tccttataca accaacttcc 960
acaagaaagt caagtcagag acaacaaaaa aacaagcaaa ggaaattttt taatttgagt 1020
tgtcttgttt gctgcataat ttatgcagta aaacactaca cataaccctt ttagcagtag 1080
agcaatggtt gaccgtgtgc ttagcttctt ttattttatt tttttatcag caaagaataa 1140
ataaaataaa atgagacact tcagggatgt ttcaacaagc tctagactgg aattcgtcga 1200
cgagctccct atagtgagtc gtattagagg ccgacttggc caaattcgta atcatggtca 1260
tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 1320
agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 1380
cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 1440
caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 1500
tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 1560
cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 1620
aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 1680
gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 1740
agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 1800
cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 1860
cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 1920
ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 1980
gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 2040
tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 2100
acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 2160
tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 2220
attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 2280
gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 2340
ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 2400
taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 2460
ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 2520
ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 2580
gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 2640
ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 2700
gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 2760
tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 2820
atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 2880
gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 2940
tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 3000
atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 3060
agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 3120
ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 3180
tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 3240
aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 3300
tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 3360
aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt 3420
agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 3480
agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 3540
tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg 3600
cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga 3660
tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 3720
caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg 3780
ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt 3840
aacaaaatat taacaaaata ttaacgttta caatttccca ttcgccattc aggctgcgca 3900
actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg 3960
gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta 4020
aaacgacggc cagtgccaag ctgacttggt cagcggccgc tctagaacta gtggatccta 4080
gcctaagtac gtactcaaaa tgccaacaaa taaaaaaaaa gttgctttaa taatgccaaa 4140
acaaattaat aaaacactta caacaccgga ttttttttaa ttaaaatgtg ccatttagga 4200
taaatagtta atatttttaa taattattta aaaagccgta tctactaaaa tgatttttat 4260
ttggttgaaa atattaatat gtttaaatca acacaatcta tcaaaattaa actaaaaaaa 4320
aaataagtgt acgtggttaa cattagtaca gtaatataag aggaaaatga gaaattaaga 4380
aattgaaagc gagtctaatt tttaaattat gaacctgcat atataaaagg aaagaaagaa 4440
tccaggaaga aaagaaatga aaccatgcat ggtcccctcg tcatcacgag tttctgccat 4500
ttgcaataga aacactgaaa cacctttctc tttgtcactt aattgagatg ccgaagccac 4560
ctcacaccat gaacttcatg aggtgtagca cccaaggctt ccatagccat gcatactgaa 4620
gaatgtctca agctcagcac cctacttctg tgacgtgtcc ctcattcacc ttcctctctt 4680
ccctataaat aaccacgcct caggttctcc gcttcacaac tcaaacattc tctccattgg 4740
tccttaaaca ctcatcagtc atcaccatgg ccaagctagt tttttccctt tgttttctgc 4800
ttttcagtgg ctgctgcttc gctttcagtt ccagagagca gcctcagcaa aacgagtgcc 4860
agatccaaaa actcaatgcc ctcaaaccgg ataaccgtat agagtcagaa ggagggctca 4920
ttgagacatg gaaccctaac aacaagccat tccagtgtgc cggtgttgcc ctctctcgct 4980
gcaccctcaa ccgcaacgcc cttcgtagac cttcctacac caacggtccc caggaaatct 5040
acatccaaca aggtaagggt atttttggca tgatataccc gggttgtcct agcacatttg 5100
aagagcctca gcaacctcaa caaagaggac aaagcagcag accacaagac cgtcaccaga 5160
agatctataa cttcagagag ggtgatttga tcgcagtgcc tactggtgtt gcatggtgga 5220
tgtacaacaa tgaagacact cctgttgttg ccgtttctat tattgacacc aacagcttgg 5280
agaaccagct cgaccagatg cctaggagat tctatcttgc tgggaaccaa gagcaagagt 5340
ttctaaaata tcagcaagag caaggaggtc atcaaagcca gaaaggaaag catcagcaag 5400
aagaagaaaa cgaaggaggc agcatattga gtggcttcac cctggaattc ttggaacatg 5460
cattcagcgt ggacaagcag atagcgaaaa acctacaagg agagaacgaa ggggaagaca 5520
agggagccat tgtgacagtg aaaggaggtc tgagcgtgtt aaaaccaccc acggacgagc 5580
agcaacaaag accccaggaa gaggaagaag aagaagagga tgagaagcca cagtgcaagg 5640
gtaaagacaa acactgccaa cgcccccgag gaagccaaag caaaagcaga agaaatggca 5700
ttgacgagac catatgcacc atgagacttc gccacaacat tggccagact tcatcacctg 5760
acatctacaa ccctcaagcc ggtagcgtca caaccgccac cagccttgac ttcccagccc 5820
tctcgtggct cagactcagt gctgagtttg gatctctccg caagaatgca atgttcgtgc 5880
cacactacaa cctgaacgcg aacagcataa tatacgcatt gaatggacgg gcattgatac 5940
aagtggtgaa ttgcaacggt gagagagtgt ttgatggaga gctgcaagag ggacgggtgc 6000
tgatcgtgcc acaaaacttt gtggtggctg caagatcaca gagtgacaac ttcgagtatg 6060
tgtcattcaa gaccaatgat acacccatga tcggcactct tgcaggggca aactcattgt 6120
tgaacgcatt accagaggaa gtgattcagc acactttcaa cctaaaaagc cagcaggcca 6180
ggcagataaa gaacaacaac cctttcaagt tcctggttcc acctcaggag tctcagaaga 6240
gagctgtggc ttagggtac 6259
<210>46
<211>5267
<212>DNA
<213>人工序列
<220>
<223>质粒pKS120
<400>46
atctgatcaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 60
ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 120
cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 180
gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 240
tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 300
agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 360
tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 420
cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg 480
ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 540
ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 600
ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 660
ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 720
cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 780
gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 840
atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 900
ttttggtcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc 960
gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt 1020
gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg 1080
ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata 1140
tggacatatt gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca 1200
tacgatttag gtgacactat agaacggcgc gccaagcttg gatccgtcga cggcgcgccc 1260
gatcatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc 1320
ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg 1380
gctttgttag cagccggatc gatccaagct gtacctcact attcctttgc cctcggacga 1440
gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg 1500
gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt 1560
gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga 1620
tagagttggt caagaccaat gcggagcata tacgcccgga gccgcggcga tcctgcaagc 1680
tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc 1740
cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca 1800
atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc 1860
acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc 1920
gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca 1980
gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat 2040
gggccgaacc cgctcgtctg gctaagatcg gccgcagcga tcgcatccat agcctccgcg 2100
accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt 2160
gcacggcggg agatgcaata ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc 2220
ggaatcggga gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca 2280
tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca 2340
cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc 2400
agaaacttct cgacagacgt cgcggtgagt tcaggctttt ccatgggtat atctccttct 2460
taaagttaaa caaaattatt tctagaggga aaccgttgtg gtctccctat agtgagtcgt 2520
attaatttcg cgggatcgag atcgatccaa ttccaatccc acaaaaatct gagcttaaca 2580
gcacagttgc tcctctcaga gcagaatcgg gtattcaaca ccctcatatc aactactacg 2640
ttgtgtataa cggtccacat gccggtatat acgatgactg gggttgtaca aaggcggcaa 2700
caaacggcgt tcccggagtt gcacacaaga aatttgccac tattacagag gcaagagcag 2760
cagctgacgc gtacacaaca agtcagcaaa cagacaggtt gaacttcatc cccaaaggag 2820
aagctcaact caagcccaag agctttgcta aggccctaac aagcccacca aagcaaaaag 2880
cccactggct cacgctagga accaaaaggc ccagcagtga tccagcccca aaagagatct 2940
cctttgcccc ggagattaca atggacgatt tcctctatct ttacgatcta ggaaggaagt 3000
tcgaaggtga aggtgacgac actatgttca ccactgataa tgagaaggtt agcctcttca 3060
atttcagaaa gaatgctgac ccacagatgg ttagagaggc ctacgcagca ggtctcatca 3120
agacgatcta cccgagtaac aatctccagg agatcaaata ccttcccaag aaggttaaag 3180
atgcagtcaa aagattcagg actaattgca tcaagaacac agagaaagac atatttctca 3240
agatcagaag tactattcca gtatggacga ttcaaggctt gcttcataaa ccaaggcaag 3300
taatagagat tggagtctct aaaaaggtag ttcctactga atctaaggcc atgcatggag 3360
tctaagattc aaatcgagga tctaacagaa ctcgccgtga agactggcga acagttcata 3420
cagagtcttt tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac 3480
actctggtct actccaaaaa tgtcaaagat acagtctcag aagaccaaag ggctattgag 3540
acttttcaac aaaggataat ttcgggaaac ctcctcggat tccattgccc agctatctgt 3600
cacttcatcg aaaggacagt agaaaaggaa ggtggctcct acaaatgcca tcattgcgat 3660
aaaggaaagg ctatcattca agatgcctct gccgacagtg gtcccaaaga tggaccccca 3720
cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat 3780
tgatgtgaca tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac 3840
ccttcctcta tataaggaag ttcatttcat ttggagagga cacgctcgag ctcatttctc 3900
tattacttca gccataacaa aagaactctt ttctcttctt attaaaccat gaaaaagcct 3960
gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac 4020
ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt 4080
ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat 4140
cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc 4200
gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct 4260
gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg 4320
gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac 4380
actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact 4440
gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg 4500
gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc 4560
ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat 4620
tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag 4680
cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg 4740
tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat 4800
gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc 4860
gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta 4920
ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga atagtgaggt 4980
acctaaagaa ggagtgcgtc gaagcagatc gttcaaacat ttggcaataa agtttcttaa 5040
gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta 5100
agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta 5160
gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg 5220
ataaattatc gcgcgcggtg tcatctatgt tactagatcg atgtcga 5267
<210>47
<211>8643
<212>DNA
<213>人工序列
<220>
<223>质粒pKS242
<400>47
tcgacggcgc gcccgatcat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga 60
cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca gcagccaact 120
cagcttcctt tcgggctttg ttagcagccg gatcgatcca agctgtacct cactattcct 180
ttgccctcgg acgagtgctg gggcgtcggt ttccactatc ggcgagtact tctacacagc 240
catcggtcca gacggccgcg cttctgcggg cgatttgtgt acgcccgaca gtcccggctc 300
cggatcggac gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg aaattgccgt 360
caaccaagct ctgatagagt tggtcaagac caatgcggag catatacgcc cggagccgcg 420
gcgatcctgc aagctccgga tgcctccgct cgaagtagcg cgtctgctgc tccatacaag 480
ccaaccacgg cctccagaag aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg 540
cctcgctcca gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca ttgttggagc 600
cgaaatccgc gtgcacgagg tgccggactt cggggcagtc ctcggcccaa agcatcagct 660
catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt tgccagtgat 720
acacatgggg atcagcaatc gcgcatatga aatcacgcca tgtagtgtat tgaccgattc 780
cttgcggtcc gaatgggccg aacccgctcg tctggctaag atcggccgca gcgatcgcat 840
ccatagcctc cgcgaccggc tgcagaacag cgggcagttc ggtttcaggc aggtcttgca 900
acgtgacacc ctgtgcacgg cgggagatgc aataggtcag gctctcgctg aattccccaa 960
tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa gtgccgataa acataacgat 1020
ctttgtagaa accatcggcg cagctattta cccgcaggac atatccacgc cctcctacat 1080
cgaagctgaa agcacgagat tcttcgccct ccgagagctg catcaggtcg gagacgctgt 1140
cgaacttttc gatcagaaac ttctcgacag acgtcgcggt gagttcaggc ttttccatgg 1200
gtatatctcc ttcttaaagt taaacaaaat tatttctaga gggaaaccgt tgtggtctcc 1260
ctatagtgag tcgtattaat ttcgcgggat cgagatcgat ccaattccaa tcccacaaaa 1320
atctgagctt aacagcacag ttgctcctct cagagcagaa tcgggtattc aacaccctca 1380
tatcaactac tacgttgtgt ataacggtcc acatgccggt atatacgatg actggggttg 1440
tacaaaggcg gcaacaaacg gcgttcccgg agttgcacac aagaaatttg ccactattac 1500
agaggcaaga gcagcagctg acgcgtacac aacaagtcag caaacagaca ggttgaactt 1560
catccccaaa ggagaagctc aactcaagcc caagagcttt gctaaggccc taacaagccc 1620
accaaagcaa aaagcccact ggctcacgct aggaaccaaa aggcccagca gtgatccagc 1680
cccaaaagag atctcctttg ccccggagat tacaatggac gatttcctct atctttacga 1740
tctaggaagg aagttcgaag gtgaaggtga cgacactatg ttcaccactg ataatgagaa 1800
ggttagcctc ttcaatttca gaaagaatgc tgacccacag atggttagag aggcctacgc 1860
agcaggtctc atcaagacga tctacccgag taacaatctc caggagatca aataccttcc 1920
caagaaggtt aaagatgcag tcaaaagatt caggactaat tgcatcaaga acacagagaa 1980
agacatattt ctcaagatca gaagtactat tccagtatgg acgattcaag gcttgcttca 2040
taaaccaagg caagtaatag agattggagt ctctaaaaag gtagttccta ctgaatctaa 2100
ggccatgcat ggagtctaag attcaaatcg aggatctaac agaactcgcc gtgaagactg 2160
gcgaacagtt catacagagt cttttacgac tcaatgacaa gaagaaaatc ttcgtcaaca 2220
tggtggagca cgacactctg gtctactcca aaaatgtcaa agatacagtc tcagaagacc 2280
aaagggctat tgagactttt caacaaagga taatttcggg aaacctcctc ggattccatt 2340
gcccagctat ctgtcacttc atcgaaagga cagtagaaaa ggaaggtggc tcctacaaat 2400
gccatcattg cgataaagga aaggctatca ttcaagatgc ctctgccgac agtggtccca 2460
aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt 2520
caaagcaagt ggattgatgt gacatctcca ctgacgtaag ggatgacgca caatcccact 2580
atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag aggacacgct 2640
cgagctcatt tctctattac ttcagccata acaaaagaac tcttttctct tcttattaaa 2700
ccatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg 2760
acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg 2820
atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag 2880
atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca 2940
ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt 3000
tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg 3060
atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag 3120
gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt 3180
atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg 3240
agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg 3300
gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg 3360
cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg 3420
cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc 3480
cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg 3540
acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg 3600
gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg 3660
gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa 3720
aggaatagtg aggtacctaa agaaggagtg cgtcgaagca gatcgttcaa acatttggca 3780
ataaagtttc ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct 3840
gttgaattac gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg 3900
ggtttttatg attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata 3960
gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct atgttactag atcgatgtcg 4020
aatctgatca acctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 4080
gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 4140
gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 4200
ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 4260
ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 4320
cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 4380
ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 4440
tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc 4500
gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 4560
tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 4620
gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 4680
tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag 4740
ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 4800
agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 4860
gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 4920
attttggtca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg 4980
cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct 5040
tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc 5100
gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat 5160
atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac 5220
atacgattta ggtgacacta tagaacggcg cgccaagctt ggatcctagc ctaagtacgt 5280
actcaaaatg ccaacaaata aaaaaaaagt tgctttaata atgccaaaac aaattaataa 5340
aacacttaca acaccggatt ttttttaatt aaaatgtgcc atttaggata aatagttaat 5400
atttttaata attatttaaa aagccgtatc tactaaaatg atttttattt ggttgaaaat 5460
attaatatgt ttaaatcaac acaatctatc aaaattaaac taaaaaaaaa ataagtgtac 5520
gtggttaaca ttagtacagt aatataagag gaaaatgaga aattaagaaa ttgaaagcga 5580
gtctaatttt taaattatga acctgcatat ataaaaggaa agaaagaatc caggaagaaa 5640
agaaatgaaa ccatgcatgg tcccctcgtc atcacgagtt tctgccattt gcaatagaaa 5700
cactgaaaca cctttctctt tgtcacttaa ttgagatgcc gaagccacct cacaccatga 5760
acttcatgag gtgtagcacc caaggcttcc atagccatgc atactgaaga atgtctcaag 5820
ctcagcaccc tacttctgtg acgtgtccct cattcacctt cctctcttcc ctataaataa 5880
ccacgcctca ggttctccgc ttcacaactc aaacattctc tccattggtc cttaaacact 5940
catcagtcat caccatggcc aagctagttt tttccctttg ttttctgctt ttcagtggct 6000
gctgcttcgc tttcagttcc agagagcagc ctcagcaaaa cgagtgccag atccaaaaac 6060
tcaatgccct caaaccggat aaccgtatag agtcagaagg agggctcatt gagacatgga 6120
accctaacaa caagccattc cagtgtgccg gtgttgccct ctctcgctgc accctcaacc 6180
gcaacgccct tcgtagacct tcctacacca acggtcccca ggaaatctac atccaacaag 6240
gtaagggtat ttttggcatg atatacccgg gttgtcctag cacatttgaa gagcctcagc 6300
aacctcaaca aagaggacaa agcagcagac cacaagaccg tcaccagaag atctataact 6360
tcagagaggg tgatttgatc gcagtgccta ctggtgttgc atggtggatg tacaacaatg 6420
aagacactcc tgttgttgcc gtttctatta ttgacaccaa cagcttggag aaccagctcg 6480
accagatgcc taggagattc tatcttgctg ggaaccaaga gcaagagttt ctaaaatatc 6540
agcaagagca aggaggtcat caaagccaga aaggaaagca tcagcaagaa gaagaaaacg 6600
aaggaggcag catattgagt ggcttcaccc tggaattctt ggaacatgca ttcagcgtgg 6660
acaagcagat agcgaaaaac ctacaaggag agaacgaagg ggaagacaag ggagccattg 6720
tgacagtgaa aggaggtctg agcgtgttaa aaccacccac ggacgagcag caacaaagac 6780
cccaggaaga ggaagaagaa gaagaggatg agaagccaca gtgcaagggt aaagacaaac 6840
actgccaacg cccccgagga agccaaagca aaagcagaag aaatggcatt gacgagacca 6900
tatgcaccat gagacttcgc cacaacattg gccagacttc atcacctgac atctacaacc 6960
ctcaagccgg tagcgtcaca accgccacca gccttgactt cccagccctc tcgtggctca 7020
gactcagtgc tgagtttgga tctctccgca agaatgcaat gttcgtgcca cactacaacc 7080
tgaacgcgaa cagcataata tacgcattga atggacgggc attgatacaa gtggtgaatt 7140
gcaacggtga gagagtgttt gatggagagc tgcaagaggg acgggtgctg atcgtgccac 7200
aaaactttgt ggtggctgca agatcacaga gtgacaactt cgagtatgtg tcattcaaga 7260
ccaatgatac acccatgatc ggcactcttg caggggcaaa ctcattgttg aacgcattac 7320
cagaggaagt gattcagcac actttcaacc taaaaagcca gcaggccagg cagataaaga 7380
acaacaaccc tttcaagttc ctggttccac ctcaggagtc tcagaagaga gctgtggctt 7440
agggtacctc tagggtggcg gccgcaagta tgaactaaaa tgcatgtagg tgtaagagct 7500
catggagagc atggaatatt gtatccgacc atgtaacagt ataataactg agctccatct 7560
cacttcttct atgaataaac aaaggatgtt atgatatatt aacactctat ctatgcacct 7620
tattgttcta tgataaattt cctcttatta ttataaatca tctgaatcgt gacggcttat 7680
ggaatgcttc aaatagtaca aaaacaaatg tgtactataa gactttctaa acaattctaa 7740
ccttagcatt gtgaacgaga cataagtgtt aagaagacat aacaattata atggaagaag 7800
tttgtctcca tttatatatt atatattacc cacttatgta ttatattagg atgttaagga 7860
gacataacaa ttataaagag agaagtttgt atccatttat atattatata ctacccattt 7920
atatattata cttatccact tatttaatgt ctttataagg tttgatccat gatatttcta 7980
atattttagt tgatatgtat atgaaagggt actatttgaa ctctcttact ctgtataaag 8040
gttggatcat ccttaaagtg ggtctattta attttattgc ttcttacaga taaaaaaaaa 8100
attatgagtt ggtttgataa aatattgaag gatttaaaat aataataaat aacatataat 8160
atatgtatat aaatttatta taatataaca tttatctata aaaaagtaaa tattgtcata 8220
aatctataca atcgtttagc cttgctggac gaatctcaat tatttaaacg agagtaaaca 8280
tatttgactt tttggttatt taacaaatta ttatttaaca ctatatgaaa tttttttttt 8340
tatcagcaaa gaataaaatt aaattaagaa ggacaatggt gtcccaatcc ttatacaacc 8400
aacttccaca agaaagtcaa gtcagagaca acaaaaaaac aagcaaagga aattttttaa 8460
tttgagttgt cttgtttgct gcataattta tgcagtaaaa cactacacat aaccctttta 8520
gcagtagagc aatggttgac cgtgtgctta gcttctttta ttttattttt ttatcagcaa 8580
agaataaata aaataaaatg agacacttca gggatgtttc aacaagctct agactggaat 8640
tcg 8643
<210>48
<211>8720
<212>DNA
<213>人工序列
<220>
<223>质粒pK8349
<400>48
ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacaagctc tagactggaa ttcgtcgacg gcgcgcccga 1200
tcatccggat atagttcctc ctttcagcaa aaaacccctc aagacccgtt tagaggcccc 1260
aaggggttat gctagttatt gctcagcggt ggcagcagcc aactcagctt cctttcgggc 1320
tttgttagca gccggatcga tccaagctgt acctcactat tcctttgccc tcggacgagt 1380
gctggggcgt cggtttccac tatcggcgag tacttctaca cagccatcgg tccagacggc 1440
cgcgcttctg cgggcgattt gtgtacgccc gacagtcccg gctccggatc ggacgattgc 1500
gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca agctctgata 1560
gagttggtca agaccaatgc ggagcatata cgcccggagc cgcggcgatc ctgcaagctc 1620
cggatgcctc cgctcgaagt agcgcgtctg ctgctccata caagccaacc acggcctcca 1680
gaagaagatg ttggcgacct cgtattggga atccccgaac atcgcctcgc tccagtcaat 1740
gaccgctgtt atgcggccat tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac 1800
gaggtgccgg acttcggggc agtcctcggc ccaaagcatc agctcatcga gagcctgcgc 1860
gacggacgca ctgacggtgt cgtccatcac agtttgccag tgatacacat ggggatcagc 1920
aatcgcgcat atgaaatcac gccatgtagt gtattgaccg attccttgcg gtccgaatgg 1980
gccgaacccg ctcgtctggc taagatcggc cgcagcgatc gcatccatag cctccgcgac 2040
cggctgcaga acagcgggca gttcggtttc aggcaggtct tgcaacgtga caccctgtgc 2100
acggcgggag atgcaatagg tcaggctctc gctgaattcc ccaatgtcaa gcacttccgg 2160
aatcgggagc gcggccgatg caaagtgccg ataaacataa cgatctttgt agaaaccatc 2220
ggcgcagcta tttacccgca ggacatatcc acgccctcct acatcgaagc tgaaagcacg 2280
agattcttcg ccctccgaga gctgcatcag gtcggagacg ctgtcgaact tttcgatcag 2340
aaacttctcg acagacgtcg cggtgagttc aggcttttcc atgggtatat ctccttctta 2400
aagttaaaca aaattatttc tagagggaaa ccgttgtggt ctccctatag tgagtcgtat 2460
taatttcgcg ggatcgagat cgatccaatt ccaatcccac aaaaatctga gcttaacagc 2520
acagttgctc ctctcagagc agaatcgggt attcaacacc ctcatatcaa ctactacgtt 2580
gtgtataacg gtccacatgc cggtatatac gatgactggg gttgtacaaa ggcggcaaca 2640
aacggcgttc ccggagttgc acacaagaaa tttgccacta ttacagaggc aagagcagca 2700
gctgacgcgt acacaacaag tcagcaaaca gacaggttga acttcatccc caaaggagaa 2760
gctcaactca agcccaagag ctttgctaag gccctaacaa gcccaccaaa gcaaaaagcc 2820
cactggctca cgctaggaac caaaaggccc agcagtgatc cagccccaaa agagatctcc 2880
tttgccccgg agattacaat ggacgatttc ctctatcttt acgatctagg aaggaagttc 2940
gaaggtgaag gtgacgacac tatgttcacc actgataatg agaaggttag cctcttcaat 3000
ttcagaaaga atgctgaccc acagatggtt agagaggcct acgcagcagg tctcatcaag 3060
acgatctacc cgagtaacaa tctccaggag atcaaatacc ttcccaagaa ggttaaagat 3120
gcagtcaaaa gattcaggac taattgcatc aagaacacag agaaagacat atttctcaag 3180
atcagaagta ctattccagt atggacgatt caaggcttgc ttcataaacc aaggcaagta 3240
atagagattg gagtctctaa aaaggtagtt cctactgaat ctaaggccat gcatggagtc 3300
taagattcaa atcgaggatc taacagaact cgccgtgaag actggcgaac agttcataca 3360
gagtctttta cgactcaatg acaagaagaa aatcttcgtc aacatggtgg agcacgacac 3420
tctggtctac tccaaaaatg tcaaagatac agtctcagaa gaccaaaggg ctattgagac 3480
ttttcaacaa aggataattt cgggaaacct cctcggattc cattgcccag ctatctgtca 3540
cttcatcgaa aggacagtag aaaaggaagg tggctcctac aaatgccatc attgcgataa 3600
aggaaaggct atcattcaag atgcctctgc cgacagtggt cccaaagatg gacccccacc 3660
cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg 3720
atgtgacatc tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc 3780
ttcctctata taaggaagtt catttcattt ggagaggaca cgctcgagct catttctcta 3840
ttacttcagc cataacaaaa gaactctttt ctcttcttat taaaccatga aaaagcctga 3900
actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct 3960
gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg 4020
atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg 4080
gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga 4140
gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga 4200
aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc 4260
cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac 4320
tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt 4380
gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc 4440
cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct 4500
gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc 4560
ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca 4620
gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta 4680
tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga 4740
tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg 4800
gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact 4860
cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agtgaggtac 4920
ctaaagaagg agtgcgtcga agcagatcgt tcaaacattt ggcaataaag tttcttaaga 4980
ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag 5040
catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt tatgattaga 5100
gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc aaactaggat 5160
aaattatcgc gcgcggtgtc atctatgtta ctagatcgat gtcgaatctg atcaacctgc 5220
attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 5280
cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 5340
caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 5400
caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 5460
ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 5520
cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 5580
ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 5640
tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 5700
gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 5760
ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 5820
ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 5880
gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 5940
aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 6000
tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 6060
ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgacat 6120
taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg 6180
gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg 6240
ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc 6300
ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatggac atattgtcgt 6360
tagaacgcgg ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac 6420
actatagaac ggcgcgccaa gcttggatcc tagcctaagt acgtactcaa aatgccaaca 6480
aataaaaaaa aagttgcttt aataatgcca aaacaaatta ataaaacact tacaacaccg 6540
gatttttttt aattaaaatg tgccatttag gataaatagt taatattttt aataattatt 6600
taaaaagccg tatctactaa aatgattttt atttggttga aaatattaat atgtttaaat 6660
caacacaatc tatcaaaatt aaactaaaaa aaaaataagt gtacgtggtt aacattagta 6720
cagtaatata agaggaaaat gagaaattaa gaaattgaaa gcgagtctaa tttttaaatt 6780
atgaacctgc atatataaaa ggaaagaaag aatccaggaa gaaaagaaat gaaaccatgc 6840
atggtcccct cgtcatcacg agtttctgcc atttgcaata gaaacactga aacacctttc 6900
tctttgtcac ttaattgaga tgccgaagcc acctcacacc atgaacttca tgaggtgtag 6960
cacccaaggc ttccatagcc atgcatactg aagaatgtct caagctcagc accctacttc 7020
tgtgacgtgt ccctcattca ccttcctctc ttccctataa ataaccacgc ctcaggttct 7080
ccgcttcaca actcaaacat tctctccatt ggtccttaaa cactcatcag tcatcaccat 7140
ggaggtccga cgacgaaaga tagacgtgct caaggcccag aaaaacggct acgaatcggg 7200
cccaccatct cgacaatcgt cgcagccctc ctcaagagca tcgtccagaa cccgcaacaa 7260
acactcctcg tccaccctgt cgctcagcgg actgaccatg aaagtccaga agaaacctgc 7320
gggacccccg gcgaactcca aaacgccatt cctacacatc aagcccgtgc acacgtgctg 7380
ctccacatca atgctttcgc gcgattatga cggctccaac cccagcttca agggcttcaa 7440
aaacatcggc atgatcattc tcattgtggg aaatctacgg ctcgcattcg aaaactacct 7500
caaatacggc atttccaacc cgttcttcga ccccaaaatt actccttccg agtggcagct 7560
ctcaggcttg ctcatagtcg tggcctacgc acatatcctc atggcctacg ctattgagag 7620
cgctgccaag ctgctgttcc tctctagcaa acaccactac atggccgtgg ggcttctgca 7680
taccatgaac actttgtcgt ccatctcgtt gctgtcctac gtcgtctact actacctgcc 7740
caaccccgtg gcaggcacaa tagtcgagtt tgtggccgtt attctgtctc tcaaactcgc 7800
ctcatacgcc ctcactaact cggatctccg aaaagccgca attcatgccc agaagctcga 7860
caagacgcaa gacgataacg aaaaggaatc cacctcgtct tcctcttctt cagatgacgc 7920
agagactttg gcagacattg acgtcattcc tgcatactac gcacagctgc cctaccccca 7980
gaatgtgacg ctgtcgaacc tgctgtactt ctggtttgct cccacactgg tctaccagcc 8040
cgtgtacccc aagacggagc gtattcgacc caagcacgtg atccgaaacc tgtttgagct 8100
cgtctctctg tgcatgctta ttcagtttct catcttccag tacgcctacc ccatcatgca 8160
gtcgtgtctg gctctgttct tccagcccaa gctcgattat gccaacatct ccgagcgcct 8220
catgaagttg gcctccgtgt ctatgatggt ctggctcatt ggattctacg ctttcttcca 8280
gaacggtctc aatcttattg ccgagctcac ctgttttgga aacagaacct tctaccagca 8340
gtggtggaat tcccgctcca ttggccagta ctggactcta tggaacaagc cagtcaacca 8400
gtactttaga caccacgtct acgtgcctct tctcgctcgg ggcatgtcgc ggttcaatgc 8460
gtcggtggtg gttttctttt tctccgccgt catccatgaa ctgcttgtcg gcatccccac 8520
tcacaacatc atcggagccg ccttcttcgg catgatgtcg caggtgcctc tgatcatggc 8580
tactgagaac cttcagcata ttaactcctc tctgggcccc ttccttggca actgtgcatt 8640
ctggttcacc tttttcctgg gacaacccac ttgtgcattc ctttactatt tggcctacaa 8700
ctacaagcag aaccagtagc 8720
<210>49
<211>5280
<212>DNA
<213>人工序列
<220>
<223>质粒pKS121/BS
<220>
<221>misc_feature
<222>(4090)..(4090)
<223>n为a,c,g,或t
<400>49
ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct 60
tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 120
acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 180
tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 240
tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 300
cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 360
actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 420
gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 480
ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 540
acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 600
ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 660
cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 720
tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 780
gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 840
ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 900
acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 960
gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 1020
ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 1080
tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 1140
gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 1200
tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 1260
ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 1320
taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 1380
cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 1440
gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 1500
gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 1560
tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 1620
gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 1680
ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 1740
ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 1800
cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 1860
ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 1920
gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 1980
ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 2040
ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 2100
tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 2160
ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 2220
cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 2280
tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 2340
tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 2400
gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 2460
gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 2520
atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg 2580
atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa 2640
aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct 2700
gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 2760
agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 2820
ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac 2880
cgggcccccc ctcgaggtcg acggtatcga taagcttgat atcgaattcc tgcagcccgg 2940
gggatccact agttctagag cggcgcgccg tcgacggata taatgagccg taaacaaaga 3000
tgattaagta gtaattaata cgtactagta aaagtggcaa aagataacga gaaagaacca 3060
atttctttgc attcggcctt agcggaaggc atatataagc tttgattatt ttatttagtg 3120
taatgatttc gtacaaccaa agcatttatt tagtactctc acacttgtgt cgcggccgct 3180
tggggggcta tggaagactt tcttagttag ttgtgtgaat aagcaatgtt gggagaatcg 3240
ggactactta taggatagga ataaaacaga aaagtattaa gtgctaatga aatatttaga 3300
ctgataatta aaatcttcac gtatgtccac ttgatataaa aacgtcagga ataaaggaag 3360
tacagtagaa tttaaaggta ctctttttat atatacccgt gttctctttt tggctagcta 3420
gttgcataaa aaataatcta tatttttatc attattttaa atatcttatg agatggtaaa 3480
tatttatcat aatttttttt actattattt attatttgtg tgtgtaatac atatagaagt 3540
taattacaaa ttttatttac tttttcatta ttttgatatg attcaccatt aatttagtgt 3600
tattatttat aatagttcat tttaatcttt ttgtatatat tatgcgtgca gtactttttt 3660
cctacatata actactatta cattttattt atataatatt tttattaatg aattttcgtg 3720
ataatatgta atattgttca ttattatttc agatttttta aaaatatttg tgttattatt 3780
tatgaaatat gtaatttttt tagtatttga ttttatgatg ataaagtgtt ctaaattcaa 3840
aagaaggggg aaagcgtaaa cattaaaaaa cgtcatcaaa caaaaacaaa atcttgttaa 3900
taaagataaa actgtttgtt ttgatcactg ttatttcgta atataaaaac attatttata 3960
tttatattgt tgacaaccaa atttgcctat caaatctaac caatataatg catgcgtggc 4020
aggtaatgta ctaccatgaa cttaagtcat gacataataa accgtgaatc tgaccaatgc 4080
atgtacctan ctaaattgta tttgtgacac gaagcaaatg attcaattca caatggagat 4140
gggaaacaaa taatgaagaa cccagaacta agaaagcttt tctgaaaaat aaaataaagg 4200
caatgtcaaa agtatactgc atcatcagtc cagaaagcac atgatatttt tttatcagta 4260
tcaatgcagc tagttttatt ttacaatatc gatatagcta gtttaaatat attgcagcta 4320
gatttataaa tatttgtgtt attatttatc atttgtgtaa tcctgttttt agtattttag 4380
tttatatatg atgataatgt attccaaatt taaaagaagg gaaataaatt taaacaagaa 4440
aaaaagtcat caaacaaaaa acaaatgaaa gggtggaaag atgttaccat gtaatgtgaa 4500
tgttacagta tttcttttat tatagagtta acaaattaac taatatgatt ttgttaataa 4560
tgataaaata ttttttttat tattatttca taatataaaa atagtttact taatataaaa 4620
aaaattctat cgttcacaac aaagttggcc acctaattta accatgcatg tacccatgga 4680
ccatattagg taaccatcaa acctgatgaa gagataaaga gatgaagact taagtcataa 4740
cacaaaacca taaaaaacaa aaatacaatc aaccgtcaat ctgaccaatg catgaaaaag 4800
ctgcaatagt gagtggcgac acaaagcaca tgattttctt acaacggaga taaaaccaaa 4860
aaaatatttc atgaacaacc tagaacaaat aaagctttta tataataaat atataaataa 4920
ataaaggcta tggaataata tacttcaata tatttggatt aaataaattg ttggcggggt 4980
tgatatattt atacacacct aaagtcactt caatctcatt ttcacttaac ttttattttt 5040
tttttctttt tatttatcat aaagagaata ttgataatat actttttaac atatttttat 5100
gacatttttt attggtgaaa acttattaaa aatcataaat tttgtaagtt agatttattt 5160
aaagagttcc tcttcttatt ttaaattttt taataaattt ttaaataact aaaatttgtg 5220
ttaaaaatgt taaaaaagtg tgttattaac ccttctcttc gaggatccaa gcttggcgcg 5280
<210>50
<211>5968
<212>DNA
<213>人工序列
<220>
<223>质粒pDs-Red in pKS121/BS
<220>
<221>misc_feature
<222>(5052)..(5052)
<223>n为a,c,g,或t
<400>50
catggcctcc tccgaggacg tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg 60
ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg 120
cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat 180
cctgtccccc cagttccagt acggctccaa ggtgtacgtg aagcaccccg ccgacatccc 240
cgactacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga 300
ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggct ccttcatcta 360
caaggtgaag ttcatcggcg tgaacttccc ctccgacggc cccgtaatgc agaagaagac 420
tatgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga 480
gatccacaag gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagtccat 540
ctacatggcc aagaagcccg tgcagctgcc cggctactac tacgtggact ccaagctgga 600
catcacctcc cacaacgagg actacaccat cgtggagcag tacgagcgcg ccgagggccg 660
ccaccacctg ttcctgtagc ggccggccgc gacacaagtg tgagagtact aaataaatgc 720
tttggttgta cgaaatcatt acactaaata aaataatcaa agcttatata tgccttccgc 780
taaggccgaa tgcaaagaaa ttggttcttt ctcgttatct tttgccactt ttactagtac 840
gtattaatta ctacttaatc atctttgttt acggctcatt atatccgtcg acggcgcgcc 900
gctctagaac tagtggatcc cccgggctgc aggaattcga tatcaagctt atcgataccg 960
tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc 1020
tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat 1080
cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat 1140
cgcccttccc aacagttgcg cagcctgaat ggcgaatgga aattgtaagc gttaatattt 1200
tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa 1260
tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag 1320
tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg 1380
tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga 1440
ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg 1500
gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg 1560
cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc 1620
cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 1680
tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 1740
ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc 1800
ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 1860
aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg 1920
gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag 1980
ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc 2040
gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta 2100
cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 2160
cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca 2220
acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 2280
caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat 2340
taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg 2400
ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 2460
aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 2520
agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 2580
atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag 2640
tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 2700
tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 2760
gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 2820
taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 2880
aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 2940
ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 3000
catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 3060
ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 3120
ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 3180
agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 3240
taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 3300
atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 3360
cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 3420
ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 3480
accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca 3540
gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc 3600
gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg 3660
agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta 3720
tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca 3780
gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg 3840
agctccaccg cggtggcggc ccgcgccaag cttggatcct cgaagagaag ggttaataac 3900
acactttttt aacattttta acacaaattt tagttattta aaaatttatt aaaaaattta 3960
aaataagaag aggaactctt taaataaatc taacttacaa aatttatgat ttttaataag 4020
ttttcaccaa taaaaaatgt cataaaaata tgttaaaaag tatattatca atattctctt 4080
tatgataaat aaaaagaaaa aaaaaataaa agttaagtga aaatgagatt gaagtgactt 4140
taggtgtgta taaatatatc aaccccgcca acaatttatt taatccaaat atattgaagt 4200
atattattcc atagccttta tttatttata tatttattat ataaaagctt tatttgttct 4260
aggttgttca tgaaatattt ttttggtttt atctccgttg taagaaaatc atgtgctttg 4320
tgtcgccact cactattgca gctttttcat gcattggtca gattgacggt tgattgtatt 4380
tttgtttttt atggttttgt gttatgactt aagtcttcat ctctttatct cttcatcagg 4440
tttgatggtt acctaatatg gtccatgggt acatgcatgg ttaaattagg tggccaactt 4500
tgttgtgaac gatagaattt tttttatatt aagtaaacta tttttatatt atgaaataat 4560
aataaaaaaa atattttatc attattaaca aaatcatatt agttaatttg ttaactctat 4620
aataaaagaa atactgtaac attcacatta catggtaaca tctttccacc ctttcatttg 4680
ttttttgttt gatgactttt tttcttgttt aaatttattt cccttctttt aaatttggaa 4740
tacattatca tcatatataa actaaaatac taaaaacagg attacacaaa tgataaataa 4800
taacacaaat atttataaat ctagctgcaa tatatttaaa ctagctatat cgatattgta 4860
aaataaaact agctgcattg atactgataa aaaaatatca tgtgctttct ggactgatga 4920
tgcagtatac ttttgacatt gcctttattt tatttttcag aaaagctttc ttagttctgg 4980
gttcttcatt atttgtttcc catctccatt gtgaattgaa tcatttgctt cgtgtcacaa 5040
atacaattta gntaggtaca tgcattggtc agattcacgg tttattatgt catgacttaa 5100
gttcatggta gtacattacc tgccacgcat gcattatatt ggttagattt gataggcaaa 5160
tttggttgtc aacaatataa atataaataa tgtttttata ttacgaaata acagtgatca 5220
aaacaaacag ttttatcttt attaacaaga ttttgttttt gtttgatgac gttttttaat 5280
gtttacgctt tcccccttct tttgaattta gaacacttta tcatcataaa atcaaatact 5340
aaaaaaatta catatttcat aaataataac acaaatattt ttaaaaaatc tgaaataata 5400
atgaacaata ttacatatta tcacgaaaat tcattaataa aaatattata taaataaaat 5460
gtaatagtag ttatatgtag gaaaaaagta ctgcacgcat aatatataca aaaagattaa 5520
aatgaactat tataaataat aacactaaat taatggtgaa tcatatcaaa ataatgaaaa 5580
agtaaataaa atttgtaatt aacttctata tgtattacac acacaaataa taaataatag 5640
taaaaaaaat tatgataaat atttaccatc tcataagata tttaaaataa tgataaaaat 5700
atagattatt ttttatgcaa ctagctagcc aaaaagagaa cacgggtata tataaaaaga 5760
gtacctttaa attctactgt acttccttta ttcctgacgt ttttatatca agtggacata 5820
cgtgaagatt ttaattatca gtctaaatat ttcattagca cttaatactt ttctgtttta 5880
ttcctatcct ataagtagtc ccgattctcc caacattgct tattcacaca actaactaag 5940
aaagtcttcc atagcccccc aagcggcc 5968
<210>51
<211>10058
<212>DNA
<213>人工序列
<220>
<223>质粒pKS332
<220>
<221>misc_feature
<222>(1178)..(1178)
<223>n为a,c,g,或t
<400>51
gatcctcgaa gagaagggtt aataacacac ttttttaaca tttttaacac aaattttagt 60
tatttaaaaa tttattaaaa aatttaaaat aagaagagga actctttaaa taaatctaac 120
ttacaaaatt tatgattttt aataagtttt caccaataaa aaatgtcata aaaatatgtt 180
aaaaagtata ttatcaatat tctctttatg ataaataaaa agaaaaaaaa aataaaagtt 240
aagtgaaaat gagattgaag tgactttagg tgtgtataaa tatatcaacc ccgccaacaa 300
tttatttaat ccaaatatat tgaagtatat tattccatag cctttattta tttatatatt 360
tattatataa aagctttatt tgttctaggt tgttcatgaa atattttttt ggttttatct 420
ccgttgtaag aaaatcatgt gctttgtgtc gccactcact attgcagctt tttcatgcat 480
tggtcagatt gacggttgat tgtatttttg ttttttatgg ttttgtgtta tgacttaagt 540
cttcatctct ttatctcttc atcaggtttg atggttacct aatatggtcc atgggtacat 600
gcatggttaa attaggtggc caactttgtt gtgaacgata gaattttttt tatattaagt 660
aaactatttt tatattatga aataataata aaaaaaatat tttatcatta ttaacaaaat 720
catattagtt aatttgttaa ctctataata aaagaaatac tgtaacattc acattacatg 780
gtaacatctt tccacccttt catttgtttt ttgtttgatg actttttttc ttgtttaaat 840
ttatttccct tcttttaaat ttggaataca ttatcatcat atataaacta aaatactaaa 900
aacaggatta cacaaatgat aaataataac acaaatattt ataaatctag ctgcaatata 960
tttaaactag ctatatcgat attgtaaaat aaaactagct gcattgatac tgataaaaaa 1020
atatcatgtg ctttctggac tgatgatgca gtatactttt gacattgcct ttattttatt 1080
tttcagaaaa gctttcttag ttctgggttc ttcattattt gtttcccatc tccattgtga 1140
attgaatcat ttgcttcgtg tcacaaatac aatttagnta ggtacatgca ttggtcagat 1200
tcacggttta ttatgtcatg acttaagttc atggtagtac attacctgcc acgcatgcat 1260
tatattggtt agatttgata ggcaaatttg gttgtcaaca atataaatat aaataatgtt 1320
tttatattac gaaataacag tgatcaaaac aaacagtttt atctttatta acaagatttt 1380
gtttttgttt gatgacgttt tttaatgttt acgctttccc ccttcttttg aatttagaac 1440
actttatcat cataaaatca aatactaaaa aaattacata tttcataaat aataacacaa 1500
atatttttaa aaaatctgaa ataataatga acaatattac atattatcac gaaaattcat 1560
taataaaaat attatataaa taaaatgtaa tagtagttat atgtaggaaa aaagtactgc 1620
acgcataata tatacaaaaa gattaaaatg aactattata aataataaca ctaaattaat 1680
ggtgaatcat atcaaaataa tgaaaaagta aataaaattt gtaattaact tctatatgta 1740
ttacacacac aaataataaa taatagtaaa aaaaattatg ataaatattt accatctcat 1800
aagatattta aaataatgat aaaaatatag attatttttt atgcaactag ctagccaaaa 1860
agagaacacg ggtatatata aaaagagtac ctttaaattc tactgtactt cctttattcc 1920
tgacgttttt atatcaagtg gacatacgtg aagattttaa ttatcagtct aaatatttca 1980
ttagcactta atacttttct gttttattcc tatcctataa gtagtcccga ttctcccaac 2040
attgcttatt cacacaacta actaagaaag tcttccatag ccccccaagc ggcccatggc 2100
ctcctccgag gacgtcatca aggagttcat gcgcttcaag gtgcgcatgg agggctccgt 2160
gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca 2220
gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc 2280
cccccagttc cagtacggct ccaaggtgta cgtgaagcac cccgccgaca tccccgacta 2340
caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg 2400
cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggctccttca tctacaaggt 2460
gaagttcatc ggcgtgaact tcccctccga cggccccgta atgcagaaga agactatggg 2520
ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca 2580
caaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaagt ccatctacat 2640
ggccaagaag cccgtgcagc tgcccggcta ctactacgtg gactccaagc tggacatcac 2700
ctcccacaac gaggactaca ccatcgtgga gcagtacgag cgcgccgagg gccgccacca 2760
cctgttcctg tagcggccgg ccgcgacaca agtgtgagag tactaaataa atgctttggt 2820
tgtacgaaat cattacacta aataaaataa tcaaagctta tatatgcctt ccgctaaggc 2880
cgaatgcaaa gaaattggtt ctttctcgtt atcttttgcc acttttacta gtacgtatta 2940
attactactt aatcatcttt gtttacggct cattatatcc gtcgacggcg cgccgctcta 3000
gaactagtgg atccgtcgac ggcgcgcccg atcatccgga tatagttcct cctttcagca 3060
aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat tgctcagcgg 3120
tggcagcagc caactcagct tcctttcggg ctttgttagc agccggatcg atccaagctg 3180
tacctcacta ttcctttgcc ctcggacgag tgctggggcg tcggtttcca ctatcggcga 3240
gtacttctac acagccatcg gtccagacgg ccgcgcttct gcgggcgatt tgtgtacgcc 3300
cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc caagctgcat 3360
catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg cggagcatat 3420
acgcccggag ccgcggcgat cctgcaagct ccggatgcct ccgctcgaag tagcgcgtct 3480
gctgctccat acaagccaac cacggcctcc agaagaagat gttggcgacc tcgtattggg 3540
aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca ttgtccgtca 3600
ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg cagtcctcgg 3660
cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg tcgtccatca 3720
cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca cgccatgtag 3780
tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg ctaagatcgg 3840
ccgcagcgat cgcatccata gcctccgcga ccggctgcag aacagcgggc agttcggttt 3900
caggcaggtc ttgcaacgtg acaccctgtg cacggcggga gatgcaatag gtcaggctct 3960
cgctgaattc cccaatgtca agcacttccg gaatcgggag cgcggccgat gcaaagtgcc 4020
gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc aggacatatc 4080
cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag agctgcatca 4140
ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc gcggtgagtt 4200
caggcttttc catgggtata tctccttctt aaagttaaac aaaattattt ctagagggaa 4260
accgttgtgg tctccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatccaat 4320
tccaatccca caaaaatctg agcttaacag cacagttgct cctctcagag cagaatcggg 4380
tattcaacac cctcatatca actactacgt tgtgtataac ggtccacatg ccggtatata 4440
cgatgactgg ggttgtacaa aggcggcaac aaacggcgtt cccggagttg cacacaagaa 4500
atttgccact attacagagg caagagcagc agctgacgcg tacacaacaa gtcagcaaac 4560
agacaggttg aacttcatcc ccaaaggaga agctcaactc aagcccaaga gctttgctaa 4620
ggccctaaca agcccaccaa agcaaaaagc ccactggctc acgctaggaa ccaaaaggcc 4680
cagcagtgat ccagccccaa aagagatctc ctttgccccg gagattacaa tggacgattt 4740
cctctatctt tacgatctag gaaggaagtt cgaaggtgaa ggtgacgaca ctatgttcac 4800
cactgataat gagaaggtta gcctcttcaa tttcagaaag aatgctgacc cacagatggt 4860
tagagaggcc tacgcagcag gtctcatcaa gacgatctac ccgagtaaca atctccagga 4920
gatcaaatac cttcccaaga aggttaaaga tgcagtcaaa agattcagga ctaattgcat 4980
caagaacaca gagaaagaca tatttctcaa gatcagaagt actattccag tatggacgat 5040
tcaaggcttg cttcataaac caaggcaagt aatagagatt ggagtctcta aaaaggtagt 5100
tcctactgaa tctaaggcca tgcatggagt ctaagattca aatcgaggat ctaacagaac 5160
tcgccgtgaa gactggcgaa cagttcatac agagtctttt acgactcaat gacaagaaga 5220
aaatcttcgt caacatggtg gagcacgaca ctctggtcta ctccaaaaat gtcaaagata 5280
cagtctcaga agaccaaagg gctattgaga cttttcaaca aaggataatt tcgggaaacc 5340
tcctcggatt ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag 5400
gtggctccta caaatgccat cattgcgata aaggaaaggc tatcattcaa gatgcctctg 5460
ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg 5520
ttccaaccac gtcttcaaag caagtggatt gatgtgacat ctccactgac gtaagggatg 5580
acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt 5640
tggagaggac acgctcgagc tcatttctct attacttcag ccataacaaa agaactcttt 5700
tctcttctta ttaaaccatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc 5760
tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc 5820
gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg 5880
atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc 5940
cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg 6000
cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg 6060
tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc 6120
cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg 6180
ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg 6240
cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg 6300
tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca 6360
ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct 6420
ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg 6480
agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct 6540
atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg 6600
caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg 6660
ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca 6720
ctcgtccgag ggcaaaggaa tagtgaggta cctaaagaag gagtgcgtcg aagcagatcg 6780
ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat 6840
tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac 6900
gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat 6960
agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt 7020
actagatcga tgtcgaatct gatcaacctg cattaatgaa tcggccaacg cgcggggaga 7080
ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 7140
gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 7200
tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 7260
aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 7320
aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 7380
ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 7440
tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc 7500
agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 7560
gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 7620
tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 7680
acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 7740
tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 7800
caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 7860
aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 7920
aactcacgtt aagggatttt ggtcatgaca ttaacctata aaaataggcg tatcacgagg 7980
ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg 8040
gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg 8100
tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta 8160
ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc 8220
ttatgtatca tacacatacg atttaggtga cactatagaa cggcgcgcca agcttttgat 8280
ccatgccctt catttgccgc ttattaatta atttggtaac agtccgtact aatcagttac 8340
ttatccttcc cccatcataa ttaatcttgg tagtctcgaa tgccacaaca ctgactagtc 8400
tcttggatca taagaaaaag ccaaggaaca aaagaagaca aaacacaatg agagtatcct 8460
ttgcatagca atgtctaagt tcataaaatt caaacaaaaa cgcaatcaca cacagtggac 8520
atcacttatc cactagctga tcaggatcgc cgcgtcaaga aaaaaaaact ggaccccaaa 8580
agccatgcac aacaacacgt actcacaaag gtgtcaatcg agcagcccaa aacattcacc 8640
aactcaaccc atcatgagcc ctcacatttg ttgtttctaa cccaacctca aactcgtatt 8700
ctcttccgcc acctcatttt tgtttatttc aacacccgtc aaactgcatg ccaccccgtg 8760
gccaaatgtc catgcatgtt aacaagacct atgactataa atagctgcaa tctcggccca 8820
ggttttcatc atcaagaacc agttcaatat cctagtacac cgtattaaag aatttaagat 8880
atactgcggc cgcaagtatg aactaaaatg catgtaggtg taagagctca tggagagcat 8940
ggaatattgt atccgaccat gtaacagtat aataactgag ctccatctca cttcttctat 9000
gaataaacaa aggatgttat gatatattaa cactctatct atgcacctta ttgttctatg 9060
ataaatttcc tcttattatt ataaatcatc tgaatcgtga cggcttatgg aatgcttcaa 9120
atagtacaaa aacaaatgtg tactataaga ctttctaaac aattctaacc ttagcattgt 9180
gaacgagaca taagtgttaa gaagacataa caattataat ggaagaagtt tgtctccatt 9240
tatatattat atattaccca cttatgtatt atattaggat gttaaggaga cataacaatt 9300
ataaagagag aagtttgtat ccatttatat attatatact acccatttat atattatact 9360
tatccactta tttaatgtct ttataaggtt tgatccatga tatttctaat attttagttg 9420
atatgtatat gaaagggtac tatttgaact ctcttactct gtataaaggt tggatcatcc 9480
ttaaagtggg tctatttaat tttattgctt cttacagata aaaaaaaaat tatgagttgg 9540
tttgataaaa tattgaagga tttaaaataa taataaataa catataatat atgtatataa 9600
atttattata atataacatt tatctataaa aaagtaaata ttgtcataaa tctatacaat 9660
cgtttagcct tgctggacga atctcaatta tttaaacgag agtaaacata tttgactttt 9720
tggttattta acaaattatt atttaacact atatgaaatt ttttttttta tcagcaaaga 9780
ataaaattaa attaagaagg acaatggtgt cccaatcctt atacaaccaa cttccacaag 9840
aaagtcaagt cagagacaac aaaaaaacaa gcaaaggaaa ttttttaatt tgagttgtct 9900
tgtttgctgc ataatttatg cagtaaaaca ctacacataa cccttttagc agtagagcaa 9960
tggttgaccg tgtgcttagc ttcttttatt ttattttttt atcagcaaag aataaataaa 10020
ataaaatgag acacttcagg gatgtttcaa caagcttg 10058
<210>52
<211>11611
<212>DNA
<213>人工序列
<220>
<223>质粒pKS362
<220>
<221>misc_feature
<222>(2349)..(2349)
<223>n为a,c,g,或t
<400>52
ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacaagctt ggatcctcga agagaagggt taataacaca 1200
cttttttaac atttttaaca caaattttag ttatttaaaa atttattaaa aaatttaaaa 1260
taagaagagg aactctttaa ataaatctaa cttacaaaat ttatgatttt taataagttt 1320
tcaccaataa aaaatgtcat aaaaatatgt taaaaagtat attatcaata ttctctttat 1380
gataaataaa aagaaaaaaa aaataaaagt taagtgaaaa tgagattgaa gtgactttag 1440
gtgtgtataa atatatcaac cccgccaaca atttatttaa tccaaatata ttgaagtata 1500
ttattccata gcctttattt atttatatat ttattatata aaagctttat ttgttctagg 1560
ttgttcatga aatatttttt tggttttatc tccgttgtaa gaaaatcatg tgctttgtgt 1620
cgccactcac tattgcagct ttttcatgca ttggtcagat tgacggttga ttgtattttt 1680
gttttttatg gttttgtgtt atgacttaag tcttcatctc tttatctctt catcaggttt 1740
gatggttacc taatatggtc catgggtaca tgcatggtta aattaggtgg ccaactttgt 1800
tgtgaacgat agaatttttt ttatattaag taaactattt ttatattatg aaataataat 1860
aaaaaaaata ttttatcatt attaacaaaa tcatattagt taatttgtta actctataat 1920
aaaagaaata ctgtaacatt cacattacat ggtaacatct ttccaccctt tcatttgttt 1980
tttgtttgat gacttttttt cttgtttaaa tttatttccc ttcttttaaa tttggaatac 2040
attatcatca tatataaact aaaatactaa aaacaggatt acacaaatga taaataataa 2100
cacaaatatt tataaatcta gctgcaatat atttaaacta gctatatcga tattgtaaaa 2160
taaaactagc tgcattgata ctgataaaaa aatatcatgt gctttctgga ctgatgatgc 2220
agtatacttt tgacattgcc tttattttat ttttcagaaa agctttctta gttctgggtt 2280
cttcattatt tgtttcccat ctccattgtg aattgaatca tttgcttcgt gtcacaaata 2340
caatttagnt aggtacatgc attggtcaga ttcacggttt attatgtcat gacttaagtt 2400
catggtagta cattacctgc cacgcatgca ttatattggt tagatttgat aggcaaattt 2460
ggttgtcaac aatataaata taaataatgt ttttatatta cgaaataaca gtgatcaaaa 2520
caaacagttt tatctttatt aacaagattt tgtttttgtt tgatgacgtt ttttaatgtt 2580
tacgctttcc cccttctttt gaatttagaa cactttatca tcataaaatc aaatactaaa 2640
aaaattacat atttcataaa taataacaca aatattttta aaaaatctga aataataatg 2700
aacaatatta catattatca cgaaaattca ttaataaaaa tattatataa ataaaatgta 2760
atagtagtta tatgtaggaa aaaagtactg cacgcataat atatacaaaa agattaaaat 2820
gaactattat aaataataac actaaattaa tggtgaatca tatcaaaata atgaaaaagt 2880
aaataaaatt tgtaattaac ttctatatgt attacacaca caaataataa ataatagtaa 2940
aaaaaattat gataaatatt taccatctca taagatattt aaaataatga taaaaatata 3000
gattattttt tatgcaacta gctagccaaa aagagaacac gggtatatat aaaaagagta 3060
cctttaaatt ctactgtact tcctttattc ctgacgtttt tatatcaagt ggacatacgt 3120
gaagatttta attatcagtc taaatatttc attagcactt aatacttttc tgttttattc 3180
ctatcctata agtagtcccg attctcccaa cattgcttat tcacacaact aactaagaaa 3240
gtcttccata gccccccaag cggcccatgg cctcctccga ggacgtcatc aaggagttca 3300
tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca cgagttcgag atcgagggcg 3360
agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg 3420
gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt 3480
acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca 3540
agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct 3600
ccctgcagga cggctccttc atctacaagg tgaagttcat cggcgtgaac ttcccctccg 3660
acggccccgt aatgcagaag aagactatgg gctgggaggc ctccaccgag cgcctgtacc 3720
cccgcgacgg cgtgctgaag ggcgagatcc acaaggccct gaagctgaag gacggcggcc 3780
actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag ctgcccggct 3840
actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac accatcgtgg 3900
agcagtacga gcgcgccgag ggccgccacc acctgttcct gtagcggccg gccgcgacac 3960
aagtgtgaga gtactaaata aatgctttgg ttgtacgaaa tcattacact aaataaaata 4020
atcaaagctt atatatgcct tccgctaagg ccgaatgcaa agaaattggt tctttctcgt 4080
tatcttttgc cacttttact agtacgtatt aattactact taatcatctt tgtttacggc 4140
tcattatatc cgtcgacggc gcgccgctct agaactagtg gatccgtcga cggcgcgccc 4200
gatcatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc 4260
ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg 4320
gctttgttag cagccggatc gatccaagct gtacctcact attcctttgc cctcggacga 4380
gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg 4440
gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt 4500
gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga 4560
tagagttggt caagaccaat gcggagcata tacgcccgga gccgcggcga tcctgcaagc 4620
tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc 4680
cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca 4740
atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc 4800
acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc 4860
gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca 4920
gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat 4980
gggccgaacc cgctcgtctg gctaagatcg gccgcagcga tcgcatccat agcctccgcg 5040
accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt 5100
gcacggcggg agatgcaata ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc 5160
ggaatcggga gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca 5220
tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca 5280
cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc 5340
agaaacttct cgacagacgt cgcggtgagt tcaggctttt ccatgggtat atctccttct 5400
taaagttaaa caaaattatt tctagaggga aaccgttgtg gtctccctat agtgagtcgt 5460
attaatttcg cgggatcgag atcgatccaa ttccaatccc acaaaaatct gagcttaaca 5520
gcacagttgc tcctctcaga gcagaatcgg gtattcaaca ccctcatatc aactactacg 5580
ttgtgtataa cggtccacat gccggtatat acgatgactg gggttgtaca aaggcggcaa 5640
caaacggcgt tcccggagtt gcacacaaga aatttgccac tattacagag gcaagagcag 5700
cagctgacgc gtacacaaca agtcagcaaa cagacaggtt gaacttcatc cccaaaggag 5760
aagctcaact caagcccaag agctttgcta aggccctaac aagcccacca aagcaaaaag 5820
cccactggct cacgctagga accaaaaggc ccagcagtga tccagcccca aaagagatct 5880
cctttgcccc ggagattaca atggacgatt tcctctatct ttacgatcta ggaaggaagt 5940
tcgaaggtga aggtgacgac actatgttca ccactgataa tgagaaggtt agcctcttca 6000
atttcagaaa gaatgctgac ccacagatgg ttagagaggc ctacgcagca ggtctcatca 6060
agacgatcta cccgagtaac aatctccagg agatcaaata ccttcccaag aaggttaaag 6120
atgcagtcaa aagattcagg actaattgca tcaagaacac agagaaagac atatttctca 6180
agatcagaag tactattcca gtatggacga ttcaaggctt gcttcataaa ccaaggcaag 6240
taatagagat tggagtctct aaaaaggtag ttcctactga atctaaggcc atgcatggag 6300
tctaagattc aaatcgagga tctaacagaa ctcgccgtga agactggcga acagttcata 6360
cagagtcttt tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac 6420
actctggtct actccaaaaa tgtcaaagat acagtctcag aagaccaaag ggctattgag 6480
acttttcaac aaaggataat ttcgggaaac ctcctcggat tccattgccc agctatctgt 6540
cacttcatcg aaaggacagt agaaaaggaa ggtggctcct acaaatgcca tcattgcgat 6600
aaaggaaagg ctatcattca agatgcctct gccgacagtg gtcccaaaga tggaccccca 6660
cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat 6720
tgatgtgaca tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac 6780
ccttcctcta tataaggaag ttcatttcat ttggagagga cacgctcgag ctcatttctc 6840
tattacttca gccataacaa aagaactctt ttctcttctt attaaaccat gaaaaagcct 6900
gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac 6960
ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt 7020
ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat 7080
cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc 7140
gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct 7200
gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg 7260
gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac 7320
actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact 7380
gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg 7440
gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc 7500
ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat 7560
tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag 7620
cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg 7680
tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat 7740
gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc 7800
gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta 7860
ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga atagtgaggt 7920
acctaaagaa ggagtgcgtc gaagcagatc gttcaaacat ttggcaataa agtttcttaa 7980
gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta 8040
agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta 8100
gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg 8160
ataaattatc gcgcgcggtg tcatctatgt tactagatcg atgtcgaatc tgatcaacct 8220
gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 8280
ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 8340
ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 8400
agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 8460
taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 8520
cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 8580
tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 8640
gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 8700
gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 8760
tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 8820
gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 8880
cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 8940
aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 9000
tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 9060
ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgac 9120
attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 9180
cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 9240
tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 9300
gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc 9360
gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg 9420
acactataga acggcgcgcc aagcttttga tccatgccct tcatttgccg cttattaatt 9480
aatttggtaa cagtccgtac taatcagtta cttatccttc ccccatcata attaatcttg 9540
gtagtctcga atgccacaac actgactagt ctcttggatc ataagaaaaa gccaaggaac 9600
aaaagaagac aaaacacaat gagagtatcc tttgcatagc aatgtctaag ttcataaaat 9660
tcaaacaaaa acgcaatcac acacagtgga catcacttat ccactagctg atcaggatcg 9720
ccgcgtcaag aaaaaaaaac tggaccccaa aagccatgca caacaacacg tactcacaaa 9780
ggtgtcaatc gagcagccca aaacattcac caactcaacc catcatgagc cctcacattt 9840
gttgtttcta acccaacctc aaactcgtat tctcttccgc cacctcattt ttgtttattt 9900
caacacccgt caaactgcat gccaccccgt ggccaaatgt ccatgcatgt taacaagacc 9960
tatgactata aatagctgca atctcggccc aggttttcat catcaagaac cagttcaata 10020
tcctagtaca ccgtattaaa gaatttaaga tatactgcgg ccgcatgact atcgactcac 10080
aatactacaa gtcgcgagac aaaaacgaca cggcacccaa aatcgcggga atccgatatg 10140
ccccgctatc gacaccatta ctcaaccgat gtgagacctt ctctctggtc tggcacattt 10200
tcagcattcc cactttcctc acaattttca tgctatgctg cgcaattcca ctgctctggc 10260
catttgtgat tgcgtatgta gtgtacgctg ttaaagacga ctccccgtcc aacggaggag 10320
tggtcaagcg atactcgcct atttcaagaa acttcttcat ctggaagctc tttggccgct 10380
acttccccat aactctgcac aagacggtgg atctggagcc cacgcacaca tactaccctc 10440
tggacgtcca ggagtatcac ctgattgctg agagatactg gccgcagaac aagtacctcc 10500
gagcaatcat caccaccatc gagtactttc tgcccgcctt catgaaacgg tctctttcta 10560
tcaacgagca ggagcagcct gccgagcgag atcctctcct gtctcccgtt tctcccagct 10620
ctccgggttc tcaacctgac aagtggatta accacgacag cagatatagc cgtggagaat 10680
catctggctc caacggccac gcctcgggct ccgaacttaa cggcaacggc aacaacggca 10740
ccactaaccg acgacctttg tcgtccgcct ctgctggctc cactgcatct gattccacgc 10800
ttcttaacgg gtccctcaac tcctacgcca accagatcat tggcgaaaac gacccacagc 10860
tgtcgcccac aaaactcaag cccactggca gaaaatacat cttcggctac cacccccacg 10920
gcattatcgg catgggagcc tttggtggaa ttgccaccga gggagctgga tggtccaagc 10980
tctttccggg catccctgtt tctcttatga ctctcaccaa caacttccga gtgcctctct 11040
acagagagta cctcatgagt ctgggagtcg cttctgtctc caagaagtcc tgcaaggccc 11100
tcctcaagcg aaaccagtct atctgcattg tcgttggtgg agcacaggaa agtcttctgg 11160
ccagacccgg tgtcatggac ctggtgctac tcaagcgaaa gggttttgtt cgacttggta 11220
tggaggtcgg aaatgtcgcc cttgttccca tcatggcctt tggtgagaac gacctctatg 11280
accaggttag caacgacaag tcgtccaagc tgtaccgatt ccagcagttt gtcaagaact 11340
tccttggatt cacccttcct ttgatgcatg cccgaggcgt cttcaactac gatgtcggtc 11400
ttgtccccta caggcgaccc gtcaacattg tggttggttc ccccattgac ttgccttatc 11460
tcccacaccc caccgacgaa gaagtgtccg aataccacga ccgatacatc gccgagctgc 11520
agcgaatcta caacgagcac aaggatgaat atttcatcga ttggaccgag gagggcaaag 11580
gagccccaga gttccgaatg attgagtaag c 11611
<210>53
<211>1422
<212>DNA
<213>大豆
<400>53
ggatccaact aaaaaaagct ctcaaattac attttgagtt gtttcaggtt ccattgcctt 60
attgctaaaa ctccaactaa aataacaaat agcacatgca ggtgcaaaca acacgttact 120
ctgatgaagg tgatgtgcct ctagcagtct agcttatgag gctcgctgct tatcaacgat 180
tcatcattcc ccaagacgtg tacgcagatt aaacaatgga caaaacttca atcgattata 240
gaataataat tttaacagtg ccgacttttt tctgtaaaca aaaggccaga atcatatcgc 300
acatcatctt gaatgcagtg tcgagtttgg accatttgag tacaaagcca atattgaatg 360
atttttcgat tttacatgtg tgaatcagac aaaagtgcat gcaatcactt gcaagtaaat 420
taaggatact aatctattcc tttcatttta tatgctccac ttttatataa aaaaatatac 480
attattatat atgcattatt aattattgca gtattatgct attggtttta tggccctgct 540
aaataaccta aatgagtcta actattgcat atgaatcaaa tgaaggaaga atcatgatct 600
aaacctgagt acccaatgca ataaaatgcg tcctattacc taaacttcaa acacacattg 660
ccatcggacg tataaattaa tgcatataga ttattttgag aaaagaaaac atcaaaagct 720
ctaaaacttc ttttaacttt gaaataagct gataaaaata cgctttaaat caactgtgtg 780
ctgtatataa gctgcaattt cacattttac caaaccgaaa caagaatggt aacagtgagg 840
caaaaatttg aaaaatgtcc tacttcacat tcacatcaaa ttaattacaa ctaaataaat 900
aaacatcgtg attcaagcag taatgaaagt cgaaatcaga tagaatatac acgtttaaca 960
tcaattgaat ttttttttaa atggatatat acaagtttac tattttatat ataatgaaaa 1020
ttcattttgt gttagcacaa aacttacaga aagagataaa ttttaaataa agagaattat 1080
atccaatttt ataatccaaa ataatcaaat taaagaatat tggctagata gaccggcttt 1140
ttcactgccc ctgctggata atgaaaattc atatcaaaac aatacagaag ttctagttta 1200
ataataaaaa agttggcaaa ctgtcattcc ctgttggttt ttaagccaaa tcacaattca 1260
attacgtatc agaaattaat ttaaaccaaa tatatagcta cgagggaact tcttcagtca 1320
ttactagcta gctcactaat cactatatat acgacatgct acaagtgaag tgaccatatc 1380
ttaatttcaa atcataaaat tcttccacca agttgcggcc gc 1422
<210>54
<211>41
<212>DNA
<213>人工序列
<220>
<223>引物oP34-1
<400>54
cgcggatcca actaaaaaaa gctctcaaat tacattttga g 41
<210>55
<211>44
<212>DNA
<213>人工序列
<220>
<223>引物oP34-2
<400>55
gaattcgcgg ccgcaacttg gtggaagaat tttatgattt gaaa 44
<210>56
<211>7466
<212>DNA
<213>人工序列
<220>
<223>质粒pZBL115
<400>56
gatccgtcga cggcgcgccc gatcatccgg atatagttcc tcctttcagc aaaaaacccc 60
tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag 120
ccaactcagc ttcctttcgg gctttgttag cagccggatc gatccaagct gtacctcact 180
attcctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta 240
cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc 300
cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat 360
tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga 420
gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca 480
tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga 540
acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt 600
tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca 660
tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc 720
agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac 780
cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga 840
tcgcatccat agcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt 900
cttgcaacgt gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaatt 960
ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat 1020
aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc 1080
ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga 1140
cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt 1200
ccatgggtat atctccttct taaagttaaa caaaattatt tctagaggga aaccgttgtg 1260
gtctccctat agtgagtcgt attaatttcg cgggatcgag atcgatccaa ttccaatccc 1320
acaaaaatct gagcttaaca gcacagttgc tcctctcaga gcagaatcgg gtattcaaca 1380
ccctcatatc aactactacg ttgtgtataa cggtccacat gccggtatat acgatgactg 1440
gggttgtaca aaggcggcaa caaacggcgt tcccggagtt gcacacaaga aatttgccac 1500
tattacagag gcaagagcag cagctgacgc gtacacaaca agtcagcaaa cagacaggtt 1560
gaacttcatc cccaaaggag aagctcaact caagcccaag agctttgcta aggccctaac 1620
aagcccacca aagcaaaaag cccactggct cacgctagga accaaaaggc ccagcagtga 1680
tccagcccca aaagagatct cctttgcccc ggagattaca atggacgatt tcctctatct 1740
ttacgatcta ggaaggaagt tcgaaggtga aggtgacgac actatgttca ccactgataa 1800
tgagaaggtt agcctcttca atttcagaaa gaatgctgac ccacagatgg ttagagaggc 1860
ctacgcagca ggtctcatca agacgatcta cccgagtaac aatctccagg agatcaaata 1920
ccttcccaag aaggttaaag atgcagtcaa aagattcagg actaattgca tcaagaacac 1980
agagaaagac atatttctca agatcagaag tactattcca gtatggacga ttcaaggctt 2040
gcttcataaa ccaaggcaag taatagagat tggagtctct aaaaaggtag ttcctactga 2100
atctaaggcc atgcatggag tctaagattc aaatcgagga tctaacagaa ctcgccgtga 2160
agactggcga acagttcata cagagtcttt tacgactcaa tgacaagaag aaaatcttcg 2220
tcaacatggt ggagcacgac actctggtct actccaaaaa tgtcaaagat acagtctcag 2280
aagaccaaag ggctattgag acttttcaac aaaggataat ttcgggaaac ctcctcggat 2340
tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct 2400
acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatgcctct gccgacagtg 2460
gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca 2520
cgtcttcaaa gcaagtggat tgatgtgaca tctccactga cgtaagggat gacgcacaat 2580
cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga 2640
cacgctcgag ctcatttctc tattacttca gccataacaa aagaactctt ttctcttctt 2700
attaaaccat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa 2760
agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca 2820
gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct 2880
acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc 2940
ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg 3000
tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg 3060
ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac 3120
cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc 3180
atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc 3240
tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg 3300
atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga 3360
gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt 3420
ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag 3480
gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct 3540
tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc 3600
gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga 3660
ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga 3720
gggcaaagga atagtgaggt acctaaagaa ggagtgcgtc gaagcagatc gttcaaacat 3780
ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata 3840
atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat 3900
gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa 3960
aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg 4020
atgtcgaatc tgatcaacct gcattaatga atcggccaac gcgcggggag aggcggtttg 4080
cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 4140
cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 4200
aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 4260
gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 4320
tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 4380
agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 4440
ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg 4500
taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 4560
gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 4620
gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 4680
ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 4740
ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 4800
gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 4860
caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 4920
taagggattt tggtcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt 4980
ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 5040
acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 5100
gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg 5160
caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc 5220
atacacatac gatttaggtg acactataga acggcgcgcc aagcttgttg aaacatccct 5280
gaagtgtctc attttatttt atttattctt tgctgataaa aaaataaaat aaaagaagct 5340
aagcacacgg tcaaccattg ctctactgct aaaagggtta tgtgtagtgt tttactgcat 5400
aaattatgca gcaaacaaga caactcaaat taaaaaattt cctttgcttg tttttttgtt 5460
gtctctgact tgactttctt gtggaagttg gttgtataag gattgggaca ccattgtcct 5520
tcttaattta attttattct ttgctgataa aaaaaaaaat ttcatatagt gttaaataat 5580
aatttgttaa ataaccaaaa agtcaaatat gtttactctc gtttaaataa ttgagattcg 5640
tccagcaagg ctaaacgatt gtatagattt atgacaatat ttactttttt atagataaat 5700
gttatattat aataaattta tatacatata ttatatgtta tttattatta ttttaaatcc 5760
ttcaatattt tatcaaacca actcataatt ttttttttat ctgtaagaag caataaaatt 5820
aaatagaccc actttaagga tgatccaacc tttatacaga gtaagagagt tcaaatagta 5880
ccctttcata tacatatcaa ctaaaatatt agaaatatca tggatcaaac cttataaaga 5940
cattaaataa gtggataagt ataatatata aatgggtagt atataatata taaatggata 6000
caaacttctc tctttataat tgttatgtct ccttaacatc ctaatataat acataagtgg 6060
gtaatatata atatataaat ggagacaaac ttcttccatt ataattgtta tgtcttctta 6120
acacttatgt ctcgttcaca atgctaaggt tagaattgtt tagaaagtct tatagtacac 6180
atttgttttt gtactatttg aagcattcca taagccgtca cgattcagat gatttataat 6240
aataagagga aatttatcat agaacaataa ggtgcataga tagagtgtta atatatcata 6300
acatcctttg tttattcata gaagaagtga gatggagctc agttattata ctgttacatg 6360
gtcggataca atattccatg ctctccatga gctcttacac ctacatgcat tttagttcat 6420
acttgcggcc gctctagccc ggccgcctcc agctttcttt cctccctgca ttttcaaaga 6480
tcaattattt cctttctttg tttctcactt aggttctttc ttctttcaac agcacaccgc 6540
atgctaccaa ataaatagat ttaggtgcgg cgctatcttt tgcacacgtg tcagtgcagt 6600
acgcgtggca agtttctgag gattattgga ctaatcgtct tacacgtaat tgccgcgtgt 6660
ctgtttgcaa cgagcttcaa cacagtgtta gggacacgtg gcaatcaacc actcgtgcaa 6720
taggggctta cataaacggt cgattaatct aattggggat gaaattgact tctcttatcc 6780
cttccaaatt gtctctccgt tatcccttcc aaatgatgtt aagatttgta ttaatattta 6840
aaatgatatc attttgaaag aataatttga aaaaggataa taaaaagtta aatcaaatgc 6900
gaaatatact aaaagttatt gattaaagtt attgattaaa attattgcta atcatctctc 6960
aactctcaaa tgtttctctt taattctatg caacttaagc tctaattaat agcacttact 7020
agtagttaga ggcagataaa aacatgaaga aaaaaaaaac tttctttatc aattgataca 7080
agaatatgat cctttattta tattaaagtg gaagtgacat ttagcctatc gcacgtagtg 7140
ttgaaccatt cctcaattga aagagaatgt gaagggtgaa ttgaaaagaa aaaagagaga 7200
tggaaaggaa gaatttgaaa tagagataga acaacaaagt ttagctcttt gccttcgtac 7260
atatatttta aaaatattat ttttcagtac aaaaatcgca ataatgtcac actatgattt 7320
atgtatgaca ttgttgtaag ttggtgtggc gtaagtactt gattaaatta tttatcaaat 7380
gcaatttgca aaacaatttt gaatgaattt gtttaaaaag tattgctgtt aaatttttta 7440
tcttagaaga attgagggat cgggcg 7466
<210>57
<211>7841
<212>DNA
<213>人工序列
<220>
<223>质粒pJS89
<400>57
gatccgtcga cggcgcgccc gatcatccgg atatagttcc tcctttcagc aaaaaacccc 60
tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag 120
ccaactcagc ttcctttcgg gctttgttag cagccggatc gatccaagct gtacctcact 180
attcctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta 240
cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc 300
cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat 360
tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga 420
gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca 480
tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga 540
acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt 600
tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca 660
tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc 720
agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac 780
cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga 840
tcgcatccat agcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt 900
cttgcaacgt gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaatt 960
ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat 1020
aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc 1080
ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga 1140
cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt 1200
ccatgggtat atctccttct taaagttaaa caaaattatt tctagaggga aaccgttgtg 1260
gtctccctat agtgagtcgt attaatttcg cgggatcgag atcgatccaa ttccaatccc 1320
acaaaaatct gagcttaaca gcacagttgc tcctctcaga gcagaatcgg gtattcaaca 1380
ccctcatatc aactactacg ttgtgtataa cggtccacat gccggtatat acgatgactg 1440
gggttgtaca aaggcggcaa caaacggcgt tcccggagtt gcacacaaga aatttgccac 1500
tattacagag gcaagagcag cagctgacgc gtacacaaca agtcagcaaa cagacaggtt 1560
gaacttcatc cccaaaggag aagctcaact caagcccaag agctttgcta aggccctaac 1620
aagcccacca aagcaaaaag cccactggct cacgctagga accaaaaggc ccagcagtga 1680
tccagcccca aaagagatct cctttgcccc ggagattaca atggacgatt tcctctatct 1740
ttacgatcta ggaaggaagt tcgaaggtga aggtgacgac actatgttca ccactgataa 1800
tgagaaggtt agcctcttca atttcagaaa gaatgctgac ccacagatgg ttagagaggc 1860
ctacgcagca ggtctcatca agacgatcta cccgagtaac aatctccagg agatcaaata 1920
ccttcccaag aaggttaaag atgcagtcaa aagattcagg actaattgca tcaagaacac 1980
agagaaagac atatttctca agatcagaag tactattcca gtatggacga ttcaaggctt 2040
gcttcataaa ccaaggcaag taatagagat tggagtctct aaaaaggtag ttcctactga 2100
atctaaggcc atgcatggag tctaagattc aaatcgagga tctaacagaa ctcgccgtga 2160
agactggcga acagttcata cagagtcttt tacgactcaa tgacaagaag aaaatcttcg 2220
tcaacatggt ggagcacgac actctggtct actccaaaaa tgtcaaagat acagtctcag 2280
aagaccaaag ggctattgag acttttcaac aaaggataat ttcgggaaac ctcctcggat 2340
tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct 2400
acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatgcctct gccgacagtg 2460
gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca 2520
cgtcttcaaa gcaagtggat tgatgtgaca tctccactga cgtaagggat gacgcacaat 2580
cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga 2640
cacgctcgag ctcatttctc tattacttca gccataacaa aagaactctt ttctcttctt 2700
attaaaccat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa 2760
agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca 2820
gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct 2880
acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc 2940
ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg 3000
tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg 3060
ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac 3120
cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc 3180
atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc 3240
tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg 3300
atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga 3360
gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt 3420
ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag 3480
gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct 3540
tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc 3600
gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga 3660
ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga 3720
gggcaaagga atagtgaggt acctaaagaa ggagtgcgtc gaagcagatc gttcaaacat 3780
ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata 3840
atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat 3900
gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa 3960
aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg 4020
atgtcgaatc tgatcaacct gcattaatga atcggccaac gcgcggggag aggcggtttg 4080
cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 4140
cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 4200
aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 4260
gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 4320
tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 4380
agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 4440
ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg 4500
taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 4560
gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 4620
gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 4680
ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 4740
ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 4800
gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 4860
caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 4920
taagggattt tggtcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt 4980
ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 5040
acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 5100
gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg 5160
caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc 5220
atacacatac gatttaggtg acactataga acggcgcgcc aagcttgttg aaacatccct 5280
gaagtgtctc attttatttt atttattctt tgctgataaa aaaataaaat aaaagaagct 5340
aagcacacgg tcaaccattg ctctactgct aaaagggtta tgtgtagtgt tttactgcat 5400
aaattatgca gcaaacaaga caactcaaat taaaaaattt cctttgcttg tttttttgtt 5460
gtctctgact tgactttctt gtggaagttg gttgtataag gattgggaca ccattgtcct 5520
tcttaattta attttattct ttgctgataa aaaaaaaaat ttcatatagt gttaaataat 5580
aatttgttaa ataaccaaaa agtcaaatat gtttactctc gtttaaataa ttgagattcg 5640
tccagcaagg ctaaacgatt gtatagattt atgacaatat ttactttttt atagataaat 5700
gttatattat aataaattta tatacatata ttatatgtta tttattatta ttttaaatcc 5760
ttcaatattt tatcaaacca actcataatt ttttttttat ctgtaagaag caataaaatt 5820
aaatagaccc actttaagga tgatccaacc tttatacaga gtaagagagt tcaaatagta 5880
ccctttcata tacatatcaa ctaaaatatt agaaatatca tggatcaaac cttataaaga 5940
cattaaataa gtggataagt ataatatata aatgggtagt atataatata taaatggata 6000
caaacttctc tctttataat tgttatgtct ccttaacatc ctaatataat acataagtgg 6060
gtaatatata atatataaat ggagacaaac ttcttccatt ataattgtta tgtcttctta 6120
acacttatgt ctcgttcaca atgctaaggt tagaattgtt tagaaagtct tatagtacac 6180
atttgttttt gtactatttg aagcattcca taagccgtca cgattcagat gatttataat 6240
aataagagga aatttatcat agaacaataa ggtgcataga tagagtgtta atatatcata 6300
acatcctttg tttattcata gaagaagtga gatggagctc agttattata ctgttacatg 6360
gtcggataca atattccatg ctctccatga gctcttacac ctacatgcat tttagttcat 6420
acttgcggcc gcaacttggt ggaagaattt tatgatttga aattaagata tggtcacttc 6480
acttgtagca tgtcgtatat atagtgatta gtgagctagc tagtaatgac tgaagaagtt 6540
ccctcgtagc tatatatttg gtttaaatta atttctgata cgtaattgaa ttgtgatttg 6600
gcttaaaaac caacagggaa tgacagtttg ccaacttttt tattattaaa ctagaacttc 6660
tgtattgttt tgatatgaat tttcattatc cagcaggggc agtgaaaaag ccggtctatc 6720
tagccaatat tctttaattt gattattttg gattataaaa ttggatataa ttctctttat 6780
ttaaaattta tctctttctg taagttttgt gctaacacaa aatgaatttt cattatatat 6840
aaaatagtaa acttgtatat atccatttaa aaaaaaattc aattgatgtt aaacgtgtat 6900
attctatctg atttcgactt tcattactgc ttgaatcacg atgtttattt atttagttgt 6960
aattaatttg atgtgaatgt gaagtaggac atttttcaaa tttttgcctc actgttacca 7020
ttcttgtttc ggtttggtaa aatgtgaaat tgcagcttat atacagcaca cagttgattt 7080
aaagcgtatt tttatcagct tatttcaaag ttaaaagaag ttttagagct tttgatgttt 7140
tcttttctca aaataatcta tatgcattaa tttatacgtc cgatggcaat gtgtgtttga 7200
agtttaggta ataggacgca ttttattgca ttgggtactc aggtttagat catgattctt 7260
ccttcatttg attcatatgc aatagttaga ctcatttagg ttatttagca gggccataaa 7320
accaatagca taatactgca ataattaata atgcatatat aataatgtat atttttttat 7380
ataaaagtgg agcatataaa atgaaaggaa tagattagta tccttaattt acttgcaagt 7440
gattgcatgc acttttgtct gattcacaca tgtaaaatcg aaaaatcatt caatattggc 7500
tttgtactca aatggtccaa actcgacact gcattcaaga tgatgtgcga tatgattctg 7560
gccttttgtt tacagaaaaa agtcggcact gttaaaatta ttattctata atcgattgaa 7620
gttttgtcca ttgtttaatc tgcgtacacg tcttggggaa tgatgaatcg ttgataagca 7680
gcgagcctca taagctagac tgctagaggc acatcacctt catcagagta acgtgttgtt 7740
tgcacctgca tgtgctattt gttattttag ttggagtttt agcaataagg caatggaacc 7800
tgaaacaact caaaatgtaa tttgagagct ttttttagtt g 7841
<210>58
<211>1390
<212>DNA
<213>Morteriella alpina
<400>58
gcggccgcat ggctgctgct cccagtgtga ggacgtttac tcgggccgag gttttgaatg 60
ccgaggctct gaatgagggc aagaaggatg ccgaggcacc cttcttgatg atcatcgaca 120
acaaggtgta cgatgtccgc gagttcgtcc ctgatcatcc cggtggaagt gtgattctca 180
cgcacgttgg caaggacggc actgacgtct ttgacacttt tcaccccgag gctgcttggg 240
agactcttgc caacttttac gttggtgata ttgacgagag cgaccgcgat atcaagaatg 300
atgactttgc ggccgaggtc cgcaagctgc gtaccttgtt ccagtctctt ggttactacg 360
attcttccaa ggcatactac gccttcaagg tctcgttcaa cctctgcatc tggggtttgt 420
cgacggtcat tgtggccaag tggggccaga cctcgaccct cgccaacgtg ctctcggctg 480
cgcttttggg tctgttctgg cagcagtgcg gatggttggc tcacgacttt ttgcatcacc 540
aggtcttcca ggaccgtttc tggggtgatc ttttcggcgc cttcttggga ggtgtctgcc 600
agggcttctc gtcctcgtgg tggaaggaca agcacaacac tcaccacgcc gcccccaacg 660
tccacggcga ggatcccgac attgacaccc accctctgtt gacctggagt gagcatgcgt 720
tggagatgtt ctcggatgtc ccagatgagg agctgacccg catgtggtcg cgtttcatgg 780
tcctgaacca gacctggttt tacttcccca ttctctcgtt tgcccgtctc tcctggtgcc 840
tccagtccat tctctttgtg ctgcctaacg gtcaggccca caagccctcg ggcgcgcgtg 900
tgcccatctc gttggtcgag cagctgtcgc ttgcgatgca ctggacctgg tacctcgcca 960
ccatgttcct gttcatcaag gatcccgtca acatgctggt gtactttttg gtgtcgcagg 1020
cggtgtgcgg aaacttgttg gcgatcgtgt tctcgctcaa ccacaacggt atgcctgtga 1080
tctcgaagga ggaggcggtc gatatggatt tcttcacgaa gcagatcatc acgggtcgtg 1140
atgtccaccc gggtctattt gccaactggt tcacgggtgg attgaactat cagatcgagc 1200
accacttgtt cccttcgatg cctcgccaca acttttcaaa gatccagcct gctgtcgaga 1260
ccctgtgcaa aaagtacaat gtccgatacc acaccaccgg tatgatcgag ggaactgcag 1320
aggtctttag ccgtctgaac gaggtctcca aggctgcctc caagatgggt aaggcgcagt 1380
aagcggccgc 1390
<210>59
<211>9223
<212>DNA
<213>人工序列
<220>
<223>质粒pJS93
<400>59
gatccgtcga cggcgcgccc gatcatccgg atatagttcc tcctttcagc aaaaaacccc 60
tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag 120
ccaactcagc ttcctttcgg gctttgttag cagccggatc gatccaagct gtacctcact 180
attcctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta 240
cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc 300
cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat 360
tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga 420
gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca 480
tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga 540
acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt 600
tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca 660
tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc 720
agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac 780
cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga 840
tcgcatccat agcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt 900
cttgcaacgt gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaatt 960
ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat 1020
aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc 1080
ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga 1140
cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt 1200
ccatgggtat atctccttct taaagttaaa caaaattatt tctagaggga aaccgttgtg 1260
gtctccctat agtgagtcgt attaatttcg cgggatcgag atcgatccaa ttccaatccc 1320
acaaaaatct gagcttaaca gcacagttgc tcctctcaga gcagaatcgg gtattcaaca 1380
ccctcatatc aactactacg ttgtgtataa cggtccacat gccggtatat acgatgactg 1440
gggttgtaca aaggcggcaa caaacggcgt tcccggagtt gcacacaaga aatttgccac 1500
tattacagag gcaagagcag cagctgacgc gtacacaaca agtcagcaaa cagacaggtt 1560
gaacttcatc cccaaaggag aagctcaact caagcccaag agctttgcta aggccctaac 1620
aagcccacca aagcaaaaag cccactggct cacgctagga accaaaaggc ccagcagtga 1680
tccagcccca aaagagatct cctttgcccc ggagattaca atggacgatt tcctctatct 1740
ttacgatcta ggaaggaagt tcgaaggtga aggtgacgac actatgttca ccactgataa 1800
tgagaaggtt agcctcttca atttcagaaa gaatgctgac ccacagatgg ttagagaggc 1860
ctacgcagca ggtctcatca agacgatcta cccgagtaac aatctccagg agatcaaata 1920
ccttcccaag aaggttaaag atgcagtcaa aagattcagg actaattgca tcaagaacac 1980
agagaaagac atatttctca agatcagaag tactattcca gtatggacga ttcaaggctt 2040
gcttcataaa ccaaggcaag taatagagat tggagtctct aaaaaggtag ttcctactga 2100
atctaaggcc atgcatggag tctaagattc aaatcgagga tctaacagaa ctcgccgtga 2160
agactggcga acagttcata cagagtcttt tacgactcaa tgacaagaag aaaatcttcg 2220
tcaacatggt ggagcacgac actctggtct actccaaaaa tgtcaaagat acagtctcag 2280
aagaccaaag ggctattgag acttttcaac aaaggataat ttcgggaaac ctcctcggat 2340
tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct 2400
acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatgcctct gccgacagtg 2460
gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca 2520
cgtcttcaaa gcaagtggat tgatgtgaca tctccactga cgtaagggat gacgcacaat 2580
cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga 2640
cacgctcgag ctcatttctc tattacttca gccataacaa aagaactctt ttctcttctt 2700
attaaaccat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa 2760
agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca 2820
gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct 2880
acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc 2940
ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg 3000
tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg 3060
ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac 3120
cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc 3180
atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc 3240
tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg 3300
atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga 3360
gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt 3420
ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag 3480
gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct 3540
tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc 3600
gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga 3660
ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga 3720
gggcaaagga atagtgaggt acctaaagaa ggagtgcgtc gaagcagatc gttcaaacat 3780
ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata 3840
atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat 3900
gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa 3960
aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg 4020
atgtcgaatc tgatcaacct gcattaatga atcggccaac gcgcggggag aggcggtttg 4080
cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 4140
cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 4200
aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 4260
gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 4320
tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 4380
agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 4440
ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg 4500
taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 4560
gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 4620
gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 4680
ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 4740
ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 4800
gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 4860
caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 4920
taagggattt tggtcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt 4980
ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 5040
acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 5100
gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg 5160
caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc 5220
atacacatac gatttaggtg acactataga acggcgcgcc aagcttgttg aaacatccct 5280
gaagtgtctc attttatttt atttattctt tgctgataaa aaaataaaat aaaagaagct 5340
aagcacacgg tcaaccattg ctctactgct aaaagggtta tgtgtagtgt tttactgcat 5400
aaattatgca gcaaacaaga caactcaaat taaaaaattt cctttgcttg tttttttgtt 5460
gtctctgact tgactttctt gtggaagttg gttgtataag gattgggaca ccattgtcct 5520
tcttaattta attttattct ttgctgataa aaaaaaaaat ttcatatagt gttaaataat 5580
aatttgttaa ataaccaaaa agtcaaatat gtttactctc gtttaaataa ttgagattcg 5640
tccagcaagg ctaaacgatt gtatagattt atgacaatat ttactttttt atagataaat 5700
gttatattat aataaattta tatacatata ttatatgtta tttattatta ttttaaatcc 5760
ttcaatattt tatcaaacca actcataatt ttttttttat ctgtaagaag caataaaatt 5820
aaatagaccc actttaagga tgatccaacc tttatacaga gtaagagagt tcaaatagta 5880
ccctttcata tacatatcaa ctaaaatatt agaaatatca tggatcaaac cttataaaga 5940
cattaaataa gtggataagt ataatatata aatgggtagt atataatata taaatggata 6000
caaacttctc tctttataat tgttatgtct ccttaacatc ctaatataat acataagtgg 6060
gtaatatata atatataaat ggagacaaac ttcttccatt ataattgtta tgtcttctta 6120
acacttatgt ctcgttcaca atgctaaggt tagaattgtt tagaaagtct tatagtacac 6180
atttgttttt gtactatttg aagcattcca taagccgtca cgattcagat gatttataat 6240
aataagagga aatttatcat agaacaataa ggtgcataga tagagtgtta atatatcata 6300
acatcctttg tttattcata gaagaagtga gatggagctc agttattata ctgttacatg 6360
gtcggataca atattccatg ctctccatga gctcttacac ctacatgcat tttagttcat 6420
acttgcggcc gcttactgcg ccttacccat cttggaggca gccttggaga cctcgttcag 6480
acggctaaag acctctgcag ttccctcgat cataccggtg gtgtggtatc ggacattgta 6540
ctttttgcac agggtctcga cagcaggctg gatctttgaa aagttgtggc gaggcatcga 6600
agggaacaag tggtgctcga tctgatagtt caatccaccc gtgaaccagt tggcaaatag 6660
acccgggtgg acatcacgac ccgtgatgat ctgcttcgtg aagaaatcca tatcgaccgc 6720
ctcctccttc gagatcacag gcataccgtt gtggttgagc gagaacacga tcgccaacaa 6780
gtttccgcac accgcctgcg acaccaaaaa gtacaccagc atgttgacgg gatccttgat 6840
gaacaggaac atggtggcga ggtaccaggt ccagtgcatc gcaagcgaca gctgctcgac 6900
caacgagatg ggcacacgcg cgcccgaggg cttgtgggcc tgaccgttag gcagcacaaa 6960
gagaatggac tggaggcacc aggagagacg ggcaaacgag agaatgggga agtaaaacca 7020
ggtctggttc aggaccatga aacgcgacca catgcgggtc agctcctcat ctgggacatc 7080
cgagaacatc tccaacgcat gctcactcca ggtcaacaga gggtgggtgt caatgtcggg 7140
atcctcgccg tggacgttgg gggcggcgtg gtgagtgttg tgcttgtcct tccaccacga 7200
ggacgagaag ccctggcaga cacctcccaa gaaggcgccg aaaagatcac cccagaaacg 7260
gtcctggaag acctggtgat gcaaaaagtc gtgagccaac catccgcact gctgccagaa 7320
cagacccaaa agcgcagccg agagcacgtt ggcgagggtc gaggtctggc cccacttggc 7380
cacaatgacc gtcgacaaac cccagatgca gaggttgaac gagaccttga aggcgtagta 7440
tgccttggaa gaatcgtagt aaccaagaga ctggaacaag gtacgcagct tgcggacctc 7500
ggccgcaaag tcatcattct tgatatcgcg gtcgctctcg tcaatatcac caacgtaaaa 7560
gttggcaaga gtctcccaag cagcctcggg gtgaaaagtg tcaaagacgt cagtgccgtc 7620
cttgccaacg tgcgtgagaa tcacacttcc accgggatga tcagggacga actcgcggac 7680
atcgtacacc ttgttgtcga tgatcatcaa gaagggtgcc tcggcatcct tcttgccctc 7740
attcagagcc tcggcattca aaacctcggc ccgagtaaac gtcctcacac tgggagcagc 7800
agccatgcgg ccgcaacttg gtggaagaat tttatgattt gaaattaaga tatggtcact 7860
tcacttgtag catgtcgtat atatagtgat tagtgagcta gctagtaatg actgaagaag 7920
ttccctcgta gctatatatt tggtttaaat taatttctga tacgtaattg aattgtgatt 7980
tggcttaaaa accaacaggg aatgacagtt tgccaacttt tttattatta aactagaact 8040
tctgtattgt tttgatatga attttcatta tccagcaggg gcagtgaaaa agccggtcta 8100
tctagccaat attctttaat ttgattattt tggattataa aattggatat aattctcttt 8160
atttaaaatt tatctctttc tgtaagtttt gtgctaacac aaaatgaatt ttcattatat 8220
ataaaatagt aaacttgtat atatccattt aaaaaaaaat tcaattgatg ttaaacgtgt 8280
atattctatc tgatttcgac tttcattact gcttgaatca cgatgtttat ttatttagtt 8340
gtaattaatt tgatgtgaat gtgaagtagg acatttttca aatttttgcc tcactgttac 8400
cattcttgtt tcggtttggt aaaatgtgaa attgcagctt atatacagca cacagttgat 8460
ttaaagcgta tttttatcag cttatttcaa agttaaaaga agttttagag cttttgatgt 8520
tttcttttct caaaataatc tatatgcatt aatttatacg tccgatggca atgtgtgttt 8580
gaagtttagg taataggacg cattttattg cattgggtac tcaggtttag atcatgattc 8640
ttccttcatt tgattcatat gcaatagtta gactcattta ggttatttag cagggccata 8700
aaaccaatag cataatactg caataattaa taatgcatat ataataatgt atattttttt 8760
atataaaagt ggagcatata aaatgaaagg aatagattag tatccttaat ttacttgcaa 8820
gtgattgcat gcacttttgt ctgattcaca catgtaaaat cgaaaaatca ttcaatattg 8880
gctttgtact caaatggtcc aaactcgaca ctgcattcaa gatgatgtgc gatatgattc 8940
tggccttttg tttacagaaa aaagtcggca ctgttaaaat tattattcta taatcgattg 9000
aagttttgtc cattgtttaa tctgcgtaca cgtcttgggg aatgatgaat cgttgataag 9060
cagcgagcct cataagctag actgctagag gcacatcacc ttcatcagag taacgtgttg 9120
tttgcacctg catgtgctat ttgttatttt agttggagtt ttagcaataa ggcaatggaa 9180
cctgaaacaa ctcaaaatgt aatttgagag ctttttttag ttg 9223
<210>60
<211>7472
<212>DNA
<213>人工序列
<220>
<223>质粒pKS127
<400>60
gatccgtcga cggcgcgccc gatcatccgg atatagttcc tcctttcagc aaaaaacccc 60
tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag 120
ccaactcagc ttcctttcgg gctttgttag cagccggatc gatccaagct gtacctcact 180
attcctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta 240
cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc 300
cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat 360
tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga 420
gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca 480
tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga 540
acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt 600
tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca 660
tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc 720
agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac 780
cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga 840
tcgcatccat agcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt 900
cttgcaacgt gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaatt 960
ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat 1020
aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc 1080
ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga 1140
cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt 1200
ccatgggtat atctccttct taaagttaaa caaaattatt tctagaggga aaccgttgtg 1260
gtctccctat agtgagtcgt attaatttcg cgggatcgag atcgatccaa ttccaatccc 1320
acaaaaatct gagcttaaca gcacagttgc tcctctcaga gcagaatcgg gtattcaaca 1380
ccctcatatc aactactacg ttgtgtataa cggtccacat gccggtatat acgatgactg 1440
gggttgtaca aaggcggcaa caaacggcgt tcccggagtt gcacacaaga aatttgccac 1500
tattacagag gcaagagcag cagctgacgc gtacacaaca agtcagcaaa cagacaggtt 1560
gaacttcatc cccaaaggag aagctcaact caagcccaag agctttgcta aggccctaac 1620
aagcccacca aagcaaaaag cccactggct cacgctagga accaaaaggc ccagcagtga 1680
tccagcccca aaagagatct cctttgcccc ggagattaca atggacgatt tcctctatct 1740
ttacgatcta ggaaggaagt tcgaaggtga aggtgacgac actatgttca ccactgataa 1800
tgagaaggtt agcctcttca atttcagaaa gaatgctgac ccacagatgg ttagagaggc 1860
ctacgcagca ggtctcatca agacgatcta cccgagtaac aatctccagg agatcaaata 1920
ccttcccaag aaggttaaag atgcagtcaa aagattcagg actaattgca tcaagaacac 1980
agagaaagac atatttctca agatcagaag tactattcca gtatggacga ttcaaggctt 2040
gcttcataaa ccaaggcaag taatagagat tggagtctct aaaaaggtag ttcctactga 2100
atctaaggcc atgcatggag tctaagattc aaatcgagga tctaacagaa ctcgccgtga 2160
agactggcga acagttcata cagagtcttt tacgactcaa tgacaagaag aaaatcttcg 2220
tcaacatggt ggagcacgac actctggtct actccaaaaa tgtcaaagat acagtctcag 2280
aagaccaaag ggctattgag acttttcaac aaaggataat ttcgggaaac ctcctcggat 2340
tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct 2400
acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatgcctct gccgacagtg 2460
gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca 2520
cgtcttcaaa gcaagtggat tgatgtgaca tctccactga cgtaagggat gacgcacaat 2580
cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga 2640
cacgctcgag ctcatttctc tattacttca gccataacaa aagaactctt ttctcttctt 2700
attaaaccat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa 2760
agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca 2820
gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct 2880
acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc 2940
ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg 3000
tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg 3060
ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac 3120
cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc 3180
atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc 3240
tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg 3300
atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga 3360
gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt 3420
ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag 3480
gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct 3540
tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc 3600
gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga 3660
ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga 3720
gggcaaagga atagtgaggt acctaaagaa ggagtgcgtc gaagcagatc gttcaaacat 3780
ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata 3840
atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat 3900
gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa 3960
aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg 4020
atgtcgaatc tgatcaacct gcattaatga atcggccaac gcgcggggag aggcggtttg 4080
cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 4140
cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 4200
aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 4260
gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 4320
tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 4380
agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 4440
ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg 4500
taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 4560
gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 4620
gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 4680
ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 4740
ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 4800
gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 4860
caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 4920
taagggattt tggtcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt 4980
ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 5040
acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 5100
gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg 5160
caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc 5220
atacacatac gatttaggtg acactataga acggcgcgcc aagcttggat ccgttgaaac 5280
atccctgaag tgtctcattt tattttattt attctttgct gataaaaaaa taaaataaaa 5340
gaagctaagc acacggtcaa ccattgcttt actgctaaaa gggttatgtg tagtgtttta 5400
ctgcataaat tatgcagcaa acaagacaac tcaaattaaa aaatttcctt tgcttgtttt 5460
tttgttgtct ctgacttgac tttcttgtgg aagttggttg tataaggatt gggacaccat 5520
tgtcctcctt aatttaattt tattctttgc tgataaaaaa aaaaatttca tatagtgtta 5580
aataataatt tgttaaataa ccaaaaagtc aaatatgttt actctcgttt aaataattga 5640
gattcgtcca gcaaggctaa acgattgtat agatttatga caatatttac ttttttatag 5700
ataaatgtta tattataata aatttatata catatattat atgttattta ttattatttt 5760
aaatccttca atattttatc aaaccaactc ataatttttt ttttatctgt aagaagcaat 5820
aaaattaaat agacccactt taaggatgat ccaaccttta tacagagtaa gagagttcaa 5880
atagtaccct ttcatataca tatcaactaa aatattagaa atatcatgga tcaaacctta 5940
taaagacatt aaataagtgg ataagtataa tatataaatg ggtagtatat aatatataaa 6000
tggatacaaa cttctctctt tataattgtt atgtctcctt aacatcctaa tataatacat 6060
aagtgggtaa tatataatat ataaatggag acaaacttct tccattataa ttgttatgtc 6120
ttcttaacac ttatgtctcg ttcacaatgc taaagttaga attgtttaga aagtcttata 6180
gtacacattt gtttttgtac tatttgaagc attccataag ccgtcacgat tcagatgatt 6240
tataataata agaggaaatt tatcatagaa caataaggtg catagataga gtgttaatat 6300
atcataacat cctttgttta ttcatagaag aagtgagatg gagctcagtt attatactgt 6360
tacatggtcg gatacaatat tccatgctct ccatgagctc ttacacctac gtgcatttta 6420
gttcatactt gcggccgctc tagcccggcc gcctccagct ttctttcctc cctgcatttt 6480
caaagatcaa ttatttcctt tctttgtttc tcacttaggt tctttcttct ttcaacagca 6540
caccgcatgc taccaaataa atagatttag gtgcggcgct atcttttgca cacgtgtcag 6600
tgcagtacgc gtggcaagtt tctgaggatt attggactaa tcgtcttaca cgtaattgcc 6660
gcgtgtctgt ttgcaacgag cttcaacaca gtgttaggga cacgtggcaa tcaaccactc 6720
gtgcaatagg ggcttacata aacggtcgat taatctaatt ggggatgaaa ttgacttctc 6780
ttatcccttc caaattgtct ctccgttatc ccttccaaat gatgttaaga tttgtattaa 6840
tatttaaaat gatatcattt tgaaagaata atttgaaaaa ggataataaa aagttaaatc 6900
aaatgcgaaa tatactaaaa gttattgatt aaagttattg attaaaatta ttgctaatca 6960
tctctcaact ctcaaatgtt tctctttaat tctatgcaac ttaagctcta attaatagca 7020
cttactagta gttagaggca gataaaaaca tgaagaaaaa aaaaactttc tttatcaatt 7080
gatacaagaa tatgatcctt tatttatatt aaagtggaag tgacatttag cctatcgcac 7140
gtagtgttga accattcctc aattgaaaga gaatgtgaag ggtgaattga aaagaaaaaa 7200
gagagatgga aaggaagaat ttgaaataga gatagaacaa caaagtttag ctctttgcct 7260
tcgtacatat attttaaaaa tattattttt cagtacaaaa atcgcaataa tgtcacacta 7320
tgatttatgt atgacattgt tgtaagttgg tgtggcgtaa gtacttgatt aaattattta 7380
tcaaatgcaa tttgcaaaac aattttgaat gaatttgttt aaaaagtatt gctgttaaat 7440
tttttatctt agaagaattg agggatcggg cg 7472
<210>61
<211>7847
<212>DNA
<213>人工序列
<220>
<223>质粒pKS343
<400>61
ggccgcaagt atgaactaaa atgcacgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta actttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaagg aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtaaag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacggatcc aagcttggcg cgccgttcta tagtgtcacc 1200
taaatcgtat gtgtatgata cataaggtta tgtattaatt gtagccgcgt tctaacgaca 1260
atatgtccat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc 1320
cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg 1380
cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat 1440
caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca 1500
tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 1560
tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 1620
aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 1680
aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt 1740
taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 1800
taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 1860
agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 1920
tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca 1980
cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 2040
agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 2100
gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 2160
aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 2220
tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag 2280
ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg 2340
aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcaggt 2400
tgatcagatt cgacatcgat ctagtaacat agatgacacc gcgcgcgata atttatccta 2460
gtttgcgcgc tatattttgt tttctatcgc gtattaaatg tataattgcg ggactctaat 2520
cataaaaacc catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac 2580
gtaattcaac agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa 2640
gaaactttat tgccaaatgt ttgaacgatc tgcttcgacg cactccttct ttaggtacct 2700
cactattcct ttgccctcgg acgagtgctg gggcgtcggt ttccactatc ggcgagtact 2760
tctacacagc catcggtcca gacggccgcg cttctgcggg cgatttgtgt acgcccgaca 2820
gtcccggctc cggatcggac gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg 2880
aaattgccgt caaccaagct ctgatagagt tggtcaagac caatgcggag catatacgcc 2940
cggagccgcg gcgatcctgc aagctccgga tgcctccgct cgaagtagcg cgtctgctgc 3000
tccatacaag ccaaccacgg cctccagaag aagatgttgg cgacctcgta ttgggaatcc 3060
ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca 3120
ttgttggagc cgaaatccgc gtgcacgagg tgccggactt cggggcagtc ctcggcccaa 3180
agcatcagct catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt 3240
tgccagtgat acacatgggg atcagcaatc gcgcatatga aatcacgcca tgtagtgtat 3300
tgaccgattc cttgcggtcc gaatgggccg aacccgctcg tctggctaag atcggccgca 3360
gcgatcgcat ccatggcctc cgcgaccggc tgcagaacag cgggcagttc ggtttcaggc 3420
aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc aataggtcag gctctcgctg 3480
aattccccaa tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa gtgccgataa 3540
acataacgat ctttgtagaa accatcggcg cagctattta cccgcaggac atatccacgc 3600
cctcctacat cgaagctgaa agcacgagat tcttcgccct ccgagagctg catcaggtcg 3660
gagacgctgt cgaacttttc gatcagaaac ttctcgacag acgtcgcggt gagttcaggc 3720
tttttcatgg tttaataaga agagaaaaga gttcttttgt tatggctgaa gtaatagaga 3780
aatgagctcg agcgtgtcct ctccaaatga aatgaacttc cttatataga ggaagggtct 3840
tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatgtc acatcaatcc 3900
acttgctttg aagacgtggt tggaacgtct tctttttcca cgatgctcct cgtgggtggg 3960
ggtccatctt tgggaccact gtcggcagag gcatcttgaa tgatagcctt tcctttatcg 4020
caatgatggc atttgtagga gccaccttcc ttttctactg tcctttcgat gaagtgacag 4080
atagctgggc aatggaatcc gaggaggttt cccgaaatta tcctttgttg aaaagtctca 4140
atagcccttt ggtcttctga gactgtatct ttgacatttt tggagtagac cagagtgtcg 4200
tgctccacca tgttgacgaa gattttcttc ttgtcattga gtcgtaaaag actctgtatg 4260
aactgttcgc cagtcttcac ggcgagttct gttagatcct cgatttgaat cttagactcc 4320
atgcatggcc ttagattcag taggaactac ctttttagag actccaatct ctattacttg 4380
ccttggttta tgaagcaagc cttgaatcgt ccatactgga atagtacttc tgatcttgag 4440
aaatatgtct ttctctgtgt tcttgatgca attagtcctg aatcttttga ctgcatcttt 4500
aaccttcttg ggaaggtatt tgatctcctg gagattgtta ctcgggtaga tcgtcttgat 4560
gagacctgct gcgtaggcct ctctaaccat ctgtgggtca gcattctttc tgaaattgaa 4620
gaggctaacc ttctcattat cagtggtgaa catagtgtcg tcaccttcac cttcgaactt 4680
ccttcctaga tcgtaaagat agaggaaatc gtccattgta atctccgggg caaaggagat 4740
ctcttttggg gctggatcac tgctgggcct tttggttcct agcgtgagcc agtgggcttt 4800
ttgctttggt gggcttgtta gggccttagc aaagctcttg ggcttgagtt gagcttctcc 4860
tttggggatg aagttcaacc tgtctgtttg ctgacttgtt gtgtacgcgt cagctgctgc 4920
tcttgcctct gtaatagtgg caaatttctt gtgtgcaact ccgggaacgc cgtttgttgc 4980
cgcctttgta caaccccagt catcgtatat accggcatgt ggaccgttat acacaacgta 5040
gtagttgata tgagggtgtt gaatacccga ttctgctctg agaggagcaa ctgtgctgtt 5100
aagctcagat ttttgtggga ttggaattgg atcgatctcg atcccgcgaa attaatacga 5160
ctcactatag ggagaccaca acggtttccc tctagaaata attttgttta actttaagaa 5220
ggagatatac ccatggaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc 5280
gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct 5340
ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt 5400
ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa 5460
gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag 5520
ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg 5580
gaggctatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc 5640
ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat 5700
ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag 5760
gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac 5820
gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac 5880
tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg 5940
ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt 6000
gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag 6060
agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc 6120
gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc 6180
tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt 6240
ccgagggcaa aggaatagtg aggtacagct tggatcgatc cggctgctaa caaagcccga 6300
aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc 6360
tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atgatcgggc 6420
gcgccgtcga cggatccaac taaaaaaagc tctcaaatta cattttgagt tgtttcaggt 6480
tccattgcct tattgctaaa actccaacta aaataacaaa tagcacatgc aggtgcaaac 6540
aacacgttac tctgatgaag gtgatgtgcc tctagcagtc tagcttatga ggctcgctgc 6600
ttatcaacga ttcatcattc cccaagacgt gtacgcagat taaacaatgg acaaaacttc 6660
aatcgattat agaataataa ttttaacagt gccgactttt ttctgtaaac aaaaggccag 6720
aatcatatcg cacatcatct tgaatgcagt gtcgagtttg gaccatttga gtacaaagcc 6780
aatattgaat gatttttcga ttttacatgt gtgaatcaga caaaagtgca tgcaatcact 6840
tgcaagtaaa ttaaggatac taatctattc ctttcatttt atatgctcca cttttatata 6900
aaaaaatata cattattata tatgcattat taattattgc agtattatgc tattggtttt 6960
atggccctgc taaataacct aaatgagtct aactattgca tatgaatcaa atgaaggaag 7020
aatcatgatc taaacctgag tacccaatgc aataaaatgc gtcctattac ctaaacttca 7080
aacacacatt gccatcggac gtataaatta atgcatatag attattttga gaaaagaaaa 7140
catcaaaagc tctaaaactt cttttaactt tgaaataagc tgataaaaat acgctttaaa 7200
tcaactgtgt gctgtatata agctgcaatt tcacatttta ccaaaccgaa acaagaatgg 7260
taacagtgag gcaaaaattt gaaaaatgtc ctacttcaca ttcacatcaa attaattaca 7320
actaaataaa taaacatcgt gattcaagca gtaatgaaag tcgaaatcag atagaatata 7380
cacgtttaac atcaattgaa ttttttttta aatggatata tacaagttta ctattttata 7440
tataatgaaa attcattttg tgttagcaca aaacttacag aaagagataa attttaaata 7500
aagagaatta tatccaattt tataatccaa aataatcaaa ttaaagaata ttggctagat 7560
agaccggctt tttcactgcc cctgctggat aatgaaaatt catatcaaaa caatacagaa 7620
gttctagttt aataataaaa aagttggcaa actgtcattc cctgttggtt tttaagccaa 7680
atcacaattc aattacgtat cagaaattaa tttaaaccaa atatatagct acgagggaac 7740
ttcttcagtc attactagct agctcactaa tcactatata tacgacatgc tacaagtgaa 7800
gtgaccatat cttaatttca aatcataaaa ttcttccacc aagttgc 7847
<210>62
<211>10866
<212>DNA
<213>人工序列
<220>
<223>质粒pKS352
<220>
<221>misc_feature
<222>(9689)..(9689)
<223>n为a,c,g,或t
<400>62
agcttggcgc gccgttctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat 60
gtattaattg tagccgcgtt ctaacgacaa tatgtccata tggtgcactc tcagtacaat 120
ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc 180
ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag 240
ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt 300
gatacgccta tttttatagg ttaatgtcat gaccaaaatc ccttaacgtg agttttcgtt 360
ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 420
gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 480
ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 540
aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 600
gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 660
gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 720
aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 780
cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 840
tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 900
ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 960
atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 1020
cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt 1080
ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga 1140
gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc 1200
cgcgcgttgg ccgattcatt aatgcaggtt gatcagattc gacatcgatc tagtaacata 1260
gatgacaccg cgcgcgataa tttatcctag tttgcgcgct atattttgtt ttctatcgcg 1320
tattaaatgt ataattgcgg gactctaatc ataaaaaccc atctcataaa taacgtcatg 1380
cattacatgt taattattac atgcttaacg taattcaaca gaaattatat gataatcatc 1440
gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt tgaacgatct 1500
gcttcgacgc actccttctt taggtacctc actattcctt tgccctcgga cgagtgctgg 1560
ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag acggccgcgc 1620
ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg attgcgtcgc 1680
atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc tgatagagtt 1740
ggtcaagacc aatgcggagc atatacgccc ggagccgcgg cgatcctgca agctccggat 1800
gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc ctccagaaga 1860
agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg 1920
ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt 1980
gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg 2040
acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga tcagcaatcg 2100
cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg aatgggccga 2160
acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catggcctcc gcgaccggct 2220
gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc 2280
gggagatgca ataggtcagg ctctcgctga attccccaat gtcaagcact tccggaatcg 2340
ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc 2400
agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt 2460
cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact 2520
tctcgacaga cgtcgcggtg agttcaggct ttttcatggt ttaataagaa gagaaaagag 2580
ttcttttgtt atggctgaag taatagagaa atgagctcga gcgtgtcctc tccaaatgaa 2640
atgaacttcc ttatatagag gaagggtctt gcgaaggata gtgggattgt gcgtcatccc 2700
ttacgtcagt ggagatgtca catcaatcca cttgctttga agacgtggtt ggaacgtctt 2760
ctttttccac gatgctcctc gtgggtgggg gtccatcttt gggaccactg tcggcagagg 2820
catcttgaat gatagccttt cctttatcgc aatgatggca tttgtaggag ccaccttcct 2880
tttctactgt cctttcgatg aagtgacaga tagctgggca atggaatccg aggaggtttc 2940
ccgaaattat cctttgttga aaagtctcaa tagccctttg gtcttctgag actgtatctt 3000
tgacattttt ggagtagacc agagtgtcgt gctccaccat gttgacgaag attttcttct 3060
tgtcattgag tcgtaaaaga ctctgtatga actgttcgcc agtcttcacg gcgagttctg 3120
ttagatcctc gatttgaatc ttagactcca tgcatggcct tagattcagt aggaactacc 3180
tttttagaga ctccaatctc tattacttgc cttggtttat gaagcaagcc ttgaatcgtc 3240
catactggaa tagtacttct gatcttgaga aatatgtctt tctctgtgtt cttgatgcaa 3300
ttagtcctga atcttttgac tgcatcttta accttcttgg gaaggtattt gatctcctgg 3360
agattgttac tcgggtagat cgtcttgatg agacctgctg cgtaggcctc tctaaccatc 3420
tgtgggtcag cattctttct gaaattgaag aggctaacct tctcattatc agtggtgaac 3480
atagtgtcgt caccttcacc ttcgaacttc cttcctagat cgtaaagata gaggaaatcg 3540
tccattgtaa tctccggggc aaaggagatc tcttttgggg ctggatcact gctgggcctt 3600
ttggttccta gcgtgagcca gtgggctttt tgctttggtg ggcttgttag ggccttagca 3660
aagctcttgg gcttgagttg agcttctcct ttggggatga agttcaacct gtctgtttgc 3720
tgacttgttg tgtacgcgtc agctgctgct cttgcctctg taatagtggc aaatttcttg 3780
tgtgcaactc cgggaacgcc gtttgttgcc gcctttgtac aaccccagtc atcgtatata 3840
ccggcatgtg gaccgttata cacaacgtag tagttgatat gagggtgttg aatacccgat 3900
tctgctctga gaggagcaac tgtgctgtta agctcagatt tttgtgggat tggaattgga 3960
tcgatctcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa cggtttccct 4020
ctagaaataa ttttgtttaa ctttaagaag gagatatacc catggaaaag cctgaactca 4080
ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc 4140
agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg 4200
tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact 4260
ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc 4320
tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg 4380
aactgcccgc tgttctgcag ccggtcgcgg aggctatgga tgcgatcgct gcggccgatc 4440
ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat 4500
ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg 4560
acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg 4620
actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg 4680
acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat 4740
acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc 4800
gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc 4860
tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag 4920
cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta 4980
cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg 5040
atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagtga ggtacagctt 5100
ggatcgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg 5160
agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga 5220
aaggaggaac tatatccgga tgatcgggcg cgccgtcgac ggatccaact aaaaaaagct 5280
ctcaaattac attttgagtt gtttcaggtt ccattgcctt attgctaaaa ctccaactaa 5340
aataacaaat agcacatgca ggtgcaaaca acacgttact ctgatgaagg tgatgtgcct 5400
ctagcagtct agcttatgag gctcgctgct tatcaacgat tcatcattcc ccaagacgtg 5460
tacgcagatt aaacaatgga caaaacttca atcgattata gaataataat tttaacagtg 5520
ccgacttttt tctgtaaaca aaaggccaga atcatatcgc acatcatctt gaatgcagtg 5580
tcgagtttgg accatttgag tacaaagcca atattgaatg atttttcgat tttacatgtg 5640
tgaatcagac aaaagtgcat gcaatcactt gcaagtaaat taaggatact aatctattcc 5700
tttcatttta tatgctccac ttttatataa aaaaatatac attattatat atgcattatt 5760
aattattgca gtattatgct attggtttta tggccctgct aaataaccta aatgagtcta 5820
actattgcat atgaatcaaa tgaaggaaga atcatgatct aaacctgagt acccaatgca 5880
ataaaatgcg tcctattacc taaacttcaa acacacattg ccatcggacg tataaattaa 5940
tgcatataga ttattttgag aaaagaaaac atcaaaagct ctaaaacttc ttttaacttt 6000
gaaataagct gataaaaata cgctttaaat caactgtgtg ctgtatataa gctgcaattt 6060
cacattttac caaaccgaaa caagaatggt aacagtgagg caaaaatttg aaaaatgtcc 6120
tacttcacat tcacatcaaa ttaattacaa ctaaataaat aaacatcgtg attcaagcag 6180
taatgaaagt cgaaatcaga tagaatatac acgtttaaca tcaattgaat ttttttttaa 6240
atggatatat acaagtttac tattttatat ataatgaaaa ttcattttgt gttagcacaa 6300
aacttacaga aagagataaa ttttaaataa agagaattat atccaatttt ataatccaaa 6360
ataatcaaat taaagaatat tggctagata gaccggcttt ttcactgccc ctgctggata 6420
atgaaaattc atatcaaaac aatacagaag ttctagttta ataataaaaa agttggcaaa 6480
ctgtcattcc ctgttggttt ttaagccaaa tcacaattca attacgtatc agaaattaat 6540
ttaaaccaaa tatatagcta cgagggaact tcttcagtca ttactagcta gctcactaat 6600
cactatatat acgacatgct acaagtgaag tgaccatatc ttaatttcaa atcataaaat 6660
tcttccacca agttgcggcc gcaagtatga actaaaatgc acgtaggtgt aagagctcat 6720
ggagagcatg gaatattgta tccgaccatg taacagtata ataactgagc tccatctcac 6780
ttcttctatg aataaacaaa ggatgttatg atatattaac actctatcta tgcaccttat 6840
tgttctatga taaatttcct cttattatta taaatcatct gaatcgtgac ggcttatgga 6900
atgcttcaaa tagtacaaaa acaaatgtgt actataagac tttctaaaca attctaactt 6960
tagcattgtg aacgagacat aagtgttaag aagacataac aattataatg gaagaagttt 7020
gtctccattt atatattata tattacccac ttatgtatta tattaggatg ttaaggagac 7080
ataacaatta taaagagaga agtttgtatc catttatata ttatatacta cccatttata 7140
tattatactt atccacttat ttaatgtctt tataaggttt gatccatgat atttctaata 7200
ttttagttga tatgtatatg aaagggtact atttgaactc tcttactctg tataaaggtt 7260
ggatcatcct taaagtgggt ctatttaatt ttattgcttc ttacagataa aaaaaaaatt 7320
atgagttggt ttgataaaat attgaaggat ttaaaataat aataaataac atataatata 7380
tgtatataaa tttattataa tataacattt atctataaaa aagtaaatat tgtcataaat 7440
ctatacaatc gtttagcctt gctggacgaa tctcaattat ttaaacgaga gtaaacatat 7500
ttgacttttt ggttatttaa caaattatta tttaacacta tatgaaattt ttttttttat 7560
cagcaaagaa taaaattaaa ttaaggagga caatggtgtc ccaatcctta tacaaccaac 7620
ttccacaaga aagtcaagtc agagacaaca aaaaaacaag caaaggaaat tttttaattt 7680
gagttgtctt gtttgctgca taatttatgc agtaaaacac tacacataac ccttttagca 7740
gtaaagcaat ggttgaccgt gtgcttagct tcttttattt tattttttta tcagcaaaga 7800
ataaataaaa taaaatgaga cacttcaggg atgtttcaac ggatccaagc tgatccacta 7860
gttctagagc ggcccgcgcc gtcgacggat ataatgagcc gtaaacaaag atgattaagt 7920
agtaattaat acgtactagt aaaagtggca aaagataacg agaaagaacc aatttctttg 7980
cattcggcct tagcggaagg catatataag ctttgattat tttatttagt gtaatgattt 8040
cgtacaacca aagcatttat ttagtactct cacacttgtg tcgcggccgg ccgctacagg 8100
aacaggtggt ggcggccctc ggcgcgctcg tactgctcca cgatggtgta gtcctcgttg 8160
tgggaggtga tgtccagctt ggagtccacg tagtagtagc cgggcagctg cacgggcttc 8220
ttggccatgt agatggactt gaactccacc aggtagtggc cgccgtcctt cagcttcagg 8280
gccttgtgga tctcgccctt cagcacgccg tcgcgggggt acaggcgctc ggtggaggcc 8340
tcccagccca tagtcttctt ctgcattacg gggccgtcgg aggggaagtt cacgccgatg 8400
aacttcacct tgtagatgaa ggagccgtcc tgcagggagg agtcctgggt cacggtcacc 8460
acgccgccgt cctcgaagtt catcacgcgc tcccacttga agccctcggg gaaggacagc 8520
ttcttgtagt cggggatgtc ggcggggtgc ttcacgtaca ccttggagcc gtactggaac 8580
tggggggaca ggatgtccca ggcgaagggc agggggccgc ccttggtcac cttcagcttg 8640
gcggtctggg tgccctcgta ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg 8700
ccgttcacgg agccctccat gcgcaccttg aagcgcatga actccttgat gacgtcctcg 8760
gaggaggcca tgggccgctt ggggggctat ggaagacttt cttagttagt tgtgtgaata 8820
agcaatgttg ggagaatcgg gactacttat aggataggaa taaaacagaa aagtattaag 8880
tgctaatgaa atatttagac tgataattaa aatcttcacg tatgtccact tgatataaaa 8940
acgtcaggaa taaaggaagt acagtagaat ttaaaggtac tctttttata tatacccgtg 9000
ttctcttttt ggctagctag ttgcataaaa aataatctat atttttatca ttattttaaa 9060
tatcttatga gatggtaaat atttatcata atttttttta ctattattta ttatttgtgt 9120
gtgtaataca tatagaagtt aattacaaat tttatttact ttttcattat tttgatatga 9180
ttcaccatta atttagtgtt attatttata atagttcatt ttaatctttt tgtatatatt 9240
atgcgtgcag tacttttttc ctacatataa ctactattac attttattta tataatattt 9300
ttattaatga attttcgtga taatatgtaa tattgttcat tattatttca gattttttaa 9360
aaatatttgt gttattattt atgaaatatg taattttttt agtatttgat tttatgatga 9420
taaagtgttc taaattcaaa agaaggggga aagcgtaaac attaaaaaac gtcatcaaac 9480
aaaaacaaaa tcttgttaat aaagataaaa ctgtttgttt tgatcactgt tatttcgtaa 9540
tataaaaaca ttatttatat ttatattgtt gacaaccaaa tttgcctatc aaatctaacc 9600
aatataatgc atgcgtggca ggtaatgtac taccatgaac ttaagtcatg acataataaa 9660
ccgtgaatct gaccaatgca tgtacctanc taaattgtat ttgtgacacg aagcaaatga 9720
ttcaattcac aatggagatg ggaaacaaat aatgaagaac ccagaactaa gaaagctttt 9780
ctgaaaaata aaataaaggc aatgtcaaaa gtatactgca tcatcagtcc agaaagcaca 9840
tgatattttt ttatcagtat caatgcagct agttttattt tacaatatcg atatagctag 9900
tttaaatata ttgcagctag atttataaat atttgtgtta ttatttatca tttgtgtaat 9960
cctgttttta gtattttagt ttatatatga tgataatgta ttccaaattt aaaagaaggg 10020
aaataaattt aaacaagaaa aaaagtcatc aaacaaaaaa caaatgaaag ggtggaaaga 10080
tgttaccatg taatgtgaat gttacagtat ttcttttatt atagagttaa caaattaact 10140
aatatgattt tgttaataat gataaaatat tttttttatt attatttcat aatataaaaa 10200
tagtttactt aatataaaaa aaattctatc gttcacaaca aagttggcca cctaatttaa 10260
ccatgcatgt acccatggac catattaggt aaccatcaaa cctgatgaag agataaagag 10320
atgaagactt aagtcataac acaaaaccat aaaaaacaaa aatacaatca accgtcaatc 10380
tgaccaatgc atgaaaaagc tgcaatagtg agtggcgaca caaagcacat gattttctta 10440
caacggagat aaaaccaaaa aaatatttca tgaacaacct agaacaaata aagcttttat 10500
ataataaata tataaataaa taaaggctat ggaataatat acttcaatat atttggatta 10560
aataaattgt tggcggggtt gatatattta tacacaccta aagtcacttc aatctcattt 10620
tcacttaact tttatttttt ttttcttttt atttatcata aagagaatat tgataatata 10680
ctttttaaca tatttttatg acatttttta ttggtgaaaa cttattaaaa atcataaatt 10740
ttgtaagtta gatttattta aagagttcct cttcttattt taaatttttt aataaatttt 10800
taaataacta aaatttgtgt taaaaatgtt aaaaaagtgt gttattaacc cttctcttcg 10860
aggatc 10866
<210>63
<211>12055
<212>DNA
<213>人工序列
<220>
<223>质粒pKS364
<400>63
agcttttgat ccatgccctt catttgccgc ttattaatta atttggtaac agtccgtact 60
aatcagttac ttatccttcc cccatcataa ttaatcttgg tagtctcgaa tgccacaaca 120
ctgactagtc tcttggatca taagaaaaag ccaaggaaca aaagaagaca aaacacaatg 180
agagtatcct ttgcatagca atgtctaagt tcataaaatt caaacaaaaa cgcaatcaca 240
cacagtggac atcacttatc cactagctga tcaggatcgc cgcgtcaaga aaaaaaaact 300
ggaccccaaa agccatgcac aacaacacgt actcacaaag gtgtcaatcg agcagcccaa 360
aacattcacc aactcaaccc atcatgagcc ctcacatttg ttgtttctaa cccaacctca 420
aactcgtatt ctcttccgcc acctcatttt tgtttatttc aacacccgtc aaactgcatg 480
ccaccccgtg gccaaatgtc catgcatgtt aacaagacct atgactataa atagctgcaa 540
tctcggccca ggttttcatc atcaagaacc agttcaatat cctagtacac cgtattaaag 600
aatttaagat atactgcggc cgcatgacta tcgactcaca atactacaag tcgcgagaca 660
aaaacgacac ggcacccaaa atcgcgggaa tccgatatgc cccgctatcg acaccattac 720
tcaaccgatg tgagaccttc tctctggtct ggcacatttt cagcattccc actttcctca 780
caattttcat gctatgctgc gcaattccac tgctctggcc atttgtgatt gcgtatgtag 840
tgtacgctgt taaagacgac tccccgtcca acggaggagt ggtcaagcga tactcgccta 900
tttcaagaaa cttcttcatc tggaagctct ttggccgcta cttccccata actctgcaca 960
agacggtgga tctggagccc acgcacacat actaccctct ggacgtccag gagtatcacc 1020
tgattgctga gagatactgg ccgcagaaca agtacctccg agcaatcatc accaccatcg 1080
agtactttct gcccgccttc atgaaacggt ctctttctat caacgagcag gagcagcctg 1140
ccgagcgaga tcctctcctg tctcccgttt ctcccagctc tccgggttct caacctgaca 1200
agtggattaa ccacgacagc agatatagcc gtggagaatc atctggctcc aacggccacg 1260
cctcgggctc cgaacttaac ggcaacggca acaacggcac cactaaccga cgacctttgt 1320
cgtccgcctc tgctggctcc actgcatctg attccacgct tcttaacggg tccctcaact 1380
cctacgccaa ccagatcatt ggcgaaaacg acccacagct gtcgcccaca aaactcaagc 1440
ccactggcag aaaatacatc ttcggctacc acccccacgg cattatcggc atgggagcct 1500
ttggtggaat tgccaccgag ggagctggat ggtccaagct ctttccgggc atccctgttt 1560
ctcttatgac tctcaccaac aacttccgag tgcctctcta cagagagtac ctcatgagtc 1620
tgggagtcgc ttctgtctcc aagaagtcct gcaaggccct cctcaagcga aaccagtcta 1680
tctgcattgt cgttggtgga gcacaggaaa gtcttctggc cagacccggt gtcatggacc 1740
tggtgctact caagcgaaag ggttttgttc gacttggtat ggaggtcgga aatgtcgccc 1800
ttgttcccat catggccttt ggtgagaacg acctctatga ccaggttagc aacgacaagt 1860
cgtccaagct gtaccgattc cagcagtttg tcaagaactt ccttggattc acccttcctt 1920
tgatgcatgc ccgaggcgtc ttcaactacg atgtcggtct tgtcccctac aggcgacccg 1980
tcaacattgt ggttggttcc cccattgact tgccttatct cccacacccc accgacgaag 2040
aagtgtccga ataccacgac cgatacatcg ccgagctgca gcgaatctac aacgagcaca 2100
aggatgaata tttcatcgat tggaccgagg agggcaaagg agccccagag ttccgaatga 2160
ttgagtaagc ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag 2220
catggaatat tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc 2280
tatgaataaa caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct 2340
atgataaatt tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt 2400
caaatagtac aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat 2460
tgtgaacgag acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc 2520
atttatatat tatatattac ccacttatgt attatattag gatgttaagg agacataaca 2580
attataaaga gagaagtttg tatccattta tatattatat actacccatt tatatattat 2640
acttatccac ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag 2700
ttgatatgta tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca 2760
tccttaaagt gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt 2820
tggtttgata aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata 2880
taaatttatt ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac 2940
aatcgtttag ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact 3000
ttttggttat ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa 3060
agaataaaat taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac 3120
aagaaagtca agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg 3180
tcttgtttgc tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag 3240
caatggttga ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat 3300
aaaataaaat gagacacttc agggatgttt caacaagctt ggatcctagc ctaagtacgt 3360
actcaaaatg ccaacaaata aaaaaaaagt tgctttaata atgccaaaac aaattaataa 3420
aacacttaca acaccggatt ttttttaatt aaaatgtgcc atttaggata aatagttaat 3480
atttttaata attatttaaa aagccgtatc tactaaaatg atttttattt ggttgaaaat 3540
attaatatgt ttaaatcaac acaatctatc aaaattaaac taaaaaaaaa ataagtgtac 3600
gtggttaaca ttagtacagt aatataagag gaaaatgaga aattaagaaa ttgaaagcga 3660
gtctaatttt taaattatga acctgcatat ataaaaggaa agaaagaatc caggaagaaa 3720
agaaatgaaa ccatgcatgg tcccctcgtc atcacgagtt tctgccattt gcaatagaaa 3780
cactgaaaca cctttctctt tgtcacttaa ttgagatgcc gaagccacct cacaccatga 3840
acttcatgag gtgtagcacc caaggcttcc atagccatgc atactgaaga atgtctcaag 3900
ctcagcaccc tacttctgtg acgtgtccct cattcacctt cctctcttcc ctataaataa 3960
ccacgcctca ggttctccgc ttcacaactc aaacattctc tccattggtc cttaaacact 4020
catcagtcat caccatggag gtccgacgac gaaagataga cgtgctcaag gcccagaaaa 4080
acggctacga atcgggccca ccatctcgac aatcgtcgca gccctcctca agagcatcgt 4140
ccagaacccg caacaaacac tcctcgtcca ccctgtcgct cagcggactg accatgaaag 4200
tccagaagaa acctgcggga cccccggcga actccaaaac gccattccta cacatcaagc 4260
ccgtgcacac gtgctgctcc acatcaatgc tttcgcgcga ttatgacggc tccaacccca 4320
gcttcaaggg cttcaaaaac atcggcatga tcattctcat tgtgggaaat ctacggctcg 4380
cattcgaaaa ctacctcaaa tacggcattt ccaacccgtt cttcgacccc aaaattactc 4440
cttccgagtg gcagctctca ggcttgctca tagtcgtggc ctacgcacat atcctcatgg 4500
cctacgctat tgagagcgct gccaagctgc tgttcctctc tagcaaacac cactacatgg 4560
ccgtggggct tctgcatacc atgaacactt tgtcgtccat ctcgttgctg tcctacgtcg 4620
tctactacta cctgcccaac cccgtggcag gcacaatagt cgagtttgtg gccgttattc 4680
tgtctctcaa actcgcctca tacgccctca ctaactcgga tctccgaaaa gccgcaattc 4740
atgcccagaa gctcgacaag acgcaagacg ataacgaaaa ggaatccacc tcgtcttcct 4800
cttcttcaga tgacgcagag actttggcag acattgacgt cattcctgca tactacgcac 4860
agctgcccta cccccagaat gtgacgctgt cgaacctgct gtacttctgg tttgctccca 4920
cactggtcta ccagcccgtg taccccaaga cggagcgtat tcgacccaag cacgtgatcc 4980
gaaacctgtt tgagctcgtc tctctgtgca tgcttattca gtttctcatc ttccagtacg 5040
cctaccccat catgcagtcg tgtctggctc tgttcttcca gcccaagctc gattatgcca 5100
acatctccga gcgcctcatg aagttggcct ccgtgtctat gatggtctgg ctcattggat 5160
tctacgcttt cttccagaac ggtctcaatc ttattgccga gctcacctgt tttggaaaca 5220
gaaccttcta ccagcagtgg tggaattccc gctccattgg ccagtactgg actctatgga 5280
acaagccagt caaccagtac tttagacacc acgtctacgt gcctcttctc gctcggggca 5340
tgtcgcggtt caatgcgtcg gtggtggttt tctttttctc cgccgtcatc catgaactgc 5400
ttgtcggcat ccccactcac aacatcatcg gagccgcctt cttcggcatg atgtcgcagg 5460
tgcctctgat catggctact gagaaccttc agcatattaa ctcctctctg ggccccttcc 5520
ttggcaactg tgcattctgg ttcacctttt tcctgggaca acccacttgt gcattccttt 5580
actatttggc ctacaactac aagcagaacc agtagcggcc gcaagtatga actaaaatgc 5640
atgtaggtgt aagagctcat ggagagcatg gaatattgta tccgaccatg taacagtata 5700
ataactgagc tccatctcac ttcttctatg aataaacaaa ggatgttatg atatattaac 5760
actctatcta tgcaccttat tgttctatga taaatttcct cttattatta taaatcatct 5820
gaatcgtgac ggcttatgga atgcttcaaa tagtacaaaa acaaatgtgt actataagac 5880
tttctaaaca attctaacct tagcattgtg aacgagacat aagtgttaag aagacataac 5940
aattataatg gaagaagttt gtctccattt atatattata tattacccac ttatgtatta 6000
tattaggatg ttaaggagac ataacaatta taaagagaga agtttgtatc catttatata 6060
ttatatacta cccatttata tattatactt atccacttat ttaatgtctt tataaggttt 6120
gatccatgat atttctaata ttttagttga tatgtatatg aaagggtact atttgaactc 6180
tcttactctg tataaaggtt ggatcatcct taaagtgggt ctatttaatt ttattgcttc 6240
ttacagataa aaaaaaaatt atgagttggt ttgataaaat attgaaggat ttaaaataat 6300
aataaataac atataatata tgtatataaa tttattataa tataacattt atctataaaa 6360
aagtaaatat tgtcataaat ctatacaatc gtttagcctt gctggacgaa tctcaattat 6420
ttaaacgaga gtaaacatat ttgacttttt ggttatttaa caaattatta tttaacacta 6480
tatgaaattt ttttttttat cagcaaagaa taaaattaaa ttaagaagga caatggtgtc 6540
ccaatcctta tacaaccaac ttccacaaga aagtcaagtc agagacaaca aaaaaacaag 6600
caaaggaaat tttttaattt gagttgtctt gtttgctgca taatttatgc agtaaaacac 6660
tacacataac ccttttagca gtagagcaat ggttgaccgt gtgcttagct tcttttattt 6720
tattttttta tcagcaaaga ataaataaaa taaaatgaga cacttcaggg atgtttcaac 6780
aagctctaga ctggaattcg tcgacggcgc gcccgatcat ccggatatag ttcctccttt 6840
cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc 6900
agcggtggca gcagccaact cagcttcctt tcgggctttg ttagcagccg gatcgatcca 6960
agctgtacct cactattcct ttgccctcgg acgagtgctg gggcgtcggt ttccactatc 7020
ggcgagtact tctacacagc catcggtcca gacggccgcg cttctgcggg cgatttgtgt 7080
acgcccgaca gtcccggctc cggatcggac gattgcgtcg catcgaccct gcgcccaagc 7140
tgcatcatcg aaattgccgt caaccaagct ctgatagagt tggtcaagac caatgcggag 7200
catatacgcc cggagccgcg gcgatcctgc aagctccgga tgcctccgct cgaagtagcg 7260
cgtctgctgc tccatacaag ccaaccacgg cctccagaag aagatgttgg cgacctcgta 7320
ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc ggccattgtc 7380
cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt cggggcagtc 7440
ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc 7500
catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga aatcacgcca 7560
tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg tctggctaag 7620
atcggccgca gcgatcgcat ccatagcctc cgcgaccggc tgcagaacag cgggcagttc 7680
ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc aataggtcag 7740
gctctcgctg aattccccaa tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa 7800
gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta cccgcaggac 7860
atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct ccgagagctg 7920
catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag acgtcgcggt 7980
gagttcaggc ttttccatgg gtatatctcc ttcttaaagt taaacaaaat tatttctaga 8040
gggaaaccgt tgtggtctcc ctatagtgag tcgtattaat ttcgcgggat cgagatcgat 8100
ccaattccaa tcccacaaaa atctgagctt aacagcacag ttgctcctct cagagcagaa 8160
tcgggtattc aacaccctca tatcaactac tacgttgtgt ataacggtcc acatgccggt 8220
atatacgatg actggggttg tacaaaggcg gcaacaaacg gcgttcccgg agttgcacac 8280
aagaaatttg ccactattac agaggcaaga gcagcagctg acgcgtacac aacaagtcag 8340
caaacagaca ggttgaactt catccccaaa ggagaagctc aactcaagcc caagagcttt 8400
gctaaggccc taacaagccc accaaagcaa aaagcccact ggctcacgct aggaaccaaa 8460
aggcccagca gtgatccagc cccaaaagag atctcctttg ccccggagat tacaatggac 8520
gatttcctct atctttacga tctaggaagg aagttcgaag gtgaaggtga cgacactatg 8580
ttcaccactg ataatgagaa ggttagcctc ttcaatttca gaaagaatgc tgacccacag 8640
atggttagag aggcctacgc agcaggtctc atcaagacga tctacccgag taacaatctc 8700
caggagatca aataccttcc caagaaggtt aaagatgcag tcaaaagatt caggactaat 8760
tgcatcaaga acacagagaa agacatattt ctcaagatca gaagtactat tccagtatgg 8820
acgattcaag gcttgcttca taaaccaagg caagtaatag agattggagt ctctaaaaag 8880
gtagttccta ctgaatctaa ggccatgcat ggagtctaag attcaaatcg aggatctaac 8940
agaactcgcc gtgaagactg gcgaacagtt catacagagt cttttacgac tcaatgacaa 9000
gaagaaaatc ttcgtcaaca tggtggagca cgacactctg gtctactcca aaaatgtcaa 9060
agatacagtc tcagaagacc aaagggctat tgagactttt caacaaagga taatttcggg 9120
aaacctcctc ggattccatt gcccagctat ctgtcacttc atcgaaagga cagtagaaaa 9180
ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggctatca ttcaagatgc 9240
ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga 9300
agacgttcca accacgtctt caaagcaagt ggattgatgt gacatctcca ctgacgtaag 9360
ggatgacgca caatcccact atccttcgca agacccttcc tctatataag gaagttcatt 9420
tcatttggag aggacacgct cgagctcatt tctctattac ttcagccata acaaaagaac 9480
tcttttctct tcttattaaa ccatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa 9540
gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga 9600
atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg 9660
cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc 9720
gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg 9780
ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca 9840
gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt 9900
cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc 9960
gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc 10020
cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca 10080
cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc 10140
ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt 10200
cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca 10260
tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca 10320
actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg 10380
cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag 10440
cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc 10500
cagcactcgt ccgagggcaa aggaatagtg aggtacctaa agaaggagtg cgtcgaagca 10560
gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg 10620
atgattatca tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc 10680
atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac 10740
gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct 10800
atgttactag atcgatgtcg aatctgatca acctgcatta atgaatcggc caacgcgcgg 10860
ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 10920
cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 10980
cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 11040
accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 11100
acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 11160
cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 11220
acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt 11280
atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 11340
agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 11400
acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 11460
gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg 11520
gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 11580
gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 11640
gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 11700
acgaaaactc acgttaaggg attttggtca tgacattaac ctataaaaat aggcgtatca 11760
cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc 11820
tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg 11880
gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga 11940
ttgtactgag agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca 12000
taaccttatg tatcatacac atacgattta ggtgacacta tagaacggcg cgcca 12055
<210>64
<211>1581
<212>DNA
<213>解脂耶氏酵母
<400>64
atggaggtgc gcagacgcaa gattgacgtt ctcaaggccc agaagaatgg ttacgagtcc 60
ggcccaccat ctagacagtc ctctcagcca tcctctagag cctcctccag aacacgcaac 120
aagcactcct cctccacact ttccctttcc ggcctcacca tgaaggtgca gaagaagcca 180
gctggtccac cagccaactc caagacccca ttcctccaca tcaagccagt gcacacttgc 240
tgctccacct caatgctctc cagagactac gacggctcca acccatcctt caagggcttc 300
aagaacatcg gcatgatcat cctcatcgtg ggcaatcttc gcctcgcctt cgagaactac 360
ctcaagtacg gcatctccaa ccccttcttc gaccccaaga tcacaccctc cgagtggcag 420
ctttctggac ttctcatcgt tgtggcttac gctcacatcc tcatggctta cgctatcgag 480
tccgctgcca agctcctctt cctctcctcc aagcaccact acatggccgt tggtctcctc 540
cacaccatga acaccctctc ctccatttcc ctcctctcct acgtggtgta ctactacctc 600
cccaatccag tggctggcac tatcgttgag ttcgtggccg tgatcctttc tctcaagctc 660
gcttcctacg cccttaccaa ctccgatctc agaaaggctg ccattcacgc ccagaagctc 720
gacaagacac aggacgacaa cgagaaggag tccacctcct cttcctcttc ctctgacgac 780
gccgagacac ttgccgacat cgacgttatc ccagcttact acgctcagct tccctaccca 840
cagaacgtta ccctctccaa cctcctctac ttctggttcg ctcccaccct tgtgtaccag 900
cccgtttacc caaagacaga gagaatccgc cccaagcatg tgatccgcaa cctcttcgag 960
cttgtgtctc tctgcatgct catccagttc ctcatcttcc agtacgccta ccccatcatg 1020
cagtcttgcc ttgccctctt cttccagcca aagctcgact acgccaacat ttccgagcgc 1080
ctcatgaagc tcgcctctgt gtccatgatg gtgtggctca tcggtttcta cgccttcttc 1140
cagaacggtc tcaacctcat cgccgagctc acctgcttcg gtaaccgcac attctaccag 1200
cagtggtgga actccagatc tattggccag tactggacac tctggaacaa gcccgtgaac 1260
cagtacttcc gccatcacgt ttacgttcct cttctcgcta ggggcatgtc cagattcaac 1320
gcctctgttg tggtgttctt cttctccgct gtgattcacg agctccttgt gggcattccc 1380
acacacaaca tcatcggcgc tgccttcttc ggaatgatgt cccaggtgcc acttatcatg 1440
gccaccgaga accttcagca catcaactcc tcccttggcc cattccttgg taactgcgct 1500
ttctggttca ccttcttcct tggtcagcct acctgcgcct ttctctacta ccttgcctac 1560
aactacaagc agaaccagta g 1581
<210>65
<211>526
<212>PRT
<213>解脂耶氏酵母
<400>65
Met Glu Val Arg Arg Arg Lys Ile Asp Val Leu Lys Ala Gln Lys Asn
1 5 10 15
Gly Tyr Glu Ser Gly Pro Pro Ser Arg Gln Ser Ser Gln Pro Ser Ser
20 25 30
Arg Ala Ser Ser Arg Thr Arg Asn Lys His Ser Ser Ser Thr Leu Ser
35 40 45
Leu Ser Gly Leu Thr Met Lys Val Gln Lys Lys Pro Ala Gly Pro Pro
50 55 60
Ala Asn Ser Lys Thr Pro Phe Leu His Ile Lys Pro Val His Thr Cys
65 70 75 80
Cys Ser Thr Ser Met Leu Ser Arg Asp Tyr Asp Gly Ser Asn Pro Ser
85 90 95
Phe Lys Gly Phe Lys Asn Ile Gly Met Ile Ile Leu Ile Val Gly Asn
100 105 110
Leu Arg Leu Ala Phe Glu Asn Tyr Leu Lys Tyr Gly Ile Ser Asn Pro
115 120 125
Phe Phe Asp Pro Lys Ile Thr Pro Ser Glu Trp Gln Leu Ser Gly Leu
130 135 140
Leu Ile Val Val Ala Tyr Ala His Ile Leu Met Ala Tyr Ala Ile Glu
145 150 155 160
Ser Ala Ala Lys Leu Leu Phe Leu Ser Ser Lys His His Tyr Met Ala
165 170 175
Val Gly Leu Leu His Thr Met Asn Thr Leu Ser Ser Ile Ser Leu Leu
180 185 190
Ser Tyr Val Val Tyr Tyr Tyr Leu Pro Asn Pro Val Ala Gly Thr Ile
195 200 205
Val Glu Phe Val Ala Val Ile Leu Ser Leu Lys Leu Ala Ser Tyr Ala
210 215 220
Leu Thr Asn Ser Asp Leu Arg Lys Ala Ala Ile His Ala Gln Lys Leu
225 230 235 240
Asp Lys Thr Gln Asp Asp Asn Glu Lys Glu Ser Thr Ser Ser Ser Ser
245 250 255
Ser Ser Asp Asp Ala Glu Thr Leu Ala Asp Ile Asp Val Ile Pro Ala
260 265 270
Tyr Tyr Ala Gln Leu Pro Tyr Pro Gln Asn Val Thr Leu Ser Asn Leu
275 280 285
Leu Tyr Phe Trp Phe Ala Pro Thr Leu Val Tyr Gln Pro Val Tyr Pro
290 295 300
Lys Thr Glu Arg Ile Arg Pro Lys His Val Ile Arg Asn Leu Phe Glu
305 310 315 320
Leu Val Ser Leu Cys Met Leu Ile Gln Phe Leu Ile Phe Gln Tyr Ala
325 330 335
Tyr Pro Ile Met Gln Ser Cys Leu Ala Leu Phe Phe Gln Pro Lys Leu
340 345 350
Asp Tyr Ala Asn Ile Ser Glu Arg Leu Met Lys Leu Ala Ser Val Ser
355 360 365
Met Met Val Trp Leu Ile Gly Phe Tyr Ala Phe Phe Gln Asn Gly Leu
370 375 380
Asn Leu Ile Ala Glu Leu Thr Cys Phe Gly Asn Arg Thr Phe Tyr Gln
385 390 395 400
Gln Trp Trp Asn Ser Arg Ser Ile Gly Gln Tyr Trp Thr Leu Trp Asn
405 410 415
Lys Pro Val Asn Gln Tyr Phe Arg His His Val Tyr Val Pro Leu Leu
420 425 430
Ala Arg Gly Met Ser Arg Phe Asn Ala Ser Val Val Val Phe Phe Phe
435 440 445
Ser Ala Val Ile His Glu Leu Leu Val Gly Ile Pro Thr His Asn Ile
450 455 460
Ile Gly Ala Ala Phe Phe Gly Met Met Ser Gln Val Pro Leu Ile Met
465 470 475 480
Ala Thr Glu Asn Leu Gln His Ile Asn Ser Ser Leu Gly Pro Phe Leu
485 490 495
Gly Asn Cys Ala Phe Trp Phe Thr Phe Phe Leu Gly Gln Pro Thr Cys
500 505 510
Ala Phe Leu Tyr Tyr Leu Ala Tyr Asn Tyr Lys Gln Asn Gln
515 520 525
<210>66
<211>1545
<212>DNA
<213>解脂耶氏酵母
<400>66
atgaccatcg actcccagta ctacaagtcc cgcgacaaga acgacaccgc tcccaagatt 60
gctggtatcc gctacgctcc tctgtctact ccactcttga accgctgcga gactttctct 120
ttggtgtggc acatcttctc catccctacc ttcctcacca tcttcatgct ctgctgtgcc 180
attcccttgc tctggccctt cgtgatcgct tacgttgtgt acgccgtgaa ggatgactcc 240
ccttccaacg gcggtgttgt gaagcgctat tcccctatct cccgcaactt cttcatctgg 300
aagctctttg gccgctactt ccccatcacc ctccacaaga ccgttgattt ggagcccacc 360
cacacttact atcccttgga cgtccaagag taccacctga ttgctgagcg ctattggcct 420
cagaacaagt acctgcgcgc catcattacc accatcgagt acttcctgcc cgctttcatg 480
aagcgttccc tctccatcaa cgagcaggag cagcctgctg agagagaccc tctgttgtct 540
cccgtttccc cctcttctcc tggttctcag cccgacaagt ggatcaacca tgattcccgc 600
tattccagag gtgagtcctc tggctccaat ggtcatgctt ctggctctga actgaatggc 660
aatggcaaca acggcactac caatcgccgc cctctttctt ctgcttccgc tggctctact 720
gcttctgact ctaccttgct caacggctct ctcaactcct acgccaacca gatcattggc 780
gagaacgacc ctcagttgtc tcccactaag ctcaagccta ctggcaggaa gtacatcttc 840
gggtaccatc ctcacggtat cattgggatg ggcgcctttg gtggcattgc tactgaaggt 900
gctggctggt ccaagctctt tcctggtatc cccgtttccc tcatgaccct caccaacaac 960
tttcgcgttc ctctctatcg cgagtacctc atgtccctgg gcgttgcttc cgtttctaag 1020
aagtcctgta aggccctcct caagcgtaat cagtccatct gcatcgtggt gggtggtgcc 1080
caggaatcct tgttggctag gcctggtgtt atggatctcg tgttgctgaa gcgcaagggt 1140
tttgtgcgct tgggtatgga ggtgggcaat gttgccctcg ttccaatcat ggctttcggc 1200
gagaacgatc tctacgacca ggtgtccaac gacaagtcct ccaagctgta ccgcttccag 1260
cagttcgtga agaacttcct cggcttcacc ctccctctca tgcatgccag aggtgtgttc 1320
aactacgatg ttgggctcgt tccctaccgc cgccctgtga acattgtggt tggttctcct 1380
atcgacttgc cttacctccc tcatcccacc gatgaggaag tttccgagta ccacgaccgt 1440
tacatcgctg agctccagcg catctacaac gagcacaagg acgagtactt catcgactgg 1500
actgaggagg gtaagggcgc tcctgagttt cgtatgatcg agtag 1545
<210>67
<211>514
<212>PRT
<213>解脂耶氏酵母
<400>67
Met Thr Ile Asp Ser Gln Tyr Tyr Lys Ser Arg Asp Lys Asn Asp Thr
1 5 10 15
Ala Pro Lys Ile Ala Gly Ile Arg Tyr Ala Pro Leu Ser Thr Pro Leu
20 25 30
Leu Asn Arg Cys Glu Thr Phe Ser Leu Val Trp His Ile Phe Ser Ile
35 40 45
Pro Thr Phe Leu Thr Ile Phe Met Leu Cys Cys Ala Ile Pro Leu Leu
50 55 60
Trp Pro Phe Val Ile Ala Tyr Val Val Tyr Ala Val Lys Asp Asp Ser
65 70 75 80
Pro Ser Asn Gly Gly Val Val Lys Arg Tyr Ser Pro Ile Ser Arg Asn
85 90 95
Phe Phe Ile Trp Lys Leu Phe Gly Arg Tyr Phe Pro Ile Thr Leu His
100 105 110
Lys Thr Val Asp Leu Glu Pro Thr His Thr Tyr Tyr Pro Leu Asp Val
115 120 125
Gln Glu Tyr His Leu Ile Ala Glu Arg Tyr Trp Pro Gln Asn Lys Tyr
130 135 140
Leu Arg Ala Ile Ile Thr Thr Ile Glu Tyr Phe Leu Pro Ala Phe Met
145 150 155 160
Lys Arg Ser Leu Ser Ile Asn Glu Gln Glu Gln Pro Ala Glu Arg Asp
165 170 175
Pro Leu Leu Ser Pro Val Ser Pro Ser Ser Pro Gly Ser Gln Pro Asp
180 185 190
Lys Trp Ile Asn His Asp Ser Arg Tyr Ser Arg Gly Glu Ser Ser Gly
195 200 205
Ser Asn Gly His Ala Ser Gly Ser Glu Leu Asn Gly Asn Gly Asn Asn
210 215 220
Gly Thr Thr Asn Arg Arg Pro Leu Ser Ser Ala Ser Ala Gly Ser Thr
225 230 235 240
Ala Ser Asp Ser Thr Leu Leu Asn Gly Ser Leu Asn Ser Tyr Ala Asn
245 250 255
Gln Ile Ile Gly Glu Asn Asp Pro Gln Leu Ser Pro Thr Lys Leu Lys
260 265 270
Pro Thr Gly Arg Lys Tyr Ile Phe Gly Tyr His Pro His Gly Ile Ile
275 280 285
Gly Met Gly Ala Phe Gly Gly Ile Ala Thr Glu Gly Ala Gly Trp Ser
290 295 300
Lys Leu Phe Pro Gly Ile Pro Val Ser Leu Met Thr Leu Thr Asn Asn
305 310 315 320
Phe Arg Val Pro Leu Tyr Arg Glu Tyr Leu Met Ser Leu Gly Val Ala
325 330 335
Ser Val Ser Lys Lys Ser Cys Lys Ala Leu Leu Lys Arg Asn Gln Ser
340 345 350
Ile Cys Ile Val Val Gly Gly Ala Gln Glu Ser Leu Leu Ala Arg Pro
355 360 365
Gly Val Met Asp Leu Val Leu Leu Lys Arg Lys Gly Phe Val Arg Leu
370 375 380
Gly Met Glu Val Gly Asn Val Ala Leu Val Pro Ile Met Ala Phe Gly
385 390 395 400
Glu Asn Asp Leu Tyr Asp Gln Val Ser Asn Asp Lys Ser Ser Lys Leu
405 410 415
Tyr Arg Phe Gln Gln Phe Val Lys Asn Phe Leu Gly Phe Thr Leu Pro
420 425 430
Leu Met His Ala Arg Gly Val Phe Asn Tyr Asp Val Gly Leu Val Pro
435 440 445
Tyr Arg Arg Pro Val Asn Ile Val Val Gly Ser Pro Ile Asp Leu Pro
450 455 460
Tyr Leu Pro His Pro Thr Asp Glu Glu Val Ser Glu Tyr His Asp Arg
465 470 475 480
Tyr Ile Ala Glu Leu Gln Arg Ile Tyr Asn Glu His Lys Asp Glu Tyr
485 490 495
Phe Ile Asp Trp Thr Glu Glu Gly Lys Gly Ala Pro Glu Phe Arg Met
500 505 510
Ile Glu
<210>68
<211>8638
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1234
<400>68
ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacaagctt ggcgcgccgt tctatagtgt cacctaaatc 1200
gtatgtgtat gatacataag gttatgtatt aattgtagcc gcgttctaac gacaatatgt 1260
ccatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac 1320
acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca 1380
gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga 1440
aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgacca 1500
aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 1560
gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 1620
cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 1680
ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc 1740
accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 1800
tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 1860
cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 1920
gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc 1980
ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 2040
cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 2100
tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 2160
ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 2220
ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 2280
ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 2340
gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc aggttgatcg 2400
attcgacatc gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg 2460
cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa 2520
acccatctca taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc 2580
aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt 2640
tattgccaaa tgtttgaacg atctgcttcg acgcactcct tctttaggta cctcactatt 2700
cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac 2760
agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg 2820
ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc 2880
cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc 2940
gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac 3000
aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca 3060
tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg 3120
agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca 3180
gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt 3240
gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga 3300
ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg 3360
catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt 3420
gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaattccc 3480
caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac 3540
gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta 3600
catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc 3660
tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggctttttca 3720
tggtttaata agaagagaaa agagttcttt tgttatggct gaagtaatag agaaatgagc 3780
tcgagcgtgt cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag 3840
gatagtggga ttgtgcgtca tcccttacgt cagtggagat gtcacatcaa tccacttgct 3900
ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat 3960
ctttgggacc actgtcggca gaggcatctt gaatgatagc ctttccttta tcgcaatgat 4020
ggcatttgta ggagccacct tccttttcta ctgtcctttc gatgaagtga cagatagctg 4080
ggcaatggaa tccgaggagg tttcccgaaa ttatcctttg ttgaaaagtc tcaatagccc 4140
tttggtcttc tgagactgta tctttgacat ttttggagta gaccagagtg tcgtgctcca 4200
ccatgttgac gaagattttc ttcttgtcat tgagtcgtaa aagactctgt atgaactgtt 4260
cgccagtctt cacggcgagt tctgttagat cctcgatttg aatcttagac tccatgcatg 4320
gccttagatt cagtaggaac taccttttta gagactccaa tctctattac ttgccttggt 4380
ttatgaagca agccttgaat cgtccatact ggaatagtac ttctgatctt gagaaatatg 4440
tctttctctg tgttcttgat gcaattagtc ctgaatcttt tgactgcatc tttaaccttc 4500
ttgggaaggt atttgatctc ctggagattg ttactcgggt agatcgtctt gatgagacct 4560
gctgcgtagg cctctctaac catctgtggg tcagcattct ttctgaaatt gaagaggcta 4620
accttctcat tatcagtggt gaacatagtg tcgtcacctt caccttcgaa cttccttcct 4680
agatcgtaaa gatagaggaa atcgtccatt gtaatctccg gggcaaagga gatctctttt 4740
ggggctggat cactgctggg ccttttggtt cctagcgtga gccagtgggc tttttgcttt 4800
ggtgggcttg ttagggcctt agcaaagctc ttgggcttga gttgagcttc tcctttgggg 4860
atgaagttca acctgtctgt ttgctgactt gttgtgtacg cgtcagctgc tgctcttgcc 4920
tctgtaatag tggcaaattt cttgtgtgca actccgggaa cgccgtttgt tgccgccttt 4980
gtacaacccc agtcatcgta tataccggca tgtggaccgt tatacacaac gtagtagttg 5040
atatgagggt gttgaatacc cgattctgct ctgagaggag caactgtgct gttaagctca 5100
gatttttgtg ggattggaat tggatcgatc tcgatcccgc gaaattaata cgactcacta 5160
tagggagacc acaacggttt ccctctagaa ataattttgt ttaactttaa gaaggagata 5220
tacccatgga aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt 5280
tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct 5340
tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca 5400
aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg 5460
acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca 5520
cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcta 5580
tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc 5640
aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg 5700
tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg 5760
atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt 5820
tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg 5880
aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt 5940
tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat 6000
cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg 6060
ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat 6120
ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg 6180
atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg 6240
caaaggaata gtgaggtaca gcttggatcg atccggctgc taacaaagcc cgaaaggaag 6300
ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac 6360
gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatgatcg ggcgcgccgt 6420
cgacggatcc gtacgagatc cggccggcca gatcctgcag gagatccaag cttttgatcc 6480
atgcccttca tttgccgctt attaattaat ttggtaacag tccgtactaa tcagttactt 6540
atccttcccc catcataatt aatcttggta gtctcgaatg ccacaacact gactagtctc 6600
ttggatcata agaaaaagcc aaggaacaaa agaagacaaa acacaatgag agtatccttt 6660
gcatagcaat gtctaagttc ataaaattca aacaaaaacg caatcacaca cagtggacat 6720
cacttatcca ctagctgatc aggatcgccg cgtcaagaaa aaaaaactgg accccaaaag 6780
ccatgcacaa caacacgtac tcacaaaggt gtcaatcgag cagcccaaaa cattcaccaa 6840
ctcaacccat catgagccct cacatttgtt gtttctaacc caacctcaaa ctcgtattct 6900
cttccgccac ctcatttttg tttatttcaa cacccgtcaa actgcatgcc accccgtggc 6960
caaatgtcca tgcatgttaa caagacctat gactataaat agctgcaatc tcggcccagg 7020
ttttcatcat caagaaccag ttcaatatcc tagtacaccg tattaaagaa tttaagatat 7080
actgcggccg catgactatc gactcacaat actacaagtc gcgagacaaa aacgacacgg 7140
cacccaaaat cgcgggaatc cgatatgccc cgctatcgac accattactc aaccgatgtg 7200
agaccttctc tctggtctgg cacattttca gcattcccac tttcctcaca attttcatgc 7260
tatgctgcgc aattccactg ctctggccat ttgtgattgc gtatgtagtg tacgctgtta 7320
aagacgactc cccgtccaac ggaggagtgg tcaagcgata ctcgcctatt tcaagaaact 7380
tcttcatctg gaagctcttt ggccgctact tccccataac tctgcacaag acggtggatc 7440
tggagcccac gcacacatac taccctctgg acgtccagga gtatcacctg attgctgaga 7500
gatactggcc gcagaacaag tacctccgag caatcatcac caccatcgag tactttctgc 7560
ccgccttcat gaaacggtct ctttctatca acgagcagga gcagcctgcc gagcgagatc 7620
ctctcctgtc tcccgtttct cccagctctc cgggttctca acctgacaag tggattaacc 7680
acgacagcag atatagccgt ggagaatcat ctggctccaa cggccacgcc tcgggctccg 7740
aacttaacgg caacggcaac aacggcacca ctaaccgacg acctttgtcg tccgcctctg 7800
ctggctccac tgcatctgat tccacgcttc ttaacgggtc cctcaactcc tacgccaacc 7860
agatcattgg cgaaaacgac ccacagctgt cgcccacaaa actcaagccc actggcagaa 7920
aatacatctt cggctaccac ccccacggca ttatcggcat gggagccttt ggtggaattg 7980
ccaccgaggg agctggatgg tccaagctct ttccgggcat ccctgtttct cttatgactc 8040
tcaccaacaa cttccgagtg cctctctaca gagagtacct catgagtctg ggagtcgctt 8100
ctgtctccaa gaagtcctgc aaggccctcc tcaagcgaaa ccagtctatc tgcattgtcg 8160
ttggtggagc acaggaaagt cttctggcca gacccggtgt catggacctg gtgctactca 8220
agcgaaaggg ttttgttcga cttggtatgg aggtcggaaa tgtcgccctt gttcccatca 8280
tggcctttgg tgagaacgac ctctatgacc aggttagcaa cgacaagtcg tccaagctgt 8340
accgattcca gcagtttgtc aagaacttcc ttggattcac ccttcctttg atgcatgccc 8400
gaggcgtctt caactacgat gtcggtcttg tcccctacag gcgacccgtc aacattgtgg 8460
ttggttcccc cattgacttg ccttatctcc cacaccccac cgacgaagaa gtgtccgaat 8520
accacgaccg atacatcgcc gagctgcagc gaatctacaa cgagcacaag gatgaatatt 8580
tcatcgattg gaccgaggag ggcaaaggag ccccagagtt ccgaatgatt gagtaagc 8638
<210>69
<211>21
<212>DNA
<213>人工序列
<220>
<223>引物oSGly-2
<400>69
aagcggccgc ggtgatgact g 21
<210>70
<211>32
<212>DNA
<213>人工序列
<220>
<223>引物oSGly-3
<400>70
taagcttcgt acgctagcct aagtacgtac tc 32
<210>71
<211>3673
<212>DNA
<213>人工序列
<220>
<223>质粒pPSgly32
<400>71
gggctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta 60
attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 120
acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 180
gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 240
tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 300
cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 360
gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 420
aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 480
gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 540
aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 600
gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 660
ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 720
cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 780
ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 840
actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 900
tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 960
gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 1020
ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 1080
cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 1140
ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 1200
tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 1260
agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 1320
gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 1380
ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 1440
gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 1500
cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 1560
acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 1620
cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 1680
cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 1740
ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 1800
tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 1860
atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 1920
tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 1980
actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 2040
aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 2100
ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 2160
ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 2220
cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt 2280
gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa 2340
aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa 2400
agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac 2460
gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga 2520
accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa 2580
aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc 2640
tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc 2700
cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc 2760
agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc 2820
agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg 2880
aattgggtac cgggcccccc ctcgaggtcg acggtatcga taagcttgat atcgaattcc 2940
tgcagcccgg gggatccgcc ctaagcttcg tacgctagcc taagtacgta ctcaaaatgc 3000
caacaaataa aaaaaaagtt gctttaataa tgccaaaaca aattaataaa acacttacaa 3060
caccggattt tttttaatta aaatgtgcca tttaggataa atagttaata tttttaataa 3120
ttatttaaaa agccgtatct actaaaatga tttttatttg gttgaaaata ttaatatgtt 3180
taaatcaaca caatctatca aaattaaact aaaaaaaaaa taagtgtacg tggttaacat 3240
tagtacagta atataagagg aaaatgagaa attaagaaat tgaaagcgag tctaattttt 3300
aaattatgaa cctgcatata taaaaggaaa gaaagaatcc aggaagaaaa gaaatgaaac 3360
catgcatggt cccctcgtca tcacgagttt ctgccatttg caatagaaac actgaaacac 3420
ctttctcttt gtcacttaat tgagatgccg aagccacctc acaccatgaa cttcatgagg 3480
tgtagcaccc aaggcttcca tagccatgca tactgaagaa tgtctcaagc tcagcaccct 3540
acttctgtga cgtgtccctc attcaccttc ctctcttccc tataaataac cacgcctcag 3600
gttctccgct tcacaactca aacattctct ccattggtcc ttaaacactc atcagtcatc 3660
accgcggccg ctt 3673
<210>72
<211>4171
<212>DNA
<213>人工序列
<220>
<223>质粒pKR264
<400>72
gcccggggga tccgccctaa gcttcgtacg ctagcctaag tacgtactca aaatgccaac 60
aaataaaaaa aaagttgctt taataatgcc aaaacaaatt aataaaacac ttacaacacc 120
ggattttttt taattaaaat gtgccattta ggataaatag ttaatatttt taataattat 180
ttaaaaagcc gtatctacta aaatgatttt tatttggttg aaaatattaa tatgtttaaa 240
tcaacacaat ctatcaaaat taaactaaaa aaaaaataag tgtacgtggt taacattagt 300
acagtaatat aagaggaaaa tgagaaatta agaaattgaa agcgagtcta atttttaaat 360
tatgaacctg catatataaa aggaaagaaa gaatccagga agaaaagaaa tgaaaccatg 420
catggtcccc tcgtcatcac gagtttctgc catttgcaat agaaacactg aaacaccttt 480
ctctttgtca cttaattgag atgccgaagc cacctcacac catgaacttc atgaggtgta 540
gcacccaagg cttccatagc catgcatact gaagaatgtc tcaagctcag caccctactt 600
ctgtgacgtg tccctcattc accttcctct cttccctata aataaccacg cctcaggttc 660
tccgcttcac aactcaaaca ttctctccat tggtccttaa acactcatca gtcatcaccg 720
cggccgcatt tcgcaccaaa tcaatgaaag taataatgaa aagtctgaat aagaatactt 780
aggcttagat gcctttgtta cttgtgtaaa ataacttgag tcatgtacct ttggcggaaa 840
cagaataaat aaaaggtgaa attccaatgc tctatgtata agttagtaat acttaatgtg 900
ttctacggtt gtttcaatat catcaaactc taattgaaac tttagaacca caaatctcaa 960
tcttttctta atgaaatgaa aaatcttaat tgtaccatgt ttatgttaaa caccttacaa 1020
ttggttggag aggaggacca accgatggga caacattggg agaaagagat tcaatggaga 1080
tttggatagg agaacaacat tctttttcac ttcaatacaa gatgagtgca acactaagga 1140
tatgtatgag actttcagaa gctacgacaa catagatgag tgaggtggtg attcctagca 1200
agaaagacat tagaggaagc caaaatcgaa caaggaagac atcaagggca agagacagga 1260
ccatccatct caggaaaagg agctttggga tagtccgaga agttgtacaa gaaatttttt 1320
ggagggtgag tgatgcattg ctggtgactt taactcaatc aaaattgaga aagaaagaaa 1380
agggaggggg ctcacatgtg aatagaaggg aaacgggaga attttacagt tttgatctaa 1440
tgggcatccc agctagtggt aacatattca ccatgtttaa ccttcacgta cgtctagagg 1500
atccccgggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa 1560
accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta 1620
atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat 1680
ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt 1740
gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa 1800
cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg 1860
tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga 1920
gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 1980
cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 2040
tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 2100
aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt 2160
ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg 2220
ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga 2280
tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc 2340
tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac 2400
actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg 2460
gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca 2520
acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg 2580
gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg 2640
acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg 2700
gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag 2760
ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg 2820
gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct 2880
cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac 2940
agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact 3000
catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 3060
tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 3120
cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 3180
gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 3240
taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc 3300
ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 3360
tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 3420
ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 3480
cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 3540
agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 3600
gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 3660
atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 3720
gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 3780
gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta 3840
ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt 3900
cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc 3960
cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca 4020
acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc 4080
cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg 4140
accatgatta cgccaagctt gcatgcctgc a 4171
<210>73
<211>6130
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1212
<400>73
ctagaggatc ccccgggctg caggaattca ctggccgtcg ttttacaacg tcgtgactgg 60
gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg 120
cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc 180
gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata 240
tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg 300
ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa 360
gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc 420
gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg 480
gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta 540
tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt 600
caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc 660
ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa 720
gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt 780
aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt 840
ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc 900
atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg 960
gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg 1020
gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac 1080
atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca 1140
aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta 1200
actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat 1260
aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa 1320
tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag 1380
ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat 1440
agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt 1500
tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 1560
aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga 1620
gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 1680
atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 1740
gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 1800
gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 1860
tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 1920
accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 1980
ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 2040
cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 2100
agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 2160
ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 2220
tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 2280
ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 2340
cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc 2400
gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt 2460
tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag 2520
cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg 2580
cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc 2640
tatgaccatg attacgccaa gcttgcatgc ctgcaggcta gcctaagtac gtactcaaaa 2700
tgccaacaaa taaaaaaaaa gttgctttaa taatgccaaa acaaattaat aaaacactta 2760
caacaccgga ttttttttaa ttaaaatgtg ccatttagga taaatagtta atatttttaa 2820
taattattta aaaagccgta tctactaaaa tgatttttat ttggttgaaa atattaatat 2880
gtttaaatca acacaatcta tcaaaattaa actaaaaaaa aaataagtgt acgtggttaa 2940
cattagtaca gtaatataag aggaaaatga gaaattaaga aattgaaagc gagtctaatt 3000
tttaaattat gaacctgcat atataaaagg aaagaaagaa tccaggaaga aaagaaatga 3060
aaccatgcat ggtcccctcg tcatcacgag tttctgccat ttgcaataga aacactgaaa 3120
cacctttctc tttgtcactt aattgagatg ccgaagccac ctcacaccat gaacttcatg 3180
aggtgtagca cccaaggctt ccatagccat gcatactgaa gaatgtctca agctcagcac 3240
cctacttctg tgacgtgtcc ctcattcacc ttcctctctt ccctataaat aaccacgcct 3300
caggttctcc gcttcacaac tcaaacattc tctccattgg tccttaaaca ctcatcagtc 3360
atcaccgcgg ccgcaaacca tggaggtccg acgacgaaag atagacgtgc tcaaggccca 3420
gaaaaacggc tacgaatcgg gcccaccatc tcgacaatcg tcgcagccct cctcaagagc 3480
atcgtccaga acccgcaaca aacactcctc gtccaccctg tcgctcagcg gactgaccat 3540
gaaagtccag aagaaacctg cgggaccccc ggcgaactcc aaaacgccat tcctacacat 3600
caagcccgtg cacacgtgct gctccacatc aatgctttcg cgcgattatg acggctccaa 3660
ccccagcttc aagggcttca aaaacatcgg catgatcatt ctcattgtgg gaaatctacg 3720
gctcgcattc gaaaactacc tcaaatacgg catttccaac ccgttcttcg accccaaaat 3780
tactccttcc gagtggcagc tctcaggctt gctcatagtc gtggcctacg cacatatcct 3840
catggcctac gctattgaga gcgctgccaa gctgctgttc ctctctagca aacaccacta 3900
catggccgtg gggcttctgc ataccatgaa cactttgtcg tccatctcgt tgctgtccta 3960
cgtcgtctac tactacctgc ccaaccccgt ggcaggcaca atagtcgagt ttgtggccgt 4020
tattctgtct ctcaaactcg cctcatacgc cctcactaac tcggatctcc gaaaagccgc 4080
aattcatgcc cagaagctcg acaagacgca agacgataac gaaaaggaat ccacctcgtc 4140
ttcctcttct tcagatgacg cagagacttt ggcagacatt gacgtcattc ctgcatacta 4200
cgcacagctg ccctaccccc agaatgtgac gctgtcgaac ctgctgtact tctggtttgc 4260
tcccacactg gtctaccagc ccgtgtaccc caagacggag cgtattcgac ccaagcacgt 4320
gatccgaaac ctgtttgagc tcgtctctct gtgcatgctt attcagtttc tcatcttcca 4380
gtacgcctac cccatcatgc agtcgtgtct ggctctgttc ttccagccca agctcgatta 4440
tgccaacatc tccgagcgcc tcatgaagtt ggcctccgtg tctatgatgg tctggctcat 4500
tggattctac gctttcttcc agaacggtct caatcttatt gccgagctca cctgttttgg 4560
aaacagaacc ttctaccagc agtggtggaa ttcccgctcc attggccagt actggactct 4620
atggaacaag ccagtcaacc agtactttag acaccacgtc tacgtgcctc ttctcgctcg 4680
gggcatgtcg cggttcaatg cgtcggtggt ggttttcttt ttctccgccg tcatccatga 4740
actgcttgtc ggcatcccca ctcacaacat catcggagcc gccttcttcg gcatgatgtc 4800
gcaggtgcct ctgatcatgg ctactgagaa ccttcagcat attaactcct ctctgggccc 4860
cttccttggc aactgtgcat tctggttcac ctttttcctg ggacaaccca cttgtgcatt 4920
cctttactat ttggcctaca actacaagca gaaccagtag cggccgcaag tatgaactaa 4980
aatgcatgta ggtgtaagag ctcatggaga gcatggaata ttgtatccga ccatgtaaca 5040
gtataataac tgagctccat ctcacttctt ctatgaataa acaaaggatg ttatgatata 5100
ttaacactct atctatgcac cttattgttc tatgataaat ttcctcttat tattataaat 5160
catctgaatc gtgacggctt atggaatgct tcaaatagta caaaaacaaa tgtgtactat 5220
aagactttct aaacaattct aaccttagca ttgtgaacga gacataagtg ttaagaagac 5280
ataacaatta taatggaaga agtttgtctc catttatata ttatatatta cccacttatg 5340
tattatatta ggatgttaag gagacataac aattataaag agagaagttt gtatccattt 5400
atatattata tactacccat ttatatatta tacttatcca cttatttaat gtctttataa 5460
ggtttgatcc atgatatttc taatatttta gttgatatgt atatgaaagg gtactatttg 5520
aactctctta ctctgtataa aggttggatc atccttaaag tgggtctatt taattttatt 5580
gcttcttaca gataaaaaaa aaattatgag ttggtttgat aaaatattga aggatttaaa 5640
ataataataa ataacatata atatatgtat ataaatttat tataatataa catttatcta 5700
taaaaaagta aatattgtca taaatctata caatcgttta gccttgctgg acgaatctca 5760
attatttaaa cgagagtaaa catatttgac tttttggtta tttaacaaat tattatttaa 5820
cactatatga aatttttttt tttatcagca aagaataaaa ttaaattaag aaggacaatg 5880
gtgtcccaat ccttatacaa ccaacttcca caagaaagtc aagtcagaga caacaaaaaa 5940
acaagcaaag gaaatttttt aatttgagtt gtcttgtttg ctgcataatt tatgcagtaa 6000
aacactacac ataacccttt tagcagtaga gcaatggttg accgtgtgct tagcttcttt 6060
tattttattt ttttatcagc aaagaataaa taaaataaaa tgagacactt cagggatgtt 6120
tcaacaagct 6130
<210>74
<211>5764
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1235
<400>74
ggccgcattt cgcaccaaat caatgaaagt aataatgaaa agtctgaata agaatactta 60
ggcttagatg cctttgttac ttgtgtaaaa taacttgagt catgtacctt tggcggaaac 120
agaataaata aaaggtgaaa ttccaatgct ctatgtataa gttagtaata cttaatgtgt 180
tctacggttg tttcaatatc atcaaactct aattgaaact ttagaaccac aaatctcaat 240
cttttcttaa tgaaatgaaa aatcttaatt gtaccatgtt tatgttaaac accttacaat 300
tggttggaga ggaggaccaa ccgatgggac aacattggga gaaagagatt caatggagat 360
ttggatagga gaacaacatt ctttttcact tcaatacaag atgagtgcaa cactaaggat 420
atgtatgaga ctttcagaag ctacgacaac atagatgagt gaggtggtga ttcctagcaa 480
gaaagacatt agaggaagcc aaaatcgaac aaggaagaca tcaagggcaa gagacaggac 540
catccatctc aggaaaagga gctttgggat agtccgagaa gttgtacaag aaattttttg 600
gagggtgagt gatgcattgc tggtgacttt aactcaatca aaattgagaa agaaagaaaa 660
gggagggggc tcacatgtga atagaaggga aacgggagaa ttttacagtt ttgatctaat 720
gggcatccca gctagtggta acatattcac catgtttaac cttcacgtac gtctagagga 780
tccccgggta ccgagctcga attcactggc cgtcgtttta caacgtcgtg actgggaaaa 840
ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa 900
tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 960
gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg 1020
cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac 1080
acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt 1140
gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag 1200
acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc 1260
ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 1320
ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 1380
atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt 1440
tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc 1500
tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat 1560
ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct 1620
atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca 1680
ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg 1740
catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa 1800
cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg 1860
ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga 1920
cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg 1980
cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt 2040
tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg 2100
agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc 2160
ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca 2220
gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc 2280
atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 2340
cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 2400
agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 2460
ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 2520
accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct 2580
tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 2640
cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 2700
gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 2760
gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 2820
gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 2880
cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 2940
tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 3000
ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 3060
ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 3120
taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 3180
agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc 3240
gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa 3300
cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc 3360
ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga 3420
ccatgattac gccaagcttg catgcctgca gcccggggga tccgccctaa gcttcgtacg 3480
ctagcctaag tacgtactca aaatgccaac aaataaaaaa aaagttgctt taataatgcc 3540
aaaacaaatt aataaaacac ttacaacacc ggattttttt taattaaaat gtgccattta 3600
ggataaatag ttaatatttt taataattat ttaaaaagcc gtatctacta aaatgatttt 3660
tatttggttg aaaatattaa tatgtttaaa tcaacacaat ctatcaaaat taaactaaaa 3720
aaaaaataag tgtacgtggt taacattagt acagtaatat aagaggaaaa tgagaaatta 3780
agaaattgaa agcgagtcta atttttaaat tatgaacctg catatataaa aggaaagaaa 3840
gaatccagga agaaaagaaa tgaaaccatg catggtcccc tcgtcatcac gagtttctgc 3900
catttgcaat agaaacactg aaacaccttt ctctttgtca cttaattgag atgccgaagc 3960
cacctcacac catgaacttc atgaggtgta gcacccaagg cttccatagc catgcatact 4020
gaagaatgtc tcaagctcag caccctactt ctgtgacgtg tccctcattc accttcctct 4080
cttccctata aataaccacg cctcaggttc tccgcttcac aactcaaaca ttctctccat 4140
tggtccttaa acactcatca gtcatcaccg cggccgcaaa ccatggaggt ccgacgacga 4200
aagatagacg tgctcaaggc ccagaaaaac ggctacgaat cgggcccacc atctcgacaa 4260
tcgtcgcagc cctcctcaag agcatcgtcc agaacccgca acaaacactc ctcgtccacc 4320
ctgtcgctca gcggactgac catgaaagtc cagaagaaac ctgcgggacc cccggcgaac 4380
tccaaaacgc cattcctaca catcaagccc gtgcacacgt gctgctccac atcaatgctt 4440
tcgcgcgatt atgacggctc caaccccagc ttcaagggct tcaaaaacat cggcatgatc 4500
attctcattg tgggaaatct acggctcgca ttcgaaaact acctcaaata cggcatttcc 4560
aacccgttct tcgaccccaa aattactcct tccgagtggc agctctcagg cttgctcata 4620
gtcgtggcct acgcacatat cctcatggcc tacgctattg agagcgctgc caagctgctg 4680
ttcctctcta gcaaacacca ctacatggcc gtggggcttc tgcataccat gaacactttg 4740
tcgtccatct cgttgctgtc ctacgtcgtc tactactacc tgcccaaccc cgtggcaggc 4800
acaatagtcg agtttgtggc cgttattctg tctctcaaac tcgcctcata cgccctcact 4860
aactcggatc tccgaaaagc cgcaattcat gcccagaagc tcgacaagac gcaagacgat 4920
aacgaaaagg aatccacctc gtcttcctct tcttcagatg acgcagagac tttggcagac 4980
attgacgtca ttcctgcata ctacgcacag ctgccctacc cccagaatgt gacgctgtcg 5040
aacctgctgt acttctggtt tgctcccaca ctggtctacc agcccgtgta ccccaagacg 5100
gagcgtattc gacccaagca cgtgatccga aacctgtttg agctcgtctc tctgtgcatg 5160
cttattcagt ttctcatctt ccagtacgcc taccccatca tgcagtcgtg tctggctctg 5220
ttcttccagc ccaagctcga ttatgccaac atctccgagc gcctcatgaa gttggcctcc 5280
gtgtctatga tggtctggct cattggattc tacgctttct tccagaacgg tctcaatctt 5340
attgccgagc tcacctgttt tggaaacaga accttctacc agcagtggtg gaattcccgc 5400
tccattggcc agtactggac tctatggaac aagccagtca accagtactt tagacaccac 5460
gtctacgtgc ctcttctcgc tcggggcatg tcgcggttca atgcgtcggt ggtggttttc 5520
tttttctccg ccgtcatcca tgaactgctt gtcggcatcc ccactcacaa catcatcgga 5580
gccgccttct tcggcatgat gtcgcaggtg cctctgatca tggctactga gaaccttcag 5640
catattaact cctctctggg ccccttcctt ggcaactgtg cattctggtt cacctttttc 5700
ctgggacaac ccacttgtgc attcctttac tatttggcct acaactacaa gcagaaccag 5760
tagc 5764
<210>75
<211>11693
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1236
<400>75
gtacgagatc cggccggcca gatcctgcag gagatccaag cttttgatcc atgcccttca 60
tttgccgctt attaattaat ttggtaacag tccgtactaa tcagttactt atccttcccc 120
catcataatt aatcttggta gtctcgaatg ccacaacact gactagtctc ttggatcata 180
agaaaaagcc aaggaacaaa agaagacaaa acacaatgag agtatccttt gcatagcaat 240
gtctaagttc ataaaattca aacaaaaacg caatcacaca cagtggacat cacttatcca 300
ctagctgatc aggatcgccg cgtcaagaaa aaaaaactgg accccaaaag ccatgcacaa 360
caacacgtac tcacaaaggt gtcaatcgag cagcccaaaa cattcaccaa ctcaacccat 420
catgagccct cacatttgtt gtttctaacc caacctcaaa ctcgtattct cttccgccac 480
ctcatttttg tttatttcaa cacccgtcaa actgcatgcc accccgtggc caaatgtcca 540
tgcatgttaa caagacctat gactataaat agctgcaatc tcggcccagg ttttcatcat 600
caagaaccag ttcaatatcc tagtacaccg tattaaagaa tttaagatat actgcggccg 660
catgactatc gactcacaat actacaagtc gcgagacaaa aacgacacgg cacccaaaat 720
cgcgggaatc cgatatgccc cgctatcgac accattactc aaccgatgtg agaccttctc 780
tctggtctgg cacattttca gcattcccac tttcctcaca attttcatgc tatgctgcgc 840
aattccactg ctctggccat ttgtgattgc gtatgtagtg tacgctgtta aagacgactc 900
cccgtccaac ggaggagtgg tcaagcgata ctcgcctatt tcaagaaact tcttcatctg 960
gaagctcttt ggccgctact tccccataac tctgcacaag acggtggatc tggagcccac 1020
gcacacatac taccctctgg acgtccagga gtatcacctg attgctgaga gatactggcc 1080
gcagaacaag tacctccgag caatcatcac caccatcgag tactttctgc ccgccttcat 1140
gaaacggtct ctttctatca acgagcagga gcagcctgcc gagcgagatc ctctcctgtc 1200
tcccgtttct cccagctctc cgggttctca acctgacaag tggattaacc acgacagcag 1260
atatagccgt ggagaatcat ctggctccaa cggccacgcc tcgggctccg aacttaacgg 1320
caacggcaac aacggcacca ctaaccgacg acctttgtcg tccgcctctg ctggctccac 1380
tgcatctgat tccacgcttc ttaacgggtc cctcaactcc tacgccaacc agatcattgg 1440
cgaaaacgac ccacagctgt cgcccacaaa actcaagccc actggcagaa aatacatctt 1500
cggctaccac ccccacggca ttatcggcat gggagccttt ggtggaattg ccaccgaggg 1560
agctggatgg tccaagctct ttccgggcat ccctgtttct cttatgactc tcaccaacaa 1620
cttccgagtg cctctctaca gagagtacct catgagtctg ggagtcgctt ctgtctccaa 1680
gaagtcctgc aaggccctcc tcaagcgaaa ccagtctatc tgcattgtcg ttggtggagc 1740
acaggaaagt cttctggcca gacccggtgt catggacctg gtgctactca agcgaaaggg 1800
ttttgttcga cttggtatgg aggtcggaaa tgtcgccctt gttcccatca tggcctttgg 1860
tgagaacgac ctctatgacc aggttagcaa cgacaagtcg tccaagctgt accgattcca 1920
gcagtttgtc aagaacttcc ttggattcac ccttcctttg atgcatgccc gaggcgtctt 1980
caactacgat gtcggtcttg tcccctacag gcgacccgtc aacattgtgg ttggttcccc 2040
cattgacttg ccttatctcc cacaccccac cgacgaagaa gtgtccgaat accacgaccg 2100
atacatcgcc gagctgcagc gaatctacaa cgagcacaag gatgaatatt tcatcgattg 2160
gaccgaggag ggcaaaggag ccccagagtt ccgaatgatt gagtaagcgg ccgcaagtat 2220
gaactaaaat gcatgtaggt gtaagagctc atggagagca tggaatattg tatccgacca 2280
tgtaacagta taataactga gctccatctc acttcttcta tgaataaaca aaggatgtta 2340
tgatatatta acactctatc tatgcacctt attgttctat gataaatttc ctcttattat 2400
tataaatcat ctgaatcgtg acggcttatg gaatgcttca aatagtacaa aaacaaatgt 2460
gtactataag actttctaaa caattctaac cttagcattg tgaacgagac ataagtgtta 2520
agaagacata acaattataa tggaagaagt ttgtctccat ttatatatta tatattaccc 2580
acttatgtat tatattagga tgttaaggag acataacaat tataaagaga gaagtttgta 2640
tccatttata tattatatac tacccattta tatattatac ttatccactt atttaatgtc 2700
tttataaggt ttgatccatg atatttctaa tattttagtt gatatgtata tgaaagggta 2760
ctatttgaac tctcttactc tgtataaagg ttggatcatc cttaaagtgg gtctatttaa 2820
ttttattgct tcttacagat aaaaaaaaaa ttatgagttg gtttgataaa atattgaagg 2880
atttaaaata ataataaata acatataata tatgtatata aatttattat aatataacat 2940
ttatctataa aaaagtaaat attgtcataa atctatacaa tcgtttagcc ttgctggacg 3000
aatctcaatt atttaaacga gagtaaacat atttgacttt ttggttattt aacaaattat 3060
tatttaacac tatatgaaat tttttttttt atcagcaaag aataaaatta aattaagaag 3120
gacaatggtg tcccaatcct tatacaacca acttccacaa gaaagtcaag tcagagacaa 3180
caaaaaaaca agcaaaggaa attttttaat ttgagttgtc ttgtttgctg cataatttat 3240
gcagtaaaac actacacata acccttttag cagtagagca atggttgacc gtgtgcttag 3300
cttcttttat tttatttttt tatcagcaaa gaataaataa aataaaatga gacacttcag 3360
ggatgtttca acaagcttgg cgcgccgttc tatagtgtca cctaaatcgt atgtgtatga 3420
tacataaggt tatgtattaa ttgtagccgc gttctaacga caatatgtcc atatggtgca 3480
ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac 3540
ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga 3600
ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac 3660
gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgaccaaa atcccttaac 3720
gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 3780
atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 3840
tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 3900
gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 3960
actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 4020
gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 4080
agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 4140
ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa 4200
aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 4260
cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 4320
gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 4380
cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 4440
cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 4500
gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca 4560
aaccgcctct ccccgcgcgt tggccgattc attaatgcag gttgatcgat tcgacatcga 4620
tctagtaaca tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg 4680
ttttctatcg cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata 4740
aataacgtca tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat 4800
atgataatca tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg 4860
tttgaacgat ctgcttcgac gcactccttc tttaggtacc tcactattcc tttgccctcg 4920
gacgagtgct ggggcgtcgg tttccactat cggcgagtac ttctacacag ccatcggtcc 4980
agacggccgc gcttctgcgg gcgatttgtg tacgcccgac agtcccggct ccggatcgga 5040
cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc 5100
tctgatagag ttggtcaaga ccaatgcgga gcatatacgc ccggagccgc ggcgatcctg 5160
caagctccgg atgcctccgc tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg 5220
gcctccagaa gaagatgttg gcgacctcgt attgggaatc cccgaacatc gcctcgctcc 5280
agtcaatgac cgctgttatg cggccattgt ccgtcaggac attgttggag ccgaaatccg 5340
cgtgcacgag gtgccggact tcggggcagt cctcggccca aagcatcagc tcatcgagag 5400
cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt ttgccagtga tacacatggg 5460
gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc 5520
cgaatgggcc gaacccgctc gtctggctaa gatcggccgc agcgatcgca tccatggcct 5580
ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg caggtcttgc aacgtgacac 5640
cctgtgcacg gcgggagatg caataggtca ggctctcgct gaattcccca atgtcaagca 5700
cttccggaat cgggagcgcg gccgatgcaa agtgccgata aacataacga tctttgtaga 5760
aaccatcggc gcagctattt acccgcagga catatccacg ccctcctaca tcgaagctga 5820
aagcacgaga ttcttcgccc tccgagagct gcatcaggtc ggagacgctg tcgaactttt 5880
cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg ctttttcatg gtttaataag 5940
aagagaaaag agttcttttg ttatggctga agtaatagag aaatgagctc gagcgtgtcc 6000
tctccaaatg aaatgaactt ccttatatag aggaagggtc ttgcgaagga tagtgggatt 6060
gtgcgtcatc ccttacgtca gtggagatgt cacatcaatc cacttgcttt gaagacgtgg 6120
ttggaacgtc ttctttttcc acgatgctcc tcgtgggtgg gggtccatct ttgggaccac 6180
tgtcggcaga ggcatcttga atgatagcct ttcctttatc gcaatgatgg catttgtagg 6240
agccaccttc cttttctact gtcctttcga tgaagtgaca gatagctggg caatggaatc 6300
cgaggaggtt tcccgaaatt atcctttgtt gaaaagtctc aatagccctt tggtcttctg 6360
agactgtatc tttgacattt ttggagtaga ccagagtgtc gtgctccacc atgttgacga 6420
agattttctt cttgtcattg agtcgtaaaa gactctgtat gaactgttcg ccagtcttca 6480
cggcgagttc tgttagatcc tcgatttgaa tcttagactc catgcatggc cttagattca 6540
gtaggaacta cctttttaga gactccaatc tctattactt gccttggttt atgaagcaag 6600
ccttgaatcg tccatactgg aatagtactt ctgatcttga gaaatatgtc tttctctgtg 6660
ttcttgatgc aattagtcct gaatcttttg actgcatctt taaccttctt gggaaggtat 6720
ttgatctcct ggagattgtt actcgggtag atcgtcttga tgagacctgc tgcgtaggcc 6780
tctctaacca tctgtgggtc agcattcttt ctgaaattga agaggctaac cttctcatta 6840
tcagtggtga acatagtgtc gtcaccttca ccttcgaact tccttcctag atcgtaaaga 6900
tagaggaaat cgtccattgt aatctccggg gcaaaggaga tctcttttgg ggctggatca 6960
ctgctgggcc ttttggttcc tagcgtgagc cagtgggctt tttgctttgg tgggcttgtt 7020
agggccttag caaagctctt gggcttgagt tgagcttctc ctttggggat gaagttcaac 7080
ctgtctgttt gctgacttgt tgtgtacgcg tcagctgctg ctcttgcctc tgtaatagtg 7140
gcaaatttct tgtgtgcaac tccgggaacg ccgtttgttg ccgcctttgt acaaccccag 7200
tcatcgtata taccggcatg tggaccgtta tacacaacgt agtagttgat atgagggtgt 7260
tgaatacccg attctgctct gagaggagca actgtgctgt taagctcaga tttttgtggg 7320
attggaattg gatcgatctc gatcccgcga aattaatacg actcactata gggagaccac 7380
aacggtttcc ctctagaaat aattttgttt aactttaaga aggagatata cccatggaaa 7440
agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct 7500
ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag 7560
ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg 7620
tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat 7680
tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc 7740
tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggctatg gatgcgatcg 7800
ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc 7860
aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc 7920
aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc 7980
tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca 8040
atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg 8100
gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg 8160
agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc 8220
gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt 8280
tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga 8340
ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag 8400
aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagt 8460
gaggtacagc ttggatcgat ccggctgcta acaaagcccg aaaggaagct gagttggctg 8520
ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg 8580
gttttttgct gaaaggagga actatatccg gatgatcggg cgcgccgtcg acggatccgt 8640
acgtgaaggt taaacatggt gaatatgtta ccactagctg ggatgcccat tagatcaaaa 8700
ctgtaaaatt ctcccgtttc ccttctattc acatgtgagc cccctccctt ttctttcttt 8760
ctcaattttg attgagttaa agtcaccagc aatgcatcac tcaccctcca aaaaatttct 8820
tgtacaactt ctcggactat cccaaagctc cttttcctga gatggatggt cctgtctctt 8880
gcccttgatg tcttccttgt tcgattttgg cttcctctaa tgtctttctt gctaggaatc 8940
accacctcac tcatctatgt tgtcgtagct tctgaaagtc tcatacatat ccttagtgtt 9000
gcactcatct tgtattgaag tgaaaaagaa tgttgttctc ctatccaaat ctccattgaa 9060
tctctttctc ccaatgttgt cccatcggtt ggtcctcctc tccaaccaat tgtaaggtgt 9120
ttaacataaa catggtacaa ttaagatttt tcatttcatt aagaaaagat tgagatttgt 9180
ggttctaaag tttcaattag agtttgatga tattgaaaca accgtagaac acattaagta 9240
ttactaactt atacatagag cattggaatt tcacctttta tttattctgt ttccgccaaa 9300
ggtacatgac tcaagttatt ttacacaagt aacaaaggca tctaagccta agtattctta 9360
ttcagacttt tcattattac tttcattgat ttggtgcgaa atgcggccgc tactggttct 9420
gcttgtagtt gtaggccaaa tagtaaagga atgcacaagt gggttgtccc aggaaaaagg 9480
tgaaccagaa tgcacagttg ccaaggaagg ggcccagaga ggagttaata tgctgaaggt 9540
tctcagtagc catgatcaga ggcacctgcg acatcatgcc gaagaaggcg gctccgatga 9600
tgttgtgagt ggggatgccg acaagcagtt catggatgac ggcggagaaa aagaaaacca 9660
ccaccgacgc attgaaccgc gacatgcccc gagcgagaag aggcacgtag acgtggtgtc 9720
taaagtactg gttgactggc ttgttccata gagtccagta ctggccaatg gagcgggaat 9780
tccaccactg ctggtagaag gttctgtttc caaaacaggt gagctcggca ataagattga 9840
gaccgttctg gaagaaagcg tagaatccaa tgagccagac catcatagac acggaggcca 9900
acttcatgag gcgctcggag atgttggcat aatcgagctt gggctggaag aacagagcca 9960
gacacgactg catgatgggg taggcgtact ggaagatgag aaactgaata agcatgcaca 10020
gagagacgag ctcaaacagg tttcggatca cgtgcttggg tcgaatacgc tccgtcttgg 10080
ggtacacggg ctggtagacc agtgtgggag caaaccagaa gtacagcagg ttcgacagcg 10140
tcacattctg ggggtagggc agctgtgcgt agtatgcagg aatgacgtca atgtctgcca 10200
aagtctctgc gtcatctgaa gaagaggaag acgaggtgga ttccttttcg ttatcgtctt 10260
gcgtcttgtc gagcttctgg gcatgaattg cggcttttcg gagatccgag ttagtgaggg 10320
cgtatgaggc gagtttgaga gacagaataa cggccacaaa ctcgactatt gtgcctgcca 10380
cggggttggg caggtagtag tagacgacgt aggacagcaa cgagatggac gacaaagtgt 10440
tcatggtatg cagaagcccc acggccatgt agtggtgttt gctagagagg aacagcagct 10500
tggcagcgct ctcaatagcg taggccatga ggatatgtgc gtaggccacg actatgagca 10560
agcctgagag ctgccactcg gaaggagtaa ttttggggtc gaagaacggg ttggaaatgc 10620
cgtatttgag gtagttttcg aatgcgagcc gtagatttcc cacaatgaga atgatcatgc 10680
cgatgttttt gaagcccttg aagctggggt tggagccgtc ataatcgcgc gaaagcattg 10740
atgtggagca gcacgtgtgc acgggcttga tgtgtaggaa tggcgttttg gagttcgccg 10800
ggggtcccgc aggtttcttc tggactttca tggtcagtcc gctgagcgac agggtggacg 10860
aggagtgttt gttgcgggtt ctggacgatg ctcttgagga gggctgcgac gattgtcgag 10920
atggtgggcc cgattcgtag ccgtttttct gggccttgag cacgtctatc tttcgtcgtc 10980
ggacctccat ggtttgcggc cgcggtgatg actgatgagt gtttaaggac caatggagag 11040
aatgtttgag ttgtgaagcg gagaacctga ggcgtggtta tttataggga agagaggaag 11100
gtgaatgagg gacacgtcac agaagtaggg tgctgagctt gagacattct tcagtatgca 11160
tggctatgga agccttgggt gctacacctc atgaagttca tggtgtgagg tggcttcggc 11220
atctcaatta agtgacaaag agaaaggtgt ttcagtgttt ctattgcaaa tggcagaaac 11280
tcgtgatgac gaggggacca tgcatggttt catttctttt cttcctggat tctttctttc 11340
cttttatata tgcaggttca taatttaaaa attagactcg ctttcaattt cttaatttct 11400
cattttcctc ttatattact gtactaatgt taaccacgta cacttatttt ttttttagtt 11460
taattttgat agattgtgtt gatttaaaca tattaatatt ttcaaccaaa taaaaatcat 11520
tttagtagat acggcttttt aaataattat taaaaatatt aactatttat cctaaatggc 11580
acattttaat taaaaaaaat ccggtgttgt aagtgtttta ttaatttgtt ttggcattat 11640
taaagcaact ttttttttat ttgttggcat tttgagtacg tacttaggct agc 11693
<210>76
<211>34
<212>DNA
<213>人工序列
<220>
<223>引物oYDG2-1
<400>76
agcggccgca acatgactat cgactcacaa tact 34
<210>77
<211>25
<212>DNA
<213>人工序列
<220>
<223>引物oYDG2-2
<400>77
tgcggccgct tactcaatca ttcgg 25
<210>78
<211>5079
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1254
<400>78
aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgatgca 60
tagcttgagt attctaacgc gtcacctaaa tagcttggcg taatcatggt catagctgtt 120
tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 180
gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 240
gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 300
ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 360
ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 420
cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaagcccag 480
gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 540
tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 600
ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 660
atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 720
gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 780
tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 840
cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 900
cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 960
tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 1020
cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 1080
cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 1140
gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 1200
gatcctttta aattaaaaat gaagttttag cacgtgtcag tcctgctcct cggccacgaa 1260
gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc 1320
gatctcggtc atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca 1380
ctcggcgtac agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac 1440
cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa 1500
gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc 1560
ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc aacttggcca tggtggccct 1620
cctcacgtgc tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 1680
tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 1740
acctgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa 1800
ttgtaagcgt taataattca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 1860
gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 1920
tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc 1980
cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 2040
gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg 2100
aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 2160
ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 2220
caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 2280
tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 2340
cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 2400
gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg 2460
gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 2520
cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 2580
gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga 2640
tcagagcttg atcccctgcg ccatcagatc cttggcggcg agaaagccat ccagtttact 2700
ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct 2760
gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt 2820
ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt 2880
cagcaccgtt tctgcggact ggctttctac gtgaaaagga tctaggtgaa gatccttttt 2940
gataatctca tgcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg 3000
tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg 3060
gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt 3120
tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc 3180
cccggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt 3240
ttataggtgt aaaccttaaa ctgccgtacg tataggctgc gcaactgttg ggaagggcga 3300
tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 3360
ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa 3420
ttgtaatacg actcactata gggcgaattg ggccctctag atgcatgctc gagcggccgc 3480
cagtgtgatg gatatctgca gaattcagga gcggccgcaa catgactatc gactcacaat 3540
actacaagtc gcgagacaaa aacgacacgg cacccaaaat cgcgggaatc cgatatgccc 3600
cgctatcgac accattactc aaccgatgtg agaccttctc tctggtctgg cacattttca 3660
gcattcccac tttcctcaca attttcatgc tatgctgcgc aattccactg ctctggccat 3720
ttgtgattgc gtatgtagtg tacgctgtta aagacgactc cccgtccaac ggaggagtgg 3780
tcaagcgata ctcgcctatt tcaagaaact tcttcatctg gaagctcttt ggccgctact 3840
tccccataac tctgcacaag acggtggatc tggagcccac gcacacatac taccctctgg 3900
acgtccagga gtatcacctg attgctgaga gatactggcc gcagaacaag tacctccgag 3960
caatcatcac caccatcgag tactttctgc ccgccttcat gaaacggtct ctttctatca 4020
acgagcagga gcagcctgcc gagcgagatc ctctcctgtc tcccgtttct cccagctctc 4080
cgggttctca acctgacaag tggattaacc acgacagcag atatagccgt ggagaatcat 4140
ctggctccaa cggccacgcc tcgggctccg aacttaacgg caacggcaac aacggcacca 4200
ctaaccgacg acctttgtcg tccgcctctg ctggctccac tgcatctgat tccacgcttc 4260
ttaacgggtc cctcaactcc tacgccaacc agatcattgg cgaaaacgac ccacagctgt 4320
cgcccacaaa actcaagccc actggcagaa aatacatctt cggctaccac ccccacggca 4380
ttatcggcat gggagccttt ggtggaattg ccaccgaggg agctggatgg tccaagctct 4440
ttccgggcat ccctgtttct cttatgactc tcaccaacaa cttccgagtg cctctctaca 4500
gagagtacct catgagtctg ggagtcgctt ctgtctccaa gaagtcctgc aaggccctcc 4560
tcaagcgaaa ccagtctatc tgcattgtcg ttggtggagc acaggaaagt cttctggcca 4620
gacccggtgt catggacctg gtgctactca agcgaaaggg ttttgttcga cttggtatgg 4680
aggtcggaaa tgtcgccctt gttcccatca tggcctttgg tgagaacgac ctctatgacc 4740
aggttagcaa cgacaagtcg tccaagctgt accgattcca gcagtttgtc aagaacttcc 4800
ttggattcac ccttcctttg atgcatgccc gaggcgtctt caactacgat gtcggtcttg 4860
tcccctacag gcgacccgtc aacattgtgg ttggttcccc cattgacttg ccttatctcc 4920
cacaccccac cgacgaagaa gtgtccgaat accacgaccg atacatcgcc gagctgcagc 4980
gaatctacaa cgagcacaag gatgaatatt tcatcgattg gaccgaggag ggcaaaggag 5040
ccccagagtt ccgaatgatt gagtaagcgg ccgcacctg 5079
<210>79
<211>34
<212>DNA
<213>人工序列
<220>
<223>引物YlD2_Y326F-5
<400>79
cttccgagtg cctctcttca gagagtacct catg 34
<210>80
<211>34
<212>DNA
<213>人工序列
<220>
<223>引物YlD2_Y326F-3
<400>80
catgaggtac tctctgaaga gaggcactcg gaag 34
<210>81
<211>5079
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1254_Y326F
<400>81
aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgatgca 60
tagcttgagt attctaacgc gtcacctaaa tagcttggcg taatcatggt catagctgtt 120
tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 180
gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 240
gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 300
ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 360
ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 420
cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaagcccag 480
gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 540
tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 600
ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 660
atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 720
gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 780
tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 840
cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 900
cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 960
tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 1020
cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 1080
cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 1140
gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 1200
gatcctttta aattaaaaat gaagttttag cacgtgtcag tcctgctcct cggccacgaa 1260
gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc 1320
gatctcggtc atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca 1380
ctcggcgtac agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac 1440
cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa 1500
gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc 1560
ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc aacttggcca tggtggccct 1620
cctcacgtgc tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 1680
tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 1740
acctgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa 1800
ttgtaagcgt taataattca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 1860
gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 1920
tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc 1980
cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 2040
gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg 2100
aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 2160
ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 2220
caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 2280
tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 2340
cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 2400
gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg 2460
gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 2520
cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 2580
gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga 2640
tcagagcttg atcccctgcg ccatcagatc cttggcggcg agaaagccat ccagtttact 2700
ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct 2760
gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt 2820
ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt 2880
cagcaccgtt tctgcggact ggctttctac gtgaaaagga tctaggtgaa gatccttttt 2940
gataatctca tgcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg 3000
tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg 3060
gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt 3120
tcacgggaga ctttatctga eagcagacgt gcactggcca gggggatcac catccgtcgc 3180
cccggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt 3240
ttataggtgt aaaccttaaa ctgccgtacg tataggctgc gcaactgttg ggaagggcga 3300
tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 3360
ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa 3420
ttgtaatacg actcactata gggcgaattg ggccctctag atgcatgctc gagcggccgc 3480
cagtgtgatg gatatctgca gaattcagga gcggccgcaa catgactatc gactcacaat 3540
actacaagtc gcgagacaaa aacgacacgg cacccaaaat cgcgggaatc cgatatgccc 3600
cgctatcgac accattactc aaccgatgtg agaccttctc tctggtctgg cacattttca 3660
gcattcccac tttcctcaca attttcatgc tatgctgcgc aattccactg ctctggccat 3720
ttgtgattgc gtatgtagtg tacgctgtta aagacgactc cccgtccaac ggaggagtgg 3780
tcaagcgata ctcgcctatt tcaagaaact tcttcatctg gaagctcttt ggccgctact 3840
tccccataac tctgcacaag acggtggatc tggagcccac gcacacatac taccctctgg 3900
acgtccagga gtatcacctg attgctgaga gatactggcc gcagaacaag tacctccgag 3960
caatcatcac caccatcgag tactttctgc ccgccttcat gaaacggtct ctttctatca 4020
acgagcagga gcagcctgcc gagcgagatc ctctcctgtc tcccgtttct cccagctctc 4080
cgggttctca acctgacaag tggattaacc acgacagcag atatagccgt ggagaatcat 4140
ctggctccaa cggccacgcc tcgggctccg aacttaacgg caacggcaac aacggcacca 4200
ctaaccgacg acctttgtcg tccgcctctg ctggctccac tgcatctgat tccacgcttc 4260
ttaacgggtc cctcaactcc tacgccaacc agatcattgg cgaaaacgac ccacagctgt 4320
cgcccacaaa actcaagccc actggcagaa aatacatctt cggctaccac ccccacggca 4380
ttatcggcat gggagccttt ggtggaattg ccaccgaggg agctggatgg tccaagctct 4440
ttccgggcat ccctgtttct cttatgactc tcaccaacaa cttccgagtg cctctcttca 4500
gagagtacct catgagtctg ggagtcgctt ctgtctccaa gaagtcctgc aaggccctcc 4560
tcaagcgaaa ccagtctatc tgcattgtcg ttggtggagc acaggaaagt cttctggcca 4620
gacccggtgt catggacctg gtgctactca agcgaaaggg ttttgttcga cttggtatgg 4680
aggtcggaaa tgtcgccctt gttcccatca tggcctttgg tgagaacgac ctctatgacc 4740
aggttagcaa cgacaagtcg tccaagctgt accgattcca gcagtttgtc aagaacttcc 4800
ttggattcac ccttcctttg atgcatgccc gaggcgtctt caactacgat gtcggtcttg 4860
tcccctacag gcgacccgtc aacattgtgg ttggttcccc cattgacttg ccttatctcc 4920
cacaccccac cgacgaagaa gtgtccgaat accacgaccg atacatcgcc gagctgcagc 4980
gaatctacaa cgagcacaag gatgaatatt tcatcgattg gaccgaggag ggcaaaggag 5040
ccccagagtt ccgaatgatt gagtaagcgg ccgcacctg 5079
<210>82
<211>1545
<212>DNA
<213>解脂耶氏酵母
<400>82
atgactatcg actcacaata ctacaagtcg cgagacaaaa acgacacggc acccaaaatc 60
gcgggaatcc gatatgcccc gctatcgaca ccattactca accgatgtga gaccttctct 120
ctggtctggc acattttcag cattcccact ttcctcacaa ttttcatgct atgctgcgca 180
attccactgc tctggccatt tgtgattgcg tatgtagtgt acgctgttaa agacgactcc 240
ccgtccaacg gaggagtggt caagcgatac tcgcctattt caagaaactt cttcatctgg 300
aagctctttg gccgctactt ccccataact ctgcacaaga cggtggatct ggagcccacg 360
cacacatact accctctgga cgtccaggag tatcacctga ttgctgagag atactggccg 420
cagaacaagt acctccgagc aatcatcacc accatcgagt actttctgcc cgccttcatg 480
aaacggtctc tttctatcaa cgagcaggag cagcctgccg agcgagatcc tctcctgtct 540
cccgtttctc ccagctctcc gggttctcaa cctgacaagt ggattaacca cgacagcaga 600
tatagccgtg gagaatcatc tggctccaac ggccacgcct cgggctccga acttaacggc 660
aacggcaaca acggcaccac taaccgacga cctttgtcgt ccgcctctgc tggctccact 720
gcatctgatt ccacgcttct taacgggtcc ctcaactcct acgccaacca gatcattggc 780
gaaaacgacc cacagctgtc gcccacaaaa ctcaagccca ctggcagaaa atacatcttc 840
ggctaccacc cccacggcat tatcggcatg ggagcctttg gtggaattgc caccgaggga 900
gctggatggt ccaagctctt tccgggcatc cctgtttctc ttatgactct caccaacaac 960
ttccgagtgc ctctcttcag agagtacctc atgagtctgg gagtcgcttc tgtctccaag 1020
aagtcctgca aggccctcct caagcgaaac cagtctatct gcattgtcgt tggtggagca 1080
caggaaagtc ttctggccag acccggtgtc atggacctgg tgctactcaa gcgaaagggt 1140
tttgttcgac ttggtatgga ggtcggaaat gtcgcccttg ttcccatcat ggcctttggt 1200
gagaacgacc tctatgacca ggttagcaac gacaagtcgt ccaagctgta ccgattccag 1260
cagtttgtca agaacttcct tggattcacc cttcctttga tgcatgcccg aggcgtcttc 1320
aactacgatg tcggtcttgt cccctacagg cgacccgtca acattgtggt tggttccccc 1380
attgacttgc cttatctccc acaccccacc gacgaagaag tgtccgaata ccacgaccga 1440
tacatcgccg agctgcagcg aatctacaac gagcacaagg atgaatattt catcgattgg 1500
accgaggagg gcaaaggagc cccagagttc cgaatgattg agtaa 1545
<210>83
<211>514
<212>PRT
<213>解脂耶氏酵母
<400>83
Met Thr Ile Asp Ser Gln Tyr Tyr Lys Ser Arg Asp Lys Asn Asp Thr
1 5 10 15
Ala Pro Lys Ile Ala Gly Ile Arg Tyr Ala Pro Leu Ser Thr Pro Leu
20 25 30
Leu Asn Arg Cys Glu Thr Phe Ser Leu Val Trp His Ile Phe Ser Ile
35 40 45
Pro Thr Phe Leu Thr Ile Phe Met Leu Cys Cys Ala Ile Pro Leu Leu
50 55 60
Trp Pro Phe Val Ile Ala Tyr Val Val Tyr Ala Val Lys Asp Asp Ser
65 70 75 80
Pro Ser Asn Gly Gly Val Val Lys Arg Tyr Ser Pro Ile Ser Arg Asn
85 90 95
Phe Phe Ile Trp Lys Leu Phe Gly Arg Tyr Phe Pro Ile Thr Leu His
100 105 110
Lys Thr Val Asp Leu Glu Pro Thr His Thr Tyr Tyr Pro Leu Asp Val
115 120 125
Gln Glu Tyr His Leu Ile Ala Glu Arg Tyr Trp Pro Gln Asn Lys Tyr
130 135 140
Leu Arg Ala Ile Ile Thr Thr Ile Glu Tyr Phe Leu Pro Ala Phe Met
145 150 155 160
Lys Arg Ser Leu Ser Ile Asn Glu Gln Glu Gln Pro Ala Glu Arg Asp
165 170 175
Pro Leu Leu Ser Pro Val Ser Pro Ser Ser Pro Gly Ser Gln Pro Asp
180 185 190
Lys Trp Ile Asn His Asp Ser Arg Tyr Ser Arg Gly Glu Ser Ser Gly
195 200 205
Ser Asn Gly His Ala Ser Gly Ser Glu Leu Asn Gly Asn Gly Asn Asn
210 215 220
Gly Thr Thr Asn Arg Arg Pro Leu Ser Ser Ala Ser Ala Gly Ser Thr
225 230 235 240
Ala Ser Asp Ser Thr Leu Leu Asn Gly Ser Leu Asn Ser Tyr Ala Asn
245 250 255
Gln Ile Ile Gly Glu Asn Asp Pro Gln Leu Ser Pro Thr Lys Leu Lys
260 265 270
Pro Thr Gly Arg Lys Tyr Ile Phe Gly Tyr His Pro His Gly Ile Ile
275 280 285
Gly Met Gly Ala Phe Gly Gly Ile Ala Thr Glu Gly Ala Gly Trp Ser
290 295 300
Lys Leu Phe Pro Gly Ile Pro Val Ser Leu Met Thr Leu Thr Asn Asn
305 310 315 320
Phe Arg Val Pro Leu Phe Arg Glu Tyr Leu Met Ser Leu Gly Val Ala
325 330 335
Ser Val Ser Lys Lys Ser Cys Lys Ala Leu Leu Lys Arg Asn Gln Ser
340 345 350
Ile Cys Ile Val Val Gly Gly Ala Gln Glu Ser Leu Leu Ala Arg Pro
355 360 365
Gly Val Met Asp Leu Val Leu Leu Lys Arg Lys Gly Phe Val Arg Leu
370 375 380
Gly Met Glu Val Gly Asn Val Ala Leu Val Pro Ile Met Ala Phe Gly
385 390 395 400
Glu Asn Asp Leu Tyr Asp Gln Val Ser Asn Asp Lys Ser Ser Lys Leu
405 410 415
Tyr Arg Phe Gln Gln Phe Val Lys Asn Phe Leu Gly Phe Thr Leu Pro
420 425 430
Leu Met His Ala Arg Gly Val Phe Asn Tyr Asp Val Gly Leu Val Pro
435 440 445
Tyr Arg Arg Pro Val Asn Ile Val Val Gly Ser Pro Ile Asp Leu Pro
450 455 460
Tyr Leu Pro His Pro Thr Asp Glu Glu Val Ser Glu Tyr His Asp Arg
465 470 475 480
Tyr Ile Ala Glu Leu Gln Arg Ile Tyr Asn Glu His Lys Asp Glu Tyr
485 490 495
Phe Ile Asp Trp Thr Glu Glu Gly Lys Gly Ala Pro Glu Phe Arg Met
500 505 510
Ile Glu
<210>84
<211>34
<212>DNA
<213>人工序列
<220>
<223>引物YlD2_Y326L-5
<400>84
cttccgagtg cctctcctca gagagtacct catg 34
<210>85
<211>34
<212>DNA
<213>人工序列
<220>
<223>引物YlD2_Y326L-3
<400>85
catgaggtac tctctgagga gaggcactcg gaag 34
<210>86
<211>5079
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1254_Y326L
<400>86
aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgatgca 60
tagcttgagt attctaacgc gtcacctaaa tagcttggcg taatcatggt catagctgtt 120
tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 180
gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 240
gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 300
ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 360
ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 420
cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaagcccag 480
gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 540
tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 600
ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 660
atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 720
gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 780
tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 840
cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 900
cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 960
tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 1020
cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 1080
cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 1140
gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 1200
gatcctttta aattaaaaat gaagttttag cacgtgtcag tcctgctcct cggccacgaa 1260
gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc 1320
gatctcggtc atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca 1380
ctcggcgtac agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac 1440
cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa 1500
gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc 1560
ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc aacttggcca tggtggccct 1620
cctcacgtgc tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 1680
tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 1740
acctgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa 1800
ttgtaagcgt taataattca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 1860
gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 1920
tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc 1980
cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 2040
gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg 2100
aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 2160
ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 2220
caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 2280
tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 2340
cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 2400
gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg 2460
gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 2520
cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 2580
gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga 2640
tcagagcttg atcccctgcg ccatcagatc cttggcggcg agaaagccat ccagtttact 2700
ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct 2760
gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt 2820
ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt 2880
cagcaccgtt tctgcggact ggctttctac gtgaaaagga tctaggtgaa gatccttttt 2940
gataatctca tgcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg 3000
tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg 3060
gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt 3120
tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc 3180
cccggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt 3240
ttataggtgt aaaccttaaa ctgccgtacg tataggctgc gcaactgttg ggaagggcga 3300
tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 3360
ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa 3420
ttgtaatacg actcactata gggcgaattg ggccctctag atgcatgctc gagcggccgc 3480
cagtgtgatg gatatctgca gaattcagga gcggccgcaa catgactatc gactcacaat 3540
actacaagtc gcgagacaaa aacgacacgg cacccaaaat cgcgggaatc cgatatgccc 3600
cgctatcgac accattactc aaccgatgtg agaccttctc tctggtctgg cacattttca 3660
gcattcccac tttcctcaca attttcatgc tatgctgcgc aattccactg ctctggccat 3720
ttgtgattgc gtatgtagtg tacgctgtta aagacgactc cccgtccaac ggaggagtgg 3780
tcaagcgata ctcgcctatt tcaagaaact tcttcatctg gaagctcttt ggccgctact 3840
tccccataac tctgcacaag acggtggatc tggagcccac gcacacatac taccctctgg 3900
acgtccagga gtatcacctg attgctgaga gatactggcc gcagaacaag tacctccgag 3960
caatcatcac caccatcgag tactttctgc ccgccttcat gaaacggtct ctttctatca 4020
acgagcagga gcagcctgcc gagcgagatc ctctcctgtc tcccgtttct cccagctctc 4080
cgggttctca acctgacaag tggattaacc acgacagcag atatagccgt ggagaatcat 4140
ctggctccaa cggccacgcc tcgggctccg aacttaacgg caacggcaac aacggcacca 4200
ctaaccgacg acctttgtcg tccgcctctg ctggctccac tgcatctgat tccacgcttc 4260
ttaacgggtc cctcaactcc tacgccaacc agatcattgg cgaaaacgac ccacagctgt 4320
cgcccacaaa actcaagccc actggcagaa aatacatctt cggctaccac ccccacggca 4380
ttatcggcat gggagccttt ggtggaattg ccaccgaggg agctggatgg tccaagctct 4440
ttccgggcat ccctgtttct cttatgactc tcaccaacaa cttccgagtg cctctcctca 4500
gagagtacct catgagtctg ggagtcgctt ctgtctccaa gaagtcctgc aaggccctcc 4560
tcaagcgaaa ccagtctatc tgcattgtcg ttggtggagc acaggaaagt cttctggcca 4620
gacccggtgt catggacctg gtgctactca agcgaaaggg ttttgttcga cttggtatgg 4680
aggtcggaaa tgtcgccctt gttcccatca tggcctttgg tgagaacgac ctctatgacc 4740
aggttagcaa cgacaagtcg tccaagctgt accgattcca gcagtttgtc aagaacttcc 4800
ttggattcac ccttcctttg atgcatgccc gaggcgtctt caactacgat gtcggtcttg 4860
tcccctacag gcgacccgtc aacattgtgg ttggttcccc cattgacttg ccttatctcc 4920
cacaccccac cgacgaagaa gtgtccgaat accacgaccg atacatcgcc gagctgcagc 4980
gaatctacaa cgagcacaag gatgaatatt tcatcgattg gaccgaggag ggcaaaggag 5040
ccccagagtt ccgaatgatt gagtaagcgg ccgcacctg 5079
<210>87
<211>1545
<212>DNA
<213>解脂耶氏酵母
<400>87
atgactatcg actcacaata ctacaagtcg cgagacaaaa acgacacggc acccaaaatc 60
gcgggaatcc gatatgcccc gctatcgaca ccattactca accgatgtga gaccttctct 120
ctggtctggc acattttcag cattcccact ttcctcacaa ttttcatgct atgctgcgca 180
attccactgc tctggccatt tgtgattgcg tatgtagtgt acgctgttaa agacgactcc 240
ccgtccaacg gaggagtggt caagcgatac tcgcctattt caagaaactt cttcatctgg 300
aagctctttg gccgctactt ccccataact ctgcacaaga cggtggatct ggagcccacg 360
cacacatact accctctgga cgtccaggag tatcacctga ttgctgagag atactggccg 420
cagaacaagt acctccgagc aatcatcacc accatcgagt actttctgcc cgccttcatg 480
aaacggtctc tttctatcaa cgagcaggag cagcctgccg agcgagatcc tctcctgtct 540
cccgtttctc ccagctctcc gggttctcaa cctgacaagt ggattaacca cgacagcaga 600
tatagccgtg gagaatcatc tggctccaac ggccacgcct cgggctccga acttaacggc 660
aacggcaaca acggcaccac taaccgacga cctttgtcgt ccgcctctgc tggctccact 720
gcatctgatt ccacgcttct taacgggtcc ctcaactcct acgccaacca gatcattggc 780
gaaaacgacc cacagctgtc gcccacaaaa ctcaagccca ctggcagaaa atacatcttc 840
ggctaccacc cccacggcat tatcggcatg ggagcctttg gtggaattgc caccgaggga 900
gctggatggt ccaagctctt tccgggcatc cctgtttctc ttatgactct caccaacaac 960
ttccgagtgc ctctcctcag agagtacctc atgagtctgg gagtcgcttc tgtctccaag 1020
aagtcctgca aggccctcct caagcgaaac cagtctatct gcattgtcgt tggtggagca 1080
caggaaagtc ttctggccag acccggtgtc atggacctgg tgctactcaa gcgaaagggt 1140
tttgttcgac ttggtatgga ggtcggaaat gtcgcccttg ttcccatcat ggcctttggt 1200
gagaacgacc tctatgacca ggttagcaac gacaagtcgt ccaagctgta ccgattccag 1260
cagtttgtca agaacttcct tggattcacc cttcctttga tgcatgcccg aggcgtcttc 1320
aactacgatg tcggtcttgt cccctacagg cgacccgtca acattgtggt tggttccccc 1380
attgacttgc cttatctccc acaccccacc gacgaagaag tgtccgaata ccacgaccga 1440
tacatcgccg agctgcagcg aatctacaac gagcacaagg atgaatattt catcgattgg 1500
accgaggagg gcaaaggagc cccagagttc cgaatgattg agtaa 1545
<210>88
<211>514
<212>PRT
<213>解脂耶氏酵母
<400>88
Met Thr Ile Asp Ser Gln Tyr Tyr Lys Ser Arg Asp Lys Asn Asp Thr
1 5 10 15
Ala Pro Lys Ile Ala Gly Ile Arg Tyr Ala Pro Leu Ser Thr Pro Leu
20 25 30
Leu Asn Arg Cys Glu Thr Phe Ser Leu Val Trp His Ile Phe Ser Ile
35 40 45
Pro Thr Phe Leu Thr Ile Phe Met Leu Cys Cys Ala Ile Pro Leu Leu
50 55 60
Trp Pro Phe Val Ile Ala Tyr Val Val Tyr Ala Val Lys Asp Asp Ser
65 70 75 80
Pro Ser Asn Gly Gly Val Val Lys Arg Tyr Ser Pro Ile Ser Arg Asn
85 90 95
Phe Phe Ile Trp Lys Leu Phe Gly Arg Tyr Phe Pro Ile Thr Leu His
100 105 110
Lys Thr Val Asp Leu Glu Pro Thr His Thr Tyr Tyr Pro Leu Asp Val
115 120 125
Gln Glu Tyr His Leu Ile Ala Glu Arg Tyr Trp Pro Gln Asn Lys Tyr
130 135 140
Leu Arg Ala Ile Ile Thr Thr Ile Glu Tyr Phe Leu Pro Ala Phe Met
145 150 155 160
Lys Arg Ser Leu Ser Ile Asn Glu Gln Glu Gln Pro Ala Glu Arg Asp
165 170 175
Pro Leu Leu Ser Pro Val Ser Pro Ser Ser Pro Gly Ser Gln Pro Asp
180 185 190
Lys Trp Ile Asn His Asp Ser Arg Tyr Ser Arg Gly Glu Ser Ser Gly
195 200 205
Ser Asn Gly His Ala Ser Gly Ser Glu Leu Asn Gly Asn Gly Asn Asn
210 215 220
Gly Thr Thr Asn Arg Arg Pro Leu Ser Ser Ala Ser Ala Gly Ser Thr
225 230 235 240
Ala Ser Asp Ser Thr Leu Leu Asn Gly Ser Leu Asn Ser Tyr Ala Asn
245 250 255
Gln Ile Ile Gly Glu Asn Asp Pro Gln Leu Ser Pro Thr Lys Leu Lys
260 265 270
Pro Thr Gly Arg Lys Tyr Ile Phe Gly Tyr His Pro His Gly Ile Ile
275 280 285
Gly Met Gly Ala Phe Gly Gly Ile Ala Thr Glu Gly Ala Gly Trp Ser
290 295 300
Lys Leu Phe Pro Gly Ile Pro Val Ser Leu Met Thr Leu Thr Asn Asn
305 310 315 320
Phe Arg Val Pro Leu Leu Arg Glu Tyr Leu Met Ser Leu Gly Val Ala
325 330 335
Ser Val Ser Lys Lys Ser Cys Lys Ala Leu Leu Lys Arg Asn Gln Ser
340 345 350
Ile Cys Ile Val Val Gly Gly Ala Gln Glu Ser Leu Leu Ala Arg Pro
355 360 365
Gly Val Met Asp Leu Val Leu Leu Lys Arg Lys Gly Phe Val Arg Leu
370 375 380
Gly Met Glu Val Gly Asn Val Ala Leu Val Pro Ile Met Ala Phe Gly
385 390 395 400
Glu Asn Asp Leu Tyr Asp Gln Val Ser Asn Asp Lys Ser Ser Lys Leu
405 410 415
Tyr Arg Phe Gln Gln Phe Val Lys Asn Phe Leu Gly Phe Thr Leu Pro
420 425 430
Leu Met His Ala Arg Gly Val Phe Asn Tyr Asp Val Gly Leu Val Pro
435 440 445
Tyr Arg Arg Pro Val Asn Ile Val Val Gly Ser Pro Ile Asp Leu Pro
450 455 460
Tyr Leu Pro His Pro Thr Asp Glu Glu Val Ser Glu Tyr His Asp Arg
465 470 475 480
Tyr Ile Ala Glu Leu Gln Arg Ile Tyr Asn Glu His Lys Asp Glu Tyr
485 490 495
Phe Ile Asp Trp Thr Glu Glu Gly Lys Gly Ala Pro Glu Phe Arg Met
500 505 510
Ile Glu
<210>89
<211>35
<212>DNA
<213>人工序列
<220>
<223>引物YlD2_R327K-5
<400>89
ccgagtgcct ctctacaaag agtacctcat gagtc 35
<210>90
<211>35
<212>DNA
<213>人工序列
<220>
<223>引物YlD2_R327K-3
<400>90
gactcatgag gtactctttg tagagaggca ctcgg 35
<210>91
<211>5079
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1254_R327K
<400>91
aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgatgca 60
tagcttgagt attctaacgc gtcacctaaa tagcttggcg taatcatggt catagctgtt 120
tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 180
gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 240
gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 300
ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 360
ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 420
cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaagcccag 480
gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 540
tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 600
ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 660
atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 720
gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 780
tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 840
cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 900
cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 960
tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 1020
cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 1080
cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 1140
gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 1200
gatcctttta aattaaaaat gaagttttag cacgtgtcag tcctgctcct cggccacgaa 1260
gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc 1320
gatctcggtc atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca 1380
ctcggcgtac agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac 1440
cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa 1500
gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc 1560
ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc aacttggcca tggtggccct 1620
cctcacgtgc tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 1680
tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 1740
acctgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa 1800
ttgtaagcgt taataattca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 1860
gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 1920
tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc 1980
cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 2040
gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg 2100
aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 2160
ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 2220
caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 2280
tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 2340
cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 2400
gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg 2460
gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 2520
cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 2580
gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga 2640
tcagagcttg atcccctgcg ccatcagatc cttggcggcg agaaagccat ccagtttact 2700
ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct 2760
gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt 2820
ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt 2880
cagcaccgtt tctgcggact ggctttctac gtgaaaagga tctaggtgaa gatccttttt 2940
gataatctca tgcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg 3000
tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg 3060
gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt 3120
tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc 3180
cccggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt 3240
ttataggtgt aaaccttaaa ctgccgtacg tataggctgc gcaactgttg ggaagggcga 3300
tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 3360
ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa 3420
ttgtaatacg actcactata gggcgaattg ggccctctag atgcatgctc gagcggccgc 3480
cagtgtgatg gatatctgca gaattcagga gcggccgcaa catgactatc gactcacaat 3540
actacaagtc gcgagacaaa aacgacacgg cacccaaaat cgcgggaatc cgatatgccc 3600
cgctatcgac accattactc aaccgatgtg agaccttctc tctggtctgg cacattttca 3660
gcattcccac tttcctcaca attttcatgc tatgctgcgc aattccactg ctctggccat 3720
ttgtgattgc gtatgtagtg tacgctgtta aagacgactc cccgtccaac ggaggagtgg 3780
tcaagcgata ctcgcctatt tcaagaaact tcttcatctg gaagctcttt ggccgctact 3840
tccccataac tctgcacaag acggtggatc tggagcccac gcacacatac taccctctgg 3900
acgtccagga gtatcacctg attgctgaga gatactggcc gcagaacaag tacctccgag 3960
caatcatcac caccatcgag tactttctgc ccgccttcat gaaacggtct ctttctatca 4020
acgagcagga gcagcctgcc gagcgagatc ctctcctgtc tcccgtttct cccagctctc 4080
cgggttctca acctgacaag tggattaacc acgacagcag atatagccgt ggagaatcat 4140
ctggctccaa cggccacgcc tcgggctccg aacttaacgg caacggcaac aacggcacca 4200
ctaaccgacg acctttgtcg tccgcctctg ctggctccac tgcatctgat tccacgcttc 4260
ttaacgggtc cctcaactcc tacgccaacc agatcattgg cgaaaacgac ccacagctgt 4320
cgcccacaaa actcaagccc actggcagaa aatacatctt cggctaccac ccccacggca 4380
ttatcggcat gggagccttt ggtggaattg ccaccgaggg agctggatgg tccaagctct 4440
ttccgggcat ccctgtttct cttatgactc tcaccaacaa cttccgagtg cctctctaca 4500
aagagtacct catgagtctg ggagtcgctt ctgtctccaa gaagtcctgc aaggccctcc 4560
tcaagcgaaa ccagtctatc tgcattgtcg ttggtggagc acaggaaagt cttctggcca 4620
gacccggtgt catggacctg gtgctactca agcgaaaggg ttttgttcga cttggtatgg 4680
aggtcggaaa tgtcgccctt gttcccatca tggcctttgg tgagaacgac ctctatgacc 4740
aggttagcaa cgacaagtcg tccaagctgt accgattcca gcagtttgtc aagaacttcc 4800
ttggattcac ccttcctttg atgcatgccc gaggcgtctt caactacgat gtcggtcttg 4860
tcccctacag gcgacccgtc aacattgtgg ttggttcccc cattgacttg ccttatctcc 4920
cacaccccac cgacgaagaa gtgtccgaat accacgaccg atacatcgcc gagctgcagc 4980
gaatctacaa cgagcacaag gatgaatatt tcatcgattg gaccgaggag ggcaaaggag 5040
ccccagagtt ccgaatgatt gagtaagcgg ccgcacctg 5079
<210>92
<211>1545
<212>DNA
<213>解脂耶氏酵母
<400>92
atgactatcg actcacaata ctacaagtcg cgagacaaaa acgacacggc acccaaaatc 60
gcgggaatcc gatatgcccc gctatcgaca ccattactca accgatgtga gaccttctct 120
ctggtctggc acattttcag cattcccact ttcctcacaa ttttcatgct atgctgcgca 180
attccactgc tctggccatt tgtgattgcg tatgtagtgt acgctgttaa agacgactcc 240
ccgtccaacg gaggagtggt caagcgatac tcgcctattt caagaaactt cttcatctgg 300
aagctctttg gccgctactt ccccataact ctgcacaaga cggtggatct ggagcccacg 360
cacacatact accctctgga cgtccaggag tatcacctga ttgctgagag atactggccg 420
cagaacaagt acctccgagc aatcatcacc accatcgagt actttctgcc cgccttcatg 480
aaacggtctc tttctatcaa cgagcaggag cagcctgccg agcgagatcc tctcctgtct 540
cccgtttctc ccagctctcc gggttctcaa cctgacaagt ggattaacca cgacagcaga 600
tatagccgtg gagaatcatc tggctccaac ggccacgcct cgggctccga acttaacggc 660
aacggcaaca acggcaccac taaccgacga cctttgtcgt ccgcctctgc tggctccact 720
gcatctgatt ccacgcttct taacgggtcc ctcaactcct acgccaacca gatcattggc 780
gaaaacgacc cacagctgtc gcccacaaaa ctcaagccca ctggcagaaa atacatcttc 840
ggctaccacc cccacggcat tatcggcatg ggagcctttg gtggaattgc caccgaggga 900
gctggatggt ccaagctctt tccgggcatc cctgtttctc ttatgactct caccaacaac 960
ttccgagtgc ctctctacaa agagtacctc atgagtctgg gagtcgcttc tgtctccaag 1020
aagtcctgca aggccctcct caagcgaaac cagtctatct gcattgtcgt tggtggagca 1080
caggaaagtc ttctggccag acccggtgtc atggacctgg tgctactcaa gcgaaagggt 1140
tttgttcgac ttggtatgga ggtcggaaat gtcgcccttg ttcccatcat ggcctttggt 1200
gagaacgacc tctatgacca ggttagcaac gacaagtcgt ccaagctgta ccgattccag 1260
cagtttgtca agaacttcct tggattcacc cttcctttga tgcatgcccg aggcgtcttc 1320
aactacgatg tcggtcttgt cccctacagg cgacccgtca acattgtggt tggttccccc 1380
attgacttgc cttatctccc acaccccacc gacgaagaag tgtccgaata ccacgaccga 1440
tacatcgccg agctgcagcg aatctacaac gagcacaagg atgaatattt catcgattgg 1500
accgaggagg gcaaaggagc cccagagttc cgaatgattg agtaa 1545
<210>93
<211>514
<212>PRT
<213>解脂耶氏酵母
<400>93
Met Thr Ile Asp Ser Gln Tyr Tyr Lys Ser Arg Asp Lys Asn Asp Thr
1 5 10 15
Ala Pro Lys Ile Ala Gly Ile Arg Tyr Ala Pro Leu Ser Thr Pro Leu
20 25 30
Leu Asn Arg Cys Glu Thr Phe Ser Leu Val Trp His Ile Phe Ser Ile
35 40 45
Pro Thr Phe Leu Thr Ile Phe Met Leu Cys Cys Ala Ile Pro Leu Leu
50 55 60
Trp Pro Phe Val Ile Ala Tyr Val Val Tyr Ala Val Lys Asp Asp Ser
65 70 75 80
Pro Ser Asn Gly Gly Val Val Lys Arg Tyr Ser Pro Ile Ser Arg Asn
85 90 95
Phe Phe Ile Trp Lys Leu Phe Gly Arg Tyr Phe Pro Ile Thr Leu His
100 105 110
Lys Thr Val Asp Leu Glu Pro Thr His Thr Tyr Tyr Pro Leu Asp Val
115 120 125
Gln Glu Tyr His Leu Ile Ala Glu Arg Tyr Trp Pro Gln Asn Lys Tyr
130 135 140
Leu Arg Ala Ile Ile Thr Thr Ile Glu Tyr Phe Leu Pro Ala Phe Met
145 150 155 160
Lys Arg Ser Leu Ser Ile Asn Glu Gln Glu Gln Pro Ala Glu Arg Asp
165 170 175
Pro Leu Leu Ser Pro Val Ser Pro Ser Ser Pro Gly Ser Gln Pro Asp
180 185 190
Lys Trp Ile Asn His Asp Ser Arg Tyr Ser Arg Gly Glu Ser Ser Gly
195 200 205
Ser Asn Gly His Ala Ser Gly Ser Glu Leu Asn Gly Asn Gly Asn Asn
210 215 220
Gly Thr Thr Asn Arg Arg Pro Leu Ser Ser Ala Ser Ala Gly Ser Thr
225 230 235 240
Ala Ser Asp Ser Thr Leu Leu Asn Gly Ser Leu Asn Ser Tyr Ala Asn
245 250 255
Gln Ile Ile Gly Glu Asn Asp Pro Gln Leu Ser Pro Thr Lys Leu Lys
260 265 270
Pro Thr Gly Arg Lys Tyr Ile Phe Gly Tyr His Pro His Gly Ile Ile
275 280 285
Gly Met Gly Ala Phe Gly Gly Ile Ala Thr Glu Gly Ala Gly Trp Ser
290 295 300
Lys Leu Phe Pro Gly Ile Pro Val Ser Leu Met Thr Leu Thr Asn Asn
305 310 315 320
Phe Arg Val Pro Leu Tyr Lys Glu Tyr Leu Met Ser Leu Gly Val Ala
325 330 335
Ser Val Ser Lys Lys Ser Cys Lys Ala Leu Leu Lys Arg Asn Gln Ser
340 345 350
Ile Cys Ile Val Val Gly Gly Ala Gln Glu Ser Leu Leu Ala Arg Pro
355 360 365
Gly Val Met Asp Leu Val Leu Leu Lys Arg Lys Gly Phe Val Arg Leu
370 375 380
Gly Met Glu Val Gly Asn Val Ala Leu Val Pro Ile Met Ala Phe Gly
385 390 395 400
Glu Asn Asp Leu Tyr Asp Gln Val Ser Asn Asp Lys Ser Ser Lys Leu
405 410 415
Tyr Arg Phe Gln Gln Phe Val Lys Asn Phe Leu Gly Phe Thr Leu Pro
420 425 430
Leu Met His Ala Arg Gly Val Phe Asn Tyr Asp Val Gly Leu Val Pro
435 440 445
Tyr Arg Arg Pro Val Asn Ile Val Val Gly Ser Pro Ile Asp Leu Pro
450 455 460
Tyr Leu Pro His Pro Thr Asp Glu Glu Val Ser Glu Tyr His Asp Arg
465 470 475 480
Tyr Ile Ala Glu Leu Gln Arg Ile Tyr Asn Glu His Lys Asp Glu Tyr
485 490 495
Phe Ile Asp Trp Thr Glu Glu Gly Lys Gly Ala Pro Glu Phe Arg Met
500 505 510
Ile Glu
<210>94
<211>9074
<212>DNA
<213>人工序列
<220>
<223>质粒pY191
<400>94
ggccgcaggg cggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg 60
acctcgaggg ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca 120
ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 180
cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 240
ccttcccaac agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca 300
ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 360
gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 420
caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 480
cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 540
tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 600
acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 660
gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 720
ttaacgttta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 780
accgcatatc gacggtcgag gagaacttct agtatatcca catacctaat attattgcct 840
tattaaaaat ggaatcccaa caattacatc aaaatccaca ttctcttcaa aatcaattgt 900
cctgtacttc cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct 960
atttctcaac aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat 1020
atcttgaccg cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc 1080
tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac 1140
agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata 1200
cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gaggccggaa ccggcttttc 1260
atatagaata gagaagcgtt catgactaaa tgcttgcatc acaatacttg aagttgacaa 1320
tattatttaa ggacctattg ttttttccaa taggtggtta gcaatcgtct tactttctaa 1380
cttttcttac cttttacatt tcagcaatat atatatatat ttcaaggata taccattcta 1440
atgtctgccc ctatgtctgc ccctaagaag atcgtcgttt tgccaggtga ccacgttggt 1500
caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt tcgttccaat 1560
gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc tacaggtgtc 1620
ccacttccag atgaggcgct ggaagcctcc aagaaggttg atgccgtttt gttaggtgct 1680
gtggctggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt actaaaaatc 1740
cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc cgactctctt 1800
ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt tgttgtcaga 1860
gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga tggtgtcgct 1920
tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat ggccgctttc 1980
atggccctac aacatgagcc accattgcct atttggtcct tggataaagc taatcttttg 2040
gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga attccctaca 2100
ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa gaacccaacc 2160
cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc cgatgaagcc 2220
tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc tttgccagac 2280
aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga tttgccaaag 2340
aataaggttg accctatcgc cactatcttg tctgctgcaa tgatgttgaa attgtcattg 2400
aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt ggatgcaggt 2460
atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga tgctgtcgcc 2520
gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata 2580
aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca 2640
tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag 2700
gatagtaaag gaatacaggt aagcaaattg atactaatgg ctcaacgtga taaggaaaaa 2760
gaattgcact ttaacattaa tattgacaag gaggagggca ccacacaaaa agttaggtgt 2820
aacagaaaat catgaaacta cgattcctaa tttgatattg gaggattttc tctaaaaaaa 2880
aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa 2940
taccttcttg aagcatttcc cataatggtg aaagttccct caagaatttt actctgtcag 3000
aaacggcctt acgacgtagt cgatatggtg cactctcagt acaatctgct ctgatgccgc 3060
atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 3120
gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 3180
gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt 3240
ataggttaat gtcatgataa taatggtttc ttagtatgat ccaatatcaa aggaaatgat 3300
agcattgaag gatgagacta atccaattga ggagtggcag catatagaac agctaaaggg 3360
tagtgctgaa ggaagcatac gataccccgc atggaatggg ataatatcac aggaggtact 3420
agactacctt tcatcctaca taaatagacg catataagta cgcatttaag cataaacacg 3480
cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac 3540
gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg 3600
aaacgctttg aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcaga 3660
gcgcttttga aaaccaaaag cgctctgaag acgcactttc aaaaaaccaa aaacgcaccg 3720
gactgtaacg agctactaaa atattgcgaa taccgcttcc acaaacattg ctcaaaagta 3780
tctctttgct atatatctct gtgctatatc cctatataac ctacccatcc acctttcgct 3840
ccttgaactt gcatctaaac tcgacctcta cattttttat gtttatctct agtattactc 3900
tttagacaaa aaaattgtag taagaactat tcatagagtg aatcgaaaac aatacgaaaa 3960
tgtaaacatt tcctatacgt agtatataga gacaaaatag aagaaaccgt tcataatttt 4020
ctgaccaatg aagaatcatc aacgctatca ctttctgttc acaaagtatg cgcaatccac 4080
atcggtatag aatataatcg gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc 4140
tagtaatcag taaacgcggg aagtggagtc aggctttttt tatggaagag aaaatagaca 4200
ccaaagtagc cttcttctaa ccttaacgga cctacagtgc aaaaagttat caagagactg 4260
cattatagag cgcacaaagg agaaaaaaag taatctaaga tgctttgtta gaaaaatagc 4320
gctctcggga tgcatttttg tagaacaaaa aagaagtata gattctttgt tggtaaaata 4380
gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa 4440
ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa gcacagattc ttcgttggta 4500
aaatagcgct ttcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg 4560
aaaaattagc gctctcgcgt tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt 4620
caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 4680
attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 4740
aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 4800
tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 4860
agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 4920
gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 4980
cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 5040
agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 5100
taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 5160
tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 5220
taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 5280
acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 5340
ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 5400
cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 5460
agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 5520
tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 5580
agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 5640
tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 5700
ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 5760
tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 5820
aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 5880
tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 5940
agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 6000
taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 6060
caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6120
agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 6180
aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 6240
gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 6300
tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 6360
gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 6420
ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 6480
ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 6540
aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 6600
aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 6660
atgtgagtta cctcactcat taggcacccc aggctttaca ctttatgctt ccggctccta 6720
tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 6780
acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggga 6840
tttcgaaact aagttcttgg tgttttaaaa ctaaaaaaaa gactaactat aaaagtagaa 6900
tttaagaagt ttaagaaata gatttacaga attacaatca atacctaccg tctttatata 6960
cttattagtc aagtagggga ataatttcag ggaactggtt tcaacctttt ttttcagctt 7020
tttccaaatc agagagagca gaaggtaata gaaggtgtaa gaaaatgaga tagatacatg 7080
cgtgggtcaa ttgccttgtg tcatcattta ctccaggcag gttgcatcac tccattgagg 7140
ttgtgcccgt tttttgcctg tttgtgcccc tgttctctgt agttgcgcta agagaatgga 7200
cctatgaact gatggttggt gaagaaaaca atattttggt gctgggattc tttttttttc 7260
tggatgccag cttaaaaagc gggctccatt atatttagtg gatgccagga ataaactgtt 7320
cacccagaca cctacgatgt tatatattct gtgtaacccg ccccctattt tgggcatgta 7380
cgggttacag cagaattaaa aggctaattt tttgactaaa taaagttagg aaaatcacta 7440
ctattaatta tttacgtatt ctttgaaatg gcagtattga taatgataaa ctcgaaatca 7500
ctagtggatc cgcccagcgg ccgcaacatg actatcgact cacaatacta caagtcgcga 7560
gacaaaaacg acacggcacc caaaatcgcg ggaatccgat atgccccgct atcgacacca 7620
ttactcaacc gatgtgagac cttctctctg gtctggcaca ttttcagcat tcccactttc 7680
ctcacaattt tcatgctatg ctgcgcaatt ccactgctct ggccatttgt gattgcgtat 7740
gtagtgtacg ctgttaaaga cgactccccg tccaacggag gagtggtcaa gcgatactcg 7800
cctatttcaa gaaacttctt catctggaag ctctttggcc gctacttccc cataactctg 7860
cacaagacgg tggatctgga gcccacgcac acatactacc ctctggacgt ccaggagtat 7920
cacctgattg ctgagagata ctggccgcag aacaagtacc tccgagcaat catcaccacc 7980
atcgagtact ttctgcccgc cttcatgaaa cggtctcttt ctatcaacga gcaggagcag 8040
cctgccgagc gagatcctct cctgtctccc gtttctccca gctctccggg ttctcaacct 8100
gacaagtgga ttaaccacga cagcagatat agccgtggag aatcatctgg ctccaacggc 8160
cacgcctcgg gctccgaact taacggcaac ggcaacaacg gcaccactaa ccgacgacct 8220
ttgtcgtccg cctctgctgg ctccactgca tctgattcca cgcttcttaa cgggtccctc 8280
aactcctacg ccaaccagat cattggcgaa aacgacccac agctgtcgcc cacaaaactc 8340
aagcccactg gcagaaaata catcttcggc taccaccccc acggcattat cggcatggga 8400
gcctttggtg gaattgccac cgagggagct ggatggtcca agctctttcc gggcatccct 8460
gtttctctta tgactctcac caacaacttc cgagtgcctc tctacagaga gtacctcatg 8520
agtctgggag tcgcttctgt ctccaagaag tcctgcaagg ccctcctcaa gcgaaaccag 8580
tctatctgca ttgtcgttgg tggagcacag gaaagtcttc tggccagacc cggtgtcatg 8640
gacctggtgc tactcaagcg aaagggtttt gttcgacttg gtatggaggt cggaaatgtc 8700
gcccttgttc ccatcatggc ctttggtgag aacgacctct atgaccaggt tagcaacgac 8760
aagtcgtcca agctgtaccg attccagcag tttgtcaaga acttccttgg attcaccctt 8820
cctttgatgc atgcccgagg cgtcttcaac tacgatgtcg gtcttgtccc ctacaggcga 8880
cccgtcaaca ttgtggttgg ttcccccatt gacttgcctt atctcccaca ccccaccgac 8940
gaagaagtgt ccgaatacca cgaccgatac atcgccgagc tgcagcgaat ctacaacgag 9000
cacaaggatg aatatttcat cgattggacc gaggagggca aaggagcccc agagttccga 9060
atgattgagt aagc 9074
<210>95
<211>9074
<212>DNA
<213>人工序列
<220>
<223>质粒pY192
<400>95
ggccgcaggg cggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg 60
acctcgaggg ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca 120
ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 180
cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 240
ccttcccaac agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca 300
ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 360
gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 420
caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 480
cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 540
tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 600
acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 660
gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 720
ttaacgttta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 780
accgcatatc gacggtcgag gagaacttct agtatatcca catacctaat attattgcct 840
tattaaaaat ggaatcccaa caattacatc aaaatccaca ttctcttcaa aatcaattgt 900
cctgtacttc cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct 960
atttctcaac aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat 1020
atcttgaccg cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc 1080
tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac 1140
agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata 1200
cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gaggccggaa ccggcttttc 1260
atatagaata gagaagcgtt catgactaaa tgcttgcatc acaatacttg aagttgacaa 1320
tattatttaa ggacctattg ttttttccaa taggtggtta gcaatcgtct tactttctaa 1380
cttttcttac cttttacatt tcagcaatat atatatatat ttcaaggata taccattcta 1440
atgtctgccc ctatgtctgc ccctaagaag atcgtcgttt tgccaggtga ccacgttggt 1500
caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt tcgttccaat 1560
gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc tacaggtgtc 1620
ccacttccag atgaggcgct ggaagcctcc aagaaggttg atgccgtttt gttaggtgct 1680
gtggctggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt actaaaaatc 1740
cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc cgactctctt 1800
ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt tgttgtcaga 1860
gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga tggtgtcgct 1920
tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat ggccgctttc 1980
atggccctac aacatgagcc accattgcct atttggtcct tggataaagc taatcttttg 2040
gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga attccctaca 2100
ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa gaacccaacc 2160
cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc cgatgaagcc 2220
tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc tttgccagac 2280
aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga tttgccaaag 2340
aataaggttg accctatcgc cactatcttg tctgctgcaa tgatgttgaa attgtcattg 2400
aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt ggatgcaggt 2460
atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga tgctgtcgcc 2520
gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata 2580
aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca 2640
tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag 2700
gatagtaaag gaatacaggt aagcaaattg atactaatgg ctcaacgtga taaggaaaaa 2760
gaattgcact ttaacattaa tattgacaag gaggagggca ccacacaaaa agttaggtgt 2820
aacagaaaat catgaaacta cgattcctaa tttgatattg gaggattttc tctaaaaaaa 2880
aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa 2940
taccttcttg aagcatttcc cataatggtg aaagttccct caagaatttt actctgtcag 3000
aaacggcctt acgacgtagt cgatatggtg cactctcagt acaatctgct ctgatgccgc 3060
atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 3120
gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 3180
gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt 3240
ataggttaat gtcatgataa taatggtttc ttagtatgat ccaatatcaa aggaaatgat 3300
agcattgaag gatgagacta atccaattga ggagtggcag catatagaac agctaaaggg 3360
tagtgctgaa ggaagcatac gataccccgc atggaatggg ataatatcac aggaggtact 3420
agactacctt tcatcctaca taaatagacg catataagta cgcatttaag cataaacacg 3480
cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac 3540
gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg 3600
aaacgctttg aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcaga 3660
gcgcttttga aaaccaaaag cgctctgaag acgcactttc aaaaaaccaa aaacgcaccg 3720
gactgtaacg agctactaaa atattgcgaa taccgcttcc acaaacattg ctcaaaagta 3780
tctctttgct atatatctct gtgctatatc cctatataac ctacccatcc acctttcgct 3840
ccttgaactt gcatctaaac tcgacctcta cattttttat gtttatctct agtattactc 3900
tttagacaaa aaaattgtag taagaactat tcatagagtg aatcgaaaac aatacgaaaa 3960
tgtaaacatt tcctatacgt agtatataga gacaaaatag aagaaaccgt tcataatttt 4020
ctgaccaatg aagaatcatc aacgctatca ctttctgttc acaaagtatg cgcaatccac 4080
atcggtatag aatataatcg gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc 4140
tagtaatcag taaacgcggg aagtggagtc aggctttttt tatggaagag aaaatagaca 4200
ccaaagtagc cttcttctaa ccttaacgga cctacagtgc aaaaagttat caagagactg 4260
cattatagag cgcacaaagg agaaaaaaag taatctaaga tgctttgtta gaaaaatagc 4320
gctctcggga tgcatttttg tagaacaaaa aagaagtata gattctttgt tggtaaaata 4380
gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa 4440
ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa gcacagattc ttcgttggta 4500
aaatagcgct ttcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg 4560
aaaaattagc gctctcgcgt tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt 4620
caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 4680
attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 4740
aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 4800
tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 4860
agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 4920
gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 4980
cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 5040
agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 5100
taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 5160
tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 5220
taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 5280
acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 5340
ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 5400
cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 5460
agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 5520
tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 5580
agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 5640
tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 5700
ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 5760
tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 5820
aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 5880
tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 5940
agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 6000
taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 6060
caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6120
agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 6180
aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 6240
gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 6300
tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 6360
gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 6420
ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 6480
ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 6540
aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 6600
aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 6660
atgtgagtta cctcactcat taggcacccc aggctttaca ctttatgctt ccggctccta 6720
tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 6780
acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggga 6840
tttcgaaact aagttcttgg tgttttaaaa ctaaaaaaaa gactaactat aaaagtagaa 6900
tttaagaagt ttaagaaata gatttacaga attacaatca atacctaccg tctttatata 6960
cttattagtc aagtagggga ataatttcag ggaactggtt tcaacctttt ttttcagctt 7020
tttccaaatc agagagagca gaaggtaata gaaggtgtaa gaaaatgaga tagatacatg 7080
cgtgggtcaa ttgccttgtg tcatcattta ctccaggcag gttgcatcac tccattgagg 7140
ttgtgcccgt tttttgcctg tttgtgcccc tgttctctgt agttgcgcta agagaatgga 7200
cctatgaact gatggttggt gaagaaaaca atattttggt gctgggattc tttttttttc 7260
tggatgccag cttaaaaagc gggctccatt atatttagtg gatgccagga ataaactgtt 7320
cacccagaca cctacgatgt tatatattct gtgtaacccg ccccctattt tgggcatgta 7380
cgggttacag cagaattaaa aggctaattt tttgactaaa taaagttagg aaaatcacta 7440
ctattaatta tttacgtatt ctttgaaatg gcagtattga taatgataaa ctcgaaatca 7500
ctagtggatc cgcccagcgg ccgcaacatg actatcgact cacaatacta caagtcgcga 7560
gacaaaaacg acacggcacc caaaatcgcg ggaatccgat atgccccgct atcgacacca 7620
ttactcaacc gatgtgagac cttctctctg gtctggcaca ttttcagcat tcccactttc 7680
ctcacaattt tcatgctatg ctgcgcaatt ccactgctct ggccatttgt gattgcgtat 7740
gtagtgtacg ctgttaaaga cgactccccg tccaacggag gagtggtcaa gcgatactcg 7800
cctatttcaa gaaacttctt catctggaag ctctttggcc gctacttccc cataactctg 7860
cacaagacgg tggatctgga gcccacgcac acatactacc ctctggacgt ccaggagtat 7920
cacctgattg ctgagagata ctggccgcag aacaagtacc tccgagcaat catcaccacc 7980
atcgagtact ttctgcccgc cttcatgaaa cggtctcttt ctatcaacga gcaggagcag 8040
cctgccgagc gagatcctct cctgtctccc gtttctccca gctctccggg ttctcaacct 8100
gacaagtgga ttaaccacga cagcagatat agccgtggag aatcatctgg ctccaacggc 8160
cacgcctcgg gctccgaact taacggcaac ggcaacaacg gcaccactaa ccgacgacct 8220
ttgtcgtccg cctctgctgg ctccactgca tctgattcca cgcttcttaa cgggtccctc 8280
aactcctacg ccaaccagat cattggcgaa aacgacccac agctgtcgcc cacaaaactc 8340
aagcccactg gcagaaaata catcttcggc taccaccccc acggcattat cggcatggga 8400
gcctttggtg gaattgccac cgagggagct ggatggtcca agctctttcc gggcatccct 8460
gtttctctta tgactctcac caacaacttc cgagtgcctc tcttcagaga gtacctcatg 8520
agtctgggag tcgcttctgt ctccaagaag tcctgcaagg ccctcctcaa gcgaaaccag 8580
tctatctgca ttgtcgttgg tggagcacag gaaagtcttc tggccagacc cggtgtcatg 8640
gacctggtgc tactcaagcg aaagggtttt gttcgacttg gtatggaggt cggaaatgtc 8700
gcccttgttc ccatcatggc ctttggtgag aacgacctct atgaccaggt tagcaacgac 8760
aagtcgtcca agctgtaccg attccagcag tttgtcaaga acttccttgg attcaccctt 8820
cctttgatgc atgcccgagg cgtcttcaac tacgatgtcg gtcttgtccc ctacaggcga 8880
cccgtcaaca ttgtggttgg ttcccccatt gacttgcctt atctcccaca ccccaccgac 8940
gaagaagtgt ccgaatacca cgaccgatac atcgccgagc tgcagcgaat ctacaacgag 9000
cacaaggatg aatatttcat cgattggacc gaggagggca aaggagcccc agagttccga 9060
atgattgagt aagc 9074
<210>96
<211>9074
<212>DNA
<213>人工序列
<220>
<223>质粒pY193
<400>96
ggccgcaggg cggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg 60
acctcgaggg ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca 120
ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 180
cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 240
ccttcccaac agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca 300
ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 360
gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 420
caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 480
cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 540
tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 600
acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 660
gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 720
ttaacgttta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 780
accgcatatc gacggtcgag gagaacttct agtatatcca catacctaat attattgcct 840
tattaaaaat ggaatcccaa caattacatc aaaatccaca ttctcttcaa aatcaattgt 900
cctgtacttc cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct 960
atttctcaac aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat 1020
atcttgaccg cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc 1080
tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac 1140
agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata 1200
cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gaggccggaa ccggcttttc 1260
atatagaata gagaagcgtt catgactaaa tgcttgcatc acaatacttg aagttgacaa 1320
tattatttaa ggacctattg ttttttccaa taggtggtta gcaatcgtct tactttctaa 1380
cttttcttac cttttacatt tcagcaatat atatatatat ttcaaggata taccattcta 1440
atgtctgccc ctatgtctgc ccctaagaag atcgtcgttt tgccaggtga ccacgttggt 1500
caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt tcgttccaat 1560
gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc tacaggtgtc 1620
ccacttccag atgaggcgct ggaagcctcc aagaaggttg atgccgtttt gttaggtgct 1680
gtggctggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt actaaaaatc 1740
cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc cgactctctt 1800
ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt tgttgtcaga 1860
gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga tggtgtcgct 1920
tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat ggccgctttc 1980
atggccctac aacatgagcc accattgcct atttggtcct tggataaagc taatcttttg 2040
gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga attccctaca 2100
ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa gaacccaacc 2160
cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc cgatgaagcc 2220
tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc tttgccagac 2280
aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga tttgccaaag 2340
aataaggttg accctatcgc cactatcttg tctgctgcaa tgatgttgaa attgtcattg 2400
aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt ggatgcaggt 2460
atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga tgctgtcgcc 2520
gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata 2580
aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca 2640
tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag 2700
gatagtaaag gaatacaggt aagcaaattg atactaatgg ctcaacgtga taaggaaaaa 2760
gaattgcact ttaacattaa tattgacaag gaggagggca ccacacaaaa agttaggtgt 2820
aacagaaaat catgaaacta cgattcctaa tttgatattg gaggattttc tctaaaaaaa 2880
aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa 2940
taccttcttg aagcatttcc cataatggtg aaagttccct caagaatttt actctgtcag 3000
aaacggcctt acgacgtagt cgatatggtg cactctcagt acaatctgct ctgatgccgc 3060
atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 3120
gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 3180
gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt 3240
ataggttaat gtcatgataa taatggtttc ttagtatgat ccaatatcaa aggaaatgat 3300
agcattgaag gatgagacta atccaattga ggagtggcag catatagaac agctaaaggg 3360
tagtgctgaa ggaagcatac gataccccgc atggaatggg ataatatcac aggaggtact 3420
agactacctt tcatcctaca taaatagacg catataagta cgcatttaag cataaacacg 3480
cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac 3540
gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg 3600
aaacgctttg aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcaga 3660
gcgcttttga aaaccaaaag cgctctgaag acgcactttc aaaaaaccaa aaacgcaccg 3720
gactgtaacg agctactaaa atattgcgaa taccgcttcc acaaacattg ctcaaaagta 3780
tctctttgct atatatctct gtgctatatc cctatataac ctacccatcc acctttcgct 3840
ccttgaactt gcatctaaac tcgacctcta cattttttat gtttatctct agtattactc 3900
tttagacaaa aaaattgtag taagaactat tcatagagtg aatcgaaaac aatacgaaaa 3960
tgtaaacatt tcctatacgt agtatataga gacaaaatag aagaaaccgt tcataatttt 4020
ctgaccaatg aagaatcatc aacgctatca ctttctgttc acaaagtatg cgcaatccac 4080
atcggtatag aatataatcg gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc 4140
tagtaatcag taaacgcggg aagtggagtc aggctttttt tatggaagag aaaatagaca 4200
ccaaagtagc cttcttctaa ccttaacgga cctacagtgc aaaaagttat caagagactg 4260
cattatagag cgcacaaagg agaaaaaaag taatctaaga tgctttgtta gaaaaatagc 4320
gctctcggga tgcatttttg tagaacaaaa aagaagtata gattctttgt tggtaaaata 4380
gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa 4440
ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa gcacagattc ttcgttggta 4500
aaatagcgct ttcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg 4560
aaaaattagc gctctcgcgt tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt 4620
caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 4680
attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 4740
aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 4800
tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 4860
agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 4920
gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 4980
cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 5040
agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 5100
taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 5160
tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 5220
taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 5280
acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 5340
ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 5400
cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 5460
agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 5520
tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 5580
agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 5640
tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 5700
ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 5760
tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 5820
aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 5880
tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 5940
agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 6000
taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 6060
caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6120
agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 6180
aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 6240
gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 6300
tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 6360
gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 6420
ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 6480
ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 6540
aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 6600
aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 6660
atgtgagtta cctcactcat taggcacccc aggctttaca ctttatgctt ccggctccta 6720
tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 6780
acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggga 6840
tttcgaaact aagttcttgg tgttttaaaa ctaaaaaaaa gactaactat aaaagtagaa 6900
tttaagaagt ttaagaaata gatttacaga attacaatca atacctaccg tctttatata 6960
cttattagtc aagtagggga ataatttcag ggaactggtt tcaacctttt ttttcagctt 7020
tttccaaatc agagagagca gaaggtaata gaaggtgtaa gaaaatgaga tagatacatg 7080
cgtgggtcaa ttgccttgtg tcatcattta ctccaggcag gttgcatcac tccattgagg 7140
ttgtgcccgt tttttgcctg tttgtgcccc tgttctctgt agttgcgcta agagaatgga 7200
cctatgaact gatggttggt gaagaaaaca atattttggt gctgggattc tttttttttc 7260
tggatgccag cttaaaaagc gggctccatt atatttagtg gatgccagga ataaactgtt 7320
cacccagaca cctacgatgt tatatattct gtgtaacccg ccccctattt tgggcatgta 7380
cgggttacag cagaattaaa aggctaattt tttgactaaa taaagttagg aaaatcacta 7440
ctattaatta tttacgtatt ctttgaaatg gcagtattga taatgataaa ctcgaaatca 7500
ctagtggatc cgcccagcgg ccgcaacatg actatcgact cacaatacta caagtcgcga 7560
gacaaaaacg acacggcacc caaaatcgcg ggaatccgat atgccccgct atcgacacca 7620
ttactcaacc gatgtgagac cttctctctg gtctggcaca ttttcagcat tcccactttc 7680
ctcacaattt tcatgctatg ctgcgcaatt ccactgctct ggccatttgt gattgcgtat 7740
gtagtgtacg ctgttaaaga cgactccccg tccaacggag gagtggtcaa gcgatactcg 7800
cctatttcaa gaaacttctt catctggaag ctctttggcc gctacttccc cataactctg 7860
cacaagacgg tggatctgga gcccacgcac acatactacc ctctggacgt ccaggagtat 7920
cacctgattg ctgagagata ctggccgcag aacaagtacc tccgagcaat catcaccacc 7980
atcgagtact ttctgcccgc cttcatgaaa cggtctcttt ctatcaacga gcaggagcag 8040
cctgccgagc gagatcctct cctgtctccc gtttctccca gctctccggg ttctcaacct 8100
gacaagtgga ttaaccacga cagcagatat agccgtggag aatcatctgg ctccaacggc 8160
cacgcctcgg gctccgaact taacggcaac ggcaacaacg gcaccactaa ccgacgacct 8220
ttgtcgtccg cctctgctgg ctccactgca tctgattcca cgcttcttaa cgggtccctc 8280
aactcctacg ccaaccagat cattggcgaa aacgacccac agctgtcgcc cacaaaactc 8340
aagcccactg gcagaaaata catcttcggc taccaccccc acggcattat cggcatggga 8400
gcctttggtg gaattgccac cgagggagct ggatggtcca agctctttcc gggcatccct 8460
gtttctctta tgactctcac caacaacttc cgagtgcctc tcctcagaga gtacctcatg 8520
agtctgggag tcgcttctgt ctccaagaag tcctgcaagg ccctcctcaa gcgaaaccag 8580
tctatctgca ttgtcgttgg tggagcacag gaaagtcttc tggccagacc cggtgtcatg 8640
gacctggtgc tactcaagcg aaagggtttt gttcgacttg gtatggaggt cggaaatgtc 8700
gcccttgttc ccatcatggc ctttggtgag aacgacctct atgaccaggt tagcaacgac 8760
aagtcgtcca agctgtaccg attccagcag tttgtcaaga acttccttgg attcaccctt 8820
cctttgatgc atgcccgagg cgtcttcaac tacgatgtcg gtcttgtccc ctacaggcga 8880
cccgtcaaca ttgtggttgg ttcccccatt gacttgcctt atctcccaca ccccaccgac 8940
gaagaagtgt ccgaatacca cgaccgatac atcgccgagc tgcagcgaat ctacaacgag 9000
cacaaggatg aatatttcat cgattggacc gaggagggca aaggagcccc agagttccga 9060
atgattgagt aagc 9074
<210>97
<211>9074
<212>DNA
<213>人工序列
<220>
<223>质粒pY194
<400>97
ggccgcaggg cggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg 60
acctcgaggg ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca 120
ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 180
cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 240
ccttcccaac agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca 300
ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 360
gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 420
caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 480
cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 540
tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 600
acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 660
gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 720
ttaacgttta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 780
accgcatatc gacggtcgag gagaacttct agtatatcca catacctaat attattgcct 840
tattaaaaat ggaatcccaa caattacatc aaaatccaca ttctcttcaa aatcaattgt 900
cctgtacttc cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct 960
atttctcaac aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat 1020
atcttgaccg cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc 1080
tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac 1140
agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata 1200
cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gaggccggaa ccggcttttc 1260
atatagaata gagaagcgtt catgactaaa tgcttgcatc acaatacttg aagttgacaa 1320
tattatttaa ggacctattg ttttttccaa taggtggtta gcaatcgtct tactttctaa 1380
cttttcttac cttttacatt tcagcaatat atatatatat ttcaaggata taccattcta 1440
atgtctgccc ctatgtctgc ccctaagaag atcgtcgttt tgccaggtga ccacgttggt 1500
caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt tcgttccaat 1560
gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc tacaggtgtc 1620
ccacttccag atgaggcgct ggaagcctcc aagaaggttg atgccgtttt gttaggtgct 1680
gtggctggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt actaaaaatc 1740
cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc cgactctctt 1800
ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt tgttgtcaga 1860
gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga tggtgtcgct 1920
tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat ggccgctttc 1980
atggccctac aacatgagcc accattgcct atttggtcct tggataaagc taatcttttg 2040
gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga attccctaca 2100
ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa gaacccaacc 2160
cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc cgatgaagcc 2220
tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc tttgccagac 2280
aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga tttgccaaag 2340
aataaggttg accctatcgc cactatcttg tctgctgcaa tgatgttgaa attgtcattg 2400
aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt ggatgcaggt 2460
atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga tgctgtcgcc 2520
gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata 2580
aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca 2640
tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag 2700
gatagtaaag gaatacaggt aagcaaattg atactaatgg ctcaacgtga taaggaaaaa 2760
gaattgcact ttaacattaa tattgacaag gaggagggca ccacacaaaa agttaggtgt 2820
aacagaaaat catgaaacta cgattcctaa tttgatattg gaggattttc tctaaaaaaa 2880
aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa 2940
taccttcttg aagcatttcc cataatggtg aaagttccct caagaatttt actctgtcag 3000
aaacggcctt acgacgtagt cgatatggtg cactctcagt acaatctgct ctgatgccgc 3060
atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 3120
gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 3180
gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt 3240
ataggttaat gtcatgataa taatggtttc ttagtatgat ccaatatcaa aggaaatgat 3300
agcattgaag gatgagacta atccaattga ggagtggcag catatagaac agctaaaggg 3360
tagtgctgaa ggaagcatac gataccccgc atggaatggg ataatatcac aggaggtact 3420
agactacctt tcatcctaca taaatagacg catataagta cgcatttaag cataaacacg 3480
cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac 3540
gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg 3600
aaacgctttg aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcaga 3660
gcgcttttga aaaccaaaag cgctctgaag acgcactttc aaaaaaccaa aaacgcaccg 3720
gactgtaacg agctactaaa atattgcgaa taccgcttcc acaaacattg ctcaaaagta 3780
tctctttgct atatatctct gtgctatatc cctatataac ctacccatcc acctttcgct 3840
ccttgaactt gcatctaaac tcgacctcta cattttttat gtttatctct agtattactc 3900
tttagacaaa aaaattgtag taagaactat tcatagagtg aatcgaaaac aatacgaaaa 3960
tgtaaacatt tcctatacgt agtatataga gacaaaatag aagaaaccgt tcataatttt 4020
ctgaccaatg aagaatcatc aacgctatca ctttctgttc acaaagtatg cgcaatccac 4080
atcggtatag aatataatcg gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc 4140
tagtaatcag taaacgcggg aagtggagtc aggctttttt tatggaagag aaaatagaca 4200
ccaaagtagc cttcttctaa ccttaacgga cctacagtgc aaaaagttat caagagactg 4260
cattatagag cgcacaaagg agaaaaaaag taatctaaga tgctttgtta gaaaaatagc 4320
gctctcggga tgcatttttg tagaacaaaa aagaagtata gattctttgt tggtaaaata 4380
gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa 4440
ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa gcacagattc ttcgttggta 4500
aaatagcgct ttcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg 4560
aaaaattagc gctctcgcgt tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt 4620
caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 4680
attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 4740
aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 4800
tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 4860
agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 4920
gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 4980
cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 5040
agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 5100
taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 5160
tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 5220
taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 5280
acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 5340
ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 5400
cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 5460
agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 5520
tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 5580
agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 5640
tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 5700
ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 5760
tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 5820
aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 5880
tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 5940
agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 6000
taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 6060
caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6120
agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 6180
aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 6240
gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 6300
tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 6360
gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 6420
ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 6480
ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 6540
aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 6600
aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 6660
atgtgagtta cctcactcat taggcacccc aggctttaca ctttatgctt ccggctccta 6720
tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 6780
acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggga 6840
tttcgaaact aagttcttgg tgttttaaaa ctaaaaaaaa gactaactat aaaagtagaa 6900
tttaagaagt ttaagaaata gatttacaga attacaatca atacctaccg tctttatata 6960
cttattagtc aagtagggga ataatttcag ggaactggtt tcaacctttt ttttcagctt 7020
tttccaaatc agagagagca gaaggtaata gaaggtgtaa gaaaatgaga tagatacatg 7080
cgtgggtcaa ttgccttgtg tcatcattta ctccaggcag gttgcatcac tccattgagg 7140
ttgtgcccgt tttttgcctg tttgtgcccc tgttctctgt agttgcgcta agagaatgga 7200
cctatgaact gatggttggt gaagaaaaca atattttggt gctgggattc tttttttttc 7260
tggatgccag cttaaaaagc gggctccatt atatttagtg gatgccagga ataaactgtt 7320
cacccagaca cctacgatgt tatatattct gtgtaacccg ccccctattt tgggcatgta 7380
cgggttacag cagaattaaa aggctaattt tttgactaaa taaagttagg aaaatcacta 7440
ctattaatta tttacgtatt ctttgaaatg gcagtattga taatgataaa ctcgaaatca 7500
ctagtggatc cgcccagcgg ccgcaacatg actatcgact cacaatacta caagtcgcga 7560
gacaaaaacg acacggcacc caaaatcgcg ggaatccgat atgccccgct atcgacacca 7620
ttactcaacc gatgtgagac cttctctctg gtctggcaca ttttcagcat tcccactttc 7680
ctcacaattt tcatgctatg ctgcgcaatt ccactgctct ggccatttgt gattgcgtat 7740
gtagtgtacg ctgttaaaga cgactccccg tccaacggag gagtggtcaa gcgatactcg 7800
cctatttcaa gaaacttctt catctggaag ctctttggcc gctacttccc cataactctg 7860
cacaagacgg tggatctgga gcccacgcac acatactacc ctctggacgt ccaggagtat 7920
cacctgattg ctgagagata ctggccgcag aacaagtacc tccgagcaat catcaccacc 7980
atcgagtact ttctgcccgc cttcatgaaa cggtctcttt ctatcaacga gcaggagcag 8040
cctgccgagc gagatcctct cctgtctccc gtttctccca gctctccggg ttctcaacct 8100
gacaagtgga ttaaccacga cagcagatat agccgtggag aatcatctgg ctccaacggc 8160
cacgcctcgg gctccgaact taacggcaac ggcaacaacg gcaccactaa ccgacgacct 8220
ttgtcgtccg cctctgctgg ctccactgca tctgattcca cgcttcttaa cgggtccctc 8280
aactcctacg ccaaccagat cattggcgaa aacgacccac agctgtcgcc cacaaaactc 8340
aagcccactg gcagaaaata catcttcggc taccaccccc acggcattat cggcatggga 8400
gcctttggtg gaattgccac cgagggagct ggatggtcca agctctttcc gggcatccct 8460
gtttctctta tgactctcac caacaacttc cgagtgcctc tctacaaaga gtacctcatg 8520
agtctgggag tcgcttctgt ctccaagaag tcctgcaagg ccctcctcaa gcgaaaccag 8580
tctatctgca ttgtcgttgg tggagcacag gaaagtcttc tggccagacc cggtgtcatg 8640
gacctggtgc tactcaagcg aaagggtttt gttcgacttg gtatggaggt cggaaatgtc 8700
gcccttgttc ccatcatggc ctttggtgag aacgacctct atgaccaggt tagcaacgac 8760
aagtcgtcca agctgtaccg attccagcag tttgtcaaga acttccttgg attcaccctt 8820
cctttgatgc atgcccgagg cgtcttcaac tacgatgtcg gtcttgtccc ctacaggcga 8880
cccgtcaaca ttgtggttgg ttcccccatt gacttgcctt atctcccaca ccccaccgac 8940
gaagaagtgt ccgaatacca cgaccgatac atcgccgagc tgcagcgaat ctacaacgag 9000
cacaaggatg aatatttcat cgattggacc gaggagggca aaggagcccc agagttccga 9060
atgattgagt aagc 9074
<210>98
<211>8641
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1256
<400>98
ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacaagctt ggcgcgccgt tctatagtgt cacctaaatc 1200
gtatgtgtat gatacataag gttatgtatt aattgtagcc gcgttctaac gacaatatgt 1260
ccatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac 1320
acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca 1380
gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga 1440
aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgacca 1500
aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 1560
gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 1620
cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 1680
ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc 1740
accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 1800
tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 1860
cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 1920
gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc 1980
ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 2040
cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 2100
tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 2160
ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 2220
ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 2280
ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 2340
gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc aggttgatcg 2400
attcgacatc gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg 2460
cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa 2520
acccatctca taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc 2580
aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt 2640
tattgccaaa tgtttgaacg atctgcttcg acgcactcct tctttaggta cctcactatt 2700
cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac 2760
agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg 2820
ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc 2880
cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc 2940
gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac 3000
aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca 3060
tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg 3120
agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca 3180
gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt 3240
gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga 3300
ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg 3360
catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt 3420
gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaattccc 3480
caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac 3540
gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta 3600
catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc 3660
tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggctttttca 3720
tggtttaata agaagagaaa agagttcttt tgttatggct gaagtaatag agaaatgagc 3780
tcgagcgtgt cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag 3840
gatagtggga ttgtgcgtca tcccttacgt cagtggagat gtcacatcaa tccacttgct 3900
ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat 3960
ctttgggacc actgtcggca gaggcatctt gaatgatagc ctttccttta tcgcaatgat 4020
ggcatttgta ggagccacct tccttttcta ctgtcctttc gatgaagtga cagatagctg 4080
ggcaatggaa tccgaggagg tttcccgaaa ttatcctttg ttgaaaagtc tcaatagccc 4140
tttggtcttc tgagactgta tctttgacat ttttggagta gaccagagtg tcgtgctcca 4200
ccatgttgac gaagattttc ttcttgtcat tgagtcgtaa aagactctgt atgaactgtt 4260
cgccagtctt cacggcgagt tctgttagat cctcgatttg aatcttagac tccatgcatg 4320
gccttagatt cagtaggaac taccttttta gagactccaa tctctattac ttgccttggt 4380
ttatgaagca agccttgaat cgtccatact ggaatagtac ttctgatctt gagaaatatg 4440
tctttctctg tgttcttgat gcaattagtc ctgaatcttt tgactgcatc tttaaccttc 4500
ttgggaaggt atttgatctc ctggagattg ttactcgggt agatcgtctt gatgagacct 4560
gctgcgtagg cctctctaac catctgtggg tcagcattct ttctgaaatt gaagaggcta 4620
accttctcat tatcagtggt gaacatagtg tcgtcacctt caccttcgaa cttccttcct 4680
agatcgtaaa gatagaggaa atcgtccatt gtaatctccg gggcaaagga gatctctttt 4740
ggggctggat cactgctggg ccttttggtt cctagcgtga gccagtgggc tttttgcttt 4800
ggtgggcttg ttagggcctt agcaaagctc ttgggcttga gttgagcttc tcctttgggg 4860
atgaagttca acctgtctgt ttgctgactt gttgtgtacg cgtcagctgc tgctcttgcc 4920
tctgtaatag tggcaaattt cttgtgtgca actccgggaa cgccgtttgt tgccgccttt 4980
gtacaacccc agtcatcgta tataccggca tgtggaccgt tatacacaac gtagtagttg 5040
atatgagggt gttgaatacc cgattctgct ctgagaggag caactgtgct gttaagctca 5100
gatttttgtg ggattggaat tggatcgatc tcgatcccgc gaaattaata cgactcacta 5160
tagggagacc acaacggttt ccctctagaa ataattttgt ttaactttaa gaaggagata 5220
tacccatgga aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt 5280
tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct 5340
tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca 5400
aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg 5460
acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca 5520
cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcta 5580
tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc 5640
aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg 5700
tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg 5760
atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt 5820
tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg 5880
aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt 5940
tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat 6000
cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg 6060
ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat 6120
ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg 6180
atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg 6240
caaaggaata gtgaggtaca gcttggatcg atccggctgc taacaaagcc cgaaaggaag 6300
ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac 6360
gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatgatcg ggcgcgccgt 6420
cgacggatcc gtacgagatc cggccggcca gatcctgcag gagatccaag cttttgatcc 6480
atgcccttca tttgccgctt attaattaat ttggtaacag tccgtactaa tcagttactt 6540
atccttcccc catcataatt aatcttggta gtctcgaatg ccacaacact gactagtctc 6600
ttggatcata agaaaaagcc aaggaacaaa agaagacaaa acacaatgag agtatccttt 6660
gcatagcaat gtctaagttc ataaaattca aacaaaaacg caatcacaca cagtggacat 6720
cacttatcca ctagctgatc aggatcgccg cgtcaagaaa aaaaaactgg accccaaaag 6780
ccatgcacaa caacacgtac tcacaaaggt gtcaatcgag cagcccaaaa cattcaccaa 6840
ctcaacccat catgagccct cacatttgtt gtttctaacc caacctcaaa ctcgtattct 6900
cttccgccac ctcatttttg tttatttcaa cacccgtcaa actgcatgcc accccgtggc 6960
caaatgtcca tgcatgttaa caagacctat gactataaat agctgcaatc tcggcccagg 7020
ttttcatcat caagaaccag ttcaatatcc tagtacaccg tattaaagaa tttaagatat 7080
actgcggccg caacatgact atcgactcac aatactacaa gtcgcgagac aaaaacgaca 7140
cggcacccaa aatcgcggga atccgatatg ccccgctatc gacaccatta ctcaaccgat 7200
gtgagacctt ctctctggtc tggcacattt tcagcattcc cactttcctc acaattttca 7260
tgctatgctg cgcaattcca ctgctctggc catttgtgat tgcgtatgta gtgtacgctg 7320
ttaaagacga ctccccgtcc aacggaggag tggtcaagcg atactcgcct atttcaagaa 7380
acttcttcat ctggaagctc tttggccgct acttccccat aactctgcac aagacggtgg 7440
atctggagcc cacgcacaca tactaccctc tggacgtcca ggagtatcac ctgattgctg 7500
agagatactg gccgcagaac aagtacctcc gagcaatcat caccaccatc gagtactttc 7560
tgcccgcctt catgaaacgg tctctttcta tcaacgagca ggagcagcct gccgagcgag 7620
atcctctcct gtctcccgtt tctcccagct ctccgggttc tcaacctgac aagtggatta 7680
accacgacag cagatatagc cgtggagaat catctggctc caacggccac gcctcgggct 7740
ccgaacttaa cggcaacggc aacaacggca ccactaaccg acgacctttg tcgtccgcct 7800
ctgctggctc cactgcatct gattccacgc ttcttaacgg gtccctcaac tcctacgcca 7860
accagatcat tggcgaaaac gacccacagc tgtcgcccac aaaactcaag cccactggca 7920
gaaaatacat cttcggctac cacccccacg gcattatcgg catgggagcc tttggtggaa 7980
ttgccaccga gggagctgga tggtccaagc tctttccggg catccctgtt tctcttatga 8040
ctctcaccaa caacttccga gtgcctctct acagagagta cctcatgagt ctgggagtcg 8100
cttctgtctc caagaagtcc tgcaaggccc tcctcaagcg aaaccagtct atctgcattg 8160
tcgttggtgg agcacaggaa agtcttctgg ccagacccgg tgtcatggac ctggtgctac 8220
tcaagcgaaa gggttttgtt cgacttggta tggaggtcgg aaatgtcgcc cttgttccca 8280
tcatggcctt tggtgagaac gacctctatg accaggttag caacgacaag tcgtccaagc 8340
tgtaccgatt ccagcagttt gtcaagaact tccttggatt cacccttcct ttgatgcatg 8400
cccgaggcgt cttcaactac gatgtcggtc ttgtccccta caggcgaccc gtcaacattg 8460
tggttggttc ccccattgac ttgccttatc tcccacaccc caccgacgaa gaagtgtccg 8520
aataccacga ccgatacatc gccgagctgc agcgaatcta caacgagcac aaggatgaat 8580
atttcatcga ttggaccgag gagggcaaag gagccccaga gttccgaatg attgagtaag 8640
c 8641
<210>99
<211>8641
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1277
<400>99
ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacaagctt ggcgcgccgt tctatagtgt cacctaaatc 1200
gtatgtgtat gatacataag gttatgtatt aattgtagcc gcgttctaac gacaatatgt 1260
ccatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac 1320
acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca 1380
gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga 1440
aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgacca 1500
aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 1560
gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 1620
cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 1680
ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc 1740
accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 1800
tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 1860
cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 1920
gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc 1980
ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 2040
cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 2100
tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 2160
ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 2220
ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 2280
ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 2340
gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc aggttgatcg 2400
attcgacatc gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg 2460
cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa 2520
acccatctca taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc 2580
aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt 2640
tattgccaaa tgtttgaacg atctgcttcg acgcactcct tctttaggta cctcactatt 2700
cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac 2760
agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg 2820
ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc 2880
cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc 2940
gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac 3000
aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca 3060
tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg 3120
agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca 3180
gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt 3240
gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga 3300
ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg 3360
catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt 3420
gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaattccc 3480
caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac 3540
gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta 3600
catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc 3660
tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggctttttca 3720
tggtttaata agaagagaaa agagttcttt tgttatggct gaagtaatag agaaatgagc 3780
tcgagcgtgt cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag 3840
gatagtggga ttgtgcgtca tcccttacgt cagtggagat gtcacatcaa tccacttgct 3900
ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat 3960
ctttgggacc actgtcggca gaggcatctt gaatgatagc ctttccttta tcgcaatgat 4020
ggcatttgta ggagccacct tccttttcta ctgtcctttc gatgaagtga cagatagctg 4080
ggcaatggaa tccgaggagg tttcccgaaa ttatcctttg ttgaaaagtc tcaatagccc 4140
tttggtcttc tgagactgta tctttgacat ttttggagta gaccagagtg tcgtgctcca 4200
ccatgttgac gaagattttc ttcttgtcat tgagtcgtaa aagactctgt atgaactgtt 4260
cgccagtctt cacggcgagt tctgttagat cctcgatttg aatcttagac tccatgcatg 4320
gccttagatt cagtaggaac taccttttta gagactccaa tctctattac ttgccttggt 4380
ttatgaagca agccttgaat cgtccatact ggaatagtac ttctgatctt gagaaatatg 4440
tctttctctg tgttcttgat gcaattagtc ctgaatcttt tgactgcatc tttaaccttc 4500
ttgggaaggt atttgatctc ctggagattg ttactcgggt agatcgtctt gatgagacct 4560
gctgcgtagg cctctctaac catctgtggg tcagcattct ttctgaaatt gaagaggcta 4620
accttctcat tatcagtggt gaacatagtg tcgtcacctt caccttcgaa cttccttcct 4680
agatcgtaaa gatagaggaa atcgtccatt gtaatctccg gggcaaagga gatctctttt 4740
ggggctggat cactgctggg ccttttggtt cctagcgtga gccagtgggc tttttgcttt 4800
ggtgggcttg ttagggcctt agcaaagctc ttgggcttga gttgagcttc tcctttgggg 4860
atgaagttca acctgtctgt ttgctgactt gttgtgtacg cgtcagctgc tgctcttgcc 4920
tctgtaatag tggcaaattt cttgtgtgca actccgggaa cgccgtttgt tgccgccttt 4980
gtacaacccc agtcatcgta tataccggca tgtggaccgt tatacacaac gtagtagttg 5040
atatgagggt gttgaatacc cgattctgct ctgagaggag caactgtgct gttaagctca 5100
gatttttgtg ggattggaat tggatcgatc tcgatcccgc gaaattaata cgactcacta 5160
tagggagacc acaacggttt ccctctagaa ataattttgt ttaactttaa gaaggagata 5220
tacccatgga aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt 5280
tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct 5340
tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca 5400
aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg 5460
acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca 5520
cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcta 5580
tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc 5640
aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg 5700
tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg 5760
atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt 5820
tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg 5880
aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt 5940
tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat 6000
cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg 6060
ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat 6120
ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg 6180
atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg 6240
caaaggaata gtgaggtaca gcttggatcg atccggctgc taacaaagcc cgaaaggaag 6300
ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac 6360
gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatgatcg ggcgcgccgt 6420
cgacggatcc gtacgagatc cggccggcca gatcctgcag gagatccaag cttttgatcc 6480
atgcccttca tttgccgctt attaattaat ttggtaacag tccgtactaa tcagttactt 6540
atccttcccc catcataatt aatcttggta gtctcgaatg ccacaacact gactagtctc 6600
ttggatcata agaaaaagcc aaggaacaaa agaagacaaa acacaatgag agtatccttt 6660
gcatagcaat gtctaagttc ataaaattca aacaaaaacg caatcacaca cagtggacat 6720
cacttatcca ctagctgatc aggatcgccg cgtcaagaaa aaaaaactgg accccaaaag 6780
ccatgcacaa caacacgtac tcacaaaggt gtcaatcgag cagcccaaaa cattcaccaa 6840
ctcaacccat catgagccct cacatttgtt gtttctaacc caacctcaaa ctcgtattct 6900
cttccgccac ctcatttttg tttatttcaa cacccgtcaa actgcatgcc accccgtggc 6960
caaatgtcca tgcatgttaa caagacctat gactataaat agctgcaatc tcggcccagg 7020
ttttcatcat caagaaccag ttcaatatcc tagtacaccg tattaaagaa tttaagatat 7080
actgcggccg caacatgact atcgactcac aatactacaa gtcgcgagac aaaaacgaca 7140
cggcacccaa aatcgcggga atccgatatg ccccgctatc gacaccatta ctcaaccgat 7200
gtgagacctt ctctctggtc tggcacattt tcagcattcc cactttcctc acaattttca 7260
tgctatgctg cgcaattcca ctgctctggc catttgtgat tgcgtatgta gtgtacgctg 7320
ttaaagacga ctccccgtcc aacggaggag tggtcaagcg atactcgcct atttcaagaa 7380
acttcttcat ctggaagctc tttggccgct acttccccat aactctgcac aagacggtgg 7440
atctggagcc cacgcacaca tactaccctc tggacgtcca ggagtatcac ctgattgctg 7500
agagatactg gccgcagaac aagtacctcc gagcaatcat caccaccatc gagtactttc 7560
tgcccgcctt catgaaacgg tctctttcta tcaacgagca ggagcagcct gccgagcgag 7620
atcctctcct gtctcccgtt tctcccagct ctccgggttc tcaacctgac aagtggatta 7680
accacgacag cagatatagc cgtggagaat catctggctc caacggccac gcctcgggct 7740
ccgaacttaa cggcaacggc aacaacggca ccactaaccg acgacctttg tcgtccgcct 7800
ctgctggctc cactgcatct gattccacgc ttcttaacgg gtccctcaac tcctacgcca 7860
accagatcat tggcgaaaac gacccacagc tgtcgcccac aaaactcaag cccactggca 7920
gaaaatacat cttcggctac cacccccacg gcattatcgg catgggagcc tttggtggaa 7980
ttgccaccga gggagctgga tggtccaagc tctttccggg catccctgtt tctcttatga 8040
ctctcaccaa caacttccga gtgcctctct tcagagagta cctcatgagt ctgggagtcg 8100
cttctgtctc caagaagtcc tgcaaggccc tcctcaagcg aaaccagtct atctgcattg 8160
tcgttggtgg agcacaggaa agtcttctgg ccagacccgg tgtcatggac ctggtgctac 8220
tcaagcgaaa gggttttgtt cgacttggta tggaggtcgg aaatgtcgcc cttgttccca 8280
tcatggcctt tggtgagaac gacctctatg accaggttag caacgacaag tcgtccaagc 8340
tgtaccgatt ccagcagttt gtcaagaact tccttggatt cacccttcct ttgatgcatg 8400
cccgaggcgt cttcaactac gatgtcggtc ttgtccccta caggcgaccc gtcaacattg 8460
tggttggttc ccccattgac ttgccttatc tcccacaccc caccgacgaa gaagtgtccg 8520
aataccacga ccgatacatc gccgagctgc agcgaatcta caacgagcac aaggatgaat 8580
atttcatcga ttggaccgag gagggcaaag gagccccaga gttccgaatg attgagtaag 8640
c 8641
<210>100
<211>8641
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1278
<400>100
ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacaagctt ggcgcgccgt tctatagtgt cacctaaatc 1200
gtatgtgtat gatacataag gttatgtatt aattgtagcc gcgttctaac gacaatatgt 1260
ccatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac 1320
acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca 1380
gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga 1440
aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgacca 1500
aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 1560
gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 1620
cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 1680
ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc 1740
accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 1800
tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 1860
cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 1920
gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc 1980
ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 2040
cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 2100
tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 2160
ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 2220
ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 2280
ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 2340
gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc aggttgatcg 2400
attcgacatc gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg 2460
cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa 2520
acccatctca taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc 2580
aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt 2640
tattgccaaa tgtttgaacg atctgcttcg acgcactcct tctttaggta cctcactatt 2700
cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac 2760
agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg 2820
ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc 2880
cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc 2940
gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac 3000
aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca 3060
tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg 3120
agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca 3180
gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt 3240
gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga 3300
ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg 3360
catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt 3420
gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaattccc 3480
caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac 3540
gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta 3600
catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc 3660
tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggctttttca 3720
tggtttaata agaagagaaa agagttcttt tgttatggct gaagtaatag agaaatgagc 3780
tcgagcgtgt cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag 3840
gatagtggga ttgtgcgtca tcccttacgt cagtggagat gtcacatcaa tccacttgct 3900
ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat 3960
ctttgggacc actgtcggca gaggcatctt gaatgatagc ctttccttta tcgcaatgat 4020
ggcatttgta ggagccacct tccttttcta ctgtcctttc gatgaagtga cagatagctg 4080
ggcaatggaa tccgaggagg tttcccgaaa ttatcctttg ttgaaaagtc tcaatagccc 4140
tttggtcttc tgagactgta tctttgacat ttttggagta gaccagagtg tcgtgctcca 4200
ccatgttgac gaagattttc ttcttgtcat tgagtcgtaa aagactctgt atgaactgtt 4260
cgccagtctt cacggcgagt tctgttagat cctcgatttg aatcttagac tccatgcatg 4320
gccttagatt cagtaggaac taccttttta gagactccaa tctctattac ttgccttggt 4380
ttatgaagca agccttgaat cgtccatact ggaatagtac ttctgatctt gagaaatatg 4440
tctttctctg tgttcttgat gcaattagtc ctgaatcttt tgactgcatc tttaaccttc 4500
ttgggaaggt atttgatctc ctggagattg ttactcgggt agatcgtctt gatgagacct 4560
gctgcgtagg cctctctaac catctgtggg tcagcattct ttctgaaatt gaagaggcta 4620
accttctcat tatcagtggt gaacatagtg tcgtcacctt caccttcgaa cttccttcct 4680
agatcgtaaa gatagaggaa atcgtccatt gtaatctccg gggcaaagga gatctctttt 4740
ggggctggat cactgctggg ccttttggtt cctagcgtga gccagtgggc tttttgcttt 4800
ggtgggcttg ttagggcctt agcaaagctc ttgggcttga gttgagcttc tcctttgggg 4860
atgaagttca acctgtctgt ttgctgactt gttgtgtacg cgtcagctgc tgctcttgcc 4920
tctgtaatag tggcaaattt cttgtgtgca actccgggaa cgccgtttgt tgccgccttt 4980
gtacaacccc agtcatcgta tataccggca tgtggaccgt tatacacaac gtagtagttg 5040
atatgagggt gttgaatacc cgattctgct ctgagaggag caactgtgct gttaagctca 5100
gatttttgtg ggattggaat tggatcgatc tcgatcccgc gaaattaata cgactcacta 5160
tagggagacc acaacggttt ccctctagaa ataattttgt ttaactttaa gaaggagata 5220
tacccatgga aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt 5280
tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct 5340
tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca 5400
aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg 5460
acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca 5520
cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcta 5580
tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc 5640
aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg 5700
tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg 5760
atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt 5820
tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg 5880
aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt 5940
tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat 6000
cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg 6060
ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat 6120
ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg 6180
atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg 6240
caaaggaata gtgaggtaca gcttggatcg atccggctgc taacaaagcc cgaaaggaag 6300
ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac 6360
gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatgatcg ggcgcgccgt 6420
cgacggatcc gtacgagatc cggccggcca gatcctgcag gagatccaag cttttgatcc 6480
atgcccttca tttgccgctt attaattaat ttggtaacag tccgtactaa tcagttactt 6540
atccttcccc catcataatt aatcttggta gtctcgaatg ccacaacact gactagtctc 6600
ttggatcata agaaaaagcc aaggaacaaa agaagacaaa acacaatgag agtatccttt 6660
gcatagcaat gtctaagttc ataaaattca aacaaaaacg caatcacaca cagtggacat 6720
cacttatcca ctagctgatc aggatcgccg cgtcaagaaa aaaaaactgg accccaaaag 6780
ccatgcacaa caacacgtac tcacaaaggt gtcaatcgag cagcccaaaa cattcaccaa 6840
ctcaacccat catgagccct cacatttgtt gtttctaacc caacctcaaa ctcgtattct 6900
cttccgccac ctcatttttg tttatttcaa cacccgtcaa actgcatgcc accccgtggc 6960
caaatgtcca tgcatgttaa caagacctat gactataaat agctgcaatc tcggcccagg 7020
ttttcatcat caagaaccag ttcaatatcc tagtacaccg tattaaagaa tttaagatat 7080
actgcggccg caacatgact atcgactcac aatactacaa gtcgcgagac aaaaacgaca 7140
cggcacccaa aatcgcggga atccgatatg ccccgctatc gacaccatta ctcaaccgat 7200
gtgagacctt ctctctggtc tggcacattt tcagcattcc cactttcctc acaattttca 7260
tgctatgctg cgcaattcca ctgctctggc catttgtgat tgcgtatgta gtgtacgctg 7320
ttaaagacga ctccccgtcc aacggaggag tggtcaagcg atactcgcct atttcaagaa 7380
acttcttcat ctggaagctc tttggccgct acttccccat aactctgcac aagacggtgg 7440
atctggagcc cacgcacaca tactaccctc tggacgtcca ggagtatcac ctgattgctg 7500
agagatactg gccgcagaac aagtacctcc gagcaatcat caccaccatc gagtactttc 7560
tgcccgcctt catgaaacgg tctctttcta tcaacgagca ggagcagcct gccgagcgag 7620
atcctctcct gtctcccgtt tctcccagct ctccgggttc tcaacctgac aagtggatta 7680
accacgacag cagatatagc cgtggagaat catctggctc caacggccac gcctcgggct 7740
ccgaacttaa cggcaacggc aacaacggca ccactaaccg acgacctttg tcgtccgcct 7800
ctgctggctc cactgcatct gattccacgc ttcttaacgg gtccctcaac tcctacgcca 7860
accagatcat tggcgaaaac gacccacagc tgtcgcccac aaaactcaag cccactggca 7920
gaaaatacat cttcggctac cacccccacg gcattatcgg catgggagcc tttggtggaa 7980
ttgccaccga gggagctgga tggtccaagc tctttccggg catccctgtt tctcttatga 8040
ctctcaccaa caacttccga gtgcctctcc tcagagagta cctcatgagt ctgggagtcg 8100
cttctgtctc caagaagtcc tgcaaggccc tcctcaagcg aaaccagtct atctgcattg 8160
tcgttggtgg agcacaggaa agtcttctgg ccagacccgg tgtcatggac ctggtgctac 8220
tcaagcgaaa gggttttgtt cgacttggta tggaggtcgg aaatgtcgcc cttgttccca 8280
tcatggcctt tggtgagaac gacctctatg accaggttag caacgacaag tcgtccaagc 8340
tgtaccgatt ccagcagttt gtcaagaact tccttggatt cacccttcct ttgatgcatg 8400
cccgaggcgt cttcaactac gatgtcggtc ttgtccccta caggcgaccc gtcaacattg 8460
tggttggttc ccccattgac ttgccttatc tcccacaccc caccgacgaa gaagtgtccg 8520
aataccacga ccgatacatc gccgagctgc agcgaatcta caacgagcac aaggatgaat 8580
atttcatcga ttggaccgag gagggcaaag gagccccaga gttccgaatg attgagtaag 8640
c 8641
<210>101
<211>11647
<212>DNA
<213>人工序列
<220>
<223>质粒pKS392
<220>
<221>misc_feature
<222>(2349)..(2349)
<223>n为a,c,g,或t
<400>101
ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacaagctt ggatcctcga agagaagggt taataacaca 1200
cttttttaac atttttaaca caaattttag ttatttaaaa atttattaaa aaatttaaaa 1260
taagaagagg aactctttaa ataaatctaa cttacaaaat ttatgatttt taataagttt 1320
tcaccaataa aaaatgtcat aaaaatatgt taaaaagtat attatcaata ttctctttat 1380
gataaataaa aagaaaaaaa aaataaaagt taagtgaaaa tgagattgaa gtgactttag 1440
gtgtgtataa atatatcaac cccgccaaca atttatttaa tccaaatata ttgaagtata 1500
ttattccata gcctttattt atttatatat ttattatata aaagctttat ttgttctagg 1560
ttgttcatga aatatttttt tggttttatc tccgttgtaa gaaaatcatg tgctttgtgt 1620
cgccactcac tattgcagct ttttcatgca ttggtcagat tgacggttga ttgtattttt 1680
gttttttatg gttttgtgtt atgacttaag tcttcatctc tttatctctt catcaggttt 1740
gatggttacc taatatggtc catgggtaca tgcatggtta aattaggtgg ccaactttgt 1800
tgtgaacgat agaatttttt ttatattaag taaactattt ttatattatg aaataataat 1860
aaaaaaaata ttttatcatt attaacaaaa tcatattagt taatttgtta actctataat 1920
aaaagaaata ctgtaacatt cacattacat ggtaacatct ttccaccctt tcatttgttt 1980
tttgtttgat gacttttttt cttgtttaaa tttatttccc ttcttttaaa tttggaatac 2040
attatcatca tatataaact aaaatactaa aaacaggatt acacaaatga taaataataa 2100
cacaaatatt tataaatcta gctgcaatat atttaaacta gctatatcga tattgtaaaa 2160
taaaactagc tgcattgata ctgataaaaa aatatcatgt gctttctgga ctgatgatgc 2220
agtatacttt tgacattgcc tttattttat ttttcagaaa agctttctta gttctgggtt 2280
cttcattatt tgtttcccat ctccattgtg aattgaatca tttgcttcgt gtcacaaata 2340
caatttagnt aggtacatgc attggtcaga ttcacggttt attatgtcat gacttaagtt 2400
catggtagta cattacctgc cacgcatgca ttatattggt tagatttgat aggcaaattt 2460
ggttgtcaac aatataaata taaataatgt ttttatatta cgaaataaca gtgatcaaaa 2520
caaacagttt tatctttatt aacaagattt tgtttttgtt tgatgacgtt ttttaatgtt 2580
tacgctttcc cccttctttt gaatttagaa cactttatca tcataaaatc aaatactaaa 2640
aaaattacat atttcataaa taataacaca aatattttta aaaaatctga aataataatg 2700
aacaatatta catattatca cgaaaattca ttaataaaaa tattatataa ataaaatgta 2760
atagtagtta tatgtaggaa aaaagtactg cacgcataat atatacaaaa agattaaaat 2820
gaactattat aaataataac actaaattaa tggtgaatca tatcaaaata atgaaaaagt 2880
aaataaaatt tgtaattaac ttctatatgt attacacaca caaataataa ataatagtaa 2940
aaaaaattat gataaatatt taccatctca taagatattt aaaataatga taaaaatata 3000
gattattttt tatgcaacta gctagccaaa aagagaacac gggtatatat aaaaagagta 3060
cctttaaatt ctactgtact tcctttattc ctgacgtttt tatatcaagt ggacatacgt 3120
gaagatttta attatcagtc taaatatttc attagcactt aatacttttc tgttttattc 3180
ctatcctata agtagtcccg attctcccaa cattgcttat tcacacaact aactaagaaa 3240
gtcttccata gccccccaag cggcccatgg cctcctccga ggacgtcatc aaggagttca 3300
tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca cgagttcgag atcgagggcg 3360
agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg 3420
gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt 3480
acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca 3540
agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct 3600
ccctgcagga cggctccttc atctacaagg tgaagttcat cggcgtgaac ttcccctccg 3660
acggccccgt aatgcagaag aagactatgg gctgggaggc ctccaccgag cgcctgtacc 3720
cccgcgacgg cgtgctgaag ggcgagatcc acaaggccct gaagctgaag gacggcggcc 3780
actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag ctgcccggct 3840
actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac accatcgtgg 3900
agcagtacga gcgcgccgag ggccgccacc acctgttcct gtagcggccg gccgcgacac 3960
aagtgtgaga gtactaaata aatgctttgg ttgtacgaaa tcattacact aaataaaata 4020
atcaaagctt atatatgcct tccgctaagg ccgaatgcaa agaaattggt tctttctcgt 4080
tatcttttgc cacttttact agtacgtatt aattactact taatcatctt tgtttacggc 4140
tcattatatc cgtcgacggc gcgccgctct agaactagtg gatccgtcga cggcgcgccc 4200
gatcatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc 4260
ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg 4320
gctttgttag cagccggatc gatccaagct gtacctcact attcctttgc cctcggacga 4380
gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg 4440
gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt 4500
gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga 4560
tagagttggt caagaccaat gcggagcata tacgcccgga gccgcggcga tcctgcaagc 4620
tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc 4680
cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca 4740
atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc 4800
acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc 4860
gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca 4920
gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat 4980
gggccgaacc cgctcgtctg gctaagatcg gccgcagcga tcgcatccat agcctccgcg 5040
accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt 5100
gcacggcggg agatgcaata ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc 5160
ggaatcggga gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca 5220
tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca 5280
cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc 5340
agaaacttct cgacagacgt cgcggtgagt tcaggctttt ccatgggtat atctccttct 5400
taaagttaaa caaaattatt tctagaggga aaccgttgtg gtctccctat agtgagtcgt 5460
attaatttcg cgggatcgag atcgatccaa ttccaatccc acaaaaatct gagcttaaca 5520
gcacagttgc tcctctcaga gcagaatcgg gtattcaaca ccctcatatc aactactacg 5580
ttgtgtataa cggtccacat gccggtatat acgatgactg gggttgtaca aaggcggcaa 5640
caaacggcgt tcccggagtt gcacacaaga aatttgccac tattacagag gcaagagcag 5700
cagctgacgc gtacacaaca agtcagcaaa cagacaggtt gaacttcatc cccaaaggag 5760
aagctcaact caagcccaag agctttgcta aggccctaac aagcccacca aagcaaaaag 5820
cccactggct cacgctagga accaaaaggc ccagcagtga tccagcccca aaagagatct 5880
cctttgcccc ggagattaca atggacgatt tcctctatct ttacgatcta ggaaggaagt 5940
tcgaaggtga aggtgacgac actatgttca ccactgataa tgagaaggtt agcctcttca 6000
atttcagaaa gaatgctgac ccacagatgg ttagagaggc ctacgcagca ggtctcatca 6060
agacgatcta cccgagtaac aatctccagg agatcaaata ccttcccaag aaggttaaag 6120
atgcagtcaa aagattcagg actaattgca tcaagaacac agagaaagac atatttctca 6180
agatcagaag tactattcca gtatggacga ttcaaggctt gcttcataaa ccaaggcaag 6240
taatagagat tggagtctct aaaaaggtag ttcctactga atctaaggcc atgcatggag 6300
tctaagattc aaatcgagga tctaacagaa ctcgccgtga agactggcga acagttcata 6360
cagagtcttt tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac 6420
actctggtct actccaaaaa tgtcaaagat acagtctcag aagaccaaag ggctattgag 6480
acttttcaac aaaggataat ttcgggaaac ctcctcggat tccattgccc agctatctgt 6540
cacttcatcg aaaggacagt agaaaaggaa ggtggctcct acaaatgcca tcattgcgat 6600
aaaggaaagg ctatcattca agatgcctct gccgacagtg gtcccaaaga tggaccccca 6660
cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat 6720
tgatgtgaca tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac 6780
ccttcctcta tataaggaag ttcatttcat ttggagagga cacgctcgag ctcatttctc 6840
tattacttca gccataacaa aagaactctt ttctcttctt attaaaccat gaaaaagcct 6900
gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac 6960
ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt 7020
ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat 7080
cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc 7140
gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct 7200
gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg 7260
gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac 7320
actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact 7380
gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg 7440
gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc 7500
ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat 7560
tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag 7620
cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg 7680
tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat 7740
gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc 7800
gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta 7860
ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga atagtgaggt 7920
acctaaagaa ggagtgcgtc gaagcagatc gttcaaacat ttggcaataa agtttcttaa 7980
gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta 8040
agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta 8100
gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg 8160
ataaattatc gcgcgcggtg tcatctatgt tactagatcg atgtcgaatc tgatcaacct 8220
gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 8280
ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 8340
ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 8400
agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 8460
taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 8520
cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 8580
tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 8640
gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 8700
gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 8760
tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 8820
gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 8880
cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 8940
aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 9000
tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 9060
ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgac 9120
attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 9180
cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 9240
tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 9300
gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc 9360
gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg 9420
acactataga acggcgcgcc aagcttttga tccatgccct tcatttgccg cttattaatt 9480
aatttggtaa cagtccgtac taatcagtta cttatccttc ccccatcata attaatcttg 9540
gtagtctcga atgccacaac actgactagt ctcttggatc ataagaaaaa gccaaggaac 9600
aaaagaagac aaaacacaat gagagtatcc tttgcatagc aatgtctaag ttcataaaat 9660
tcaaacaaaa acgcaatcac acacagtgga catcacttat ccactagctg atcaggatcg 9720
ccgcgtcaag aaaaaaaaac tggaccccaa aagccatgca caacaacacg tactcacaaa 9780
ggtgtcaatc gagcagccca aaacattcac caactcaacc catcatgagc cctcacattt 9840
gttgtttcta acccaacctc aaactcgtat tctcttccgc cacctcattt ttgtttattt 9900
caacacccgt caaactgcat gccaccccgt ggccaaatgt ccatgcatgt taacaagacc 9960
tatgactata aatagctgca atctcggccc aggttttcat catcaagaac cagttcaata 10020
tcctagtaca ccgtattaaa gaatttaaga tatactgcgg ccgcatggag gtgcgcagac 10080
gcaagattga cgttctcaag gcccagaaga atggttacga gtccggccca ccatctagac 10140
agtcctctca gccatcctct agagcctcct ccagaacacg caacaagcac tcctcctcca 10200
cactttccct ttccggcctc accatgaagg tgcagaagaa gccagctggt ccaccagcca 10260
actccaagac cccattcctc cacatcaagc cagtgcacac ttgctgctcc acctcaatgc 10320
tctccagaga ctacgacggc tccaacccat ccttcaaggg cttcaagaac atcggcatga 10380
tcatcctcat cgtgggcaat cttcgcctcg ccttcgagaa ctacctcaag tacggcatct 10440
ccaacccctt cttcgacccc aagatcacac cctccgagtg gcagctttct ggacttctca 10500
tcgttgtggc ttacgctcac atcctcatgg cttacgctat cgagtccgct gccaagctcc 10560
tcttcctctc ctccaagcac cactacatgg ccgttggtct cctccacacc atgaacaccc 10620
tctcctccat ttccctcctc tcctacgtgg tgtactacta cctccccaat ccagtggctg 10680
gcactatcgt tgagttcgtg gccgtgatcc tttctctcaa gctcgcttcc tacgccctta 10740
ccaactccga tctcagaaag gctgccattc acgcccagaa gctcgacaag acacaggacg 10800
acaacgagaa ggagtccacc tcctcttcct cttcctctga cgacgccgag acacttgccg 10860
acatcgacgt tatcccagct tactacgctc agcttcccta cccacagaac gttaccctct 10920
ccaacctcct ctacttctgg ttcgctccca cccttgtgta ccagcccgtt tacccaaaga 10980
cagagagaat ccgccccaag catgtgatcc gcaacctctt cgagcttgtg tctctctgca 11040
tgctcatcca gttcctcatc ttccagtacg cctaccccat catgcagtct tgccttgccc 11100
tcttcttcca gccaaagctc gactacgcca acatttccga gcgcctcatg aagctcgcct 11160
ctgtgtccat gatggtgtgg ctcatcggtt tctacgcctt cttccagaac ggtctcaacc 11220
tcatcgccga gctcacctgc ttcggtaacc gcacattcta ccagcagtgg tggaactcca 11280
gatctattgg ccagtactgg acactctgga acaagcccgt gaaccagtac ttccgccatc 11340
acgtttacgt tcctcttctc gctaggggca tgtccagatt caacgcctct gttgtggtgt 11400
tcttcttctc cgctgtgatt cacgagctcc ttgtgggcat tcccacacac aacatcatcg 11460
gcgctgcctt cttcggaatg atgtcccagg tgccacttat catggccacc gagaaccttc 11520
agcacatcaa ctcctccctt ggcccattcc ttggtaactg cgctttctgg ttcaccttct 11580
tccttggtca gcctacctgc gcctttctct actaccttgc ctacaactac aagcagaacc 11640
agtaggc 11647
<210>102
<211>11611
<212>DNA
<213>人工序列
<220>
<223>质粒pKS393
<220>
<221>misc_feature
<222>(2349)..(2349)
<223>n为a,c,g,或t
<400>102
ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacaagctt ggatcctcga agagaagggt taataacaca 1200
cttttttaac atttttaaca caaattttag ttatttaaaa atttattaaa aaatttaaaa 1260
taagaagagg aactctttaa ataaatctaa cttacaaaat ttatgatttt taataagttt 1320
tcaccaataa aaaatgtcat aaaaatatgt taaaaagtat attatcaata ttctctttat 1380
gataaataaa aagaaaaaaa aaataaaagt taagtgaaaa tgagattgaa gtgactttag 1440
gtgtgtataa atatatcaac cccgccaaca atttatttaa tccaaatata ttgaagtata 1500
ttattccata gcctttattt atttatatat ttattatata aaagctttat ttgttctagg 1560
ttgttcatga aatatttttt tggttttatc tccgttgtaa gaaaatcatg tgctttgtgt 1620
cgccactcac tattgcagct ttttcatgca ttggtcagat tgacggttga ttgtattttt 1680
gttttttatg gttttgtgtt atgacttaag tcttcatctc tttatctctt catcaggttt 1740
gatggttacc taatatggtc catgggtaca tgcatggtta aattaggtgg ccaactttgt 1800
tgtgaacgat agaatttttt ttatattaag taaactattt ttatattatg aaataataat 1860
aaaaaaaata ttttatcatt attaacaaaa tcatattagt taatttgtta actctataat 1920
aaaagaaata ctgtaacatt cacattacat ggtaacatct ttccaccctt tcatttgttt 1980
tttgtttgat gacttttttt cttgtttaaa tttatttccc ttcttttaaa tttggaatac 2040
attatcatca tatataaact aaaatactaa aaacaggatt acacaaatga taaataataa 2100
cacaaatatt tataaatcta gctgcaatat atttaaacta gctatatcga tattgtaaaa 2160
taaaactagc tgcattgata ctgataaaaa aatatcatgt gctttctgga ctgatgatgc 2220
agtatacttt tgacattgcc tttattttat ttttcagaaa agctttctta gttctgggtt 2280
cttcattatt tgtttcccat ctccattgtg aattgaatca tttgcttcgt gtcacaaata 2340
caatttagnt aggtacatgc attggtcaga ttcacggttt attatgtcat gacttaagtt 2400
catggtagta cattacctgc cacgcatgca ttatattggt tagatttgat aggcaaattt 2460
ggttgtcaac aatataaata taaataatgt ttttatatta cgaaataaca gtgatcaaaa 2520
caaacagttt tatctttatt aacaagattt tgtttttgtt tgatgacgtt ttttaatgtt 2580
tacgctttcc cccttctttt gaatttagaa cactttatca tcataaaatc aaatactaaa 2640
aaaattacat atttcataaa taataacaca aatattttta aaaaatctga aataataatg 2700
aacaatatta catattatca cgaaaattca ttaataaaaa tattatataa ataaaatgta 2760
atagtagtta tatgtaggaa aaaagtactg cacgcataat atatacaaaa agattaaaat 2820
gaactattat aaataataac actaaattaa tggtgaatca tatcaaaata atgaaaaagt 2880
aaataaaatt tgtaattaac ttctatatgt attacacaca caaataataa ataatagtaa 2940
aaaaaattat gataaatatt taccatctca taagatattt aaaataatga taaaaatata 3000
gattattttt tatgcaacta gctagccaaa aagagaacac gggtatatat aaaaagagta 3060
cctttaaatt ctactgtact tcctttattc ctgacgtttt tatatcaagt ggacatacgt 3120
gaagatttta attatcagtc taaatatttc attagcactt aatacttttc tgttttattc 3180
ctatcctata agtagtcccg attctcccaa cattgcttat tcacacaact aactaagaaa 3240
gtcttccata gccccccaag cggcccatgg cctcctccga ggacgtcatc aaggagttca 3300
tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca cgagttcgag atcgagggcg 3360
agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg 3420
gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt 3480
acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca 3540
agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct 3600
ccctgcagga cggctccttc atctacaagg tgaagttcat cggcgtgaac ttcccctccg 3660
acggccccgt aatgcagaag aagactatgg gctgggaggc ctccaccgag cgcctgtacc 3720
cccgcgacgg cgtgctgaag ggcgagatcc acaaggccct gaagctgaag gacggcggcc 3780
actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag ctgcccggct 3840
actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac accatcgtgg 3900
agcagtacga gcgcgccgag ggccgccacc acctgttcct gtagcggccg gccgcgacac 3960
aagtgtgaga gtactaaata aatgctttgg ttgtacgaaa tcattacact aaataaaata 4020
atcaaagctt atatatgcct tccgctaagg ccgaatgcaa agaaattggt tctttctcgt 4080
tatcttttgc cacttttact agtacgtatt aattactact taatcatctt tgtttacggc 4140
tcattatatc cgtcgacggc gcgccgctct agaactagtg gatccgtcga cggcgcgccc 4200
gatcatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc 4260
ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg 4320
gctttgttag cagccggatc gatccaagct gtacctcact attcctttgc cctcggacga 4380
gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg 4440
gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt 4500
gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga 4560
tagagttggt caagaccaat gcggagcata tacgcccgga gccgcggcga tcctgcaagc 4620
tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc 4680
cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca 4740
atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc 4800
acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc 4860
gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca 4920
gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat 4980
gggccgaacc cgctcgtctg gctaagatcg gccgcagcga tcgcatccat agcctccgcg 5040
accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt 5100
gcacggcggg agatgcaata ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc 5160
ggaatcggga gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca 5220
tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca 5280
cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc 5340
agaaacttct cgacagacgt cgcggtgagt tcaggctttt ccatgggtat atctccttct 5400
taaagttaaa caaaattatt tctagaggga aaccgttgtg gtctccctat agtgagtcgt 5460
attaatttcg cgggatcgag atcgatccaa ttccaatccc acaaaaatct gagcttaaca 5520
gcacagttgc tcctctcaga gcagaatcgg gtattcaaca ccctcatatc aactactacg 5580
ttgtgtataa cggtccacat gccggtatat acgatgactg gggttgtaca aaggcggcaa 5640
caaacggcgt tcccggagtt gcacacaaga aatttgccac tattacagag gcaagagcag 5700
cagctgacgc gtacacaaca agtcagcaaa cagacaggtt gaacttcatc cccaaaggag 5760
aagctcaact caagcccaag agctttgcta aggccctaac aagcccacca aagcaaaaag 5820
cccactggct cacgctagga accaaaaggc ccagcagtga tccagcccca aaagagatct 5880
cctttgcccc ggagattaca atggacgatt tcctctatct ttacgatcta ggaaggaagt 5940
tcgaaggtga aggtgacgac actatgttca ccactgataa tgagaaggtt agcctcttca 6000
atttcagaaa gaatgctgac ccacagatgg ttagagaggc ctacgcagca ggtctcatca 6060
agacgatcta cccgagtaac aatctccagg agatcaaata ccttcccaag aaggttaaag 6120
atgcagtcaa aagattcagg actaattgca tcaagaacac agagaaagac atatttctca 6180
agatcagaag tactattcca gtatggacga ttcaaggctt gcttcataaa ccaaggcaag 6240
taatagagat tggagtctct aaaaaggtag ttcctactga atctaaggcc atgcatggag 6300
tctaagattc aaatcgagga tctaacagaa ctcgccgtga agactggcga acagttcata 6360
cagagtcttt tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac 6420
actctggtct actccaaaaa tgtcaaagat acagtctcag aagaccaaag ggctattgag 6480
acttttcaac aaaggataat ttcgggaaac ctcctcggat tccattgccc agctatctgt 6540
cacttcatcg aaaggacagt agaaaaggaa ggtggctcct acaaatgcca tcattgcgat 6600
aaaggaaagg ctatcattca agatgcctct gccgacagtg gtcccaaaga tggaccccca 6660
cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat 6720
tgatgtgaca tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac 6780
ccttcctcta tataaggaag ttcatttcat ttggagagga cacgctcgag ctcatttctc 6840
tattacttca gccataacaa aagaactctt ttctcttctt attaaaccat gaaaaagcct 6900
gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac 6960
ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt 7020
ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat 7080
cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc 7140
gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct 7200
gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg 7260
gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac 7320
actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact 7380
gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg 7440
gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc 7500
ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat 7560
tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag 7620
cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg 7680
tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat 7740
gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc 7800
gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta 7860
ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga atagtgaggt 7920
acctaaagaa ggagtgcgtc gaagcagatc gttcaaacat ttggcaataa agtttcttaa 7980
gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta 8040
agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta 8100
gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg 8160
ataaattatc gcgcgcggtg tcatctatgt tactagatcg atgtcgaatc tgatcaacct 8220
gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 8280
ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 8340
ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 8400
agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 8460
taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 8520
cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 8580
tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 8640
gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 8700
gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 8760
tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 8820
gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 8880
cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 8940
aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 9000
tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 9060
ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgac 9120
attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 9180
cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 9240
tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 9300
gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc 9360
gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg 9420
acactataga acggcgcgcc aagcttttga tccatgccct tcatttgccg cttattaatt 9480
aatttggtaa cagtccgtac taatcagtta cttatccttc ccccatcata attaatcttg 9540
gtagtctcga atgccacaac actgactagt ctcttggatc ataagaaaaa gccaaggaac 9600
aaaagaagac aaaacacaat gagagtatcc tttgcatagc aatgtctaag ttcataaaat 9660
tcaaacaaaa acgcaatcac acacagtgga catcacttat ccactagctg atcaggatcg 9720
ccgcgtcaag aaaaaaaaac tggaccccaa aagccatgca caacaacacg tactcacaaa 9780
ggtgtcaatc gagcagccca aaacattcac caactcaacc catcatgagc cctcacattt 9840
gttgtttcta acccaacctc aaactcgtat tctcttccgc cacctcattt ttgtttattt 9900
caacacccgt caaactgcat gccaccccgt ggccaaatgt ccatgcatgt taacaagacc 9960
tatgactata aatagctgca atctcggccc aggttttcat catcaagaac cagttcaata 10020
tcctagtaca ccgtattaaa gaatttaaga tatactgcgg ccgcatgacc atcgactccc 10080
agtactacaa gtcccgcgac aagaacgaca ccgctcccaa gattgctggt atccgctacg 10140
ctcctctgtc tactccactc ttgaaccgct gcgagacttt ctctttggtg tggcacatct 10200
tctccatccc taccttcctc accatcttca tgctctgctg tgccattccc ttgctctggc 10260
ccttcgtgat cgcttacgtt gtgtacgccg tgaaggatga ctccccttcc aacggcggtg 10320
ttgtgaagcg ctattcccct atctcccgca acttcttcat ctggaagctc tttggccgct 10380
acttccccat caccctccac aagaccgttg atttggagcc cacccacact tactatccct 10440
tggacgtcca agagtaccac ctgattgctg agcgctattg gcctcagaac aagtacctgc 10500
gcgccatcat taccaccatc gagtacttcc tgcccgcttt catgaagcgt tccctctcca 10560
tcaacgagca ggagcagcct gctgagagag accctctgtt gtctcccgtt tccccctctt 10620
ctcctggttc tcagcccgac aagtggatca accatgattc ccgctattcc agaggtgagt 10680
cctctggctc caatggtcat gcttctggct ctgaactgaa tggcaatggc aacaacggca 10740
ctaccaatcg ccgccctctt tcttctgctt ccgctggctc tactgcttct gactctacct 10800
tgctcaacgg ctctctcaac tcctacgcca accagatcat tggcgagaac gaccctcagt 10860
tgtctcccac taagctcaag cctactggca ggaagtacat cttcgggtac catcctcacg 10920
gtatcattgg gatgggcgcc tttggtggca ttgctactga aggtgctggc tggtccaagc 10980
tctttcctgg tatccccgtt tccctcatga ccctcaccaa caactttcgc gttcctctct 11040
atcgcgagta cctcatgtcc ctgggcgttg cttccgtttc taagaagtcc tgtaaggccc 11100
tcctcaagcg taatcagtcc atctgcatcg tggtgggtgg tgcccaggaa tccttgttgg 11160
ctaggcctgg tgttatggat ctcgtgttgc tgaagcgcaa gggttttgtg cgcttgggta 11220
tggaggtggg caatgttgcc ctcgttccaa tcatggcttt cggcgagaac gatctctacg 11280
accaggtgtc caacgacaag tcctccaagc tgtaccgctt ccagcagttc gtgaagaact 11340
tcctcggctt caccctccct ctcatgcatg ccagaggtgt gttcaactac gatgttgggc 11400
tcgttcccta ccgccgccct gtgaacattg tggttggttc tcctatcgac ttgccttacc 11460
tccctcatcc caccgatgag gaagtttccg agtaccacga ccgttacatc gctgagctcc 11520
agcgcatcta caacgagcac aaggacgagt acttcatcga ctggactgag gagggtaagg 11580
gcgctcctga gtttcgtatg atcgagtagg c 11611
<210>103
<211>11649
<212>DNA
<213>人工序列
<220>
<223>质粒pKS391
<220>
<221>misc_feature
<222>(2349)..(2349)
<223>n为a,c,g,或t
<400>103
ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacaagctt ggatcctcga agagaagggt taataacaca 1200
cttttttaac atttttaaca caaattttag ttatttaaaa atttattaaa aaatttaaaa 1260
taagaagagg aactctttaa ataaatctaa cttacaaaat ttatgatttt taataagttt 1320
tcaccaataa aaaatgtcat aaaaatatgt taaaaagtat attatcaata ttctctttat 1380
gataaataaa aagaaaaaaa aaataaaagt taagtgaaaa tgagattgaa gtgactttag 1440
gtgtgtataa atatatcaac cccgccaaca atttatttaa tccaaatata ttgaagtata 1500
ttattccata gcctttattt atttatatat ttattatata aaagctttat ttgttctagg 1560
ttgttcatga aatatttttt tggttttatc tccgttgtaa gaaaatcatg tgctttgtgt 1620
cgccactcac tattgcagct ttttcatgca ttggtcagat tgacggttga ttgtattttt 1680
gttttttatg gttttgtgtt atgacttaag tcttcatctc tttatctctt catcaggttt 1740
gatggttacc taatatggtc catgggtaca tgcatggtta aattaggtgg ccaactttgt 1800
tgtgaacgat agaatttttt ttatattaag taaactattt ttatattatg aaataataat 1860
aaaaaaaata ttttatcatt attaacaaaa tcatattagt taatttgtta actctataat 1920
aaaagaaata ctgtaacatt cacattacat ggtaacatct ttccaccctt tcatttgttt 1980
tttgtttgat gacttttttt cttgtttaaa tttatttccc ttcttttaaa tttggaatac 2040
attatcatca tatataaact aaaatactaa aaacaggatt acacaaatga taaataataa 2100
cacaaatatt tataaatcta gctgcaatat atttaaacta gctatatcga tattgtaaaa 2160
taaaactagc tgcattgata ctgataaaaa aatatcatgt gctttctgga ctgatgatgc 2220
agtatacttt tgacattgcc tttattttat ttttcagaaa agctttctta gttctgggtt 2280
cttcattatt tgtttcccat ctccattgtg aattgaatca tttgcttcgt gtcacaaata 2340
caatttagnt aggtacatgc attggtcaga ttcacggttt attatgtcat gacttaagtt 2400
catggtagta cattacctgc cacgcatgca ttatattggt tagatttgat aggcaaattt 2460
ggttgtcaac aatataaata taaataatgt ttttatatta cgaaataaca gtgatcaaaa 2520
caaacagttt tatctttatt aacaagattt tgtttttgtt tgatgacgtt ttttaatgtt 2580
tacgctttcc cccttctttt gaatttagaa cactttatca tcataaaatc aaatactaaa 2640
aaaattacat atttcataaa taataacaca aatattttta aaaaatctga aataataatg 2700
aacaatatta catattatca cgaaaattca ttaataaaaa tattatataa ataaaatgta 2760
atagtagtta tatgtaggaa aaaagtactg cacgcataat atatacaaaa agattaaaat 2820
gaactattat aaataataac actaaattaa tggtgaatca tatcaaaata atgaaaaagt 2880
aaataaaatt tgtaattaac ttctatatgt attacacaca caaataataa ataatagtaa 2940
aaaaaattat gataaatatt taccatctca taagatattt aaaataatga taaaaatata 3000
gattattttt tatgcaacta gctagccaaa aagagaacac gggtatatat aaaaagagta 3060
cctttaaatt ctactgtact tcctttattc ctgacgtttt tatatcaagt ggacatacgt 3120
gaagatttta attatcagtc taaatatttc attagcactt aatacttttc tgttttattc 3180
ctatcctata agtagtcccg attctcccaa cattgcttat tcacacaact aactaagaaa 3240
gtcttccata gccccccaag cggcccatgg cctcctccga ggacgtcatc aaggagttca 3300
tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca cgagttcgag atcgagggcg 3360
agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg 3420
gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt 3480
acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca 3540
agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct 3600
ccctgcagga cggctccttc atctacaagg tgaagttcat cggcgtgaac ttcccctccg 3660
acggccccgt aatgcagaag aagactatgg gctgggaggc ctccaccgag cgcctgtacc 3720
cccgcgacgg cgtgctgaag ggcgagatcc acaaggccct gaagctgaag gacggcggcc 3780
actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag ctgcccggct 3840
actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac accatcgtgg 3900
agcagtacga gcgcgccgag ggccgccacc acctgttcct gtagcggccg gccgcgacac 3960
aagtgtgaga gtactaaata aatgctttgg ttgtacgaaa tcattacact aaataaaata 4020
atcaaagctt atatatgcct tccgctaagg ccgaatgcaa agaaattggt tctttctcgt 4080
tatcttttgc cacttttact agtacgtatt aattactact taatcatctt tgtttacggc 4140
tcattatatc cgtcgacggc gcgccgctct agaactagtg gatccgtcga cggcgcgccc 4200
gatcatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc 4260
ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg 4320
gctttgttag cagccggatc gatccaagct gtacctcact attcctttgc cctcggacga 4380
gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg 4440
gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt 4500
gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga 4560
tagagttggt caagaccaat gcggagcata tacgcccgga gccgcggcga tcctgcaagc 4620
tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc 4680
cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca 4740
atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc 4800
acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc 4860
gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca 4920
gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat 4980
gggccgaacc cgctcgtctg gctaagatcg gccgcagcga tcgcatccat agcctccgcg 5040
accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt 5100
gcacggcggg agatgcaata ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc 5160
ggaatcggga gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca 5220
tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca 5280
cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc 5340
agaaacttct cgacagacgt cgcggtgagt tcaggctttt ccatgggtat atctccttct 5400
taaagttaaa caaaattatt tctagaggga aaccgttgtg gtctccctat agtgagtcgt 5460
attaatttcg cgggatcgag atcgatccaa ttccaatccc acaaaaatct gagcttaaca 5520
gcacagttgc tcctctcaga gcagaatcgg gtattcaaca ccctcatatc aactactacg 5580
ttgtgtataa cggtccacat gccggtatat acgatgactg gggttgtaca aaggcggcaa 5640
caaacggcgt tcccggagtt gcacacaaga aatttgccac tattacagag gcaagagcag 5700
cagctgacgc gtacacaaca agtcagcaaa cagacaggtt gaacttcatc cccaaaggag 5760
aagctcaact caagcccaag agctttgcta aggccctaac aagcccacca aagcaaaaag 5820
cccactggct cacgctagga accaaaaggc ccagcagtga tccagcccca aaagagatct 5880
cctttgcccc ggagattaca atggacgatt tcctctatct ttacgatcta ggaaggaagt 5940
tcgaaggtga aggtgacgac actatgttca ccactgataa tgagaaggtt agcctcttca 6000
atttcagaaa gaatgctgac ccacagatgg ttagagaggc ctacgcagca ggtctcatca 6060
agacgatcta cccgagtaac aatctccagg agatcaaata ccttcccaag aaggttaaag 6120
atgcagtcaa aagattcagg actaattgca tcaagaacac agagaaagac atatttctca 6180
agatcagaag tactattcca gtatggacga ttcaaggctt gcttcataaa ccaaggcaag 6240
taatagagat tggagtctct aaaaaggtag ttcctactga atctaaggcc atgcatggag 6300
tctaagattc aaatcgagga tctaacagaa ctcgccgtga agactggcga acagttcata 6360
cagagtcttt tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac 6420
actctggtct actccaaaaa tgtcaaagat acagtctcag aagaccaaag ggctattgag 6480
acttttcaac aaaggataat ttcgggaaac ctcctcggat tccattgccc agctatctgt 6540
cacttcatcg aaaggacagt agaaaaggaa ggtggctcct acaaatgcca tcattgcgat 6600
aaaggaaagg ctatcattca agatgcctct gccgacagtg gtcccaaaga tggaccccca 6660
cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat 6720
tgatgtgaca tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac 6780
ccttcctcta tataaggaag ttcatttcat ttggagagga cacgctcgag ctcatttctc 6840
tattacttca gccataacaa aagaactctt ttctcttctt attaaaccat gaaaaagcct 6900
gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac 6960
ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt 7020
ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat 7080
cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc 7140
gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct 7200
gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg 7260
gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac 7320
actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact 7380
gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg 7440
gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc 7500
ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat 7560
tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag 7620
cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg 7680
tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat 7740
gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc 7800
gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta 7860
ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga atagtgaggt 7920
acctaaagaa ggagtgcgtc gaagcagatc gttcaaacat ttggcaataa agtttcttaa 7980
gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta 8040
agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta 8100
gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg 8160
ataaattatc gcgcgcggtg tcatctatgt tactagatcg atgtcgaatc tgatcaacct 8220
gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 8280
ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 8340
ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 8400
agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 8460
taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 8520
cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 8580
tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 8640
gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 8700
gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 8760
tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 8820
gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 8880
cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 8940
aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 9000
tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 9060
ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgac 9120
attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 9180
cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 9240
tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 9300
gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc 9360
gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg 9420
acactataga acggcgcgcc aagcttttga tccatgccct tcatttgccg cttattaatt 9480
aatttggtaa cagtccgtac taatcagtta cttatccttc ccccatcata attaatcttg 9540
gtagtctcga atgccacaac actgactagt ctcttggatc ataagaaaaa gccaaggaac 9600
aaaagaagac aaaacacaat gagagtatcc tttgcatagc aatgtctaag ttcataaaat 9660
tcaaacaaaa acgcaatcac acacagtgga catcacttat ccactagctg atcaggaccg 9720
ccgcgtcaag aaaaaaaaac tggaccccaa aagccatgca caacaacacg tactcacaaa 9780
ggtgtcaatc gagcagccca aaacattcac caactcaacc catcatgagc cctcacattt 9840
gttgtttcta acccaacctc aaactcgtat tctcttccgc cacctcattt ttgtttattt 9900
caacacccgt caaactgcat gccaccccgt ggccaaatgt ccatgcatgt taacaagacc 9960
tatgactata aatagctgca atctcggccc aggttttcat catcaagaac cagttcaata 10020
tcctagtaca ccgtattaaa gaatttaaga tatactgcgg cccatggagg tccgacgacg 10080
aaagatagac gtgctcaagg cccagaaaaa cggctacgaa tcgggcccac catctcgaca 10140
atcgtcgcag ccctcctcaa gagcatcgtc cagaacccgc aacaaacact cctcgtccac 10200
cctgtcgctc agcggactga ccatgaaagt ccagaagaaa cctgcgggac ccccggcgaa 10260
ctccaaaacg ccattcctac acatcaagcc cgtgcacacg tgctgctcca catcaatgct 10320
ttcgcgcgat tatgacggct ccaaccccag cttcaagggc ttcaaaaaca tcggcatgat 10380
cattctcatt gtgggaaatc tacggctcgc attcgaaaac tacctcaaat acggcatttc 10440
caacccgttc ttcgacccca aaattactcc ttccgagtgg cagctctcag gcttgctcat 10500
agtcgtggcc tacgcacata tcctcatggc ctacgctatt gagagcgctg ccaagctgct 10560
gttcctctct agcaaacacc actacatggc cgtggggctt ctgcatacca tgaacacttt 10620
gtcgtccatc tcgttgctgt cctacgtcgt ctactactac ctgcccaacc ccgtggcagg 10680
cacaatagtc gagtttgtgg ccgttattct gtctctcaaa ctcgcctcat acgccctcac 10740
taactcggat ctccgaaaag ccgcaattca tgcccagaag ctcgacaaga cgcaagacga 10800
taacgaaaag gaatccacct cgtcttcctc ttcttcagat gacgcagaga ctttggcaga 10860
cattgacgtc attcctgcat actacgcaca gctgccctac ccccagaatg tgacgctgtc 10920
gaacctgctg tacttctggt ttgctcccac actggtctac cagcccgtgt accccaagac 10980
ggagcgtatt cgacccaagc acgtgatccg aaacctgttt gagctcgtct ctctgtgcat 11040
gcttattcag tttctcatct tccagtacgc ctaccccatc atgcagtcgt gtctggctct 11100
gttcttccag cccaagctcg attatgccaa catctccgag cgcctcatga agttggcctc 11160
cgtgtctatg atggtctggc tcattggatt ctacgctttc ttccagaacg gtctcaatct 11220
tattgccgag ctcacctgtt ttggaaacag aaccttctac cagcagtggt ggaattcccg 11280
ctccattggc cagtactgga ctctatggaa caagccagtc aaccagtact ttagacacca 11340
cgtctacgtg cctcttctcg ctcggggcat gtcgcggttc aatgcgtcgg tggtggtttt 11400
ctttttctcc gccgtcatcc atgaactgct tgtcggcatc cccactcaca acatcatcgg 11460
agccgccttc ttcggcatga tgtcgcaggt gcctctgatc atggctactg agaaccttca 11520
gcatattaac tcctctctgg gccccttcct tggcaactgt gcattctggt tcaccttttt 11580
cctgggacaa cccacttgtg cattccttta ctatttggcc tacaactaca agcagaacca 11640
gtagcggcc 11649
<210>104
<211>5303
<212>DNA
<213>人工序列
<220>
<223>质粒pKR278
<400>104
agcttggatc tcctgcagga tctggccggc cggatctcgt acggatccgt cgacggcgcg 60
cccgatcatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag 120
gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt 180
cgggctttgt tagcagccgg atcgatccaa gctgtacctc actattcctt tgccctcgga 240
cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag 300
acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg 360
attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc 420
tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccgcgg cgatcctgca 480
agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc 540
ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag 600
tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg 660
tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc 720
tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga 780
tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg 840
aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catagcctcc 900
gcgaccggct gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc 960
tgtgcacggc gggagatgca ataggtcagg ctctcgctga attccccaat gtcaagcact 1020
tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa 1080
ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa 1140
gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg 1200
atcagaaact tctcgacaga cgtcgcggtg agttcaggct tttccatggg tatatctcct 1260
tcttaaagtt aaacaaaatt atttctagag ggaaaccgtt gtggtctccc tatagtgagt 1320
cgtattaatt tcgcgggatc gagatcgatc caattccaat cccacaaaaa tctgagctta 1380
acagcacagt tgctcctctc agagcagaat cgggtattca acaccctcat atcaactact 1440
acgttgtgta taacggtcca catgccggta tatacgatga ctggggttgt acaaaggcgg 1500
caacaaacgg cgttcccgga gttgcacaca agaaatttgc cactattaca gaggcaagag 1560
cagcagctga cgcgtacaca acaagtcagc aaacagacag gttgaacttc atccccaaag 1620
gagaagctca actcaagccc aagagctttg ctaaggccct aacaagccca ccaaagcaaa 1680
aagcccactg gctcacgcta ggaaccaaaa ggcccagcag tgatccagcc ccaaaagaga 1740
tctcctttgc cccggagatt acaatggacg atttcctcta tctttacgat ctaggaagga 1800
agttcgaagg tgaaggtgac gacactatgt tcaccactga taatgagaag gttagcctct 1860
tcaatttcag aaagaatgct gacccacaga tggttagaga ggcctacgca gcaggtctca 1920
tcaagacgat ctacccgagt aacaatctcc aggagatcaa ataccttccc aagaaggtta 1980
aagatgcagt caaaagattc aggactaatt gcatcaagaa cacagagaaa gacatatttc 2040
tcaagatcag aagtactatt ccagtatgga cgattcaagg cttgcttcat aaaccaaggc 2100
aagtaataga gattggagtc tctaaaaagg tagttcctac tgaatctaag gccatgcatg 2160
gagtctaaga ttcaaatcga ggatctaaca gaactcgccg tgaagactgg cgaacagttc 2220
atacagagtc ttttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac 2280
gacactctgg tctactccaa aaatgtcaaa gatacagtct cagaagacca aagggctatt 2340
gagacttttc aacaaaggat aatttcggga aacctcctcg gattccattg cccagctatc 2400
tgtcacttca tcgaaaggac agtagaaaag gaaggtggct cctacaaatg ccatcattgc 2460
gataaaggaa aggctatcat tcaagatgcc tctgccgaca gtggtcccaa agatggaccc 2520
ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg 2580
gattgatgtg acatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa 2640
gacccttcct ctatataagg aagttcattt catttggaga ggacacgctc gagctcattt 2700
ctctattact tcagccataa caaaagaact cttttctctt cttattaaac catgaaaaag 2760
cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc 2820
gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg 2880
cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt 2940
tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc 3000
agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg 3060
cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct 3120
gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa 3180
tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa 3240
actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt 3300
tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat 3360
gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg 3420
gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag 3480
cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg 3540
gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc 3600
gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact 3660
gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa 3720
gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagtga 3780
ggtacctaaa gaaggagtgc gtcgaagcag atcgttcaaa catttggcaa taaagtttct 3840
taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg 3900
ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga 3960
ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact 4020
aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcgatgtcga atcgatcaac 4080
ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 4140
gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 4200
cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 4260
tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 4320
cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 4380
aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 4440
cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 4500
gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 4560
ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 4620
cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 4680
aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 4740
tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc 4800
ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 4860
tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 4920
ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 4980
acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat 5040
gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg 5100
gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc 5160
tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg 5220
tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg 5280
tgacactata gaacggcgcg cca 5303
<210>105
<211>8358
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1274
<400>105
gtacggatcc gtcgacggcg cgcccgatca tccggatata gttcctcctt tcagcaaaaa 60
acccctcaag acccgtttag aggccccaag gggttatgct agttattgct cagcggtggc 120
agcagccaac tcagcttcct ttcgggcttt gttagcagcc ggatcgatcc aagctgtacc 180
tcactattcc tttgccctcg gacgagtgct ggggcgtcgg tttccactat cggcgagtac 240
ttctacacag ccatcggtcc agacggccgc gcttctgcgg gcgatttgtg tacgcccgac 300
agtcccggct ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc 360
gaaattgccg tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga gcatatacgc 420
ccggagccgc ggcgatcctg caagctccgg atgcctccgc tcgaagtagc gcgtctgctg 480
ctccatacaa gccaaccacg gcctccagaa gaagatgttg gcgacctcgt attgggaatc 540
cccgaacatc gcctcgctcc agtcaatgac cgctgttatg cggccattgt ccgtcaggac 600
attgttggag ccgaaatccg cgtgcacgag gtgccggact tcggggcagt cctcggccca 660
aagcatcagc tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt 720
ttgccagtga tacacatggg gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta 780
ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa gatcggccgc 840
agcgatcgca tccatagcct ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg 900
caggtcttgc aacgtgacac cctgtgcacg gcgggagatg caataggtca ggctctcgct 960
gaattcccca atgtcaagca cttccggaat cgggagcgcg gccgatgcaa agtgccgata 1020
aacataacga tctttgtaga aaccatcggc gcagctattt acccgcagga catatccacg 1080
ccctcctaca tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc 1140
ggagacgctg tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg 1200
cttttccatg ggtatatctc cttcttaaag ttaaacaaaa ttatttctag agggaaaccg 1260
ttgtggtctc cctatagtga gtcgtattaa tttcgcggga tcgagatcga tccaattcca 1320
atcccacaaa aatctgagct taacagcaca gttgctcctc tcagagcaga atcgggtatt 1380
caacaccctc atatcaacta ctacgttgtg tataacggtc cacatgccgg tatatacgat 1440
gactggggtt gtacaaaggc ggcaacaaac ggcgttcccg gagttgcaca caagaaattt 1500
gccactatta cagaggcaag agcagcagct gacgcgtaca caacaagtca gcaaacagac 1560
aggttgaact tcatccccaa aggagaagct caactcaagc ccaagagctt tgctaaggcc 1620
ctaacaagcc caccaaagca aaaagcccac tggctcacgc taggaaccaa aaggcccagc 1680
agtgatccag ccccaaaaga gatctccttt gccccggaga ttacaatgga cgatttcctc 1740
tatctttacg atctaggaag gaagttcgaa ggtgaaggtg acgacactat gttcaccact 1800
gataatgaga aggttagcct cttcaatttc agaaagaatg ctgacccaca gatggttaga 1860
gaggcctacg cagcaggtct catcaagacg atctacccga gtaacaatct ccaggagatc 1920
aaataccttc ccaagaaggt taaagatgca gtcaaaagat tcaggactaa ttgcatcaag 1980
aacacagaga aagacatatt tctcaagatc agaagtacta ttccagtatg gacgattcaa 2040
ggcttgcttc ataaaccaag gcaagtaata gagattggag tctctaaaaa ggtagttcct 2100
actgaatcta aggccatgca tggagtctaa gattcaaatc gaggatctaa cagaactcgc 2160
cgtgaagact ggcgaacagt tcatacagag tcttttacga ctcaatgaca agaagaaaat 2220
cttcgtcaac atggtggagc acgacactct ggtctactcc aaaaatgtca aagatacagt 2280
ctcagaagac caaagggcta ttgagacttt tcaacaaagg ataatttcgg gaaacctcct 2340
cggattccat tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaaggtgg 2400
ctcctacaaa tgccatcatt gcgataaagg aaaggctatc attcaagatg cctctgccga 2460
cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc 2520
aaccacgtct tcaaagcaag tggattgatg tgacatctcc actgacgtaa gggatgacgc 2580
acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga 2640
gaggacacgc tcgagctcat ttctctatta cttcagccat aacaaaagaa ctcttttctc 2700
ttcttattaa accatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat 2760
cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc 2820
tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg 2880
tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga 2940
agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca 3000
gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc 3060
ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt 3120
cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga 3180
tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca 3240
ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca 3300
cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga 3360
ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag 3420
gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct 3480
tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca 3540
gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat 3600
cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt 3660
ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg 3720
tccgagggca aaggaatagt gaggtaccta aagaaggagt gcgtcgaagc agatcgttca 3780
aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc 3840
atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta 3900
tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa 3960
aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta 4020
gatcgatgtc gaatcgatca acctgcatta atgaatcggc caacgcgcgg ggagaggcgg 4080
tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 4140
gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4200
ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4260
ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 4320
acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 4380
tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 4440
ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc 4500
ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 4560
ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 4620
actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4680
gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 4740
tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 4800
caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4860
atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4920
acgttaaggg attttggtca tgacattaac ctataaaaat aggcgtatca cgaggccctt 4980
tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 5040
ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 5100
gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag 5160
agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg 5220
tatcatacac atacgattta ggtgacacta tagaacggcg cgccaagctt ggatctcctg 5280
caggatctgg ccggccggat ctcgtacgct agcctaagta cgtactcaaa atgccaacaa 5340
ataaaaaaaa agttgcttta ataatgccaa aacaaattaa taaaacactt acaacaccgg 5400
atttttttta attaaaatgt gccatttagg ataaatagtt aatattttta ataattattt 5460
aaaaagccgt atctactaaa atgattttta tttggttgaa aatattaata tgtttaaatc 5520
aacacaatct atcaaaatta aactaaaaaa aaaataagtg tacgtggtta acattagtac 5580
agtaatataa gaggaaaatg agaaattaag aaattgaaag cgagtctaat ttttaaatta 5640
tgaacctgca tatataaaag gaaagaaaga atccaggaag aaaagaaatg aaaccatgca 5700
tggtcccctc gtcatcacga gtttctgcca tttgcaatag aaacactgaa acacctttct 5760
ctttgtcact taattgagat gccgaagcca cctcacacca tgaacttcat gaggtgtagc 5820
acccaaggct tccatagcca tgcatactga agaatgtctc aagctcagca ccctacttct 5880
gtgacgtgtc cctcattcac cttcctctct tccctataaa taaccacgcc tcaggttctc 5940
cgcttcacaa ctcaaacatt ctctccattg gtccttaaac actcatcagt catcaccgcg 6000
gccgcaaacc atggaggtcc gacgacgaaa gatagacgtg ctcaaggccc agaaaaacgg 6060
ctacgaatcg ggcccaccat ctcgacaatc gtcgcagccc tcctcaagag catcgtccag 6120
aacccgcaac aaacactcct cgtccaccct gtcgctcagc ggactgacca tgaaagtcca 6180
gaagaaacct gcgggacccc cggcgaactc caaaacgcca ttcctacaca tcaagcccgt 6240
gcacacgtgc tgctccacat caatgctttc gcgcgattat gacggctcca accccagctt 6300
caagggcttc aaaaacatcg gcatgatcat tctcattgtg ggaaatctac ggctcgcatt 6360
cgaaaactac ctcaaatacg gcatttccaa cccgttcttc gaccccaaaa ttactccttc 6420
cgagtggcag ctctcaggct tgctcatagt cgtggcctac gcacatatcc tcatggccta 6480
cgctattgag agcgctgcca agctgctgtt cctctctagc aaacaccact acatggccgt 6540
ggggcttctg cataccatga acactttgtc gtccatctcg ttgctgtcct acgtcgtcta 6600
ctactacctg cccaaccccg tggcaggcac aatagtcgag tttgtggccg ttattctgtc 6660
tctcaaactc gcctcatacg ccctcactaa ctcggatctc cgaaaagccg caattcatgc 6720
ccagaagctc gacaagacgc aagacgataa cgaaaaggaa tccacctcgt cttcctcttc 6780
ttcagatgac gcagagactt tggcagacat tgacgtcatt cctgcatact acgcacagct 6840
gccctacccc cagaatgtga cgctgtcgaa cctgctgtac ttctggtttg ctcccacact 6900
ggtctaccag cccgtgtacc ccaagacgga gcgtattcga cccaagcacg tgatccgaaa 6960
cctgtttgag ctcgtctctc tgtgcatgct tattcagttt ctcatcttcc agtacgccta 7020
ccccatcatg cagtcgtgtc tggctctgtt cttccagccc aagctcgatt atgccaacat 7080
ctccgagcgc ctcatgaagt tggcctccgt gtctatgatg gtctggctca ttggattcta 7140
cgctttcttc cagaacggtc tcaatcttat tgccgagctc acctgttttg gaaacagaac 7200
cttctaccag cagtggtgga attcccgctc cattggccag tactggactc tatggaacaa 7260
gccagtcaac cagtacttta gacaccacgt ctacgtgcct cttctcgctc ggggcatgtc 7320
gcggttcaat gcgtcggtgg tggttttctt tttctccgcc gtcatccatg aactgcttgt 7380
cggcatcccc actcacaaca tcatcggagc cgccttcttc ggcatgatgt cgcaggtgcc 7440
tctgatcatg gctactgaga accttcagca tattaactcc tctctgggcc ccttccttgg 7500
caactgtgca ttctggttca cctttttcct gggacaaccc acttgtgcat tcctttacta 7560
tttggcctac aactacaagc agaaccagta gcggccgcat ttcgcaccaa atcaatgaaa 7620
gtaataatga aaagtctgaa taagaatact taggcttaga tgcctttgtt acttgtgtaa 7680
aataacttga gtcatgtacc tttggcggaa acagaataaa taaaaggtga aattccaatg 7740
ctctatgtat aagttagtaa tacttaatgt gttctacggt tgtttcaata tcatcaaact 7800
ctaattgaaa ctttagaacc acaaatctca atcttttctt aatgaaatga aaaatcttaa 7860
ttgtaccatg tttatgttaa acaccttaca attggttgga gaggaggacc aaccgatggg 7920
acaacattgg gagaaagaga ttcaatggag atttggatag gagaacaaca ttctttttca 7980
cttcaataca agatgagtgc aacactaagg atatgtatga gactttcaga agctacgaca 8040
acatagatga gtgaggtggt gattcctagc aagaaagaca ttagaggaag ccaaaatcga 8100
acaaggaaga catcaagggc aagagacagg accatccatc tcaggaaaag gagctttggg 8160
atagtccgag aagttgtaca agaaattttt tggagggtga gtgatgcatt gctggtgact 8220
ttaactcaat caaaattgag aaagaaagaa aagggagggg gctcacatgt gaatagaagg 8280
gaaacgggag aattttacag ttttgatcta atgggcatcc cagctagtgg taacatattc 8340
accatgttta accttcac 8358
<210>106
<211>1251
<212>DNA
<213>大豆
<400>106
atggtggcaa cagctgctac ttcatcattt ttccctgtta cttcaccctc gccggactct 60
ggtggagcag gcagcaaact tggtggtggg cctgcaaacc ttggaggact aaaatccaaa 120
tctgcgtctt ctggtggctt gaaggcaaag gcgcaagccc cttcgaaaat taatggaacc 180
acagttgtta catctaaaga aagcttcaag catgatgatg atctaccttc gcctcccccc 240
agaactttta tcaaccagtt gcctgattgg agcatgcttc ttgctgctat cacaacaatt 300
ttcttggccg ctgaaaagca gtggatgatg cttgattgga agccacggcg acctgacatg 360
cttattgacc cctttgggat aggaaaaatt gttcaggatg gtcttgtgtt ccgtgaaaac 420
ttttctatta gatcatatga gattggtgct gatcgtaccg catctataga aacagtaatg 480
aaccatttgc aagaaactgc acttaatcat gttaaaagtg ctgggcttct tggtgatggc 540
tttggttcca cgccagaaat gtgcaaaaag aacttgatat gggtggttac tcggatgcag 600
gttgtggtgg aacgctatcc tacatggggt gacatagttc aagtggacac ttgggtttct 660
ggatcaggga agaatggtat gcgtcgtgat tggcttttac gtgactccaa aactggtgaa 720
atcttgacaa gagcttccag tgtttgggtc atgatgaata agctaacacg gaggctgtct 780
aaaattccag aagaagtcag acaggagata ggatcttatt ttgtggattc tgatccaatt 840
ctggaagagg ataacagaaa actgactaaa cttgacgaca acacagcgga ttatattcgt 900
accggtttaa gtcctaggtg gagtgatcta gatatcaatc agcatgtcaa caatgtgaag 960
tacattggct ggattctgga gagtgctcca cagccaatct tggagagtca tgagctttct 1020
tccatgactt tagagtatag gagagagtgt ggtagggaca gtgtgctgga ttccctgact 1080
gctgtatctg gggccgacat gggcaatcta gctcacagcg ggcatgttga gtgcaagcat 1140
ttgcttcgac tggaaaatgg tgctgagatt gtgaggggca ggactgagtg gaggcccaaa 1200
cctgtgaaca actttggtgt tgtgaaccag gttccagcag aaagcaccta a 1251
<210>107
<211>9592
<212>DNA
<213>人工序列
<220>
<223>质粒pTC4
<220>
<221>misc_feature
<222>(3725)..(3725)
<223>n为a,c,g,或t
<400>107
ggccgctaca tggccacgtg catgaagtat gaactaaaat gcatgtaggt gtaagagctc 60
atggagagca tggaatattg tatccgacca tgtaacagta taataactga gctccatctc 120
acttcttcta tgaataaaca aaggatgtta tgatatatta acactctatc tatgcacctt 180
attgttctat gataaatttc ctcttattat tataaatcat ctgaatcgtg acggcttatg 240
gaatgcttca aatagtacaa aaacaaatgt gtactataag actttctaaa caattctaac 300
tttagcattg tgaacgagac ataagtgtta agaagacata acaattataa tggaagaagt 360
ttgtctccat ttatatatta tatattaccc acttatgtat tatattagga tgttaaggag 420
acataacaat tataaagaga gaagtttgta tccatttata tattatatac tacccattta 480
tatattatac ttatccactt atttaatgtc tttataaggt ttgatccatg atatttctaa 540
tattttagtt gatatgtata tgaaagggta ctatttgaac tctcttactc tgtataaagg 600
ttggatcatc cttaaagtgg gtctatttaa ttttattgct tcttacagat aaaaaaaaaa 660
ttatgagttg gtttgataaa atattgaagg atttaaaata ataataaata ataaataaca 720
tataatatat gtatataaat ttattataat ataacattta tctataaaaa agtaaatatt 780
gtcataaatc tatacaatcg tttagccttg ctggacgact ctcaattatt taaacgagag 840
taaacatatt tgactttttg gttatttaac aaattattat ttaacactat atgaaatttt 900
ttttttttat cagcaaagaa ataaaattaa attaagaagg acaatggtgt gtcccaatcc 960
ttatacaacc aacttccaca agaaagtcaa gtcagagaca acaaaaaaac aagcaaagga 1020
aattttttaa tttgagttgt cttgtttgct gcataattta tgcagtaaaa cactacacat 1080
aaccctttta gcagtagagc aatggttgac cgtgtgctta gcttctttta ttttattttt 1140
ttatcagcaa agaataaata aaataaaatg agacacttca gggatgtttc aacaagcttt 1200
gctctagact ggaattcgtc gactctagag gatccaattc caatcccaca aaaatctgag 1260
cttaacagca cagttgctcc tctcagagca gaatcgggta ttcaacaccc tcatatcaac 1320
tactacgttg tgtataacgg tccacatgcc ggtatatacg atgactgggg ttgtacaaag 1380
gcggcaacaa acggcgttcc cggagttgca cacaagaaat ttgccactat tacagaggca 1440
agagcagcag ctgacgcgta cacaacaagt cagcaaacag acaggttgaa cttcatcccc 1500
aaaggagaag ctcaactcaa gcccaagagc tttgctaagg ccctaacaag cccaccaaag 1560
caaaaagccc actggctcac gctaggaacc aaaaggccca gcagtgatcc agccccaaaa 1620
gagatctcct ttgccccgga gattacaatg gacgatttcc tctatcttta cgatctagga 1680
aggaagttcg aaggtgaagg tgacgacact atgttcacca ctgataatga gaaggttagc 1740
ctcttcaatt tcagaaagaa tgctgaccca cagatggtta gagaggccta cgcagcaggt 1800
ctcatcaaga cgatctaccc gagtaacaat ctccaggaga tcaaatacct tcccaagaag 1860
gttaaagatg cagtcaaaag attcaggact aattgcatca agaacacaga gaaagacata 1920
tttctcaaga tcagaagtac tattccagta tggacgattc aaggcttgct tcataaacca 1980
aggcaagtaa tagagattgg agtctctaaa aaggtagttc ctactgaatc taaggccatg 2040
catggagtct aagattcaaa tcgaggatct aacagaactc gccgtgaaga ctggcgaaca 2100
gttcatacag agtcttttac gactcaatga caagaagaaa atcttcgtca acatggtgga 2160
gcacgacact ctggtctact ccaaaaatgt caaagataca gtctcagaag accaaagggc 2220
tattgagact tttcaacaaa ggataatttc gggaaacctc ctcggattcc attgcccagc 2280
tatctgtcac ttcatcgaaa ggacagtaga aaaggaaggt ggctcctaca aatgccatca 2340
ttgcgataaa ggaaaggcta tcattcaaga tgcctctgcc gacagtggtc ccaaagatgg 2400
acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca 2460
agtggattga tgtgacatct ccactgacgt aagggatgac gcacaatccc actatccttc 2520
gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacac gctcgagctc 2580
atttctctat tacttcagcc ataacaaaag aactcttttc tcttcttatt aaaccatgga 2640
aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt 2700
ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg 2760
agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta 2820
tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga 2880
attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga 2940
cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcta tggatgcgat 3000
cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg 3060
tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg 3120
gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat 3180
gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa 3240
caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt 3300
cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat 3360
ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct 3420
ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa 3480
tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg 3540
gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt 3600
agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata 3660
gtgaggtacc taatagtgag atccaacact tacgtttgca acgtccaaga gcaaatagac 3720
cacgnacgcc ggaaggttgc cgcagcgtgt ggattgcgtc tcaattctct cttgcaggaa 3780
tgcaatgatg aatatgatac tgactatgaa actttgaggg aatactgcct agcaccgtca 3840
cctcataacg tgcatcatgc atgccctgac aacatggaac atcgctattt ttctgaagaa 3900
ttatgctcgt tggaggatgt cgcggcaatt gcagctattg ccaacatcga actacccctc 3960
acgcatgcat tcatcaatat tattcatgcg gggaaaggca agattaatcc aactggcaaa 4020
tcatccagcg tgattggtaa cttcagttcc agcgacttga ttcgttttgg tgctacccac 4080
gttttcaata aggacgagat ggtggagtaa agaaggagtg cgtcgaagca gatcgttcaa 4140
acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg atgattatca 4200
tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc atgacgttat 4260
ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac gcgatagaaa 4320
acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct atgttactag 4380
atcgatcaaa cttcggtact gtgtaatgac gatgagcaat cgagaggctg actaacaaaa 4440
ggtacatcgg tcgacgagct ccctatagtg agtcgtatta gaggccgact tggccaaatt 4500
cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 4560
acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 4620
cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 4680
attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 4740
cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 4800
caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 4860
caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 4920
ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 4980
cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 5040
ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 5100
tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 5160
gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 5220
ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 5280
ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 5340
gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 5400
aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 5460
tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 5520
ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 5580
tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 5640
aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 5700
tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 5760
ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 5820
gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 5880
gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 5940
taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 6000
tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 6060
ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 6120
tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 6180
ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 6240
tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 6300
ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 6360
aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 6420
actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 6480
aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 6540
tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 6600
aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 6660
ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 6720
ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 6780
ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat 6840
ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 6900
ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 6960
gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 7020
tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 7080
ttaacgcgaa ttttaacaaa atattaacaa aatattaacg tttacaattt cccattcgcc 7140
attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca 7200
gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca 7260
gtcacgacgt tgtaaaacga cggccagtgc caagctgact tggtcagcgg ccggccgcag 7320
atttaggtga cactatagaa tatgcatcac tagtaagcta agcttgatcc atgcccttca 7380
tttgccgcta ttaattaatt tggtaacagt agtccgtact aatcagttac ttatccttcc 7440
tccatcataa ttaatcttgg tagtctcgaa tgccacaaca ctgactagtc tcttggatca 7500
taagaaaaag ccaaggaaca aaagaagaca aaacacaatg agagtatcct ttgcatagca 7560
atgtctaagt tcataaaatt caaacaaaaa cgcaatcaca cacagtggac atcacttatc 7620
cactagctga atcaggatcg ccgcgtcaag aaaaaaaaac tggaccccaa aagccatgca 7680
caacaacacg tactcacaaa ggtgtcaatc gagcagccca aaacattcac caactcaacc 7740
catcatgagc cctcacattt gttgtttcta acccaacctc aaactcgtat tctcttccgc 7800
cacctcattt ttgtttattt caacacccgt caaactgcat gccaccccgt ggccaaatgt 7860
ccatgcatgt taacaagacc tatgactata aatatctgca atctcggccc aggttttcat 7920
catcaagaac cagttcaata tcctagtaca ccgtattaaa gaatttaaga tatactaaca 7980
gcggccgctg acacaatagc ccttcttccc tgtttccagc ttttctcctt ctctctctct 8040
ccatcttctt cttcttcttc actcagtcag atccaactcc tcagataaca caagaccaaa 8100
cccgcttttt ctgcatttct agactagacg ttctaccgga gaagcgacct tagaaattca 8160
ttatggtggc aacagctgct acttcatcat ttttccctgt tacttcaccc tcgccggact 8220
ctggtggagc aggcagcaaa cttggtggtg ggcctgcaaa ccttggagga ctaaaatcca 8280
aatctgcgtc ttctggtggc ttgaaggcaa aggcgcaagc cccttcgaaa attaatggaa 8340
ccacagttgt tacatctaaa gaaagcttca agcatgatga tgatctacct tcgcctcccc 8400
ccagaacttt tatcaaccag ttgcctgatt ggagcatgct tcttgctgct atcacaacaa 8460
ttttcttggc cgctgaaaag cagtggatga tgcttgattg gaagccacgg cgacctgaca 8520
tgcttattga cccctttggg ataggaaaaa ttgttcagga tggtcttgtg ttccgtgaaa 8580
acttttctat tagatcatat gagattggtg ctgatcgtac cgcatctata gaaacagtaa 8640
tgaaccattt gcaagaaact gcacttaatc atgttaaaag tgctgggctt cttggtgatg 8700
gctttggttc cacgccagaa atgtgcaaaa agaacttgat atgggtggtt actcggatgc 8760
aggttgtggt ggaacgctat cctacatggg gtgacatagt tcaagtggac acttgggttt 8820
ctggatcagg gaagaatggt atgcgtcgtg attggctttt acgtgactcc aaaactggtg 8880
aaatcttgac aagagcttcc agtgtttggg tcatgatgaa taagctaaca cggaggctgt 8940
ctaaaattcc agaagaagtc agacaggaga taggatctta ttttgtggat tctgatccaa 9000
ttctggaaga ggataacaga aaactgacta aacttgacga caacacagcg gattatattc 9060
gtaccggttt aagtcctagg tggagtgatc tagatatcaa tcagcatgtc aacaatgtga 9120
agtacattgg ctggattctg gagagtgctc cacagccaat cttggagagt catgagcttt 9180
cttccatgac tttagagtat aggagagagt gtggtaggga cagtgtgctg gattccctga 9240
ctgctgtatc tggggccgac atgggcaatc tagctcacag cgggcatgtt gagtgcaagc 9300
atttgcttcg actggaaaat ggtgctgaga ttgtgagggg caggactgag tggaggccca 9360
aacctgtgaa caactttggt gttgtgaacc aggttccagc agaaagcacc taagatttga 9420
aatggttaac gattggagtt gcatcagtct ccttgctatg tttagactta ttctggttcc 9480
ctggggagag ttttgcttgt gtctatccaa tcaatctaca tgtctttaaa tatatacacc 9540
ttctaatttg tgatactttg gtgggtaagg gggaaaagca gcagtaaatc gc 9592
<210>108
<211>28
<212>DNA
<213>人工序列
<220>
<223>引物GmTE2 5-1
<400>108
aaacgcgtgt ggcaacagct gctacttc 28
<210>109
<211>51
<212>DNA
<213>人工序列
<220>
<223>引物GmTE2 3-1
<400>109
gctcttgtca agatttcacc gaattcttaa gttttggagt cacgtaaaag c 51
<210>110
<211>51
<212>DNA
<213>人工序列
<220>
<223>引物GmTE2 5-2
<400>110
gcttttacgt gactccaaaa cttaagaatt cggtgaaatc ttgacaagag c 51
<210>111
<211>60
<212>DNA
<213>人工序列
<220>
<223>引物GmTE2 3-2
<400>111
tcgcggccgc tgtacaggaa caattgtgat taattagtca caatctcagc accattttcc 60
<210>112
<211>4738
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1258
<400>112
aattcaggaa acgcgtgtgg caacagctgc tacttcatca tttttccctg ttacttcacc 60
ctcgccggac tctggtggag caggcagcaa acttggtggt gggcctgcaa accttggagg 120
actaaaatcc aaatctgcgt cttctggtgg cttgaaggca aaggcgcaag ccccttcgaa 180
aattaatgga accacagttg ttacatctaa agaaagcttc aagcatgatg atgatctacc 240
ttcgcctccc cccagaactt ttatcaacca gttgcctgat tggagcatgc ttcttgctgc 300
tatcacaaca attttcttgg ccgctgaaaa gcagtggatg atgcttgatt ggaagccacg 360
gcgacctgac atgcttattg acccctttgg gataggaaaa attgttcagg atggtcttgt 420
gttccgtgaa aacttttcta ttagatcata tgagattggt gctgatcgta ccgcatctat 480
agaaacagta atgaaccatt tgcaagaaac tgcacttaat catgttaaaa gtgctgggct 540
tcttggtgat ggctttggtt ccacgccaga aatgtgcaaa aagaacttga tatgggtggt 600
tactcggatg caggttgtgg tggaacgcta tcctacatgg ggtgacatag ttcaagtgga 660
cacttgggtt tctggatcag ggaagaatgg tatgcgtcgt gattggcttt tacgtgactc 720
caaaacttaa gaattcggtg aaatcttgac aagagcttcc agtgtttggg tcatgatgaa 780
taagctaaca cggaggctgt ctaaaattcc agaagaagtc agacaggaga taggatctta 840
ttttgtggat tctgatccaa ttctggaaga ggataacaga aaactgacta aacttgacga 900
cgacacagcg gattatattc gtaccggttt aagtcctagg tggagtgatc tagatatcaa 960
tcagcatgtc aacaatgtga agtacattgg ctggattctg gagagtgctc cacagccaat 1020
cttggagagt catgagcttt cttccatgac tttagagtat aggagagagt gtggtaggga 1080
cagtgtgctg gattccctga ctgctgtatc tggggccgac atgggcaatc tagctcacag 1140
cgggcatgtt gagtgcaagc atttgcttcg actggaaaat ggtgctgaga ttgtgactaa 1200
ttaatcacaa ttgttcctgt acagcggccg cgacctgaat tccagcacac tggcggccgt 1260
tactagtgga tccgagctcg gtaccaagct tgatgcatag cttgagtatt ctaacgcgtc 1320
acctaaatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 1380
tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 1440
gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 1500
tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 1560
ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 1620
cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 1680
gaaagaacat gtgagcaaaa ggccagcaaa agcccaggaa ccgtaaaaag gccgcgttgc 1740
tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 1800
agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 1860
tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 1920
cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 1980
ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 2040
ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 2100
ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 2160
ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 2220
cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 2280
gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 2340
atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 2400
ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 2460
gttttagcac gtgtcagtcc tgctcctcgg ccacgaagtg cacgcagttg ccggccgggt 2520
cgcgcagggc gaactcccgc ccccacggct gctcgccgat ctcggtcatg gccggcccgg 2580
aggcgtcccg gaagttcgtg gacacgacct ccgaccactc ggcgtacagc tcgtccaggc 2640
cgcgcaccca cacccaggcc agggtgttgt ccggcaccac ctggtcctgg accgcgctga 2700
tgaacagggt cacgtcgtcc cggaccacac cggcgaagtc gtcctccacg aagtcccggg 2760
agaacccgag ccggtcggtc cagaactcga ccgctccggc gacgtcgcgc gcggtgagca 2820
ccggaacggc actggtcaac ttggccatgg tggccctcct cacgtgctat tattgaagca 2880
tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 2940
aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgtatgcggt gtgaaatacc 3000
gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa taattcagaa 3060
gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta 3120
aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc 3180
caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga 3240
aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag 3300
atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc 3360
ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc 3420
tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg 3480
cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga 3540
caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac 3600
aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc 3660
ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg 3720
cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca 3780
gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg 3840
ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca gagcttgatc ccctgcgcca 3900
tcagatcctt ggcggcgaga aagccatcca gtttactttg cagggcttcc caaccttacc 3960
agagggcgcc ccagctggca attccggttc gcttgctgtc cataaaaccg cccagtctag 4020
ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgttttccct 4080
tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc 4140
tttctacgtg aaaaggatct aggtgaagat cctttttgat aatctcatgc ctgacattta 4200
tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc 4260
agccacttct tccccgataa cggagaccgg cacactggcc atatcggtgg tcatcatgcg 4320
ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggagactt tatctgacag 4380
cagacgtgca ctggccaggg ggatcaccat ccgtcgcccc ggcgtgtcaa taatatcact 4440
ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg 4500
ccgtacgtat aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt 4560
acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt 4620
ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg 4680
cgaattgggc cctctagatg catgctcgag cggccgccag tgtgatggat atctgcag 4738
<210>113
<211>1164
<212>DNA
<213>大豆
<400>113
atgggtctag caaaggaaac aacaatggga ggtagaggtc gtgtggccaa agtggaagtt 60
caagggaaga agcctctctc aagggttcca aacacaaagc caccattcac tgttggccaa 120
ctcaagaaag caattccacc acactgcttt cagcgctccc tcctcacttc attctcctat 180
gttgtttatg acctttcatt tgccttcatt ttctacattg ccaccaccta cttccacctc 240
cttcctcaac ccttttccct cattgcatgg ccaatctatt gggttctcca aggttgcctt 300
ctcactggtg tgtgggtgat tgctcacgag tgtggtcacc atgccttcag caagtaccaa 360
tgggttgatg atgttgtggg tttgaccctt cactcaacac ttttagtccc ttatttctca 420
tggaaaataa gccatcgccg ccatcactcc aacacaggtt cccttgaccg tgatgaagtg 480
tttgtcccaa aaccaaaatc caaagttgca tggttttcca agtacttaaa caaccctcta 540
ggaagggctg tttctcttct cgtcacactc acaatagggt ggcctatgta tttagccttc 600
aatgtctctg gtagacccta tgatagtttt gcaagccact accaccctta tgctcccata 660
tattctaacc gtgagaggct tctgatctat gtctctgatg ttgctttgtt ttctgtgact 720
tactctctct accgtgttgc aaccctgaaa gggttggttt ggctgctatg tgtttatggg 780
gtgcctttgc tcattgtgaa cggttttctt gtgactatca catatttgca gcacacacac 840
tttgccttgc ctcattacga ttcatcagaa tgggactggc tgaagggagc tttggcaact 900
atggacagag attatgggat tctgaacaag gtgtttcatc acataactga tactcatgtg 960
gctcaccatc tcttctctac aatgccacat taccatgcaa tggaggcaac caatgcaatc 1020
aagccaatat tgggtgagta ctaccaattt gatgacacac cattttacaa ggcactgtgg 1080
agagaagcga gagagtgcct ctatgtggag ccagatgaag gaacatccga gaagggcgtg 1140
tattggtaca ggaacaagta ttga 1164
<210>114
<211>10303
<212>DNA
<213>人工序列
<220>
<223>质粒pBS43
<220>
<221>misc_feature
<222>(3347)..(3347)
<223>n为a,c,g,或t
<400>114
gtcgactcta gaggatccaa ttccaatccc acaaaaatct gagcttaaca gcacagttgc 60
tcctctcaga gcagaatcgg gtattcaaca ccctcatatc aactactacg ttgtgtataa 120
cggtccacat gccggtatat acgatgactg gggttgtaca aaggcggcaa caaacggcgt 180
tcccggagtt gcacacaaga aatttgccac tattacagag gcaagagcag cagctgacgc 240
gtacacaaca agtcagcaaa cagacaggtt gaacttcatc cccaaaggag aagctcaact 300
caagcccaag agctttgcta aggccctaac aagcccacca aagcaaaaag cccactggct 360
cacgctagga accaaaaggc ccagcagtga tccagcccca aaagagatct cctttgcccc 420
ggagattaca atggacgatt tcctctatct ttacgatcta ggaaggaagt tcgaaggtga 480
aggtgacgac actatgttca ccactgataa tgagaaggtt agcctcttca atttcagaaa 540
gaatgctgac ccacagatgg ttagagaggc ctacgcagca ggtctcatca agacgatcta 600
cccgagtaac aatctccagg agatcaaata ccttcccaag aaggttaaag atgcagtcaa 660
aagattcagg actaattgca tcaagaacac agagaaagac atatttctca agatcagaag 720
tactattcca gtatggacga ttcaaggctt gcttcataaa ccaaggcaag taatagagat 780
tggagtctct aaaaaggtag ttcctactga atctaaggcc atgcatggag tctaagattc 840
aaatcgagga tctaacagaa ctcgccgtga agactggcga acagttcata cagagtcttt 900
tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac actctggtct 960
actccaaaaa tgtcaaagat acagtctcag aagaccaaag ggctattgag acttttcaac 1020
aaaggataat ttcgggaaac ctcctcggat tccattgccc agctatctgt cacttcatcg 1080
aaaggacagt agaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg 1140
ctatcattca agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga 1200
gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgaca 1260
tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta 1320
tataaggaag ttcatttcat ttggagagga cacgctcgag ctcatttctc tattacttca 1380
gccataacaa aagaactctt ttctcttctt attaaaccat ggtacgtcct gtagaaaccc 1440
caacccgtga aatcaaaaaa ctcgacggcc tgtgggcatt cagtctggat cgcgaaaact 1500
gtggaattga tcagcgttgg tgggaaagcg cgttacaaga aagccgggca attgctgtgc 1560
caggcagttt taacgatcag ttcgccgatg cagatattcg taattatgcg ggcaacgtct 1620
ggtatcagcg cgaagtcttt ataccgaaag gttgggcagg ccagcgtatc gtgctgcgtt 1680
tcgatgcggt cactcattac ggcaaagtgt gggtcaataa tcaggaagtg atggagcatc 1740
agggcggcta tacgccattt gaagccgatg tcacgccgta tgttattgcc gggaaaagtg 1800
tacgtatcac cgtttgtgtg aacaacgaac tgaactggca gactatcccg ccgggaatgg 1860
tgattaccga cgaaaacggc aagaaaaagc agtcttactt ccatgatttc tttaactatg 1920
ccggaatcca tcgcagcgta atgctctaca ccacgccgaa cacctgggtg gacgatatca 1980
ccgtggtgac gcatgtcgcg caagactgta accacgcgtc tgttgactgg caggtggtgg 2040
ccaatggtga tgtcagcgtt gaactgcgtg atgcggatca acaggtggtt gcaactggac 2100
aaggcactag cgggactttg caagtggtga atccgcacct ctggcaaccg ggtgaaggtt 2160
atctctatga actgtgcgtc acagccaaaa gccagacaga gtgtgatatc tacccgcttc 2220
gcgtcggcat ccggtcagtg gcagtgaagg gccaacagtt cctgattaac cacaaaccgt 2280
tctactttac tggctttggt cgtcatgaag atgcggactt acgtggcaaa ggattcgata 2340
acgtgctgat ggtgcacgac cacgcattaa tggactggat tggggccaac tcctaccgta 2400
cctcgcatta cccttacgct gaagagatgc tcgactgggc agatgaacat ggcatcgtgg 2460
tgattgatga aactgctgct gtcggcttta acctctcttt aggcattggt ttcgaagcgg 2520
gcaacaagcc gaaagaactg tacagcgaag aggcagtcaa cggggaaact cagcaagcgc 2580
acttacaggc gattaaagag ctgatagcgc gtgacaaaaa ccacccaagc gtggtgatgt 2640
ggagtattgc caacgaaccg gatacccgtc cgcaagtgca cgggaatatt tcgccactgg 2700
cggaagcaac gcgtaaactc gacccgacgc gtccgatcac ctgcgtcaat gtaatgttct 2760
gcgacgctca caccgatacc atcagcgatc tctttgatgt gctgtgcctg aaccgttatt 2820
acggatggta tgtccaaagc ggcgatttgg aaacggcaga gaaggtactg gaaaaagaac 2880
ttctggcctg gcaggagaaa ctgcatcagc cgattatcat caccgaatac ggcgtggata 2940
cgttagccgg gctgcactca atgtacaccg acatgtggag tgaagagtat cagtgtgcat 3000
ggctggatat gtatcaccgc gtctttgatc gcgtcagcgc cgtcgtcggt gaacaggtat 3060
ggaatttcgc cgattttgcg acctcgcaag gcatattgcg cgttggcggt aacaagaaag 3120
ggatcttcac tcgcgaccgc aaaccgaagt cggcggcttt tctgctgcaa aaacgctgga 3180
ctggcatgaa cttcggtgaa aaaccgcagc agggaggcaa acaatgaatc aacaactctc 3240
ctggcgcacc atcgtcggct acagcctcgg tggggaattc cccgggggta cctaatagtg 3300
agatccaaca cttacgtttg caacgtccaa gagcaaatag accacgnacg ccggaaggtt 3360
gccgcagcgt gtggattgcg tctcaattct ctcttgcagg aatgcaatga tgaatatgat 3420
actgactatg aaactttgag ggaatactgc ctagcaccgt cacctcataa cgtgcatcat 3480
gcatgccctg acaacatgga acatcgctat ttttctgaag aattatgctc gttggaggat 3540
gtcgcggcaa ttgcagctat tgccaacatc gaactacccc tcacgcatgc attcatcaat 3600
attattcatg cggggaaagg caagattaat ccaactggca aatcatccag cgtgattggt 3660
aacttcagtt ccagcgactt gattcgtttt ggtgctaccc acgttttcaa taaggacgag 3720
atggtggagt aaagaaggag tgcgtcgaag cagatcgttc aaacatttgg caataaagtt 3780
tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt 3840
acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta 3900
tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa 3960
actaggataa attatcgcgc gcggtgtcat ctatgttact agatcgatca aacttcggta 4020
ctgtgtaatg acgatgagca atcgagaggc tgactaacaa aaggtacatc ggtcgacgag 4080
ctccctatag tgagtcgtat tagaggccga cttggccaaa ttcgtaatca tggtcatagc 4140
tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 4200
taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 4260
cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 4320
gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 4380
tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 4440
tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4500
ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4560
agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4620
accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4680
ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 4740
gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 4800
ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 4860
gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 4920
taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 4980
tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 5040
gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 5100
cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 5160
agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 5220
cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 5280
cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 5340
ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 5400
taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 5460
tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 5520
ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 5580
atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 5640
gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 5700
tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 5760
cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 5820
taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 5880
ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 5940
ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 6000
cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 6060
ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 6120
gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 6180
gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 6240
aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg 6300
gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 6360
ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 6420
cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc 6480
tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga 6540
cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 6600
ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga 6660
tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 6720
aaatattaac aaaatattaa cgtttacaat ttcccattcg ccattcaggc tgcgcaactg 6780
ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg 6840
tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac 6900
gacggccagt gccaagctga cttggtcagc ggccgcagat ttaggtgaca ctatagaata 6960
tgcatcacta gtaagcttga tccatgccct tcatttgccg ctattaatta atttggtaac 7020
agtagtccgt actaatcagt tacttatcct tcctccatca taattaatct tggtagtctc 7080
gaatgccaca acactgacta gtctcttgga tcataagaaa aagccaagga acaaaagaag 7140
acaaaacaca atgagagtat cctttgcata gcaatgtcta agttcataaa attcaaacaa 7200
aaacgcaatc acacacagtg gacatcactt atccactagc tgaatcagga tcgccgcgtc 7260
aagaaaaaaa aactggaccc caaaagccat gcacaacaac acgtactcac aaaggtgtca 7320
atcgagcagc ccaaaacatt caccaactca acccatcatg agccctcaca tttgttgttt 7380
ctaacccaac ctcaaactcg tattctcttc cgccacctca tttttgttta tttcaacacc 7440
cgtcaaactg catgccaccc cgtggccaaa tgtccatgca tgttaacaag acctatgact 7500
ataaatatct gcaatctcgg cccaggtttt catcatcaag aaccagttca atatcctagt 7560
acaccgtatt aaagaattta agatatacta acgtagccca tcgaattccc atatactaat 7620
atttgcttgt attgatagcc cctccgttcc caagagtata aaactgcatc gaataataca 7680
agccactagg catgggtcta gcaaaggaaa caacaatggg aggtagaggt cgtgtggcca 7740
aagtggaagt tcaagggaag aagcctctct caagggttcc aaacacaaag ccaccattca 7800
ctgttggcca actcaagaaa gcaattccac cacactgctt tcagcgctcc ctcctcactt 7860
cattctccta tgttgtttat gacctttcat ttgccttcat tttctacatt gccaccacct 7920
acttccacct ccttcctcaa cccttttccc tcattgcatg gccaatctat tgggttctcc 7980
aaggttgcct tctcactggt gtgtgggtga ttgctcacga gtgtggtcac catgccttca 8040
gcaagtacca atgggttgat gatgttgtgg gtttgaccct tcactcaaca cttttagtcc 8100
cttatttctc atggaaaata agccatcgcc gccatcactc caacacaggt tcccttgacc 8160
gtgatgaagt gtttgtccca aaaccaaaat ccaaagttgc atggttttcc aagtacttaa 8220
acaaccctct aggaagggct gtttctcttc tcgtcacact cacaataggg tggcctatgt 8280
atttagcctt caatgtctct ggtagaccct atgatagttt tgcaagccac taccaccctt 8340
atgctcccat atattctaac cgtgagaggc ttctgatcta tgtctctgat gttgctttgt 8400
tttctgtgac ttactctctc taccgtgttg caaccctgaa agggttggtt tggctgctat 8460
gtgtttatgg ggtgcctttg ctcattgtga acggttttct tgtgactatc acatatttgc 8520
agcacacaca ctttgccttg cctcattacg attcatcaga atgggactgg ctgaagggag 8580
ctttggcaac tatggacaga gattatggga ttctgaacaa ggtgtttcat cacataactg 8640
atactcatgt ggctcaccat ctcttctcta caatgccaca ttaccatgca atggaggcaa 8700
ccaatgcaat caagccaata ttgggtgagt actaccaatt tgatgacaca ccattttaca 8760
aggcactgtg gagagaagcg agagagtgcc tctatgtgga gccagatgaa ggaacatccg 8820
agaagggcgt gtattggtac aggaacaagt attgatggag caaccaatgg gccatagtgg 8880
gagttatgga agttttgtca tgtattagta cataattagt agaatgttat aaataagtgg 8940
atttgccgcg taatgacttt gtgtgtattg tgaaacagct tgttgcgatc atggttataa 9000
tgtaaaaata attctggtat taattacatg tggaaagtgt tctgcttata gctttctgcc 9060
taaaaaaaaa gaattcctgc agcccgggtc tagacatggc cacgtgcatg aagtatgaac 9120
taaaatgcat gtaggtgtaa gagctcatgg agagcatgga atattgtatc cgaccatgta 9180
acagtataat aactgagctc catctcactt cttctatgaa taaacaaagg atgttatgat 9240
atattaacac tctatctatg caccttattg ttctatgata aatttcctct tattattata 9300
aatcatctga atcgtgacgg cttatggaat gcttcaaata gtacaaaaac aaatgtgtac 9360
tataagactt tctaaacaat tctaacttta gcattgtgaa cgagacataa gtgttaagaa 9420
gacataacaa ttataatgga agaagtttgt ctccatttat atattatata ttacccactt 9480
atgtattata ttaggatgtt aaggagacat aacaattata aagagagaag tttgtatcca 9540
tttatatatt atatactacc catttatata ttatacttat ccacttattt aatgtcttta 9600
taaggtttga tccatgatat ttctaatatt ttagttgata tgtatatgaa agggtactat 9660
ttgaactctc ttactctgta taaaggttgg atcatcctta aagtgggtct atttaatttt 9720
attgcttctt acagataaaa aaaaaattat gagttggttt gataaaatat tgaaggattt 9780
aaaataataa taaataataa ataacatata atatatgtat ataaatttat tataatataa 9840
catttatcta taaaaaagta aatattgtca taaatctata caatcgttta gccttgctgg 9900
acgactctca attatttaaa cgagagtaaa catatttgac tttttggtta tttaacaaat 9960
tattatttaa cactatatga aatttttttt ttttatcagc aaagaaataa aattaaatta 10020
agaaggacaa tggtgtgtcc caatccttat acaaccaact tccacaagaa agtcaagtca 10080
gagacaacaa aaaaacaagc aaaggaaatt ttttaatttg agttgtcttg tttgctgcat 10140
aatttatgca gtaaaacact acacataacc cttttagcag tagagcaatg gttgaccgtg 10200
tgcttagctt cttttatttt atttttttat cagcaaagaa taaataaaat aaaatgagac 10260
acttcaggga tgtttcaaca agctttgctc tagactggaa ttc 10303
<210>115
<211>50
<212>DNA
<213>人工序列
<220>
<223>引物GmFad2-1 5-1
<400>115
gcggccgcgg tacctaatta actaggtcta gcaaaggaaa caacaatggg 50
<210>116
<211>48
<212>DNA
<213>人工序列
<220>
<223>引物GmFad2-1 3-1
<400>116
gggacaaaca cttcatcacg agatcttagt caagggaacc tgtgttgg 48
<210>117
<211>48
<212>DNA
<213>人工序列
<220>
<223>引物GmFad2-1 5-2
<400>117
ccaacacagg ttcccttgac taagatctcg tgatgaagtg tttgtccc 48
<210>118
<211>34
<212>DNA
<213>人工序列
<220>
<223>引物GmFad2-1 3-2
<400>118
acacgcgttt aaaatattgg cttgattgca ttgg 34
<210>119
<211>4584
<212>DNA
<213>人工序列
<220>
<223>质粒PCRblunt-Fad2-1
<400>119
ttgtaatacg actcactata gggcgaattg ggccctctag atgcatgctc gagcggccgc 60
cagtgtgatg gatatctgca gaattcaggg cggccgcggt acctaattaa ctaggtctag 120
caaaggaaac aacaatggga ggtagaggtc gtgtggccaa agtggaagtt caagggaaga 180
agcctctctc aagggttcca aacacaaagc caccattcac tgttggccaa ctcaagaaag 240
caattccacc acactgcttt cagcgctccc tcctcacttc attctcctat gttgtttatg 300
acctttcatt tgccttcatt ttctacattg ccaccaccta cttccacctc cttcctcaac 360
ccttttccct cattgcatgg ccaatctatt gggttctcca aggttgcctt ctcactggtg 420
tgtgggtgat tgctcacgag tgtggtcacc atgccttcag caagtaccaa tgggttgatg 480
atgttgtggg tttgaccctt cactcaacac ttttagtccc ttatttctca tggaaaataa 540
gccatcgccg ccatcactcc aacacaggtt cccttgacta agatctcgtg atgaagtgtt 600
tgtcccaaaa ccaaaatcca aagttgcatg gttttccaag tacttaaaca accctctagg 660
aagggctgtt tctcttctcg tcacactcac aatagggtgg cctatgtatt tagccttcaa 720
tgtctctggt agaccctatg atagttttgc aagccactac cacccttatg ctcccatata 780
ttctaaccgt gagaggcttc tgatctatgt ctctgatgtt gctttgtttt ctgtgactta 840
ctctctctac cgtgttgcaa ccctgaaagg gttggtttgg ctgctatgtg tttatggggt 900
gcctttgctc attgtgaacg gttttcttgt gactatcaca tatttgcagc acacacactt 960
tgccttgcct cattacgatt catcagaatg ggactggctg aagggagctt tggcaactat 1020
ggacagagat tatgggattc tgaacaaggt gtttcatcac ataactgata ctcatgtggc 1080
tcaccatctc ttctctacaa tgccacatta ccatgcaatg gaggcaacca atgcaatcaa 1140
gccaatattt taaacgcgtg tcctgaattc cagcacactg gcggccgtta ctagtggatc 1200
cgagctcggt accaagcttg atgcatagct tgagtattct aacgcgtcac ctaaatagct 1260
tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 1320
acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 1380
tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 1440
tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 1500
cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 1560
actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 1620
gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 1680
ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 1740
acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 1800
ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 1860
cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 1920
tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 1980
gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 2040
ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 2100
acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 2160
gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 2220
ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 2280
tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 2340
gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttagcacgt 2400
gtcagtcctg ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga 2460
actcccgccc ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga 2520
agttcgtgga cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca 2580
cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca 2640
cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc 2700
ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac 2760
tggtcaactt ggccatggtg gccctcctca cgtgctatta ttgaagcatt tatcagggtt 2820
attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 2880
cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg aaataccgca cagatgcgta 2940
aggagaaaat accgcatcag gaaattgtaa gcgttaataa ttcagaagaa ctcgtcaaga 3000
aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag 3060
cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc 3120
tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt 3180
tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg 3240
ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg 3300
tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga 3360
tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt 3420
gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc 3480
cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca 3540
gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt 3600
tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac 3660
agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat 3720
agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga 3780
aacgatcctc atcctgtctc ttgatcagag cttgatcccc tgcgccatca gatccttggc 3840
ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca 3900
gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta 3960
agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc 4020
cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtgaaa 4080
aggatctagg tgaagatcct ttttgataat ctcatgcctg acatttatat tccccagaac 4140
atcaggttaa tggcgttttt gatgtcattt tcgcggtggc tgagatcagc cacttcttcc 4200
ccgataacgg agaccggcac actggccata tcggtggtca tcatgcgcca gctttcatcc 4260
ccgatatgca ccaccgggta aagttcacgg gagactttat ctgacagcag acgtgcactg 4320
gccaggggga tcaccatccg tcgccccggc gtgtcaataa tatcactctg tacatccaca 4380
aacagacgat aacggctctc tcttttatag gtgtaaacct taaactgccg tacgtatagg 4440
ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg 4500
aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga 4560
cgttgtaaaa cgacggccag tgaa 4584
<210>120
<211>5797
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1259
<400>120
cgcgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 60
atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 120
cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 180
gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 240
gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 300
ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 360
acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 420
cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 480
caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 540
gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 600
tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 660
aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 720
ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 780
cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 840
tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 900
tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 960
ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 1020
aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 1080
aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 1140
aatgaagttt tagcacgtgt cagtcctgct cctcggccac gaagtgcacg cagttgccgg 1200
ccgggtcgcg cagggcgaac tcccgccccc acggctgctc gccgatctcg gtcatggccg 1260
gcccggaggc gtcccggaag ttcgtggaca cgacctccga ccactcggcg tacagctcgt 1320
ccaggccgcg cacccacacc caggccaggg tgttgtccgg caccacctgg tcctggaccg 1380
cgctgatgaa cagggtcacg tcgtcccgga ccacaccggc gaagtcgtcc tccacgaagt 1440
cccgggagaa cccgagccgg tcggtccaga actcgaccgc tccggcgacg tcgcgcgcgg 1500
tgagcaccgg aacggcactg gtcaacttgg ccatggtggc cctcctcacg tgctattatt 1560
gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 1620
ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa 1680
ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaataatt 1740
cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata 1800
ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg 1860
gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat 1920
ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg 1980
acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg 2040
agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta 2100
cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc 2160
gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga 2220
gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca 2280
gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc 2340
gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc 2400
gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt 2460
gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca 2520
tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagagct tgatcccctg 2580
cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc 2640
ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag 2700
tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt 2760
tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga 2820
ctggctttct acgtgaaaag gatctaggtg aagatccttt ttgataatct catgcctgac 2880
atttatattc cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg 2940
agatcagcca cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc 3000
atgcgccagc tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct 3060
gacagcagac gtgcactggc cagggggatc accatccgtc gccccggcgt gtcaataata 3120
tcactctgta catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta 3180
aactgccgta cgtataggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg 3240
ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca 3300
gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta 3360
tagggcgaat tgggccctct agatgcatgc tcgagcggcc gccagtgtga tggatatctg 3420
cagaattcag ggcggccgcg gtacctaatt aactaggtct agcaaaggaa acaacaatgg 3480
gaggtagagg tcgtgtggcc aaagtggaag ttcaagggaa gaagcctctc tcaagggttc 3540
caaacacaaa gccaccattc actgttggcc aactcaagaa agcaattcca ccacactgct 3600
ttcagcgctc cctcctcact tcattctcct atgttgttta tgacctttca tttgccttca 3660
ttttctacat tgccaccacc tacttccacc tccttcctca acccttttcc ctcattgcat 3720
ggccaatcta ttgggttctc caaggttgcc ttctcactgg tgtgtgggtg attgctcacg 3780
agtgtggtca ccatgccttc agcaagtacc aatgggttga tgatgttgtg ggtttgaccc 3840
ttcactcaac acttttagtc ccttatttct catggaaaat aagccatcgc cgccatcact 3900
ccaacacagg ttcccttgac taagatctcg tgatgaagtg tttgtcccaa aaccaaaatc 3960
caaagttgca tggttttcca agtacttaaa caaccctcta ggaagggctg tttctcttct 4020
cgtcacactc acaatagggt ggcctatgta tttagccttc aatgtctctg gtagacccta 4080
tgatagtttt gcaagccact accaccctta tgctcccata tattctaacc gtgagaggct 4140
tctgatctat gtctctgatg ttgctttgtt ttctgtgact tactctctct accgtgttgc 4200
aaccctgaaa gggttggttt ggctgctatg tgtttatggg gtgcctttgc tcattgtgaa 4260
cggttttctt gtgactatca catatttgca gcacacacac tttgccttgc ctcattacga 4320
ttcatcagaa tgggactggc tgaagggagc tttggcaact atggacagag attatgggat 4380
tctgaacaag gtgtttcatc acataactga tactcatgtg gctcaccatc tcttctctac 4440
aatgccacat taccatgcaa tggaggcaac caatgcaatc aagccaatat tttaaacgcg 4500
tgtggcaaca gctgctactt catcattttt ccctgttact tcaccctcgc cggactctgg 4560
tggagcaggc agcaaacttg gtggtgggcc tgcaaacctt ggaggactaa aatccaaatc 4620
tgcgtcttct ggtggcttga aggcaaaggc gcaagcccct tcgaaaatta atggaaccac 4680
agttgttaca tctaaagaaa gcttcaagca tgatgatgat ctaccttcgc ctccccccag 4740
aacttttatc aaccagttgc ctgattggag catgcttctt gctgctatca caacaatttt 4800
cttggccgct gaaaagcagt ggatgatgct tgattggaag ccacggcgac ctgacatgct 4860
tattgacccc tttgggatag gaaaaattgt tcaggatggt cttgtgttcc gtgaaaactt 4920
ttctattaga tcatatgaga ttggtgctga tcgtaccgca tctatagaaa cagtaatgaa 4980
ccatttgcaa gaaactgcac ttaatcatgt taaaagtgct gggcttcttg gtgatggctt 5040
tggttccacg ccagaaatgt gcaaaaagaa cttgatatgg gtggttactc ggatgcaggt 5100
tgtggtggaa cgctatccta catggggtga catagttcaa gtggacactt gggtttctgg 5160
atcagggaag aatggtatgc gtcgtgattg gcttttacgt gactccaaaa cttaagaatt 5220
cggtgaaatc ttgacaagag cttccagtgt ttgggtcatg atgaataagc taacacggag 5280
gctgtctaaa attccagaag aagtcagaca ggagatagga tcttattttg tggattctga 5340
tccaattctg gaagaggata acagaaaact gactaaactt gacgacgaca cagcggatta 5400
tattcgtacc ggtttaagtc ctaggtggag tgatctagat atcaatcagc atgtcaacaa 5460
tgtgaagtac attggctgga ttctggagag tgctccacag ccaatcttgg agagtcatga 5520
gctttcttcc atgactttag agtataggag agagtgtggt agggacagtg tgctggattc 5580
cctgactgct gtatctgggg ccgacatggg caatctagct cacagcgggc atgttgagtg 5640
caagcatttg cttcgactgg aaaatggtgc tgagattgtg actaattaat cacaattgtt 5700
cctgtacagc ggccgccctg aattccagca cactggcggc cgttactagt ggatccgagc 5760
tcggtaccaa gcttgatgca tagcttgagt attctaa 5797
<210>121
<211>7590
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1261
<400>121
aattgttcct gtacagcggc cgccctgaat tccagcacac tggcggccgt tactagtgga 60
tccgagctcg gtaccaagct tgatgcatag cttgagtatt ctaacgcgtc acctaaatag 120
cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc 180
acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta 240
actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 300
gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 360
cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 420
tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 480
gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 540
ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 600
aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 660
tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 720
ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 780
gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 840
tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 900
caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 960
ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt 1020
cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 1080
ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 1140
cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 1200
gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttagcac 1260
gtgtcagtcc tgctcctcgg ccacgaagtg cacgcagttg ccggccgggt cgcgcagggc 1320
gaactcccgc ccccacggct gctcgccgat ctcggtcatg gccggcccgg aggcgtcccg 1380
gaagttcgtg gacacgacct ccgaccactc ggcgtacagc tcgtccaggc cgcgcaccca 1440
cacccaggcc agggtgttgt ccggcaccac ctggtcctgg accgcgctga tgaacagggt 1500
cacgtcgtcc cggaccacac cggcgaagtc gtcctccacg aagtcccggg agaacccgag 1560
ccggtcggtc cagaactcga ccgctccggc gacgtcgcgc gcggtgagca ccggaacggc 1620
actggtcaac ttggccatgg tggccctcct cacgtgctat tattgaagca tttatcaggg 1680
ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 1740
tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg tgaaataccg cacagatgcg 1800
taaggagaaa ataccgcatc aggaaattgt aagcgttaat aattcagaag aactcgtcaa 1860
gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga 1920
agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt 1980
cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat 2040
tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt 2100
cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt 2160
cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc 2220
gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca 2280
ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct 2340
gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca 2400
cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca 2460
gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg 2520
acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga 2580
atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc 2640
gaaacgatcc tcatcctgtc tcttgatcag agcttgatcc cctgcgccat cagatccttg 2700
gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc 2760
cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg 2820
taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag 2880
cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtga 2940
aaaggatcta ggtgaagatc ctttttgata atctcatgcc tgacatttat attccccaga 3000
acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca gccacttctt 3060
ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc cagctttcat 3120
ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc agacgtgcac 3180
tggccagggg gatcaccatc cgtcgccccg gcgtgtcaat aatatcactc tgtacatcca 3240
caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc cgtacgtata 3300
ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg 3360
cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac 3420
gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggcc 3480
ctctagatgc atgctcgagc ggccgccagt gtgatggata tctgcagaat tcagggcggc 3540
cgcggtacct aattaactag gtctagcaaa ggaaacaaca atgggaggta gaggtcgtgt 3600
ggccaaagtg gaagttcaag ggaagaagcc tctctcaagg gttccaaaca caaagccacc 3660
attcactgtt ggccaactca agaaagcaat tccaccacac tgctttcagc gctccctcct 3720
cacttcattc tcctatgttg tttatgacct ttcatttgcc ttcattttct acattgccac 3780
cacctacttc cacctccttc ctcaaccctt ttccctcatt gcatggccaa tctattgggt 3840
tctccaaggt tgccttctca ctggtgtgtg ggtgattgct cacgagtgtg gtcaccatgc 3900
cttcagcaag taccaatggg ttgatgatgt tgtgggtttg acccttcact caacactttt 3960
agtcccttat ttctcatgga aaataagcca tcgccgccat cactccaaca caggttccct 4020
tgactaagat ctcgtgatga agtgtttgtc ccaaaaccaa aatccaaagt tgcatggttt 4080
tccaagtact taaacaaccc tctaggaagg gctgtttctc ttctcgtcac actcacaata 4140
gggtggccta tgtatttagc cttcaatgtc tctggtagac cctatgatag ttttgcaagc 4200
cactaccacc cttatgctcc catatattct aaccgtgaga ggcttctgat ctatgtctct 4260
gatgttgctt tgttttctgt gacttactct ctctaccgtg ttgcaaccct gaaagggttg 4320
gtttggctgc tatgtgttta tggggtgcct ttgctcattg tgaacggttt tcttgtgact 4380
atcacatatt tgcagcacac acactttgcc ttgcctcatt acgattcatc agaatgggac 4440
tggctgaagg gagctttggc aactatggac agagattatg ggattctgaa caaggtgttt 4500
catcacataa ctgatactca tgtggctcac catctcttct ctacaatgcc acattaccat 4560
gcaatggagg caaccaatgc aatcaagcca atattttaaa cgcgtgtggc aacagctgct 4620
acttcatcat ttttccctgt tacttcaccc tcgccggact ctggtggagc aggcagcaaa 4680
cttggtggtg ggcctgcaaa ccttggagga ctaaaatcca aatctgcgtc ttctggtggc 4740
ttgaaggcaa aggcgcaagc cccttcgaaa attaatggaa ccacagttgt tacatctaaa 4800
gaaagcttca agcatgatga tgatctacct tcgcctcccc ccagaacttt tatcaaccag 4860
ttgcctgatt ggagcatgct tcttgctgct atcacaacaa ttttcttggc cgctgaaaag 4920
cagtggatga tgcttgattg gaagccacgg cgacctgaca tgcttattga cccctttggg 4980
ataggaaaaa ttgttcagga tggtcttgtg ttccgtgaaa acttttctat tagatcatat 5040
gagattggtg ctgatcgtac cgcatctata gaaacagtaa tgaaccattt gcaagaaact 5100
gcacttaatc atgttaaaag tgctgggctt cttggtgatg gctttggttc cacgccagaa 5160
atgtgcaaaa agaacttgat atgggtggtt actcggatgc aggttgtggt ggaacgctat 5220
cctacatggg gtgacatagt tcaagtggac acttgggttt ctggatcagg gaagaatggt 5280
atgcgtcgtg attggctttt acgtgactcc aaaacttaag aattcggtga aatcttgaca 5340
agagcttcca gtgtttgggt catgatgaat aagctaacac ggaggctgtc taaaattcca 5400
gaagaagtca gacaggagat aggatcttat tttgtggatt ctgatccaat tctggaagag 5460
gataacagaa aactgactaa acttgacgac gacacagcgg attatattcg taccggttta 5520
agtcctaggt ggagtgatct agatatcaat cagcatgtca acaatgtgaa gtacattggc 5580
tggattctgg agagtgctcc acagccaatc ttggagagtc atgagctttc ttccatgact 5640
ttagagtata ggagagagtg tggtagggac agtgtgctgg attccctgac tgctgtatct 5700
ggggccgaca tgggcaatct agctcacagc gggcatgttg agtgcaagca tttgcttcga 5760
ctggaaaatg gtgctgagat tgtgactaat taatcacaat tcttaagttt tggagtcacg 5820
taaaagccaa tcacgacgca taccattctt ccctgatcca gaaacccaag tgtccacttg 5880
aactatgtca ccccatgtag gatagcgttc caccacaacc tgcatccgag taaccaccca 5940
tatcaagttc tttttgcaca tttctggcgt ggaaccaaag ccatcaccaa gaagcccagc 6000
acttttaaca tgattaagtg cagtttcttg caaatggttc attactgttt ctatagatgc 6060
ggtacgatca gcaccaatct catatgatct aatagaaaag ttttcacgga acacaagacc 6120
atcctgaaca atttttccta tcccaaaggg gtcaataagc atgtcaggtc gccgtggctt 6180
ccaatcaagc atcatccact gcttttcagc ggccaagaaa attgttgtga tagcagcaag 6240
aagcatgctc caatcaggca actggttgat aaaagttctg gggggaggcg aaggtagatc 6300
atcatcatgc ttgaagcttt ctttagatgt aacaactgtg gttccattaa ttttcgaagg 6360
ggcttgcgcc tttgccttca agccaccaga agacgcagat ttggatttta gtcctccaag 6420
gtttgcaggc ccaccaccaa gtttgctgcc tgctccacca gagtccggcg agggtgaagt 6480
aacagggaaa aatgatgaag tagcagctgt tgccacacgc gtttaaaata ttggcttgat 6540
tgcattggtt gcctccattg catggtaatg tggcattgta gagaagagat ggtgagccac 6600
atgagtatca gttatgtgat gaaacacctt gttcagaatc ccataatctc tgtccatagt 6660
tgccaaagct cccttcagcc agtcccattc tgatgaatcg taatgaggca aggcaaagtg 6720
tgtgtgctgc aaatatgtga tagtcacaag aaaaccgttc acaatgagca aaggcacccc 6780
ataaacacat agcagccaaa ccaacccttt cagggttgca acacggtaga gagagtaagt 6840
cacagaaaac aaagcaacat cagagacata gatcagaagc ctctcacggt tagaatatat 6900
gggagcataa gggtggtagt ggcttgcaaa actatcatag ggtctaccag agacattgaa 6960
ggctaaatac ataggccacc ctattgtgag tgtgacgaga agagaaacag cccttcctag 7020
agggttgttt aagtacttgg aaaaccatgc aactttggat tttggttttg ggacaaacac 7080
ttcatcacga gatcttagtc aagggaacct gtgttggagt gatggcggcg atggcttatt 7140
ttccatgaga aataagggac taaaagtgtt gagtgaaggg tcaaacccac aacatcatca 7200
acccattggt acttgctgaa ggcatggtga ccacactcgt gagcaatcac ccacacacca 7260
gtgagaaggc aaccttggag aacccaatag attggccatg caatgaggga aaagggttga 7320
ggaaggaggt ggaagtaggt ggtggcaatg tagaaaatga aggcaaatga aaggtcataa 7380
acaacatagg agaatgaagt gaggagggag cgctgaaagc agtgtggtgg aattgctttc 7440
ttgagttggc caacagtgaa tggtggcttt gtgtttggaa cccttgagag aggcttcttc 7500
ccttgaactt ccactttggc cacacgacct ctacctccca ttgttgtttc ctttgctaga 7560
cctagttaat taggtaccgc ggccgccctg 7590
<210>122
<211>4993
<212>DNA
<213>人工序列
<220>
<223>质粒pKR123R
<220>
<221>misc_feature
<222>(1126)..(1126)
<223>n为a,c,g,或t
<400>122
ctagacctgc aggatataat gagccgtaaa caaagatgat taagtagtaa ttaatacgta 60
ctagtaaaag tggcaaaaga taacgagaaa gaaccaattt ctttgcattc ggccttagcg 120
gaaggcatat ataagctttg attattttat ttagtgtaat gatttcgtac aaccaaagca 180
tttatttagt actctcacac ttgtgtcgcg gccgcttggg gggctatgga agactttctt 240
agttagttgt gtgaataagc aatgttggga gaatcgggac tacttatagg ataggaataa 300
aacagaaaag tattaagtgc taatgaaata tttagactga taattaaaat cttcacgtat 360
gtccacttga tataaaaacg tcaggaataa aggaagtaca gtagaattta aaggtactct 420
ttttatatat acccgtgttc tctttttggc tagctagttg cataaaaaat aatctatatt 480
tttatcatta ttttaaatat cttatgagat ggtaaatatt tatcataatt ttttttacta 540
ttatttatta tttgtgtgtg taatacatat agaagttaat tacaaatttt atttactttt 600
tcattatttt gatatgattc accattaatt tagtgttatt atttataata gttcatttta 660
atctttttgt atatattatg cgtgcagtac ttttttccta catataacta ctattacatt 720
ttatttatat aatattttta ttaatgaatt ttcgtgataa tatgtaatat tgttcattat 780
tatttcagat tttttaaaaa tatttgtgtt attatttatg aaatatgtaa tttttttagt 840
atttgatttt atgatgataa agtgttctaa attcaaaaga agggggaaag cgtaaacatt 900
aaaaaacgtc atcaaacaaa aacaaaatct tgttaataaa gataaaactg tttgttttga 960
tcactgttat ttcgtaatat aaaaacatta tttatattta tattgttgac aaccaaattt 1020
gcctatcaaa tctaaccaat ataatgcatg cgtggcaggt aatgtactac catgaactta 1080
agtcatgaca taataaaccg tgaatctgac caatgcatgt acctanctaa attgtatttg 1140
tgacacgaag caaatgattc aattcacaat ggagatggga aacaaataat gaagaaccca 1200
gaactaagaa agcttttctg aaaaataaaa taaaggcaat gtcaaaagta tactgcatca 1260
tcagtccaga aagcacatga tattttttta tcagtatcaa tgcagctagt tttattttac 1320
aatatcgata tagctagttt aaatatattg cagctagatt tataaatatt tgtgttatta 1380
tttatcattt gtgtaatcct gtttttagta ttttagttta tatatgatga taatgtattc 1440
caaatttaaa agaagggaaa taaatttaaa caagaaaaaa agtcatcaaa caaaaaacaa 1500
atgaaagggt ggaaagatgt taccatgtaa tgtgaatgtt acagtatttc ttttattata 1560
gagttaacaa attaactaat atgattttgt taataatgat aaaatatttt ttttattatt 1620
atttcataat ataaaaatag tttacttaat ataaaaaaaa ttctatcgtt cacaacaaag 1680
ttggccacct aatttaacca tgcatgtacc catggaccat attaggtaac catcaaacct 1740
gatgaagaga taaagagatg aagacttaag tcataacaca aaaccataaa aaacaaaaat 1800
acaatcaacc gtcaatctga ccaatgcatg aaaaagctgc aatagtgagt ggcgacacaa 1860
agcacatgat tttcttacaa cggagataaa accaaaaaaa tatttcatga acaacctaga 1920
acaaataaag cttttatata ataaatatat aaataaataa aggctatgga ataatatact 1980
tcaatatatt tggattaaat aaattgttgg cggggttgat atatttatac acacctaaag 2040
tcacttcaat ctcattttca cttaactttt attttttttt tctttttatt tatcataaag 2100
agaatattga taatatactt tttaacatat ttttatgaca ttttttattg gtgaaaactt 2160
attaaaaatc ataaattttg taagttagat ttatttaaag agttcctctt cttattttaa 2220
attttttaat aaatttttaa ataactaaaa tttgtgttaa aaatgttaaa aaatgtgtta 2280
ttaacccttc tcttcgagga cgtacgtcta gagtcgacct gcaggcatgc aagcttggcg 2340
taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 2400
atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 2460
ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 2520
taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 2580
tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 2640
aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 2700
aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 2760
ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 2820
acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 2880
ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 2940
tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 3000
tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 3060
gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 3120
agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 3180
tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 3240
agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 3300
tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 3360
acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 3420
tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 3480
agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 3540
tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 3600
acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 3660
tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 3720
ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 3780
agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 3840
tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 3900
acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 3960
agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 4020
actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 4080
tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 4140
gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 4200
ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 4260
tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 4320
aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 4380
tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 4440
tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 4500
gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 4560
ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg 4620
gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg 4680
tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta 4740
ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 4800
atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc 4860
tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta 4920
acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt 4980
acccggggat cc t 4993
<210>123
<211>9036
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1266
<220>
<221>misc_feature
<222>(4081)..(4081)
<223>n为a,c,g,或t
<400>123
ggccgcgaca caagtgtgag agtactaaat aaatgctttg gttgtacgaa atcattacac 60
taaataaaat aatcaaagct tatatatgcc ttccgctaag gccgaatgca aagaaattgg 120
ttctttctcg ttatcttttg ccacttttac tagtacgtat taattactac ttaatcatct 180
ttgtttacgg ctcattatat cctgcaggtc tagaggatcc ccgggtaccg agctcgaatt 240
cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc 300
gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc 360
gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc 420
ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg 480
atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg 540
cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt 600
gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc 660
tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc 720
ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc 780
cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga 840
gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt 900
ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag 960
tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag 1020
aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta 1080
ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg 1140
agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca 1200
gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag 1260
gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc 1320
gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg 1380
tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc 1440
ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg 1500
cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg 1560
gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga 1620
cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac 1680
tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa 1740
aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca 1800
aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 1860
gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 1920
cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 1980
ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc 2040
accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 2100
tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 2160
cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 2220
gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 2280
ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 2340
cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 2400
tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 2460
ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 2520
ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 2580
ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 2640
gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg 2700
acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca 2760
ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg 2820
tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat 2880
gcctgcaggt cgactctaga cgtacgtcct cgaagagaag ggttaataac acatttttta 2940
acatttttaa cacaaatttt agttatttaa aaatttatta aaaaatttaa aataagaaga 3000
ggaactcttt aaataaatct aacttacaaa atttatgatt tttaataagt tttcaccaat 3060
aaaaaatgtc ataaaaatat gttaaaaagt atattatcaa tattctcttt atgataaata 3120
aaaagaaaaa aaaaataaaa gttaagtgaa aatgagattg aagtgacttt aggtgtgtat 3180
aaatatatca accccgccaa caatttattt aatccaaata tattgaagta tattattcca 3240
tagcctttat ttatttatat atttattata taaaagcttt atttgttcta ggttgttcat 3300
gaaatatttt tttggtttta tctccgttgt aagaaaatca tgtgctttgt gtcgccactc 3360
actattgcag ctttttcatg cattggtcag attgacggtt gattgtattt ttgtttttta 3420
tggttttgtg ttatgactta agtcttcatc tctttatctc ttcatcaggt ttgatggtta 3480
cctaatatgg tccatgggta catgcatggt taaattaggt ggccaacttt gttgtgaacg 3540
atagaatttt ttttatatta agtaaactat ttttatatta tgaaataata ataaaaaaaa 3600
tattttatca ttattaacaa aatcatatta gttaatttgt taactctata ataaaagaaa 3660
tactgtaaca ttcacattac atggtaacat ctttccaccc tttcatttgt tttttgtttg 3720
atgacttttt ttcttgttta aatttatttc ccttctttta aatttggaat acattatcat 3780
catatataaa ctaaaatact aaaaacagga ttacacaaat gataaataat aacacaaata 3840
tttataaatc tagctgcaat atatttaaac tagctatatc gatattgtaa aataaaacta 3900
gctgcattga tactgataaa aaaatatcat gtgctttctg gactgatgat gcagtatact 3960
tttgacattg cctttatttt atttttcaga aaagctttct tagttctggg ttcttcatta 4020
tttgtttccc atctccattg tgaattgaat catttgcttc gtgtcacaaa tacaatttag 4080
ntaggtacat gcattggtca gattcacggt ttattatgtc atgacttaag ttcatggtag 4140
tacattacct gccacgcatg cattatattg gttagatttg ataggcaaat ttggttgtca 4200
acaatataaa tataaataat gtttttatat tacgaaataa cagtgatcaa aacaaacagt 4260
tttatcttta ttaacaagat tttgtttttg tttgatgacg ttttttaatg tttacgcttt 4320
cccccttctt ttgaatttag aacactttat catcataaaa tcaaatacta aaaaaattac 4380
atatttcata aataataaca caaatatttt taaaaaatct gaaataataa tgaacaatat 4440
tacatattat cacgaaaatt cattaataaa aatattatat aaataaaatg taatagtagt 4500
tatatgtagg aaaaaagtac tgcacgcata atatatacaa aaagattaaa atgaactatt 4560
ataaataata acactaaatt aatggtgaat catatcaaaa taatgaaaaa gtaaataaaa 4620
tttgtaatta acttctatat gtattacaca cacaaataat aaataatagt aaaaaaaatt 4680
atgataaata tttaccatct cataagatat ttaaaataat gataaaaata tagattattt 4740
tttatgcaac tagctagcca aaaagagaac acgggtatat ataaaaagag tacctttaaa 4800
ttctactgta cttcctttat tcctgacgtt tttatatcaa gtggacatac gtgaagattt 4860
taattatcag tctaaatatt tcattagcac ttaatacttt tctgttttat tcctatccta 4920
taagtagtcc cgattctccc aacattgctt attcacacaa ctaactaaga aagtcttcca 4980
tagcccccca agcggccgcg gtacctaatt aactaggtct agcaaaggaa acaacaatgg 5040
gaggtagagg tcgtgtggcc aaagtggaag ttcaagggaa gaagcctctc tcaagggttc 5100
caaacacaaa gccaccattc actgttggcc aactcaagaa agcaattcca ccacactgct 5160
ttcagcgctc cctcctcact tcattctcct atgttgttta tgacctttca tttgccttca 5220
ttttctacat tgccaccacc tacttccacc tccttcctca acccttttcc ctcattgcat 5280
ggccaatcta ttgggttctc caaggttgcc ttctcactgg tgtgtgggtg attgctcacg 5340
agtgtggtca ccatgccttc agcaagtacc aatgggttga tgatgttgtg ggtttgaccc 5400
ttcactcaac acttttagtc ccttatttct catggaaaat aagccatcgc cgccatcact 5460
ccaacacagg ttcccttgac taagatctcg tgatgaagtg tttgtcccaa aaccaaaatc 5520
caaagttgca tggttttcca agtacttaaa caaccctcta ggaagggctg tttctcttct 5580
cgtcacactc acaatagggt ggcctatgta tttagccttc aatgtctctg gtagacccta 5640
tgatagtttt gcaagccact accaccctta tgctcccata tattctaacc gtgagaggct 5700
tctgatctat gtctctgatg ttgctttgtt ttctgtgact tactctctct accgtgttgc 5760
aaccctgaaa gggttggttt ggctgctatg tgtttatggg gtgcctttgc tcattgtgaa 5820
cggttttctt gtgactatca catatttgca gcacacacac tttgccttgc ctcattacga 5880
ttcatcagaa tgggactggc tgaagggagc tttggcaact atggacagag attatgggat 5940
tctgaacaag gtgtttcatc acataactga tactcatgtg gctcaccatc tcttctctac 6000
aatgccacat taccatgcaa tggaggcaac caatgcaatc aagccaatat tttaaacgcg 6060
tgtggcaaca gctgctactt catcattttt ccctgttact tcaccctcgc cggactctgg 6120
tggagcaggc agcaaacttg gtggtgggcc tgcaaacctt ggaggactaa aatccaaatc 6180
tgcgtcttct ggtggcttga aggcaaaggc gcaagcccct tcgaaaatta atggaaccac 6240
agttgttaca tctaaagaaa gcttcaagca tgatgatgat ctaccttcgc ctccccccag 6300
aacttttatc aaccagttgc ctgattggag catgcttctt gctgctatca caacaatttt 6360
cttggccgct gaaaagcagt ggatgatgct tgattggaag ccacggcgac ctgacatgct 6420
tattgacccc tttgggatag gaaaaattgt tcaggatggt cttgtgttcc gtgaaaactt 6480
ttctattaga tcatatgaga ttggtgctga tcgtaccgca tctatagaaa cagtaatgaa 6540
ccatttgcaa gaaactgcac ttaatcatgt taaaagtgct gggcttcttg gtgatggctt 6600
tggttccacg ccagaaatgt gcaaaaagaa cttgatatgg gtggttactc ggatgcaggt 6660
tgtggtggaa cgctatccta catggggtga catagttcaa gtggacactt gggtttctgg 6720
atcagggaag aatggtatgc gtcgtgattg gcttttacgt gactccaaaa cttaagaatt 6780
cggtgaaatc ttgacaagag cttccagtgt ttgggtcatg atgaataagc taacacggag 6840
gctgtctaaa attccagaag aagtcagaca ggagatagga tcttattttg tggattctga 6900
tccaattctg gaagaggata acagaaaact gactaaactt gacgacgaca cagcggatta 6960
tattcgtacc ggtttaagtc ctaggtggag tgatctagat atcaatcagc atgtcaacaa 7020
tgtgaagtac attggctgga ttctggagag tgctccacag ccaatcttgg agagtcatga 7080
gctttcttcc atgactttag agtataggag agagtgtggt agggacagtg tgctggattc 7140
cctgactgct gtatctgggg ccgacatggg caatctagct cacagcgggc atgttgagtg 7200
caagcatttg cttcgactgg aaaatggtgc tgagattgtg actaattaat cacaattctt 7260
aagttttgga gtcacgtaaa agccaatcac gacgcatacc attcttccct gatccagaaa 7320
cccaagtgtc cacttgaact atgtcacccc atgtaggata gcgttccacc acaacctgca 7380
tccgagtaac cacccatatc aagttctttt tgcacatttc tggcgtggaa ccaaagccat 7440
caccaagaag cccagcactt ttaacatgat taagtgcagt ttcttgcaaa tggttcatta 7500
ctgtttctat agatgcggta cgatcagcac caatctcata tgatctaata gaaaagtttt 7560
cacggaacac aagaccatcc tgaacaattt ttcctatccc aaaggggtca ataagcatgt 7620
caggtcgccg tggcttccaa tcaagcatca tccactgctt ttcagcggcc aagaaaattg 7680
ttgtgatagc agcaagaagc atgctccaat caggcaactg gttgataaaa gttctggggg 7740
gaggcgaagg tagatcatca tcatgcttga agctttcttt agatgtaaca actgtggttc 7800
cattaatttt cgaaggggct tgcgcctttg ccttcaagcc accagaagac gcagatttgg 7860
attttagtcc tccaaggttt gcaggcccac caccaagttt gctgcctgct ccaccagagt 7920
ccggcgaggg tgaagtaaca gggaaaaatg atgaagtagc agctgttgcc acacgcgttt 7980
aaaatattgg cttgattgca ttggttgcct ccattgcatg gtaatgtggc attgtagaga 8040
agagatggtg agccacatga gtatcagtta tgtgatgaaa caccttgttc agaatcccat 8100
aatctctgtc catagttgcc aaagctccct tcagccagtc ccattctgat gaatcgtaat 8160
gaggcaaggc aaagtgtgtg tgctgcaaat atgtgatagt cacaagaaaa ccgttcacaa 8220
tgagcaaagg caccccataa acacatagca gccaaaccaa ccctttcagg gttgcaacac 8280
ggtagagaga gtaagtcaca gaaaacaaag caacatcaga gacatagatc agaagcctct 8340
cacggttaga atatatggga gcataagggt ggtagtggct tgcaaaacta tcatagggtc 8400
taccagagac attgaaggct aaatacatag gccaccctat tgtgagtgtg acgagaagag 8460
aaacagccct tcctagaggg ttgtttaagt acttggaaaa ccatgcaact ttggattttg 8520
gttttgggac aaacacttca tcacgagatc ttagtcaagg gaacctgtgt tggagtgatg 8580
gcggcgatgg cttattttcc atgagaaata agggactaaa agtgttgagt gaagggtcaa 8640
acccacaaca tcatcaaccc attggtactt gctgaaggca tggtgaccac actcgtgagc 8700
aatcacccac acaccagtga gaaggcaacc ttggagaacc caatagattg gccatgcaat 8760
gagggaaaag ggttgaggaa ggaggtggaa gtaggtggtg gcaatgtaga aaatgaaggc 8820
aaatgaaagg tcataaacaa cataggagaa tgaagtgagg agggagcgct gaaagcagtg 8880
tggtggaatt gctttcttga gttggccaac agtgaatggt ggctttgtgt ttggaaccct 8940
tgagagaggc ttcttccctt gaacttccac tttggccaca cgacctctac ctcccattgt 9000
tgtttccttt gctagaccta gttaattagg taccgc 9036
<210>124
<211>11615
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1267
<220>
<221>misc_feature
<222>(6454)..(6454)
<223>n a,c,g,或t
<400>124
ggagatccaa gcttggcgcg ccgttctata gtgtcaccta aatcgtatgt gtatgataca 60
taaggttatg tattaattgt agccgcgttc taacgacaat atgtccatat ggtgcactct 120
cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc 180
tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt 240
ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa 300
gggcctcgtg atacgcctat ttttataggt taatgtcatg accaaaatcc cttaacgtga 360
gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 420
tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 480
ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 540
gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc 600
tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 660
cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 720
gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 780
actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc 840
ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 900
gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 960
atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 1020
tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 1080
tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg 1140
aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc 1200
gcctctcccc gcgcgttggc cgattcatta atgcaggttg atcgattcga catcgatcta 1260
gtaacataga tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat attttgtttt 1320
ctatcgcgta ttaaatgtat aattgcggga ctctaatcat aaaaacccat ctcataaata 1380
acgtcatgca ttacatgtta attattacat gcttaacgta attcaacaga aattatatga 1440
taatcatcgc aagaccggca acaggattca atcttaagaa actttattgc caaatgtttg 1500
aacgatctgc ttcgacgcac tccttcttta ggtacctcac tattcctttg ccctcggacg 1560
agtgctgggg cgtcggtttc cactatcggc gagtacttct acacagccat cggtccagac 1620
ggccgcgctt ctgcgggcga tttgtgtacg cccgacagtc ccggctccgg atcggacgat 1680
tgcgtcgcat cgaccctgcg cccaagctgc atcatcgaaa ttgccgtcaa ccaagctctg 1740
atagagttgg tcaagaccaa tgcggagcat atacgcccgg agccgcggcg atcctgcaag 1800
ctccggatgc ctccgctcga agtagcgcgt ctgctgctcc atacaagcca accacggcct 1860
ccagaagaag atgttggcga cctcgtattg ggaatccccg aacatcgcct cgctccagtc 1920
aatgaccgct gttatgcggc cattgtccgt caggacattg ttggagccga aatccgcgtg 1980
cacgaggtgc cggacttcgg ggcagtcctc ggcccaaagc atcagctcat cgagagcctg 2040
cgcgacggac gcactgacgg tgtcgtccat cacagtttgc cagtgataca catggggatc 2100
agcaatcgcg catatgaaat cacgccatgt agtgtattga ccgattcctt gcggtccgaa 2160
tgggccgaac ccgctcgtct ggctaagatc ggccgcagcg atcgcatcca tggcctccgc 2220
gaccggctgc agaacagcgg gcagttcggt ttcaggcagg tcttgcaacg tgacaccctg 2280
tgcacggcgg gagatgcaat aggtcaggct ctcgctgaat tccccaatgt caagcacttc 2340
cggaatcggg agcgcggccg atgcaaagtg ccgataaaca taacgatctt tgtagaaacc 2400
atcggcgcag ctatttaccc gcaggacata tccacgccct cctacatcga agctgaaagc 2460
acgagattct tcgccctccg agagctgcat caggtcggag acgctgtcga acttttcgat 2520
cagaaacttc tcgacagacg tcgcggtgag ttcaggcttt ttcatggttt aataagaaga 2580
gaaaagagtt cttttgttat ggctgaagta atagagaaat gagctcgagc gtgtcctctc 2640
caaatgaaat gaacttcctt atatagagga agggtcttgc gaaggatagt gggattgtgc 2700
gtcatccctt acgtcagtgg agatgtcaca tcaatccact tgctttgaag acgtggttgg 2760
aacgtcttct ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc 2820
ggcagaggca tcttgaatga tagcctttcc tttatcgcaa tgatggcatt tgtaggagcc 2880
accttccttt tctactgtcc tttcgatgaa gtgacagata gctgggcaat ggaatccgag 2940
gaggtttccc gaaattatcc tttgttgaaa agtctcaata gccctttggt cttctgagac 3000
tgtatctttg acatttttgg agtagaccag agtgtcgtgc tccaccatgt tgacgaagat 3060
tttcttcttg tcattgagtc gtaaaagact ctgtatgaac tgttcgccag tcttcacggc 3120
gagttctgtt agatcctcga tttgaatctt agactccatg catggcctta gattcagtag 3180
gaactacctt tttagagact ccaatctcta ttacttgcct tggtttatga agcaagcctt 3240
gaatcgtcca tactggaata gtacttctga tcttgagaaa tatgtctttc tctgtgttct 3300
tgatgcaatt agtcctgaat cttttgactg catctttaac cttcttggga aggtatttga 3360
tctcctggag attgttactc gggtagatcg tcttgatgag acctgctgcg taggcctctc 3420
taaccatctg tgggtcagca ttctttctga aattgaagag gctaaccttc tcattatcag 3480
tggtgaacat agtgtcgtca ccttcacctt cgaacttcct tcctagatcg taaagataga 3540
ggaaatcgtc cattgtaatc tccggggcaa aggagatctc ttttggggct ggatcactgc 3600
tgggcctttt ggttcctagc gtgagccagt gggctttttg ctttggtggg cttgttaggg 3660
ccttagcaaa gctcttgggc ttgagttgag cttctccttt ggggatgaag ttcaacctgt 3720
ctgtttgctg acttgttgtg tacgcgtcag ctgctgctct tgcctctgta atagtggcaa 3780
atttcttgtg tgcaactccg ggaacgccgt ttgttgccgc ctttgtacaa ccccagtcat 3840
cgtatatacc ggcatgtgga ccgttataca caacgtagta gttgatatga gggtgttgaa 3900
tacccgattc tgctctgaga ggagcaactg tgctgttaag ctcagatttt tgtgggattg 3960
gaattggatc gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg 4020
gtttccctct agaaataatt ttgtttaact ttaagaagga gatataccca tggaaaagcc 4080
tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga 4140
cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg 4200
tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta 4260
tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag 4320
cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc 4380
tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gctatggatg cgatcgctgc 4440
ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata 4500
cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac 4560
tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg 4620
ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt 4680
cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga 4740
ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca 4800
gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc 4860
gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga 4920
tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt 4980
cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt 5040
actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagtgagg 5100
tacagcttgg atcgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc 5160
caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt 5220
tttgctgaaa ggaggaacta tatccggatg atcgggcgcg ccgtcgacgg atccgtacgt 5280
cctcgaagag aagggttaat aacacatttt ttaacatttt taacacaaat tttagttatt 5340
taaaaattta ttaaaaaatt taaaataaga agaggaactc tttaaataaa tctaacttac 5400
aaaatttatg atttttaata agttttcacc aataaaaaat gtcataaaaa tatgttaaaa 5460
agtatattat caatattctc tttatgataa ataaaaagaa aaaaaaaata aaagttaagt 5520
gaaaatgaga ttgaagtgac tttaggtgtg tataaatata tcaaccccgc caacaattta 5580
tttaatccaa atatattgaa gtatattatt ccatagcctt tatttattta tatatttatt 5640
atataaaagc tttatttgtt ctaggttgtt catgaaatat ttttttggtt ttatctccgt 5700
tgtaagaaaa tcatgtgctt tgtgtcgcca ctcactattg cagctttttc atgcattggt 5760
cagattgacg gttgattgta tttttgtttt ttatggtttt gtgttatgac ttaagtcttc 5820
atctctttat ctcttcatca ggtttgatgg ttacctaata tggtccatgg gtacatgcat 5880
ggttaaatta ggtggccaac tttgttgtga acgatagaat tttttttata ttaagtaaac 5940
tatttttata ttatgaaata ataataaaaa aaatatttta tcattattaa caaaatcata 6000
ttagttaatt tgttaactct ataataaaag aaatactgta acattcacat tacatggtaa 6060
catctttcca ccctttcatt tgttttttgt ttgatgactt tttttcttgt ttaaatttat 6120
ttcccttctt ttaaatttgg aatacattat catcatatat aaactaaaat actaaaaaca 6180
ggattacaca aatgataaat aataacacaa atatttataa atctagctgc aatatattta 6240
aactagctat atcgatattg taaaataaaa ctagctgcat tgatactgat aaaaaaatat 6300
catgtgcttt ctggactgat gatgcagtat acttttgaca ttgcctttat tttatttttc 6360
agaaaagctt tcttagttct gggttcttca ttatttgttt cccatctcca ttgtgaattg 6420
aatcatttgc ttcgtgtcac aaatacaatt tagntaggta catgcattgg tcagattcac 6480
ggtttattat gtcatgactt aagttcatgg tagtacatta cctgccacgc atgcattata 6540
ttggttagat ttgataggca aatttggttg tcaacaatat aaatataaat aatgttttta 6600
tattacgaaa taacagtgat caaaacaaac agttttatct ttattaacaa gattttgttt 6660
ttgtttgatg acgtttttta atgtttacgc tttccccctt cttttgaatt tagaacactt 6720
tatcatcata aaatcaaata ctaaaaaaat tacatatttc ataaataata acacaaatat 6780
ttttaaaaaa tctgaaataa taatgaacaa tattacatat tatcacgaaa attcattaat 6840
aaaaatatta tataaataaa atgtaatagt agttatatgt aggaaaaaag tactgcacgc 6900
ataatatata caaaaagatt aaaatgaact attataaata ataacactaa attaatggtg 6960
aatcatatca aaataatgaa aaagtaaata aaatttgtaa ttaacttcta tatgtattac 7020
acacacaaat aataaataat agtaaaaaaa attatgataa atatttacca tctcataaga 7080
tatttaaaat aatgataaaa atatagatta ttttttatgc aactagctag ccaaaaagag 7140
aacacgggta tatataaaaa gagtaccttt aaattctact gtacttcctt tattcctgac 7200
gtttttatat caagtggaca tacgtgaaga ttttaattat cagtctaaat atttcattag 7260
cacttaatac ttttctgttt tattcctatc ctataagtag tcccgattct cccaacattg 7320
cttattcaca caactaacta agaaagtctt ccatagcccc ccaagcggcc gcggtaccta 7380
attaactagg tctagcaaag gaaacaacaa tgggaggtag aggtcgtgtg gccaaagtgg 7440
aagttcaagg gaagaagcct ctctcaaggg ttccaaacac aaagccacca ttcactgttg 7500
gccaactcaa gaaagcaatt ccaccacact gctttcagcg ctccctcctc acttcattct 7560
cctatgttgt ttatgacctt tcatttgcct tcattttcta cattgccacc acctacttcc 7620
acctccttcc tcaacccttt tccctcattg catggccaat ctattgggtt ctccaaggtt 7680
gccttctcac tggtgtgtgg gtgattgctc acgagtgtgg tcaccatgcc ttcagcaagt 7740
accaatgggt tgatgatgtt gtgggtttga cccttcactc aacactttta gtcccttatt 7800
tctcatggaa aataagccat cgccgccatc actccaacac aggttccctt gactaagatc 7860
tcgtgatgaa gtgtttgtcc caaaaccaaa atccaaagtt gcatggtttt ccaagtactt 7920
aaacaaccct ctaggaaggg ctgtttctct tctcgtcaca ctcacaatag ggtggcctat 7980
gtatttagcc ttcaatgtct ctggtagacc ctatgatagt tttgcaagcc actaccaccc 8040
ttatgctccc atatattcta accgtgagag gcttctgatc tatgtctctg atgttgcttt 8100
gttttctgtg acttactctc tctaccgtgt tgcaaccctg aaagggttgg tttggctgct 8160
atgtgtttat ggggtgcctt tgctcattgt gaacggtttt cttgtgacta tcacatattt 8220
gcagcacaca cactttgcct tgcctcatta cgattcatca gaatgggact ggctgaaggg 8280
agctttggca actatggaca gagattatgg gattctgaac aaggtgtttc atcacataac 8340
tgatactcat gtggctcacc atctcttctc tacaatgcca cattaccatg caatggaggc 8400
aaccaatgca atcaagccaa tattttaaac gcgtgtggca acagctgcta cttcatcatt 8460
tttccctgtt acttcaccct cgccggactc tggtggagca ggcagcaaac ttggtggtgg 8520
gcctgcaaac cttggaggac taaaatccaa atctgcgtct tctggtggct tgaaggcaaa 8580
ggcgcaagcc ccttcgaaaa ttaatggaac cacagttgtt acatctaaag aaagcttcaa 8640
gcatgatgat gatctacctt cgcctccccc cagaactttt atcaaccagt tgcctgattg 8700
gagcatgctt cttgctgcta tcacaacaat tttcttggcc gctgaaaagc agtggatgat 8760
gcttgattgg aagccacggc gacctgacat gcttattgac ccctttggga taggaaaaat 8820
tgttcaggat ggtcttgtgt tccgtgaaaa cttttctatt agatcatatg agattggtgc 8880
tgatcgtacc gcatctatag aaacagtaat gaaccatttg caagaaactg cacttaatca 8940
tgttaaaagt gctgggcttc ttggtgatgg ctttggttcc acgccagaaa tgtgcaaaaa 9000
gaacttgata tgggtggtta ctcggatgca ggttgtggtg gaacgctatc ctacatgggg 9060
tgacatagtt caagtggaca cttgggtttc tggatcaggg aagaatggta tgcgtcgtga 9120
ttggctttta cgtgactcca aaacttaaga attcggtgaa atcttgacaa gagcttccag 9180
tgtttgggtc atgatgaata agctaacacg gaggctgtct aaaattccag aagaagtcag 9240
acaggagata ggatcttatt ttgtggattc tgatccaatt ctggaagagg ataacagaaa 9300
actgactaaa cttgacgacg acacagcgga ttatattcgt accggtttaa gtcctaggtg 9360
gagtgatcta gatatcaatc agcatgtcaa caatgtgaag tacattggct ggattctgga 9420
gagtgctcca cagccaatct tggagagtca tgagctttct tccatgactt tagagtatag 9480
gagagagtgt ggtagggaca gtgtgctgga ttccctgact gctgtatctg gggccgacat 9540
gggcaatcta gctcacagcg ggcatgttga gtgcaagcat ttgcttcgac tggaaaatgg 9600
tgctgagatt gtgactaatt aatcacaatt cttaagtttt ggagtcacgt aaaagccaat 9660
cacgacgcat accattcttc cctgatccag aaacccaagt gtccacttga actatgtcac 9720
cccatgtagg atagcgttcc accacaacct gcatccgagt aaccacccat atcaagttct 9780
ttttgcacat ttctggcgtg gaaccaaagc catcaccaag aagcccagca cttttaacat 9840
gattaagtgc agtttcttgc aaatggttca ttactgtttc tatagatgcg gtacgatcag 9900
caccaatctc atatgatcta atagaaaagt tttcacggaa cacaagacca tcctgaacaa 9960
tttttcctat cccaaagggg tcaataagca tgtcaggtcg ccgtggcttc caatcaagca 10020
tcatccactg cttttcagcg gccaagaaaa ttgttgtgat agcagcaaga agcatgctcc 10080
aatcaggcaa ctggttgata aaagttctgg ggggaggcga aggtagatca tcatcatgct 10140
tgaagctttc tttagatgta acaactgtgg ttccattaat tttcgaaggg gcttgcgcct 10200
ttgccttcaa gccaccagaa gacgcagatt tggattttag tcctccaagg tttgcaggcc 10260
caccaccaag tttgctgcct gctccaccag agtccggcga gggtgaagta acagggaaaa 10320
atgatgaagt agcagctgtt gccacacgcg tttaaaatat tggcttgatt gcattggttg 10380
cctccattgc atggtaatgt ggcattgtag agaagagatg gtgagccaca tgagtatcag 10440
ttatgtgatg aaacaccttg ttcagaatcc cataatctct gtccatagtt gccaaagctc 10500
ccttcagcca gtcccattct gatgaatcgt aatgaggcaa ggcaaagtgt gtgtgctgca 10560
aatatgtgat agtcacaaga aaaccgttca caatgagcaa aggcacccca taaacacata 10620
gcagccaaac caaccctttc agggttgcaa cacggtagag agagtaagtc acagaaaaca 10680
aagcaacatc agagacatag atcagaagcc tctcacggtt agaatatatg ggagcataag 10740
ggtggtagtg gcttgcaaaa ctatcatagg gtctaccaga gacattgaag gctaaataca 10800
taggccaccc tattgtgagt gtgacgagaa gagaaacagc ccttcctaga gggttgttta 10860
agtacttgga aaaccatgca actttggatt ttggttttgg gacaaacact tcatcacgag 10920
atcttagtca agggaacctg tgttggagtg atggcggcga tggcttattt tccatgagaa 10980
ataagggact aaaagtgttg agtgaagggt caaacccaca acatcatcaa cccattggta 11040
cttgctgaag gcatggtgac cacactcgtg agcaatcacc cacacaccag tgagaaggca 11100
accttggaga acccaataga ttggccatgc aatgagggaa aagggttgag gaaggaggtg 11160
gaagtaggtg gtggcaatgt agaaaatgaa ggcaaatgaa aggtcataaa caacatagga 11220
gaatgaagtg aggagggagc gctgaaagca gtgtggtgga attgctttct tgagttggcc 11280
aacagtgaat ggtggctttg tgtttggaac ccttgagaga ggcttcttcc cttgaacttc 11340
cactttggcc acacgacctc tacctcccat tgttgtttcc tttgctagac ctagttaatt 11400
aggtaccgcg gccgcgacac aagtgtgaga gtactaaata aatgctttgg ttgtacgaaa 11460
tcattacact aaataaaata atcaaagctt atatatgcct tccgctaagg ccgaatgcaa 11520
agaaattggt tctttctcgt tatcttttgc cacttttact agtacgtatt aattactact 11580
taatcatctt tgtttacggc tcattatatc ctgca 11615
<210>125
<211>5252
<212>DNA
<213>人工序列
<220>
<223>质粒pKR457
<220>
<221>misc_feature
<222>(3872)..(3872)
<223>n为a,c,g,或t
<400>125
gtacgtgggc ggatcccccg ggctgcagga attcactggc cgtcgtttta caacgtcgtg 60
actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca 120
gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga 180
atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc 240
gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac 300
acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca 360
gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga 420
aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa 480
taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 540
gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 600
tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 660
ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 720
taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 780
gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta 840
aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc 900
gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 960
ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 1020
ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 1080
acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 1140
taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 1200
tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 1260
cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 1320
ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 1380
gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 1440
gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 1500
aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 1560
aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 1620
actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 1680
gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 1740
atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 1800
atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 1860
ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 1920
gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 1980
cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 2040
tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 2100
cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 2160
ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 2220
gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 2280
tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 2340
ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc 2400
gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg 2460
cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca 2520
gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact 2580
ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa 2640
acagctatga ccatgattac gccaagcttg catgcctgca ggtcgactcg acgtacgtcc 2700
tcgaagagaa gggttaataa cacatttttt aacattttta acacaaattt tagttattta 2760
aaaatttatt aaaaaattta aaataagaag aggaactctt taaataaatc taacttacaa 2820
aatttatgat ttttaataag ttttcaccaa taaaaaatgt cataaaaata tgttaaaaag 2880
tatattatca atattctctt tatgataaat aaaaagaaaa aaaaaataaa agttaagtga 2940
aaatgagatt gaagtgactt taggtgtgta taaatatatc aaccccgcca acaatttatt 3000
taatccaaat atattgaagt atattattcc atagccttta tttatttata tatttattat 3060
ataaaagctt tatttgttct aggttgttca tgaaatattt ttttggtttt atctccgttg 3120
taagaaaatc atgtgctttg tgtcgccact cactattgca gctttttcat gcattggtca 3180
gattgacggt tgattgtatt tttgtttttt atggttttgt gttatgactt aagtcttcat 3240
ctctttatct cttcatcagg tttgatggtt acctaatatg gtccatgggt acatgcatgg 3300
ttaaattagg tggccaactt tgttgtgaac gatagaattt tttttatatt aagtaaacta 3360
tttttatatt atgaaataat aataaaaaaa atattttatc attattaaca aaatcatatt 3420
agttaatttg ttaactctat aataaaagaa atactgtaac attcacatta catggtaaca 3480
tctttccacc ctttcatttg ttttttgttt gatgactttt tttcttgttt aaatttattt 3540
cccttctttt aaatttggaa tacattatca tcatatataa actaaaatac taaaaacagg 3600
attacacaaa tgataaataa taacacaaat atttataaat ctagctgcaa tatatttaaa 3660
ctagctatat cgatattgta aaataaaact agctgcattg atactgataa aaaaatatca 3720
tgtgctttct ggactgatga tgcagtatac ttttgacatt gcctttattt tatttttcag 3780
aaaagctttc ttagttctgg gttcttcatt atttgtttcc catctccatt gtgaattgaa 3840
tcatttgctt cgtgtcacaa atacaattta gntaggtaca tgcattggtc agattcacgg 3900
tttattatgt catgacttaa gttcatggta gtacattacc tgccacgcat gcattatatt 3960
ggttagattt gataggcaaa tttggttgtc aacaatataa atataaataa tgtttttata 4020
ttacgaaata acagtgatca aaacaaacag ttttatcttt attaacaaga ttttgttttt 4080
gtttgatgac gttttttaat gtttacgctt tcccccttct tttgaattta gaacacttta 4140
tcatcataaa atcaaatact aaaaaaatta catatttcat aaataataac acaaatattt 4200
ttaaaaaatc tgaaataata atgaacaata ttacatatta tcacgaaaat tcattaataa 4260
aaatattata taaataaaat gtaatagtag ttatatgtag gaaaaaagta ctgcacgcat 4320
aatatataca aaaagattaa aatgaactat tataaataat aacactaaat taatggtgaa 4380
tcatatcaaa ataatgaaaa agtaaataaa atttgtaatt aacttctata tgtattacac 4440
acacaaataa taaataatag taaaaaaaat tatgataaat atttaccatc tcataagata 4500
tttaaaataa tgataaaaat atagattatt ttttatgcaa ctagctagcc aaaaagagaa 4560
cacgggtata tataaaaaga gtacctttaa attctactgt acttccttta ttcctgacgt 4620
ttttatatca agtggacata cgtgaagatt ttaattatca gtctaaatat ttcattagca 4680
cttaatactt ttctgtttta ttcctatcct ataagtagtc ccgattctcc caacattgct 4740
tattcacaca actaactaag aaagtcttcc atagcccccc aagcggccgc gacacaagtg 4800
tgagagtact aaataaatgc tttggttgta cgaaatcatt acactaaata aaataatcaa 4860
agcttatata tgccttccgc taaggccgaa tgcaaagaaa ttggttcttt ctcgttatct 4920
tttgccactt ttactagtac gtattaatta ctacttaatc atctttgttt acggctcatt 4980
atatccggtc tagaggatcc aaggccgcga agttaaaagc aatgttgtca cttgtcgtac 5040
taacacatga tgtgatagtt tatgctagct agctataaca taagctgtct ctgagtgtgt 5100
tgtatattaa taaagatcat cactggtgaa tggtgatcgt gtacgtaccc tacttagtag 5160
gcaatggaag cacttagagt gtgctttgtg catggccttg cctctgtttt gagacttttg 5220
taatgttttc gagtttaaat ctttgccttt gc 5252
<210>126
<211>9295
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1264
<220>
<221>misc_feature
<222>(4340)..(4340)
<223>n为a,c,g,或t
<400>126
ggccgcgaca caagtgtgag agtactaaat aaatgctttg gttgtacgaa atcattacac 60
taaataaaat aatcaaagct tatatatgcc ttccgctaag gccgaatgca aagaaattgg 120
ttctttctcg ttatcttttg ccacttttac tagtacgtat taattactac ttaatcatct 180
ttgtttacgg ctcattatat ccggtctaga ggatccaagg ccgcgaagtt aaaagcaatg 240
ttgtcacttg tcgtactaac acatgatgtg atagtttatg ctagctagct ataacataag 300
ctgtctctga gtgtgttgta tattaataaa gatcatcact ggtgaatggt gatcgtgtac 360
gtaccctact tagtaggcaa tggaagcact tagagtgtgc tttgtgcatg gccttgcctc 420
tgttttgaga cttttgtaat gttttcgagt ttaaatcttt gcctttgcgt acgtgggcgg 480
atcccccggg ctgcaggaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc 540
ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata 600
gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc 660
gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca 720
ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac 780
ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga 840
ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac 900
gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt 960
agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 1020
aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 1080
attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 1140
cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 1200
aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 1260
ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 1320
gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact 1380
attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 1440
tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 1500
tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 1560
atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 1620
agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 1680
aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 1740
caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 1800
ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 1860
gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 1920
tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 1980
atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 2040
tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 2100
accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 2160
gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 2220
caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc 2280
tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 2340
ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 2400
tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 2460
gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 2520
tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 2580
gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 2640
gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 2700
ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 2760
ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 2820
ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag 2880
tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga 2940
ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg 3000
caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg 3060
ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc 3120
atgattacgc caagcttgca tgcctgcagg tcgactcgac gtacgtcctc gaagagaagg 3180
gttaataaca cattttttaa catttttaac acaaatttta gttatttaaa aatttattaa 3240
aaaatttaaa ataagaagag gaactcttta aataaatcta acttacaaaa tttatgattt 3300
ttaataagtt ttcaccaata aaaaatgtca taaaaatatg ttaaaaagta tattatcaat 3360
attctcttta tgataaataa aaagaaaaaa aaaataaaag ttaagtgaaa atgagattga 3420
agtgacttta ggtgtgtata aatatatcaa ccccgccaac aatttattta atccaaatat 3480
attgaagtat attattccat agcctttatt tatttatata tttattatat aaaagcttta 3540
tttgttctag gttgttcatg aaatattttt ttggttttat ctccgttgta agaaaatcat 3600
gtgctttgtg tcgccactca ctattgcagc tttttcatgc attggtcaga ttgacggttg 3660
attgtatttt tgttttttat ggttttgtgt tatgacttaa gtcttcatct ctttatctct 3720
tcatcaggtt tgatggttac ctaatatggt ccatgggtac atgcatggtt aaattaggtg 3780
gccaactttg ttgtgaacga tagaattttt tttatattaa gtaaactatt tttatattat 3840
gaaataataa taaaaaaaat attttatcat tattaacaaa atcatattag ttaatttgtt 3900
aactctataa taaaagaaat actgtaacat tcacattaca tggtaacatc tttccaccct 3960
ttcatttgtt ttttgtttga tgactttttt tcttgtttaa atttatttcc cttcttttaa 4020
atttggaata cattatcatc atatataaac taaaatacta aaaacaggat tacacaaatg 4080
ataaataata acacaaatat ttataaatct agctgcaata tatttaaact agctatatcg 4140
atattgtaaa ataaaactag ctgcattgat actgataaaa aaatatcatg tgctttctgg 4200
actgatgatg cagtatactt ttgacattgc ctttatttta tttttcagaa aagctttctt 4260
agttctgggt tcttcattat ttgtttccca tctccattgt gaattgaatc atttgcttcg 4320
tgtcacaaat acaatttagn taggtacatg cattggtcag attcacggtt tattatgtca 4380
tgacttaagt tcatggtagt acattacctg ccacgcatgc attatattgg ttagatttga 4440
taggcaaatt tggttgtcaa caatataaat ataaataatg tttttatatt acgaaataac 4500
agtgatcaaa acaaacagtt ttatctttat taacaagatt ttgtttttgt ttgatgacgt 4560
tttttaatgt ttacgctttc ccccttcttt tgaatttaga acactttatc atcataaaat 4620
caaatactaa aaaaattaca tatttcataa ataataacac aaatattttt aaaaaatctg 4680
aaataataat gaacaatatt acatattatc acgaaaattc attaataaaa atattatata 4740
aataaaatgt aatagtagtt atatgtagga aaaaagtact gcacgcataa tatatacaaa 4800
aagattaaaa tgaactatta taaataataa cactaaatta atggtgaatc atatcaaaat 4860
aatgaaaaag taaataaaat ttgtaattaa cttctatatg tattacacac acaaataata 4920
aataatagta aaaaaaatta tgataaatat ttaccatctc ataagatatt taaaataatg 4980
ataaaaatat agattatttt ttatgcaact agctagccaa aaagagaaca cgggtatata 5040
taaaaagagt acctttaaat tctactgtac ttcctttatt cctgacgttt ttatatcaag 5100
tggacatacg tgaagatttt aattatcagt ctaaatattt cattagcact taatactttt 5160
ctgttttatt cctatcctat aagtagtccc gattctccca acattgctta ttcacacaac 5220
taactaagaa agtcttccat agccccccaa gcggccgcgg tacctaatta actaggtcta 5280
gcaaaggaaa caacaatggg aggtagaggt cgtgtggcca aagtggaagt tcaagggaag 5340
aagcctctct caagggttcc aaacacaaag ccaccattca ctgttggcca actcaagaaa 5400
gcaattccac cacactgctt tcagcgctcc ctcctcactt cattctccta tgttgtttat 5460
gacctttcat ttgccttcat tttctacatt gccaccacct acttccacct ccttcctcaa 5520
cccttttccc tcattgcatg gccaatctat tgggttctcc aaggttgcct tctcactggt 5580
gtgtgggtga ttgctcacga gtgtggtcac catgccttca gcaagtacca atgggttgat 5640
gatgttgtgg gtttgaccct tcactcaaca cttttagtcc cttatttctc atggaaaata 5700
agccatcgcc gccatcactc caacacaggt tcccttgact aagatctcgt gatgaagtgt 5760
ttgtcccaaa accaaaatcc aaagttgcat ggttttccaa gtacttaaac aaccctctag 5820
gaagggctgt ttctcttctc gtcacactca caatagggtg gcctatgtat ttagccttca 5880
atgtctctgg tagaccctat gatagttttg caagccacta ccacccttat gctcccatat 5940
attctaaccg tgagaggctt ctgatctatg tctctgatgt tgctttgttt tctgtgactt 6000
actctctcta ccgtgttgca accctgaaag ggttggtttg gctgctatgt gtttatgggg 6060
tgcctttgct cattgtgaac ggttttcttg tgactatcac atatttgcag cacacacact 6120
ttgccttgcc tcattacgat tcatcagaat gggactggct gaagggagct ttggcaacta 6180
tggacagaga ttatgggatt ctgaacaagg tgtttcatca cataactgat actcatgtgg 6240
ctcaccatct cttctctaca atgccacatt accatgcaat ggaggcaacc aatgcaatca 6300
agccaatatt ttaaacgcgt gtggcaacag ctgctacttc atcatttttc cctgttactt 6360
caccctcgcc ggactctggt ggagcaggca gcaaacttgg tggtgggcct gcaaaccttg 6420
gaggactaaa atccaaatct gcgtcttctg gtggcttgaa ggcaaaggcg caagcccctt 6480
cgaaaattaa tggaaccaca gttgttacat ctaaagaaag cttcaagcat gatgatgatc 6540
taccttcgcc tccccccaga acttttatca accagttgcc tgattggagc atgcttcttg 6600
ctgctatcac aacaattttc ttggccgctg aaaagcagtg gatgatgctt gattggaagc 6660
cacggcgacc tgacatgctt attgacccct ttgggatagg aaaaattgtt caggatggtc 6720
ttgtgttccg tgaaaacttt tctattagat catatgagat tggtgctgat cgtaccgcat 6780
ctatagaaac agtaatgaac catttgcaag aaactgcact taatcatgtt aaaagtgctg 6840
ggcttcttgg tgatggcttt ggttccacgc cagaaatgtg caaaaagaac ttgatatggg 6900
tggttactcg gatgcaggtt gtggtggaac gctatcctac atggggtgac atagttcaag 6960
tggacacttg ggtttctgga tcagggaaga atggtatgcg tcgtgattgg cttttacgtg 7020
actccaaaac ttaagaattc ggtgaaatct tgacaagagc ttccagtgtt tgggtcatga 7080
tgaataagct aacacggagg ctgtctaaaa ttccagaaga agtcagacag gagataggat 7140
cttattttgt ggattctgat ccaattctgg aagaggataa cagaaaactg actaaacttg 7200
acgacgacac agcggattat attcgtaccg gtttaagtcc taggtggagt gatctagata 7260
tcaatcagca tgtcaacaat gtgaagtaca ttggctggat tctggagagt gctccacagc 7320
caatcttgga gagtcatgag ctttcttcca tgactttaga gtataggaga gagtgtggta 7380
gggacagtgt gctggattcc ctgactgctg tatctggggc cgacatgggc aatctagctc 7440
acagcgggca tgttgagtgc aagcatttgc ttcgactgga aaatggtgct gagattgtga 7500
ctaattaatc acaattctta agttttggag tcacgtaaaa gccaatcacg acgcatacca 7560
ttcttccctg atccagaaac ccaagtgtcc acttgaacta tgtcacccca tgtaggatag 7620
cgttccacca caacctgcat ccgagtaacc acccatatca agttcttttt gcacatttct 7680
ggcgtggaac caaagccatc accaagaagc ccagcacttt taacatgatt aagtgcagtt 7740
tcttgcaaat ggttcattac tgtttctata gatgcggtac gatcagcacc aatctcatat 7800
gatctaatag aaaagttttc acggaacaca agaccatcct gaacaatttt tcctatccca 7860
aaggggtcaa taagcatgtc aggtcgccgt ggcttccaat caagcatcat ccactgcttt 7920
tcagcggcca agaaaattgt tgtgatagca gcaagaagca tgctccaatc aggcaactgg 7980
ttgataaaag ttctgggggg aggcgaaggt agatcatcat catgcttgaa gctttcttta 8040
gatgtaacaa ctgtggttcc attaattttc gaaggggctt gcgcctttgc cttcaagcca 8100
ccagaagacg cagatttgga ttttagtcct ccaaggtttg caggcccacc accaagtttg 8160
ctgcctgctc caccagagtc cggcgagggt gaagtaacag ggaaaaatga tgaagtagca 8220
gctgttgcca cacgcgttta aaatattggc ttgattgcat tggttgcctc cattgcatgg 8280
taatgtggca ttgtagagaa gagatggtga gccacatgag tatcagttat gtgatgaaac 8340
accttgttca gaatcccata atctctgtcc atagttgcca aagctccctt cagccagtcc 8400
cattctgatg aatcgtaatg aggcaaggca aagtgtgtgt gctgcaaata tgtgatagtc 8460
acaagaaaac cgttcacaat gagcaaaggc accccataaa cacatagcag ccaaaccaac 8520
cctttcaggg ttgcaacacg gtagagagag taagtcacag aaaacaaagc aacatcagag 8580
acatagatca gaagcctctc acggttagaa tatatgggag cataagggtg gtagtggctt 8640
gcaaaactat catagggtct accagagaca ttgaaggcta aatacatagg ccaccctatt 8700
gtgagtgtga cgagaagaga aacagccctt cctagagggt tgtttaagta cttggaaaac 8760
catgcaactt tggattttgg ttttgggaca aacacttcat cacgagatct tagtcaaggg 8820
aacctgtgtt ggagtgatgg cggcgatggc ttattttcca tgagaaataa gggactaaaa 8880
gtgttgagtg aagggtcaaa cccacaacat catcaaccca ttggtacttg ctgaaggcat 8940
ggtgaccaca ctcgtgagca atcacccaca caccagtgag aaggcaacct tggagaaccc 9000
aatagattgg ccatgcaatg agggaaaagg gttgaggaag gaggtggaag taggtggtgg 9060
caatgtagaa aatgaaggca aatgaaaggt cataaacaac ataggagaat gaagtgagga 9120
gggagcgctg aaagcagtgt ggtggaattg ctttcttgag ttggccaaca gtgaatggtg 9180
gctttgtgtt tggaaccctt gagagaggct tcttcccttg aacttccact ttggccacac 9240
gacctctacc tcccattgtt gtttcctttg ctagacctag ttaattaggt accgc 9295
<210>127
<211>2577
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1277
<400>127
agcttggatc tcctgcagga tctggccggc cggatctcgt acggatccgt cgacggcgcg 60
cccgatcatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag 120
gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt 180
cgggctttgt tagcagccgg atcgatccaa gctgtacctc actattcctt tgccctcgga 240
cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag 300
acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg 360
attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc 420
tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccgcgg cgatcctgca 480
agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc 540
ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag 600
tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg 660
tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc 720
tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga 780
tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg 840
aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catagcctcc 900
gcgaccggct gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc 960
tgtgcacggc gggagatgca ataggtcagg ctctcgctga attccccaat gtcaagcact 1020
tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa 1080
ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa 1140
gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg 1200
atcagaaact tctcgacaga cgtcgcggtg agttcaggct tttccatggg tatatctcct 1260
tcttaaagtt aaacaaaatt atttctagag ggaaaccgtt gtggtctccc tatagtgagt 1320
cgtattaatt tcgcgggatc gagatctgat caacctgcat taatgaatcg gccaacgcgc 1380
ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 1440
ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 1500
cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 1560
gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 1620
tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 1680
ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 1740
atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag 1800
gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 1860
tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 1920
cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 1980
cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 2040
tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 2100
cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 2160
cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 2220
gaacgaaaac tcacgttaag ggattttggt catgacatta acctataaaa ataggcgtat 2280
cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca 2340
gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca 2400
gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca 2460
gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata 2520
cataacctta tgtatcatac acatacgatt taggtgacac tatagaacgg cgcgcca 2577
<210>128
<211>9219
<212>DNA
<213>人工序列
<220>
<223>质粒pKR1269
<220>
<221>misc_feature
<222>(3769)..(3769)
<223>n为a,c,g,或t
<400>128
ggatctggcc ggccggatct cgtacggatc cgtcgacggc gcgcccgatc atccggatat 60
agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc 120
tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt tgttagcagc 180
cggatcgatc caagctgtac ctcactattc ctttgccctc ggacgagtgc tggggcgtcg 240
gtttccacta tcggcgagta cttctacaca gccatcggtc cagacggccg cgcttctgcg 300
ggcgatttgt gtacgcccga cagtcccggc tccggatcgg acgattgcgt cgcatcgacc 360
ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag ctctgataga gttggtcaag 420
accaatgcgg agcatatacg cccggagccg cggcgatcct gcaagctccg gatgcctccg 480
ctcgaagtag cgcgtctgct gctccataca agccaaccac ggcctccaga agaagatgtt 540
ggcgacctcg tattgggaat ccccgaacat cgcctcgctc cagtcaatga ccgctgttat 600
gcggccattg tccgtcagga cattgttgga gccgaaatcc gcgtgcacga ggtgccggac 660
ttcggggcag tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga cggacgcact 720
gacggtgtcg tccatcacag tttgccagtg atacacatgg ggatcagcaa tcgcgcatat 780
gaaatcacgc catgtagtgt attgaccgat tccttgcggt ccgaatgggc cgaacccgct 840
cgtctggcta agatcggccg cagcgatcgc atccatagcc tccgcgaccg gctgcagaac 900
agcgggcagt tcggtttcag gcaggtcttg caacgtgaca ccctgtgcac ggcgggagat 960
gcaataggtc aggctctcgc tgaattcccc aatgtcaagc acttccggaa tcgggagcgc 1020
ggccgatgca aagtgccgat aaacataacg atctttgtag aaaccatcgg cgcagctatt 1080
tacccgcagg acatatccac gccctcctac atcgaagctg aaagcacgag attcttcgcc 1140
ctccgagagc tgcatcaggt cggagacgct gtcgaacttt tcgatcagaa acttctcgac 1200
agacgtcgcg gtgagttcag gcttttccat gggtatatct ccttcttaaa gttaaacaaa 1260
attatttcta gagggaaacc gttgtggtct ccctatagtg agtcgtatta atttcgcggg 1320
atcgagatct gatcaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 1380
gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 1440
ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 1500
acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 1560
cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 1620
caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 1680
gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 1740
tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt 1800
aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 1860
ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 1920
cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 1980
tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 2040
tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 2100
ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 2160
aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 2220
aagggatttt ggtcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 2280
tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 2340
cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 2400
ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 2460
accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca 2520
tacacatacg atttaggtga cactatagaa cggcgcgcca agcttggatc tcctgcaggt 2580
cgactcgacg tacgtcctcg aagagaaggg ttaataacac attttttaac atttttaaca 2640
caaattttag ttatttaaaa atttattaaa aaatttaaaa taagaagagg aactctttaa 2700
ataaatctaa cttacaaaat ttatgatttt taataagttt tcaccaataa aaaatgtcat 2760
aaaaatatgt taaaaagtat attatcaata ttctctttat gataaataaa aagaaaaaaa 2820
aaataaaagt taagtgaaaa tgagattgaa gtgactttag gtgtgtataa atatatcaac 2880
cccgccaaca atttatttaa tccaaatata ttgaagtata ttattccata gcctttattt 2940
atttatatat ttattatata aaagctttat ttgttctagg ttgttcatga aatatttttt 3000
tggttttatc tccgttgtaa gaaaatcatg tgctttgtgt cgccactcac tattgcagct 3060
ttttcatgca ttggtcagat tgacggttga ttgtattttt gttttttatg gttttgtgtt 3120
atgacttaag tcttcatctc tttatctctt catcaggttt gatggttacc taatatggtc 3180
catgggtaca tgcatggtta aattaggtgg ccaactttgt tgtgaacgat agaatttttt 3240
ttatattaag taaactattt ttatattatg aaataataat aaaaaaaata ttttatcatt 3300
attaacaaaa tcatattagt taatttgtta actctataat aaaagaaata ctgtaacatt 3360
cacattacat ggtaacatct ttccaccctt tcatttgttt tttgtttgat gacttttttt 3420
cttgtttaaa tttatttccc ttcttttaaa tttggaatac attatcatca tatataaact 3480
aaaatactaa aaacaggatt acacaaatga taaataataa cacaaatatt tataaatcta 3540
gctgcaatat atttaaacta gctatatcga tattgtaaaa taaaactagc tgcattgata 3600
ctgataaaaa aatatcatgt gctttctgga ctgatgatgc agtatacttt tgacattgcc 3660
tttattttat ttttcagaaa agctttctta gttctgggtt cttcattatt tgtttcccat 3720
ctccattgtg aattgaatca tttgcttcgt gtcacaaata caatttagnt aggtacatgc 3780
attggtcaga ttcacggttt attatgtcat gacttaagtt catggtagta cattacctgc 3840
cacgcatgca ttatattggt tagatttgat aggcaaattt ggttgtcaac aatataaata 3900
taaataatgt ttttatatta cgaaataaca gtgatcaaaa caaacagttt tatctttatt 3960
aacaagattt tgtttttgtt tgatgacgtt ttttaatgtt tacgctttcc cccttctttt 4020
gaatttagaa cactttatca tcataaaatc aaatactaaa aaaattacat atttcataaa 4080
taataacaca aatattttta aaaaatctga aataataatg aacaatatta catattatca 4140
cgaaaattca ttaataaaaa tattatataa ataaaatgta atagtagtta tatgtaggaa 4200
aaaagtactg cacgcataat atatacaaaa agattaaaat gaactattat aaataataac 4260
actaaattaa tggtgaatca tatcaaaata atgaaaaagt aaataaaatt tgtaattaac 4320
ttctatatgt attacacaca caaataataa ataatagtaa aaaaaattat gataaatatt 4380
taccatctca taagatattt aaaataatga taaaaatata gattattttt tatgcaacta 4440
gctagccaaa aagagaacac gggtatatat aaaaagagta cctttaaatt ctactgtact 4500
tcctttattc ctgacgtttt tatatcaagt ggacatacgt gaagatttta attatcagtc 4560
taaatatttc attagcactt aatacttttc tgttttattc ctatcctata agtagtcccg 4620
attctcccaa cattgcttat tcacacaact aactaagaaa gtcttccata gccccccaag 4680
cggccgcggt acctaattaa ctaggtctag caaaggaaac aacaatggga ggtagaggtc 4740
gtgtggccaa agtggaagtt caagggaaga agcctctctc aagggttcca aacacaaagc 4800
caccattcac tgttggccaa ctcaagaaag caattccacc acactgcttt cagcgctccc 4860
tcctcacttc attctcctat gttgtttatg acctttcatt tgccttcatt ttctacattg 4920
ccaccaccta cttccacctc cttcctcaac ccttttccct cattgcatgg ccaatctatt 4980
gggttctcca aggttgcctt ctcactggtg tgtgggtgat tgctcacgag tgtggtcacc 5040
atgccttcag caagtaccaa tgggttgatg atgttgtggg tttgaccctt cactcaacac 5100
ttttagtccc ttatttctca tggaaaataa gccatcgccg ccatcactcc aacacaggtt 5160
cccttgacta agatctcgtg atgaagtgtt tgtcccaaaa ccaaaatcca aagttgcatg 5220
gttttccaag tacttaaaca accctctagg aagggctgtt tctcttctcg tcacactcac 5280
aatagggtgg cctatgtatt tagccttcaa tgtctctggt agaccctatg atagttttgc 5340
aagccactac cacccttatg ctcccatata ttctaaccgt gagaggcttc tgatctatgt 5400
ctctgatgtt gctttgtttt ctgtgactta ctctctctac cgtgttgcaa ccctgaaagg 5460
gttggtttgg ctgctatgtg tttatggggt gcctttgctc attgtgaacg gttttcttgt 5520
gactatcaca tatttgcagc acacacactt tgccttgcct cattacgatt catcagaatg 5580
ggactggctg aagggagctt tggcaactat ggacagagat tatgggattc tgaacaaggt 5640
gtttcatcac ataactgata ctcatgtggc tcaccatctc ttctctacaa tgccacatta 5700
ccatgcaatg gaggcaacca atgcaatcaa gccaatattt taaacgcgtg tggcaacagc 5760
tgctacttca tcatttttcc ctgttacttc accctcgccg gactctggtg gagcaggcag 5820
caaacttggt ggtgggcctg caaaccttgg aggactaaaa tccaaatctg cgtcttctgg 5880
tggcttgaag gcaaaggcgc aagccccttc gaaaattaat ggaaccacag ttgttacatc 5940
taaagaaagc ttcaagcatg atgatgatct accttcgcct ccccccagaa cttttatcaa 6000
ccagttgcct gattggagca tgcttcttgc tgctatcaca acaattttct tggccgctga 6060
aaagcagtgg atgatgcttg attggaagcc acggcgacct gacatgctta ttgacccctt 6120
tgggatagga aaaattgttc aggatggtct tgtgttccgt gaaaactttt ctattagatc 6180
atatgagatt ggtgctgatc gtaccgcatc tatagaaaca gtaatgaacc atttgcaaga 6240
aactgcactt aatcatgtta aaagtgctgg gcttcttggt gatggctttg gttccacgcc 6300
agaaatgtgc aaaaagaact tgatatgggt ggttactcgg atgcaggttg tggtggaacg 6360
ctatcctaca tggggtgaca tagttcaagt ggacacttgg gtttctggat cagggaagaa 6420
tggtatgcgt cgtgattggc ttttacgtga ctccaaaact taagaattcg gtgaaatctt 6480
gacaagagct tccagtgttt gggtcatgat gaataagcta acacggaggc tgtctaaaat 6540
tccagaagaa gtcagacagg agataggatc ttattttgtg gattctgatc caattctgga 6600
agaggataac agaaaactga ctaaacttga cgacgacaca gcggattata ttcgtaccgg 6660
tttaagtcct aggtggagtg atctagatat caatcagcat gtcaacaatg tgaagtacat 6720
tggctggatt ctggagagtg ctccacagcc aatcttggag agtcatgagc tttcttccat 6780
gactttagag tataggagag agtgtggtag ggacagtgtg ctggattccc tgactgctgt 6840
atctggggcc gacatgggca atctagctca cagcgggcat gttgagtgca agcatttgct 6900
tcgactggaa aatggtgctg agattgtgac taattaatca caattcttaa gttttggagt 6960
cacgtaaaag ccaatcacga cgcataccat tcttccctga tccagaaacc caagtgtcca 7020
cttgaactat gtcaccccat gtaggatagc gttccaccac aacctgcatc cgagtaacca 7080
cccatatcaa gttctttttg cacatttctg gcgtggaacc aaagccatca ccaagaagcc 7140
cagcactttt aacatgatta agtgcagttt cttgcaaatg gttcattact gtttctatag 7200
atgcggtacg atcagcacca atctcatatg atctaataga aaagttttca cggaacacaa 7260
gaccatcctg aacaattttt cctatcccaa aggggtcaat aagcatgtca ggtcgccgtg 7320
gcttccaatc aagcatcatc cactgctttt cagcggccaa gaaaattgtt gtgatagcag 7380
caagaagcat gctccaatca ggcaactggt tgataaaagt tctgggggga ggcgaaggta 7440
gatcatcatc atgcttgaag ctttctttag atgtaacaac tgtggttcca ttaattttcg 7500
aaggggcttg cgcctttgcc ttcaagccac cagaagacgc agatttggat tttagtcctc 7560
caaggtttgc aggcccacca ccaagtttgc tgcctgctcc accagagtcc ggcgagggtg 7620
aagtaacagg gaaaaatgat gaagtagcag ctgttgccac acgcgtttaa aatattggct 7680
tgattgcatt ggttgcctcc attgcatggt aatgtggcat tgtagagaag agatggtgag 7740
ccacatgagt atcagttatg tgatgaaaca ccttgttcag aatcccataa tctctgtcca 7800
tagttgccaa agctcccttc agccagtccc attctgatga atcgtaatga ggcaaggcaa 7860
agtgtgtgtg ctgcaaatat gtgatagtca caagaaaacc gttcacaatg agcaaaggca 7920
ccccataaac acatagcagc caaaccaacc ctttcagggt tgcaacacgg tagagagagt 7980
aagtcacaga aaacaaagca acatcagaga catagatcag aagcctctca cggttagaat 8040
atatgggagc ataagggtgg tagtggcttg caaaactatc atagggtcta ccagagacat 8100
tgaaggctaa atacataggc caccctattg tgagtgtgac gagaagagaa acagcccttc 8160
ctagagggtt gtttaagtac ttggaaaacc atgcaacttt ggattttggt tttgggacaa 8220
acacttcatc acgagatctt agtcaaggga acctgtgttg gagtgatggc ggcgatggct 8280
tattttccat gagaaataag ggactaaaag tgttgagtga agggtcaaac ccacaacatc 8340
atcaacccat tggtacttgc tgaaggcatg gtgaccacac tcgtgagcaa tcacccacac 8400
accagtgaga aggcaacctt ggagaaccca atagattggc catgcaatga gggaaaaggg 8460
ttgaggaagg aggtggaagt aggtggtggc aatgtagaaa atgaaggcaa atgaaaggtc 8520
ataaacaaca taggagaatg aagtgaggag ggagcgctga aagcagtgtg gtggaattgc 8580
tttcttgagt tggccaacag tgaatggtgg ctttgtgttt ggaacccttg agagaggctt 8640
cttcccttga acttccactt tggccacacg acctctacct cccattgttg tttcctttgc 8700
tagacctagt taattaggta ccgcggccgc gacacaagtg tgagagtact aaataaatgc 8760
tttggttgta cgaaatcatt acactaaata aaataatcaa agcttatata tgccttccgc 8820
taaggccgaa tgcaaagaaa ttggttcttt ctcgttatct tttgccactt ttactagtac 8880
gtattaatta ctacttaatc atctttgttt acggctcatt atatccggtc tagaggatcc 8940
aaggccgcga agttaaaagc aatgttgtca cttgtcgtac taacacatga tgtgatagtt 9000
tatgctagct agctataaca taagctgtct ctgagtgtgt tgtatattaa taaagatcat 9060
cactggtgaa tggtgatcgt gtacgtaccc tacttagtag gcaatggaag cacttagagt 9120
gtgctttgtg catggccttg cctctgtttt gagacttttg taatgttttc gagtttaaat 9180
ctttgccttt gcgtacgtgg gcggatcccc cgggctgca 9219
Claims (42)
1.转基因大豆种子,所述转基因大豆种子的总脂肪酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量相比高出至少10%。
2.权利要求1的转基因大豆种子,其中所述种子包含具有至少一种DGAT序列的重组构建体。
3.权利要求2的转基因大豆种子,其中所述DGAT序列选自DGAT1、DGAT2和DGAT1与DGAT2的组合。
4.权利要求2或3的转基因大豆种子,其中所述DGAT序列为耶氏酵母序列。
5.用于提高大豆种子的总脂肪酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量相比高出至少10%的步骤(a)中的一个或多个转化的大豆细胞。
6.权利要求5的方法,其中所述DGAT序列选自DGAT1、DGAT2和DGAT1与DGAT2的组合。
7.权利要求5或6的方法,其中所述DGAT序列为耶氏酵母序列。
8.转基因玉米谷粒,所述转基因玉米谷粒的总脂肪酸含量与非转基因的、无转基因分离子玉米谷粒的总脂肪酸含量相比高出至少10%。
9.权利要求8的转基因玉米谷粒,其中所述谷粒包含具有至少一种DGAT序列的重组构建体。
10.权利要求9的转基因玉米谷粒,其中所述DGAT序列选自DGAT1、DGAT2和DGAT1与DGAT2的组合。
11.权利要求8或9的转基因玉米谷粒,其中所述DGAT序列为耶氏酵母序列。
12.用于提高玉米谷粒的总脂肪酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个玉米谷粒;
(b)选择总脂肪酸含量与非转基因的、无转基因分离子玉米谷粒的总脂肪酸含量相比高出至少10%的步骤(a)中的一个或多个转化的玉米谷粒。
13.权利要求12的方法,其中所述DGAT序列选自DGAT1、DGAT2和DGAT1与DGAT2的组合。
14.权利要求12或13的方法,其中所述DGAT序列为耶氏酵母序列。
15.转基因大豆种子,所述转基因大豆种子的总脂肪酸含量和油酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和油酸含量相比分别高出至少10%和高出至少25%。
16.权利要求15的转基因大豆种子,其中所述种子包含具有至少一种DGAT序列的重组构建体。
17.权利要求16的转基因大豆种子,其中所述DGAT序列选自DGAT1、DGAT2和DGAT1与DGAT2的组合。
18.权利要求16或17的转基因大豆种子,其中所述DGAT序列为耶氏酵母序列。
19.用于提高大豆种子的总脂肪酸含量和油酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和油酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和油酸含量相比分别高出至少10%和高出至少25%的步骤(a)中的一个或多个转化的大豆细胞。
20.用于提高大豆种子的总脂肪酸含量并降低其亚麻酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和亚麻酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和亚麻酸含量相比分别高出至少10%和降低至少25%的上述步骤(a)中的一个或多个转化的大豆细胞。
21.用于提高大豆种子的总脂肪酸含量并降低其亚油酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和亚油酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和亚油酸含量相比分别高出至少10%和降低至少4%的步骤(a)中的一个或多个转化的大豆细胞。
22.用于提高大豆种子的总脂肪酸含量并降低其棕榈酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和棕榈酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和棕榈酸含量相比分别高出至少10%和降低至少8%的上述步骤(a)中的一个或多个转化的大豆细胞。
23.用于提高大豆种子的总脂肪酸含量和硬脂酸含量的方法,所述方法包括:
(a)用具有至少一种DGAT序列的重组构建体转化至少一个大豆细胞;
(b)选择总脂肪酸含量和硬脂酸含量与非转基因的、无转基因分离子大豆种子的总脂肪酸含量和硬脂酸含量相比分别高出至少10%和高出至少14%的步骤(a)中的一个或多个转化的大豆细胞。
24.权利要求19至23的方法,其中所述DGAT序列选自DGAT1、DGAT2和DGAT1与DGAT2的组合。
25.权利要求19至23的方法,其中所述DGAT序列为耶氏酵母序列。
26.权利要求24的方法,其中所述DGAT序列为耶氏酵母序列。
27.由权利要求19至23的方法生产的转基因大豆。
28.由权利要求24的方法生产的转基因大豆。
29.由权利要求25的方法生产的转基因大豆。
30.由权利要求26的方法生产的转基因大豆。
31.从权利要求27的转基因大豆获得的产品和/或副产品。
32.从权利要求28的转基因大豆获得的产品和/或副产品。
33.从权利要求29的转基因大豆获得的产品和/或副产品。
34.从权利要求30的转基因大豆获得的产品和/或副产品。
35.分离的多核苷酸,所述分离的多核苷酸包括:
(a)编码具有二酰基甘油酰基转移酶活性的多肽的核苷酸序列,其中所述多肽如SEQ ID:83、88或93所示;
(b)(a)中的核苷酸序列的互补序列,其中所述互补序列与所述核苷酸序列由相同数目的核苷酸组成并且100%互补。
36.包含权利要求35的分离的核酸片段的重组DNA构建体,其中所述分离的核酸片段与至少一种调控序列可操作地连接。
37.在其基因组中包含权利要求36的重组DNA构建体的细胞。
38.权利要求37的细胞,其中所述细胞为油料种子植物细胞。
39.在其基因组中包含权利要求37的重组构建体的转基因油料种子。
40.权利要求39的油料种子,其中所述油料种子选自大豆、玉米、卡诺拉、向日葵、亚麻、棉花和红花。
41.用于提高油料种子的总脂肪酸含量的方法,所述方法包括:
(a)用权利要求37的重组构建体转化至少一个油料种子细胞;
(b)选择总脂肪酸含量高于非转基因的、无转基因分离子油料种子的总脂肪酸含量的步骤(a)中的一个或多个转化的油料种子细胞。
42.权利要求41的方法,其中所述油料种子选自大豆、玉米、卡诺拉、向日葵、亚麻、棉花和红花。
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CN104903451A (zh) * | 2012-10-30 | 2015-09-09 | 农业研究有限公司 | 改良的酰基转移酶多核苷酸、多肽、及其使用方法 |
CN104903451B (zh) * | 2012-10-30 | 2020-12-01 | 农业研究有限公司 | 改良的酰基转移酶多核苷酸、多肽、及其使用方法 |
CN104178484A (zh) * | 2013-05-21 | 2014-12-03 | 中国科学院植物研究所 | TmDGAT1b基因的启动子及其应用 |
CN104178485A (zh) * | 2013-05-21 | 2014-12-03 | 中国科学院植物研究所 | TmDGAT1a基因的启动子及其应用 |
CN104178485B (zh) * | 2013-05-21 | 2016-06-29 | 中国科学院植物研究所 | TmDGAT1a基因的启动子及其应用 |
CN104178484B (zh) * | 2013-05-21 | 2016-07-06 | 中国科学院植物研究所 | TmDGAT1b基因的启动子及其应用 |
CN110699334A (zh) * | 2019-11-08 | 2020-01-17 | 山东禹王生态食业有限公司 | 一种酰基转移酶及其应用 |
CN110699334B (zh) * | 2019-11-08 | 2021-07-30 | 山东禹王生态食业有限公司 | 一种酰基转移酶及其应用 |
CN112342235A (zh) * | 2020-11-09 | 2021-02-09 | 南京农业大学 | GmDGAT2A在提高大豆油含量并增加亚油酸含量中的应用 |
Also Published As
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US9902967B2 (en) | 2018-02-27 |
EP2147109A2 (en) | 2010-01-27 |
US20080295204A1 (en) | 2008-11-27 |
EP2147109B1 (en) | 2014-03-19 |
US20150101080A1 (en) | 2015-04-09 |
MX2009012596A (es) | 2009-12-08 |
WO2008147935A2 (en) | 2008-12-04 |
CA2685309A1 (en) | 2008-12-04 |
US8927809B2 (en) | 2015-01-06 |
BRPI0810973A2 (pt) | 2014-10-07 |
WO2008147935A3 (en) | 2009-08-13 |
CN101939434B (zh) | 2015-03-25 |
US20120058246A1 (en) | 2012-03-08 |
US8143473B2 (en) | 2012-03-27 |
CA2685309C (en) | 2019-06-25 |
EP2730656A1 (en) | 2014-05-14 |
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