下载文档原格式
常用质粒和菌株及其特性李煦常用菌株大肠杆菌菌株DH5α培养基：LB培养基培养条件：37℃，280rpm农杆菌菌株LBA4404 AGL1 GV3101培养基：YEP培养基培养条件：28℃，280rpm原核表达菌株BL21 BL21（DE3） BL21（DE3）plys培养基：LB培养基培养条件：37℃，280rpm酵母表达菌株INVSc1基因型：his3Δ1/ his3Δ1 leu2/leu2 trp-289/trp-289 ura3-52/ura3-52表型：His-，Leu-，Trp-，Ura-培养基：YPD培养基培养条件：30℃，280rpm常用抗生素的筛选浓度贮存液浓度工作浓度氨苄青霉素 100mg/ml 100mg/L卡那霉素 50mg/ml 50mg/L利福平 25mg/ml 50mg/L 常用质粒及图谱1、pCAMBIA系列载体（植物表达载体）(1)pCAMBIA1300（8958bp）细菌培养用抗生素：Kanamycin植物选择标记：hygromycin无报告基因(2)pCAMBIA1302（10549bp）细菌培养用抗生素：Kanamycin-植物选择标记：hygromycin报告基因：gfp(3)pCAMBIA1305.1（11846bp）细菌培养用抗生素：Kanamycin植物选择标记：hygromycin报告基因：gus plus(4)pCAMBIA3300（8428bp）细菌培养用抗生素：Kanamycin植物选择标记：bar无报告基因2、pROK2（植物表达载体）细菌培养用抗生素：Kanamycin植物选择标记：npt Ⅱ3、pCAMBIA1300-P35sTnos-als(pCPA)（植物表达载体）细菌培养用抗生素：Kanamycin植物选择标记：als4、pCAMBIA1300-PubiTnos-als(pCUA)（植物表达载体）细菌培养用抗生素：Kanamycin植物选择标记：als5、pCAMBIA1300-P35sTnos(pCPN)（植物表达载体）细菌培养用抗生素：Kanamycin植物选择标记：hygromycin6、pCAMBIA1300-PubiTnos(pUBI)（植物表达载体）细菌培养用抗生素：Kanamycin植物选择标记：hygromycin7、PCAMBIA3300-Pubi-MCS-Tnos（植物表达载体）细菌培养用抗生素：Kanamycin植物选择标记：bar8、pET-30a(+) (原核表达载体)细菌培养用抗生素：Kanamycin9、pYES2（酵母表达载体）细菌培养用抗生素：Ampicillin10、pGEM –T Easy 载体细菌培养用抗生素：Ampicillin。
灵芝转化体系的建立孙墨可;李晓薇;孙晓文;张曼;田娟;董玉迪;李海燕【摘要】利用TPS法提取平菇基因组,克隆甘油醛-3-磷酸脱氢酶启动子(glyceraldehyde-3-phosphate dehydrogenase,GPD),用GPD启动子替换原表达载体pCAMBIA1302中的35s启动子,构建pCAMBIA1302表达载体,利用PEG 法转化灵芝原生质体.通过酶切验证与测序结果表明GPD启动子已成功构建到表达载体pCAMBIA1302中,通过灵芝转化子的PCR检测和荧光显微镜观察证明绿色荧光蛋白(green fluorescent protein,GFP)已转入到灵芝中.该研究成功构建了灵芝转化体系,对今后开展灵芝的分子生物学研究具有重要意义.【期刊名称】《安徽农业科学》【年(卷),期】2018(046)026【总页数】4页(P91-93,107)【关键词】灵芝;表达载体;PEG;转化;GFP【作者】孙墨可;李晓薇;孙晓文;张曼;田娟;董玉迪;李海燕【作者单位】吉林省白城市农业科学院生物技术实验室,吉林白城137000;吉林农业大学生物反应器与药物开发教育部工程研究中心,吉林长春130118;吉林省镇赉县芦苇湿地管理站,吉林镇赉137300;吉林省白城市农业科学院生物技术实验室,吉林白城137000;吉林省白城市农业科学院生物技术实验室,吉林白城137000;吉林省白城市农业科学院生物技术实验室,吉林白城137000;吉林农业大学生物反应器与药物开发教育部工程研究中心,吉林长春130118【正文语种】中文【中图分类】S567.3+1灵芝是我国的传统药用真菌，在我国和东南亚国家有着悠久的应用历史[1]。
灵芝含有多糖、萜类化合物、核苷、生物碱、氨基酸多肽、微量元素等多种活性成分。
现代药理研究表明，灵芝具有保肝、抗肿瘤、抗HIV-1及HIV-1蛋白酶活性、抗组织胺释放、抑制血管紧张素、抗氧化、调节免疫及延缓衰老的作用[2-7]。
酿酒酵母表达载体pYES2,pYES2/NT,pYES2/CT,pYES3,pYES6, pYCplac22-GFP,酵母载体pAUR123,pRS303TEF,pRS304, pRS305,pRS306,pY13TEF,pY14TEF，pY15TEF，pY16TEF,酵母基因重组表达载体pUG6, pSH47,酵母单杂载体pHISi,pLacZi,pHIS2, pGAD424, 酵母双杂交系统:酿酒酵母Y187, 酿酒酵母AH109；质粒pGADT7,pGBKT7；对照质粒pGBKT7-53，pGBKT7-lam，pGADT7-T，PCL1，酿酒酵母菌株INVSc1,YM4271, AH109,Y187,Y190,毕赤酵母表达载体pPIC9K,pPIC9K-His,pPIC3.5K,pPICZalphaA,B,C,pPICZA,B,C,pGAPZαA,pAO815,pPIC9k-His,pHIL-S1,pPink hc，配套毕赤酵母Pichiapink,毕赤酵母宿主X33，KM71，KM71H，GS115，原核表达载体pQE30,31,32,40,60,61,62,等原核表达载体,包括pET系列，pET-GST，pGEX系列（含GST标签），pMAL系列pMAL-c2x,-c4x,-c4e,-c5x,-p5x,pBAD,pBADHis,pBADmycHis系列，pQE系列,pTrc99a,pTrcHis系列，pBV220,221,222,pTXB系列，pLLP-ompA,pIN-III-ompA （分泌型表达系列）,pQBI63（原核表达带荧光）pET3a, pET 3d, pET 11a, pET 12a, pET 14b, pET 15b, pET 16b, pET 17b, pET 19b, pET 20b, pET 21a,b,d, pET 22b, pET 23a, pET 23b, pET 24a,b, pET 25b, pET 26b, pET 27b, pET 28a,b, pET 29a, pET 30a, pET 31b, pET 32a, pET 35b, pET 38b, pET 39b, pET 40b, pET 41a,b pET 42a, pET 43.1a,b pET 44a, pET 49b pET302,303 pET His,pET Dsb,pET GST,pET Trx pQE2, pQE9 pQE30,31,32, pQE 40 pQE70 pQE80L pQETirs system pRSET-A pRSET-B pRSET-C pGEX4T-1,-2,-3,5x-1,6p-1,6p-2,2tk,3c pBV220,221,222 pTrcHisA,B,C pBAD24,34,43 pBAD HisA,B,C pPinPoint-Xa1,Xa2,Xa3 pMALc2x, p2x pBV220 pGEM Ex1, pGEM7ZF（+）, pTrc99A, pTwin1, pEZZ18 pkk232-8,pkk233-3,pACYC184,pBR322,pUC119 pTYB1,pTYB2,pTYB4,pTYB11 pBlueScript SK （+）,pBlueScript SK（-）pLLP ompA, pINIIIompA, pMBP-P ,pMBP-C, 大肠杆菌冷激质粒： pColdI pColdII pColdIII pColdTF 原核共表达质粒：..pACYCduet-1,pETduet-1,pCDFduet-1，pRSFduet-1 Takara公司大肠杆菌分子伴侣： pG-KJE8 pGro7 pKJE7 pGTf2 pTf16 大肠杆菌宿主细胞： DH5a JM101 JM103 JM105 JM107 JM109 JM110 Top10 Top10F BL21（DE3）HB101 ER2529 E2566 C2566 MG1655 XL-10gold XL blue M15 JF1125 K802 SG1117 BL21（AI）BL21（DE3）plysS TG1 TB1 DH5a（pir）Tuner（DE3）Bl21 codonplusRIPL Novablue （DE3）Rosetta Rosetta（DE3）Rosetta（DE3）plys Rosetta-gami（DE3）Rosetta-gamiB（DE3）, Rosetta-gamiB（DE3）plysS Orgami（DE3）OrgamiB（DE3）HMS174（DE3）植物表达/RNAi载体农杆菌pBI121,pBI121-GFP,pBI101,pBI221,pSN1301,pUN1301,pRTL2 , pRTL2-GFP , pRTL2-CFP, pRTL2-RFP , pRTL2-YFP,pCAMBIA 1300, 1301, 1302,1303,1304,1305, 1381Z,1391Z,2300, 2301,3300,3301,pCAMBIA super1300,pCAMBIAsuper1300-GFP,pPZP212,pPZP2121,pPZP212-GFP,pGDG,RNAi载体pART27,pHANNIBAL,pKANNIBAL, pFGC5941,pTCK303, pTRV1,pTRV2,T-DNA插入载体（随机突变体库）pSKI015,pSKI074,真菌ATMT载体pBIG2RHPH2-GUS-GFP,pBHt1枯草芽孢杆菌表达载体pWB980,pHT43,pHP13,pHP43,pBE2,pMUTIN4,pUB110,pE194,pMA5, pMK3,pMK4,pHT304,pHY300PLK,pBest502,pDG1363,pSG1154,pAX01, pSAS144,pDL,pDG148-stu,pDG641,pAL12,pUCX05-bgaB,pHT01，配套菌株BS 168,WB600,WB800,WB700,WB800N,1012,FZB42,1A747,广宿主质粒pVLT33RNAi基因沉默干扰敲除载体pSilencer1.0，pSilencer 2.1-U6 hygro， pSilencer 3.1-H1 hygro，pSilencer 3.1-H1 neo， pSilencer 4.1-CMV neo， pSilencer 4.1-CMV puro pMIR-REPORT Luciferase RNAi载体（oligoengine）pSuper-puro RNAi逆转录病毒载体（clontech）: RNAi-Ready pSIREN-Retro Q， RNAi-ReadypSIREN-RetroQ-ZsGreen（Luciferase shRNA Annealed Oligonucleotide）RNAi慢病毒载体（addgene）: pLKO.1..哺乳动物表达载体pcDNA3.1+/-,pcDNA4/HisMax B，pSecTag2 A，pVAX1，pBudCE4.1,pTracer CMV2，pcDNA3.1（-）/myc-His A ，pcDNA6-Myc/His B，pCEP4， pIRES，pIRESneo，pIRES hyg3，pCMV-myc，pCMV-HA，pIRES-puro3，pIRES-neo3,pCAGGS哺乳动物双杂交系统pACT，pBIND，pACT-MyoD，pBIND-Id，pG5luc,pCMV-BD, pCMV-AD, pBD-p53, pFR-luc,Cytotrap Two-Hybrid System:pSos, pSos MAFB, pMyr蜕皮激素诱导系统pIND, pVgRxR,LacSwith II哺乳动物诱导表达系统:pOPRSVI ，pOPI3CAT，pCMVLacI,GeneSwitch System:pSwitch哺乳动物表面展示系统:pDisplay, 四环素调控系统（Invitrogen）：pcDNA4/TO/Myc-His A，pcDNA4/TO/Myc-His B，pcDNA4/TO/Myc-His C，pcDNA4/TO/Myc-His/LacZ，pcDNA6/TR四环素调控系统（Clontech）：pTet-On，pTet-Off，pTRE2,pRevTRE，pRevTet-On，pRevTet-off信号通路报告载体:pGAS-TA-Luc，pSTAT3-TA-Luc, pISRE-TA-Luc, pTA-Luc，pIκB-EGFP，pNFAT-TA-Luc，pCaspase3-sensor，pAP1（PMA）-Luc;pGL4.26[luc2P/minP/Hygro],pGL4.29[luc2P/CRE/Hygro],pGL4.30[luc2P/NFA T-RE/Hygro]，pGL4.75;p53-Luc，pAP-1-Luc， pNF-κB-Luc，pSRE-Luc，pFA2-Elk1，pFC-MEKK，pFR-luc,Gateway系统（invitrogen）pcDNA6.2-GWEmGFP-miR negative, pLenti 6/TR,pcDNA 6.2-GW EmGFP-miR，乳酸菌表达载体及各种乳酸菌乳酸杆菌菌株，pNZ8148,pLEISS,pMG36e,pBBR1MCS-5,pBBR1MCS-6,pRV610,pLEM415,pHY3 00PLK,分泌型乳酸菌表达载体pVE5523，pPG611.1,pPG612.1等和乳酸杆菌菌株宿主菌NZ9000,MG1363,Lactobacillus casei 1.539,Lactobacillus casei,acidophilus NCFM,1.2,Lactobacillus sakei 23K,L.plantarum,L.rhamnosusGG,B.coagulans,Bifidobacterium bifidum,Bifidobacterium infantis,Lactococcus lactis M17,1663,Lactobacillus reuterii广宿主表达载体链球菌表达敲除载体假单胞菌表达载体pVLT33,pBBR1MCS-2,3,4,5,6, pJRD215,pJN105,pME6032,Cos载体pLAFR3,pMP2444（GFP）, pHY300PLK,pRT102,pRL1063a, 转座子载体pUT-miniTn5,pMGS100, pWHM10,pKC1139,pSET152,pOJ260,pPG611.1,pPG612.1，..腺病毒载体/慢病毒，逆转录病毒表达载体及包装包膜质粒，腺病毒系统（Stratagene）: pAdEasy-1,pShuttle-CMV,pShuttle,pAdTrack, pAdTrack-CMV, pShuttle-IRES-hrGFP-1、pShuttle-IRES-hrGFP-2、pShuttle-CMV-lacZ,pShuttle-CMV-EGFP-C,pXC1, pBHGE3, 配套大肠杆菌BJ5183,293,293T cellline 腺相关病毒系统（Stratagene）： pAAV-MCS，pAAV-RC，pHelper，pAAV-LacZ，pAAV-IRES-hrGFP，pCMV-MCS，慢病毒载体:pLVX-DsRed-Monomer-N1,pLVX-IRES-ZsGreen1,pLVX-AcGFP1-N1,Lenti6/v 5-EDST-EGFP,pWPXL, FUGW,pLentilox 3.7,RNAi-Ready pSIREN-Retro Q,RNAi-Ready pSIREN-Retro Q-ZsGreen,pSUPER.Retro-GFP/Neo,pSUPER-Retro-Neo, pSUPER.Retro-puro,PLNCX PLNCX2 pMSCV-HYG pMSCV-neo pMSCV-puro pLEGFP-C1 pLOX-CW-CRE pLOX-GFP-IRES-TK pRetroX-IRES-DsRedExpress, pLVX-IRES-mCherry质粒载体。
植物亚细胞定位（范文2篇）以下是网友分享的关于植物亚细胞定位的资料2篇，希望对您有所帮助，就爱阅读感谢您的支持。
植物亚细胞定位（1）第42卷第4期东北农业大学学报42(4):83~87植物亚细胞定位载体卡盒pCEG的构建及验证朱丹，王希，朱延明*，陈超，李勇，柏锡，才华，纪（东北农业大学生命科学学院，哈尔滨150030）巍摘要：GFP基因被广泛地应用于植物转基因以及基因功能验证研究，然而构建与GFP融合的植物表达载体，常会因酶切位点难以选择，而使得载体构建过程复杂，周期长。
以含GFP的质粒pCAMBIA-1302为基础，消除此质粒本身的多克隆位点（MCS），并在此质粒GFP基因序列前插入原核表达载体pET-32b的多克隆位点，构建了植物GFP亚细胞定位载体卡盒pCEG；采用农杆菌介导的转化方法，将此载体卡盒pCEG导入模式植物烟草叶片中进行瞬时表达，用激光共聚焦显微镜观察GFP蛋白的亚细胞定位情况，发现GFP蛋白均匀的在细胞质和细胞核中表达，证明此载体卡盒可用于植物基因的亚细胞定位分析，并为构建目的基因与GFP融合的植物表达载体提供了很多可用的酶切位点，对植物基因功能验证提供了分子操作简便的植物表达载体卡盒，具有重要的利用价值。
关键词：载体卡盒构建；亚细胞定位；GFP；烟草中图分类号：Q782文献标志码：A文章编号：1005-9369（2011）04-0083-05 Constructionandapplicationofaplantsubcellularlocalization vectorboxofpCEG/ZHUDan,WANGXi,ZHUYanming,CHEN Chao,LIYong,BAIXi,CAIHua,JIWei(CollegeofLifeSciences,NortheastAgriculturalUniversit y,Harbin150030,China)Abstract:GFPgenewaswidelyappliedinthetransgenicplants andgenesfunction’sprovingstudy.However,theconstructionofGFPfusedplantexpressionvector swasalwayscomplicatedandtimeconsumingduetothehardchoo eplasmidpCAMBIA-130 2whichincludedGFPgeneasaoriginalvector,andeliminateditso wnmultipleclonesite(MCS),andtheninsertedpET-32b’smultip leclonesiteinfrontofGFPgenesequences,namedtheeventualpla ntGFPsubcellularlocalizationvectorboxpCEG.ByAgrobacteri um-mediatedtransformationmethod,pCEGinstantaneousexp ressedintobaccoleaves,andGFPproteinsubcellularlocalization wasobservedbylaserconfocalmicroscope.Theresultdisplayedt hattheGFPproteinsexpressedinbothcytoplasmandnucleus,thi smeansthepCEGboxcanbeusedconvenientlyforplantgenessub cellularlocalizationanalysisandprovidealotofusablerestrictio nenzymecuttingsiteforplantgenesfusionedwithGFP,andthepC EGboxisconvenientforplantgenesfunctionanalysisandhasgre atusevalue.Keywords:constructionofvectorbox;subcellularlocalization ;GFP;tobacco在基因的功能研究过程中，外源重组基因表达产物的功能与其在宿主细胞中的定位有重要的关系，特别是对于真核细胞，蛋白质位于不同的细胞部位所行使的功能也不同，所以研究其在宿主细胞收稿日期：2011-01-12基金项目：国家高科技发展计划（863计划）（2008AA10Z153）；国家自然科学基金资助项目（30570990）；黑龙江省科技厅重大攻关项目（GA06B10）；黑龙江省教育厅科技项目（11521024）；黑龙江省教育厅科技项目（11521021）作者简介：朱丹（1986-），女，硕士研究生，研究方向为植物基因工程与分子生物学。
农杆菌渗透法转化烟草条件的优化作者：李晓君王绍梅谢艳兰和敏来源：《江苏农业科学》2014年第09期摘要：植物瞬时表达系统常用于研究基因表达产物的亚细胞定位和蛋白间的互作，将GFP-GUS融合蛋白植物表达载体导入农杆菌EHA105，获得工程菌，制备不同浓度的农杆菌浸染液，用注射法对烟草叶片进行转化，荧光显微镜检测烟草叶片原生质体和下表皮中GFP 的表达情况。
结果表明，浸染注射液的D600 nm 在0.3～0.7之间均具有较高的转化效率，注射后第4天至第6天为较佳观察时间。
关键词：植物瞬时表达系统；渗透法；转化；农杆菌渗透注射；烟草；绿色荧光蛋白（GFP）中图分类号： S572.01文献标志码： A文章编号：1002-1302（2014）09-0045-03收稿日期：2013-11-25基金项目：临沧师范高等专科学校高层次人才引进科研启动项目（编号：LXJ2012）；临沧师范高等专科学校校级课题（编号：LCSZL2013001 ）。
作者简介：李晓君（1985—），女，云南临沧人，博士，讲师，主要从事植物生物技术与种质创新研究。
E-mail：***************。
植物瞬时表达系统常用于基因产物的亚细胞定位、蛋白间互作、转录因子与启动子之间的互作、启动子分析等方面的研究[1-2]。
植物瞬时表达系统中外源基因的导入方法有植物病毒介导法、PEG法、电击法、基因枪法和农杆菌渗透法[2]。
农杆菌渗透法是一种比较简便的方法，通过将植物表达载体导入农杆菌，用真空法或注射法将农杆菌导入植物细胞，使得农杆菌Ti质粒上的T-DNA区整合到植物基因组中，T-DNA 区的目标基因培养几天后即可得到表达[3]。
较其他方法而言，农杆菌渗透法具有转化效率高、基因表达在完整活体内进行、可携带较大片段的目的基因等优点[2]。
原生质体是观察基因瞬时表达情况的优选材料，传统的原生质体转化方法首先要制备高浓度、高质量的质粒，经过从植物活体材料中分离纯化原生质体，在PEG的介导下将质粒DNA导入原生质体，进行原生质体培养后再检测目标基因的表达情况[4]。
测序通⽤引物名称及序列表测序通⽤引物名称及序列列表载体名称上游引物（5'primer）下游引物（3'primer）P3*FLAG-CMV CMV-30(825-854)CMV-24(1129-1152) pAc5.1/V5-His(_A,_B,_C)pAc5-5BGH rev pAcG2T pGEX-5not availablepACT T7EEV T3/EBVrevpACT2GAL4ADfor pGAL4-ADrvpACT2p17110p12584 pACYC184(BamHI-site)PBRforBam PBRrevBampACYCDuet-1(MCSI)ACYCDuetUP1Primer DuetDOWN1 pACYCDuet-1(MCSII)DuetUp2T7-Terminator pAdEasy-1not available not availablepAdTrack-CMV not available not available pAS2-1GAL4Bdfor/P24990pGAL4-BDrv/P31430pB42AD pB42ADF PB42ADRpBABE pBMN_5'pBacPAK8BAC1BAC2pBAD pBAD-F pBAD-R pBAD/HisABC pBAD-5'pBBR1MCS T3/M13rev T7/M13forpBD-GAL4pBD-GAL4-F PBD-GAL4-RpBI12135S NOSpBIND GAL4-Bdfor T3,EBVrevpBK-CMV M13F/T7M13R/T3 pBluescriptIIKS(+/-)T3/M13rev T7/M13for（优先⽤M13F）pBluescriptIISK(+/-)T3/M13rev T7/M13forpBluescriptKS(+/-)T3/M13rev T7/M13forpBluescriptSK(+/-)T3/M13rev T7/M13forpBR322(BamHI-site)PBRforBam PBRrevBampBR322(EcoRI/HindIII-sites)pBRforEco pBRrevHind pBR325(BamHI-site)PBRforBam PBRrevBampBR329(BamHI-site)PBRforBam PBRrevBampBridge(MCSI)GAL4-Bdfor3'BDpBridge(MCSII)not available not available pBS-T II T3/M13rev T7/M13forpBS（AMP）T7T3 pBSR T3T7ter pBudCE4.1(MCSI)CMV-Profor,T7c-mycrev,EBVrevpBudCE4.1(MCSII)EF-1αFwd BGHpBudCE4.1/lacZ/CAT CMV-Profor,T7c-mycrev,EBVrev pBV220pBV220F PBV220RpCAL-c T7/Tac-Profor T7-TerminatorpCAL-n T7/Tac-Profor T7-Terminator pCAMBIA1301(1300)pCAMBIA1301-5'(ECORI)/M13R pCAMBIA1301-3'(HindIII) pCAMBIA1304pCAMBIA1301-5'(ECORI)pCAMBIA1301-3'(HindIII) pCAMBIA2300M13R(-48)M13F(-47)/M13F(-49) pCANTB5E S1S6pCAT3-enhancer RVP3此载体⽆反向引物pCDFDuet-1(MCSI)ACYCDuetUP1Primer DuetDOWN1pCDFDuet-1(MCSII)DuetUP2T7TER pcDNA1.1CMV-Profor,T7pCDM8-3pcDNA2.1M13for/T7M13rev/Tac-ProforpcDNA3T7/CMV-Profor BGH rev/SP6 pcDNA3.1(+/-)pcDNA3.1F/T7/CMV-Profor BGH revpcDNA3.1/His(_A,_B,_C)T7/CMV-Profor BGH rev pcDNA3.1/myc-His(_A,_B,_C)T7/CMV-Profor BGH rev/c-mycrev pcDNA3.1/V5-His(_A,_B,_C)T7/CMV-Profor BGH rev/V5rev pcDNA3.1/Hygro(+,-)T7/CMV-Profor BGH rev/V5revpcDNA3.1/Zeo(+.-)T7BGHrevpcDNA3.3TOPOTA CMV-Profor TKPAreverse pcDNA4/HisMax(_A,_B,_C)Xpressfor BGH revpcDNA4/HisMax-TOPO Xpressfor BGH rev pcDNA4/HisMax-TOPO/lacZ Xpressfor BGH rev pcDNA4/myc-His(_A,_B,_C)T7,CMV-Profor BGH rev,c-mycrev pcDNA4/TO TREfor/CMV-Profor BGH rev pcDNA4/V5-His(_A,_B,_C)CMV-Profor,T7BGH rev,V5rev pcDNA4-TET/ON/AMP+CMV-ProforpcDNA5/FRT T7/CMV-Profor BGH rev pcDNA6/myc-His(_A,_B,_C)CMV-Profor,T7BGH rev,c-mycrev pcDNA6/V5-His(_A,_B,_C)CMV-Profor,T7BGH rev,V5rev pCEP4pCEP-F/CMV-Profor EBVrevpCI T7（17BASE）EBVrevpCI-neo T7（17BASE）EBVrev,T3pCMS-EGFP T7T3pCMS-EGFP T7EBVrev,T3pCMV-3Tag-4A T3/PFLAG-CMV-F T7 pCMV5pCMV5F PCMV5RpCMV6CMV-Profor pCMV6-3、HGHrevpCMV6-XL4CMV-Profor/T7pCMV6-3、HGHrevpCMV-HA PCMV-F,CMV-Profor,TREfor PCMV-R，EBVrev pCMV-MYC(AMP+)PCMV-F PCMV-R,EBVrevpCMVmyc/nuc CMV-Profor BGH rev,c-mycrev pcmv-scriptEXVector T3T7 pCMV-Sport6T7/M13for SP6/M13rev pCOLADuet-1(MCSI)ACYCDuetUP1Primer DuetDOWN1pCOLADuet-1(MCSII)DuetUP2T7TER pCR2.1T7,M13for M13rev pCR2.1-TOPO M13for,T7M13rev pCR3.1T7BGH rev pCR3.1-Uni T7BGH revpCR4T3/M13rev T7/M13forpCR4-Blunt T3/M13rev T7/M13forpCR4-TOPO M13rev,T3M13for,T7pCR-Blunt M13rev T7,M13forpCR-BluntII M13rev/SP6T7/M13for pCR-BluntII-TOPO M13rev/SP6M13for/T7 pCRII M13rev/SP6T7/M13for pCRII-TOPOM13rev/SP6M13for/T7pCR-ScriptSK(+)T7/M13for T3/M13revpCR-XL-TOPO M13rev T7,M13forpCS2+(MCSI)SP6EBVrev,T7?pCS2+(MCSII)T3pSG5-3pDB_GAL4cam T7PpDB_leu PDBLeu-5PDBLeu-3pDEST22DEST22F DEST22RpDEST15pGEX-5T7-TerminatorpDEST20pGEX-5pFastBac-3pDEST26TREfor/CMV-Profor T7/M13forpDisplay T7/CMV-Profor c-mycrev pDNR-LIB(MCS_A)M13for,T7pSG5-3,M13R pDonar M13F M13RpDONR201PDONR-F PDONR-RpDONR207PDONR-F PDONR-RpDONR223M13F M13RpDrive T7/M13rev SP6/M13for pDsRED1-C1(DsRed1-N-f)DsRed1-C-rpDsRed1-N1CMV-Profor RFP-Nrev,CMV-R?pDsRed2-N1(DsRed1-N-f)DsRed1-C-rpDsRed2-C1(DsRed1-N-f)DsRed1-C-r pDsRED2-C1(KAN+)pDsRED-ex-C1-F pEGFP-N-3’pDsRed-Express dsRed1_N_primer/CMV-F dsRed1_C_primer/CMV-R pDSRED-monomer-N1(KAN+)dsRed1_N_primer pDSRED-N1(KAN+)pEGFP-N-5’pDSRED-N-RpEASY-Blunt M13F T7/M13RpEASY-BluntSimple M13F T7/M13R（距离插⼊位点合适，优先安排）pEASY-T1Simple M13F M13R（距离插⼊位点合适，优先安排）pEASY-T1M13F/T7M13RpEASY-T3M13F/T7M13R/SP6pECFP-N(C)PECFP-N-5.PECFP-N-3,pEF/myc/cyto/ER/mito/nuc pEF-5Pcdna3.1R/BGH rev pEF1(4,6/myc-His)T7/pEF-5Pcdna3.1R/BGH rev pEF1(4,6/V5-His)T7/pEF-5Pcdna3.1R/BGH rev pEF1(4;6/His_A;_B;_C)T7/Xpressfor Pcdna3.1R/BGH rev pEF5/FRT/V5-D(-TOPO)T7/pEF-5Pcdna3.1R/BGH rev pEGFP M13rev EGFP-NrevpEGFP-1not available EGFP-Nrev/PEGFP-N-3' pEGFP-C PEGFP-C-5′PEGFP-C-3′pEGFP-C1EGFP-Cfor SV40-pArevpEGFP-C2EGFP-Cfor SV40-pArevpEGFP-C3EGFP-Cfor SV40-pArevpEGFP-F EGFP-Cfor SV40-pArevpEGFP-N PEGFP-N-5′PEGFP-N-3′pEGFP-N1CMV-Profor EGFP-NrevpEGFP-N2CMV-Profor EGFP-NrevpEGFP-N3CMV-Profor EGFP-NrevpENTR M13F M13R pET-11(-a,-b,-c,-d)T7T7-TerminatorpET-12(-a,-b,-c)T7T7-Terminator pET-14b T7T7-TerminatorpET-15b T7T7-TerminatorpET-16b T7T7-TerminatorpET-17b T7T7-TerminatorpET-17xb T7T7-TerminatorpET-19b T7T7-TerminatorpET-20b(+)T7T7-Terminator pET-21(-a,-b,-c,-d)(+)T7T7-Terminator pET-22b(+)T7T7-Terminator pET-23(-a,-b,-c,-d)(+)T7T7-Terminator pET-24(-a,-b,-c,-d)(+)T7T7-Terminator pET-25b(+)T7T7-TerminatorpET-26b(+)T7T7-TerminatorpET-27b(+)T7T7-TerminatorpET-28a(-b,-c)(+)T7T7-TerminatorpET-29a(-b,-c)(+)S.tag/T7T7-TerminatorpET-3(-a,-b,-c,-d)T7T7-Terminator pET-30_Ek/LIC T7T7-Terminator pET-30_Xa/LIC T7T7-Terminator pET-30a(-b,-c) (+)S.tag/T7T7-Terminator pET-31b(+)not available T7-Terminator pET-32_Ek/LIC TRXfor T7-Terminator pET-32_XA/LIC TRXfor T7-Terminator pET-32a(-b,-c)(+)S.tag/TRXfor T7-Terminator pET-33b(+)T7T7-Terminator pET-34b(+)S.tag T7-Terminator pET-35b(+)S.tag T7-Terminator pET-36b(+)S.tag T7-Terminator pET-37b(+)S.tag T7-Terminator pET-38b(+)not available T7-Terminator pET-39b(+)S.tag T7-Terminator pET-40b(+)S.tag T7-Terminator pET-41_Ek/LIC GST-Cfor T7-Terminator pET-41a(-b,-c)(+)S.tag/pGEX-5T7-Terminator pET-42a(-b,-c)(+)S.tag/pGEX-5T7-Terminator pET-43.1a(-b,-c) (+)S.tag coliDOWN pET-43.1a_EK/LIC S.tag not available pET-44a(-b,-c)(+)S.tag T7-Terminator pET-44a_EK/LIC S.tag not available pET-45b(+)T7T7-Terminator pET-46_Ek/LIC T7T7-Terminator pET-5(-a,-b,-c)T7T7-Terminator pET-52b(+)T7T7-TerminatorpET-7T7T7-Terminator pET-9(-a,-b,-c,-d)T7T7-Terminator pETcoco-1T7T7-Terminator pETDuet-1(MCSI)pETUP1DuetDOWN1 pETDuet-1(MCSII)DuetUp2T7-Terminator pET-T4T7T7-Terminator pEYFP-C1pEGFP-C-5’pEGFP-C-3' pEYFP-N1pEGFP-N-5’,CMV-Profor pEGFP-N-3’pFLAG-ATS N26for pTrcHis-rv；C24rev pFLAG-CTCN26for pTrcHis-rv；C24rev pFLAG1N26for pTrcHis-rv；C24rev pFLAG-CMV(-2)CMV-30；CMV-Profor CMV-24 pFLAG-CMV1CMV-30CMV-24pGAD424GAL4_ADfor3'ADpGADGH GAL4_ADfor3'ADpGADT7T7/GAL4_ADfor3'AD pGADT7-Rec T7/GAL4_ADfor3'AD pGADT-T T73,BD pGAD10GAL4_ADfor3'AD pGAPZa-A a-FACTOR3'AOX pGBKT7T73,BDpGBT9MATCHMAKER5'DNA-BD/GAL4BD MATCHMAKER3'DNA-BDpGEM-1T7SP6pGEM-3T7SP6 pGEM-3Zf(+)(-)SP6/M13rev T7/M13for pGEM-4T7SP6pGEM-4Z T7/M13for SP6/M13rev pGEM-5Zf(+)(-)SP6/M13rev T7/M13for pGEM-7Zf(+)(-)SP6/M13rev T7/M13for pGEM-9Zf(+)(-)T7/M13for SP6/M13rev pGEM-T T7/M13for SP6/M13rev pGEM-T_Easy T7/M13for SP6/M13rev pGene/V5-His_A(_B,_C)pGENE-5BGH rev,V5rev pGEX-2T pGEX-5pGEX-3pGEX-2TK pGEX-5pGEX-3pGEX-3X pGEX-5pGEX-3 pGEX-4T-1(-2,-3)pGEX-5pGEX-3 pGEX-5X-1(-2,-3)pGEX-5pGEX-3 pGEX-6P-1(-2,-3)pGEX-5pGEX-3 pGL2GLP1GLP2pGL3RVP3GLP2(RVP4)(没有特别说明⽤GLP2)pGL3-Basic(MCSI)RVP3(PGL3+)?GLP2(PGL3-)? pGL3-Basic(MCSII)RVP4,PLXSN-3'?pGL4pGlow(-TOPO)T7pGlow-3 pGM-T T7SP6pGWC SP6T7pHook-2T7/CMV-Profor not available pIND/V5-His_A(_B,_C)not available BGH rev,V5rev PinpointTMVector PinpointPrimer SP6pIRES(MCSI)T7pIRES-3pIRES(MCSII)not available T3,EBVrevpIRES2-EGFP CMV-Profor,pIRES2-EGFP.P5’IRESrev,pIRES2-EGFP.P3’pIRES-hrGFP-2a CMV-Profor,T3pIRES-hrGFP-3 pIRESneo2T7/CMV-Profor pIRES-3pIVEX(all)T7/pBRrevBam T7-TerminatorpLEXA pLEXA-F pLEXA-RpLNCX pLNCX-F pLNCX-RpLNCX2CMV-Profor pRevTRE-3 pLVTHM（H1启动⼦下游）H1primer SP6 pLXSN pLXSN-F pLXSN-RpMALc MalE primer M13F(-47)pMAL-c2E MalE primer M13F(-47)pMAL-C2x Pmal-c2x-F(P5’)Pmal-C2X-R(P3’)pMALd MalE primer M13F(-47)pMALe MalE primer M13F(-47)pMAL-P4X MalE primer M13F(-47)pMAL-p2X MalE primer M13F(-47)pMET A/B/C AUG1F AUG1RpMD18-T M13R(-48)ECORI M13F(-47)(HINDⅢ)pMD19-T M13F47M13R48pMD20-T M13F47M13R48pMOSBlue T7/M13rev M13forpMSCV-puro pLXSNfw pMSCVrev pMT/V5-His_A(_B,_C)Mtfor BGH rev,V5rev pNTAP T3T7pCTAP pCTAP5'primer T7pOTB7M13rev/T7M13for/SP6 pOptiVEC-TOPO TA CMV-F/CMV-Profor EMCVIRES reverse pPC86PPC86-F PPC86-R pPC97PBD-GAL4-F PBD-GAL4-RpPIC3.5K5'AOX3'AOXpPic9k(A+,Z+)5’AOX(Ppic9k-f)/a-factor3’AOX(Ppic9k-R) pPICZa5'AOX/PPICZa-F3'AOX pPROEXHTA M13R(-48)⽆反向引物pProEX-HTa(b,c)M13R48/Tac-Profor pBADrv/pTrcHis-3 pPD129.36M13F20/M13F pQE30or40or60(AMP+)PQE30-F PQE-30R pRC/CMV2/CAT CMV/T7pRECEIVER CMV-F此载体⽆反向引物pRECEIVER-M03pRECEIVER-M03F pRECEIVER-M03R pREP4pREP4forward primer EBV reverse primerpRK5SP6pSG5-3pRSET T7/Xpressfor T7ter pRSFDuet-1(MCSI)DuetUP1DuetDOWN1 pRSFDuet-1(MCSII)DuetUP2T7TERpSecTac2T7/CMV-Profor BGH rev/c-mycrevpSG5pSG5-5,/T7PSG5-R(SV40)pSHlox1T7SP6 pSHUTTLE-CMV PSHUTTLE-CMV-F PSHUTTLE-CMV-R pSI T7EBVrev pSilencer1.0-U6T7T3 pSilencer2.0-U6T7M13for pSilencer2.1-U6neo T7M13for pSilencer3.1-H1hygro M13F(-47) 3.0REV pSilencer4_1-CMVneo pSilener4.1-5'CMV-Profor pSILENCEV2.0-U6T7 2.0REV pSIneo T7EBVrev,T3Psit T7T7TpSK01-T M13F M13RpSK-CMV(KAN+)T7T3 pSOS Psos5'Psos3'pSP64SP6M13revpSP64/65SP6此载体⽆反向引物pSP65SP6M13revpSP70SP6T7pSP71SP6T7pSP72SP6T7pSP73SP6T7pSport1SP6/M13F(-47)T7/M13RpSPORT1SP6/M13for T7/M13revpSPT18SP6T7pSTBlue T7/M13rev SP6/M13for pSUPER.Basic T7/M13F M13RpSUPER T7T3pSURE-T M13F M13R pSURE-Tsimple M13F-47M13R-48pSuperCos pBRforEco T7pT3T7M13rev/T3M13for/T7pT7-7T7not availablepT7Blue?T7M13F(-47)pT7T3_18U M13rev/T7M13for/T3 pT7T3D-PAC M13rev/T7M13for/T3 pTA2M13F/T7(优先使⽤M13F）T3/M13RpT-Adv M13rev T7,M13forpTAg T7/M13rev M13for pTARGETTM pTarget.F(在T7前⾯）/T7pTarget.RpTG19-T M13F(-47)M13R(-48) pTARGETTM T7/M13F PTARGET-SEQ pThioHisA,B,andC Trx Forward Trx Reverse/M13for pTO-T7T7T7TERpTRC99A PTRC99A-F pTrcHis-rvpTrc99a-c PTRC99C-F PTRC99C-RpTRC99a-c PTRC99C-F PTRC99C-RpTrcHisB PTrcHisA-F/Xpressfor PTrcHisA-RpTrcHisA PTrcHisA-F PTrcHisA-RpT-REX-DEST30TREfor T7 pTriEx1T7/TriExup TriExdownpTriEx1.1T7/TriExup TriExdown pTriEx1.1_Hygro T7/TriExup not available pTriEx1.1_Neo T7/TriExup IRESrevpTriplEX1TriplEx-LD-5T7,TriplEx-LD-3pTriplEX2TriplEx-LD-5；pTR5‘T7,TriplEx-LD-3pTWIN-1T7T7TERpTXB1T7MxeInteinrevpTXB2T7MxeInteinrevpTXB3T7MxeInteinrevpTYB1T7InteinrevpTYB2T7InteinrevpTYB3T7InteinrevpTYB4T7InteinrevpTZ_18U T7/M13rev M13forpUB6/V5-His UB-F BGH rev,V5rev pUC13M13rev/M13rev(-48)M13for/M13for(-49)pUC18M13rev/M13rev(-48)M13for/M13for(-49) pUC18(19)/118(119)M13F(-47)M13R(-48)/PUC19(HINDⅢ)pUC19M13rev/M13rev(-48)M13for/M13for(-49)pUC57M13rev/M13rev(-48)M13for/M13for(-49)pUC8M13rev/M13rev(-48)M13for/M13for(-49)pUC9M13rev/M13rev(-48)M13for/M13for(-49)pUM-T M13F(-20)M13R(-20) pUCm-T(AMP+)M13F/T7M13RpUNI/V5-His not available BGH rev,V5revpVAX1T7T7/CMV-Profor PCDNA3.1R/BGHR pVP22/mycHis VP22-Cfor BGH rev,c-mycrev pVP22/mycHis2T7/CMV-Profor VP22-Nrev pWE15pBRrevHind T7pYES2T7pYes2.RpYES-DEST52T7/GAL1-Profor V5rev/CYC1-Terminator pYEX_4T_1Clontech pYEX_4T-3pYEX_4T_2pGEX-5pYEX_4T-4pYEX_4T_3pGEX-5pYEX_4T-4pYEX_4T_4pGEX-5pYEX_4T-4 pZeoSV2+/hu-Tyr T7not available pZERO-1M13rev/SP6T7/M13forpZERO-2M13rev/SP6M13for/T7pZome-1-C Zome-F Zome-RpZome-1-N Zome-F Zome-R YCp50(BamHI-site)PBRforBam PBRrevBam YEp13(BamHI-site)PBRforBam PBRrevBam YEp24(BamHI-site)PBRforBam PBRrevBam YIp5(BamHI-site)PBRforBam PBRrevBam RNAi-ReadypSIREN-RetroQ-U6Zs Green U6菌PCDB T7BGH 天为时代TA载体T7T3 m13（+）T7？T3？pMSCV-neo pMSCV-5'pMSCV-3'引物名称序列(5'-3')M13R CAG GAA ACA GCT ATG ACCM13F TGT AAA ACG ACG GCC AGTM13F(-47)CGC CAG GGT TTT CCC AGT CAC GAC M13R(-48)AGC GGA TAA CAA TTT CAC ACA GGA M13(-96)CCC TCA TAG TTA GCG TAA CGSP6ATT TAG GTG ACA CTA TAGT7TAA TAC GAC TCA CTA TAG GGT7terminator TGC TAG TTA TTG CTC AGC GGT3ATT AAC CCT CAC TAA AGG GA pGEX-4T-5'GGG CTG GCA AGC CAC GTT TGG TG pGEX-4T-3'CCG GGA GCT GCA TGT GTC AGA GG GLp1TGT ATC TTA TGG TAC TGT AAC TG GLp2CTT TAT GTT TTT GGC GTC TTC CARVp3CTA GCA AAA TAG GCT GTC CCRVp4GAC GAT AGT CAT GCC CCG CG pcDNA3.1R TAG AAG GCA CAG TCG AGG PinPoint primer CGT GAC GCG GTG CAG GGC G pCMV-F TCT AAA AGC TGC GGA ATT GT pCMV-R TCCAAACTCATCAATGTATCpTRC99C-F:TTG CGC CGA CAT CAT AACpTRC99C-R:CTGCGTTCTGATTTAATCTGpCEP-F:AGA GCT CGT TTA GTG AAC CGEBV-R:GTG GTT TGT CCA AAC TCA TC pIRES2-EGFP.P5’:GTA GGC GTG TAC GGT GGG AG pIRES2-EGFP.P3’:AAC GCA CAC CGG CCT TAT TC3'AD:AGA TGG TGC ACG ATG CAC AG CMV-F CGC AAA TGG GCG GTA GGC GTGS1CAA CGT GAA AAA ATT ATT ATT CGC S6GTA AAT GAA TTT TCT GTA GTA GG 5`AOX1GAC TGG TTC CAA TTG ACA AGC3`AOX1GCA AAT GGC ATT CTG ACA TCCα-Factor TAC TAT TGC CAG CAT TGC TGC GAL4AD TAC CAC TAC AAT GGA TGpACT2-R GTGCACGATGCACAGTTGAApB42ADF:CCA GCC TCT TGC TGA GTG GAG ATG pB42ADR:AAG CCG ACA ACC TTG ATT GGA G。
载体质粒图谱和多克隆位点信息载体简介载体序列LOCUS AF234298 10549 bp DNA circular SYN 24-APR-2000 DEFINITION Binary vector pCAMBIA-1302, complete sequence.ACCESSION AF234298VERSION AF234298.1 GI:7638073KEYWORDS .SOURCE Binary vector pCAMBIA-1302ORGANISM Binary vector pCAMBIA-1302other sequences; artificial sequences; vectors.REFERENCE 1 (sites)AUTHORS Hajdukiewicz,P., Svab,Z. and Maliga,P.TITLE The small, versatile pPZP family of Agrobacterium binary vectors for plant transformationJOURNAL Plant Mol. Biol. 25 (6), 989-994 (1994)PUBMED 7919218REFERENCE 2 (bases 1 to 10549)AUTHORS Roberts,C., Rajagopal,S., Smith,L.M., Nguyen,T.A., Yang,W.,Nugrohu,S., Ravi,K.S., Vijayachandra,K., Harcourt,R.L.,Dransfield,L., Desamero,N., Slamet,I., Hadjukiewicz,P., Svab,Z., Maliga,P., Mayer,J.E., Keese,P.K., Kilian,A. and Jefferson,R.A.TITLE A comprehensive set of modular vectors for advanced manipulations and efficient transformation of plantsJOURNAL UnpublishedREMARK Full description of constructsREFERENCE 3 (bases 1 to 10549)AUTHORS Roberts,C., Rajagopal,S., Smith,L.M., Nguyen,T.A., Yang,W.,Nugrohu,S., Ravi,K.S., Vijayachandra,K., Harcourt,R.L.,Dransfield,L., Desamero,N., Slamet,I., Hadjukiewicz,P., Svab,Z., Maliga,P., Mayer,J.E., Keese,P.K., Kilian,A. and Jefferson,R.A.TITLE Direct SubmissionJOURNAL Submitted (15-FEB-2000) CAMBIA, Clunies Ross St, Black Mountain / GPO Box 3200, Canberra, ACT 2601, AustraliaFEATURES Location/Qualifierssource 1..10549/organism="Binary vector pCAMBIA-1302"/mol_type="other DNA"/db_xref="taxon:118390"CDS 2..757/note="mgfp5* His6"/codon_start=1/product="mGFP5*, hexaHis tagged"/protein_id="AAF65344.1"/db_xref="GI:7638075"/translation="MVDLTSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYG KLTLKFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTI FFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADK QKNGIKANFKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEK RDHMVLLEFVTAAGITHGMDELYKASHHHHHHV"misc_feature 786..1038/note="nos (nopaline synthase) 3'UTR (polyA signal)"misc_feature 1076..1101/note="right border T-DNA repeat"misc_feature complement(2142..3142)/note="STA region from pVS1 plasmid"rep_origin complement(3735..4735)/note="pVS1-REP; replication origin from pVS1"misc_feature complement(5145..5405)/note="bom site from pBR322"rep_origin complement(5545..5825)/note="pBR322 origin of replication"CDS complement(6116..6910)/note="aadA (kanamycin resistance) gene amplified frompIG121Hm"/codon_start=1/product="aminoglycoside phosphotransferase"/protein_id="AAF65345.1"/db_xref="GI:7638076"/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLK MTDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEY EDEQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWE EDTPFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKW YDIAFCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"misc_feature 7335..7360/note="left border repeat from C58 T-DNA"misc_feature 7427..7635/note="CaMV 3'UTR (polyA signal)"CDS complement(7651..8676)/note="hptII (hygromycin resistance) gene"/codon_start=1/product="hygromycin phosphotransferase"/protein_id="AAF65346.1"/db_xref="GI:7638077"/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQD LPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWS EAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQ SLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPS TRPRAKK"promoter complement(8712..9492)/note="CaMV35S2; CaMV 35S promoter, duplicated"promoter 9640..9694/note="PlacZ; lacZ promoter"CDS 9722..9955/codon_start=1/product="LacZ alpha fragment"/protein_id="AAF65343.1"/db_xref="GI:7638074"/translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLN RLAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"misc_feature 9736..9792/note="pUC18 MCS; polylinker"promoter 10006..10543/note="CaMV35S; 35S promoter from CaMV"ORIGIN1 catggtagat ctgactagta aaggagaaga acttttcact ggagttgtcc caattcttgt 61 tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga 121 tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc 181 gtggccaaca cttgtcacta ctttctctta tggtgttcaa tgcttttcaa gatacccaga 241 tcatatgaag cggcacgact tcttcaagag cgccatgcct gagggatacg tgcaggagag 301 gaccatcttc ttcaaggacg acgggaacta caagacacgt gctgaagtca agtttgaggg 361 agacaccctc gtcaacagga tcgagcttaa gggaatcgat ttcaaggagg acggaaacat 421 cctcggccac aagttggaat acaactacaa ctcccacaac gtatacatca tggccgacaa 481 gcaaaagaac ggcatcaaag ccaacttcaa gacccgccac aacatcgaag acggcggcgt 541 gcaactcgct gatcattatc aacaaaatac tccaattggc gatggccctg tccttttacc 601 agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga 661 ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact 721 atacaaagct agccaccacc accaccacca cgtgtgaatt ggtgaccagc tcgaatttcc 781 ccgatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg 841 cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat 901 gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat 961 acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat 1021 ctatgttact agatcgggaa ttaaactatc agtgtttgac aggatatatt ggcgggtaaa 1081 cctaagagaa aagagcgttt attagaataa cggatattta aaagggcgtg aaaaggttta 1141 tccgttcgtc catttgtatg tgcatgccaa ccacagggtt cccctcggga tcaaagtact 1201 ttgatccaac ccctccgctg ctatagtgca gtcggcttct gacgttcagt gcagccgtct 1261 tctgaaaacg acatgtcgca caagtcctaa gttacgcgac aggctgccgc cctgcccttt 1321 tcctggcgtt ttcttgtcgc gtgttttagt cgcataaagt agaatacttg cgactagaac 1381 cggagacatt acgccatgaa caagagcgcc gccgctggcc tgctgggcta tgcccgcgtc 1441 agcaccgacg accaggactt gaccaaccaa cgggccgaac tgcacgcggc cggctgcacc1501 aagctgtttt ccgagaagat caccggcacc aggcgcgacc gcccggagct ggccaggatg 1561 cttgaccacc tacgccctgg cgacgttgtg acagtgacca ggctagaccg cctggcccgc 1621 agcacccgcg acctactgga cattgccgag cgcatccagg aggccggcgc gggcctgcgt 1681 agcctggcag agccgtgggc cgacaccacc acgccggccg gccgcatggt gttgaccgtg 1741 ttcgccggca ttgccgagtt cgagcgttcc ctaatcatcg accgcacccg gagcgggcgc 1801 gaggccgcca aggcccgagg cgtgaagttt ggcccccgcc ctaccctcac cccggcacag 1861 atcgcgcacg cccgcgagct gatcgaccag gaaggccgca ccgtgaaaga ggcggctgca 1921 ctgcttggcg tgcatcgctc gaccctgtac cgcgcacttg agcgcagcga ggaagtgacg 1981 cccaccgagg ccaggcggcg cggtgccttc cgtgaggacg cattgaccga ggccgacgcc 2041 ctggcggccg ccgagaatga acgccaagag gaacaagcat gaaaccgcac caggacggcc 2101 aggacgaacc gtttttcatt accgaagaga tcgaggcgga gatgatcgcg gccgggtacg 2161 tgttcgagcc gcccgcgcac gtctcaaccg tgcggctgca tgaaatcctg gccggtttgt 2221 ctgatgccaa gctggcggcc tggccggcca gcttggccgc tgaagaaacc gagcgccgcc 2281 gtctaaaaag gtgatgtgta tttgagtaaa acagcttgcg tcatgcggtc gctgcgtata 2341 tgatgcgatg agtaaataaa caaatacgca aggggaacgc atgaaggtta tcgctgtact 2401 taaccagaaa ggcgggtcag gcaagacgac catcgcaacc catctagccc gcgccctgca 2461 actcgccggg gccgatgttc tgttagtcga ttccgatccc cagggcagtg cccgcgattg 2521 ggcggccgtg cgggaagatc aaccgctaac cgttgtcggc atcgaccgcc cgacgattga 2581 ccgcgacgtg aaggccatcg gccggcgcga cttcgtagtg atcgacggag cgccccaggc 2641 ggcggacttg gctgtgtccg cgatcaaggc agccgacttc gtgctgattc cggtgcagcc 2701 aagcccttac gacatatggg ccaccgccga cctggtggag ctggttaagc agcgcattga 2761 ggtcacggat ggaaggctac aagcggcctt tgtcgtgtcg cgggcgatca aaggcacgcg 2821 catcggcggt gaggttgccg aggcgctggc cgggtacgag ctgcccattc ttgagtcccg 2881 tatcacgcag cgcgtgagct acccaggcac tgccgccgcc ggcacaaccg ttcttgaatc 2941 agaacccgag ggcgacgctg cccgcgaggt ccaggcgctg gccgctgaaa ttaaatcaaa3001 actcatttga gttaatgagg taaagagaaa atgagcaaaa gcacaaacac gctaagtgcc 3061 ggccgtccga gcgcacgcag cagcaaggct gcaacgttgg ccagcctggc agacacgcca 3121 gccatgaagc gggtcaactt tcagttgccg gcggaggatc acaccaagct gaagatgtac 3181 gcggtacgcc aaggcaagac cattaccgag ctgctatctg aatacatcgc gcagctacca 3241 gagtaaatga gcaaatgaat aaatgagtag atgaatttta gcggctaaag gaggcggcat 3301 ggaaaatcaa gaacaaccag gcaccgacgc cgtggaatgc cccatgtgtg gaggaacggg 3361 cggttggcca ggcgtaagcg gctgggttgt ctgccggccc tgcaatggca ctggaacccc 3421 caagcccgag gaatcggcgt gacggtcgca aaccatccgg cccggtacaa atcggcgcgg 3481 cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca 3541 tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag 3601 aatcccggca accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg 3661 agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca 3721 tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc 3781 gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg 3841 tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat 3901 accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac 3961 tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca 4021 ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc 4081 tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa 4141 ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag 4201 aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca 4261 tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt 4321 tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca 4381 ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg 4441 ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg4501 ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc 4561 gaaaaggtct ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga 4621 accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag 4681 tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta 4741 aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc 4801 tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc 4861 ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc 4921 gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accctgcctc 4981 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca 5041 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt 5101 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc 5161 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac 5221 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg 5281 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 5341 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 5401 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 5461 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 5521 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 5581 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 5641 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 5701 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 5761 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 5821 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 5881 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 5941 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc6001 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 6061 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgcattct aggtactaaa 6121 acaattcatc cagtaaaata taatatttta ttttctccca atcaggcttg atccccagta 6181 agtcaaaaaa tagctcgaca tactgttctt ccccgatatc ctccctgatc gaccggacgc 6241 agaaggcaat gtcataccac ttgtccgccc tgccgcttct cccaagatca ataaagccac 6301 ttactttgcc atctttcaca aagatgttgc tgtctcccag gtcgccgtgg gaaaagacaa 6361 gttcctcttc gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga tctttaaatg 6421 gagtgtcttc ttcccagttt tcgcaatcca catcggccag atcgttattc agtaagtaat 6481 ccaattcggc taagcggctg tctaagctat tcgtataggg acaatccgat atgtcgatgg 6541 agtgaaagag cctgatgcac tccgcataca gctcgataat cttttcaggg ctttgttcat 6601 cttcatactc ttccgagcaa aggacgccat cggcctcact catgagcaga ttgctccagc 6661 catcatgccg ttcaaagtgc aggacctttg gaacaggcag ctttccttcc agccatagca 6721 tcatgtcctt ttcccgttcc acatcatagg tggtcccttt ataccggctg tccgtcattt 6781 ttaaatatag gttttcattt tctcccacca gcttatatac cttagcagga gacattcctt 6841 ccgtatcttt tacgcagcgg tatttttcga tcagtttttt caattccggt gatattctca 6901 ttttagccat ttattatttc cttcctcttt tctacagtat ttaaagatac cccaagaagc 6961 taattataac aagacgaact ccaattcact gttccttgca ttctaaaacc ttaaatacca 7021 gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat aacatagtat cgacggagcc 7081 gattttgaaa ccgcggtgat cacaggcagc aacgctctgt catcgttaca atcaacatgc 7141 taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt tcttccgaat 7201 agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct gacgccgtcc 7261 cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc 7321 tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt 7381 aataacacat tgcggacgtt tttaatgtac tgaattaacg ccgaattaat tcgggggatc 7441 tggattttag tactggattt tggttttagg aattagaaat tttattgata gaagtatttt7501 acaaatacaa atacatacta agggtttctt atatgctcaa cacatgagcg aaaccctata 7561 ggaaccctaa ttcccttatc tgggaactac tcacacatta ttatggagaa actcgagctt 7621 gtcgatcgac agatccggtc ggcatctact ctatttcttt gccctcggac gagtgctggg 7681 gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga cggccgcgct 7741 tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga ttgcgtcgca 7801 tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct gatagagttg 7861 gtcaagacca atgcggagca tatacgcccg gagtcgtggc gatcctgcaa gctccggatg 7921 cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc tccagaagaa 7981 gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt caatgaccgc 8041 tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt gcacgaggtg 8101 ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct gcgcgacgga 8161 cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat cagcaatcgc 8221 gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga atgggccgaa 8281 cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atagcctccg cgaccggttg 8341 tagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg 8401 ggagatgcaa taggtcaggc tctcgctaaa ctccccaatg tcaagcactt ccggaatcgg 8461 gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac catcggcgca 8521 gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag cacgagattc 8581 ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga tcagaaactt 8641 ctcgacagac gtcgcggtga gttcaggctt tttcatatct cattgccccc cgggatctgc 8701 gaaagctcga gagagataga tttgtagaga gagactggtg atttcagcgt gtcctctcca 8761 aatgaaatga acttccttat atagaggaag gtcttgcgaa ggatagtggg attgtgcgtc 8821 atcccttacg tcagtggaga tatcacatca atccacttgc tttgaagacg tggttggaac 8881 gtcttctttt tccacgatgc tcctcgtggg tgggggtcca tctttgggac cactgtcggc 8941 agaggcatct tgaacgatag cctttccttt atcgcaatga tggcatttgt aggtgccacc9001 ttccttttct actgtccttt tgatgaagtg acagatagct gggcaatgga atccgaggag 9061 gtttcccgat attacccttt gttgaaaagt ctcaatagcc ctttggtctt ctgagactgt 9121 atctttgata ttcttggagt agacgagagt gtcgtgctcc accatgttat cacatcaatc 9181 cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc acgatgctcc tcgtgggtgg 9241 gggtccatct ttgggaccac tgtcggcaga ggcatcttga acgatagcct ttcctttatc 9301 gcaatgatgg catttgtagg tgccaccttc cttttctact gtccttttga tgaagtgaca 9361 gatagctggg caatggaatc cgaggaggtt tcccgatatt accctttgtt gaaaagtctc 9421 aatagccctt tggtcttctg agactgtatc tttgatattc ttggagtaga cgagagtgtc 9481 gtgctccacc atgttggcaa gctgctctag ccaatacgca aaccgcctct ccccgcgcgt 9541 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag 9601 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg 9661 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc 9721 tatgaccatg attacgaatt cgagctcggt acccggggat cctctagagt cgacctgcag 9781 gcatgcaagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 9841 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga 9901 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gctagagcag 9961 cttgagcttg gatcagattg tcgtttcccg ccttcagttt agcttcatgg agtcaaagat 10021 tcaaatagag gacctaacag aactcgccgt aaagactggc gaacagttca tacagagtct 10081 cttacgactc aatgacaaga agaaaatctt cgtcaacatg gtggagcacg acacacttgt 10141 ctactccaaa aatatcaaag atacagtctc agaagaccaa agggcaattg agacttttca 10201 acaaagggta atatccggaa acctcctcgg attccattgc ccagctatct gtcactttat 10261 tgtgaagata gtggaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa 10321 ggccatcgtt gaagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag 10381 gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga 10441 tatctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc10501 tatataagga agttcatttc atttggagag aacacggggg actcttgac //。
