pGEX4T1大肠杆菌表达载体说明

pGEX-4T-1编号载体名称北京华越洋生物VECT4940pGEX-­‐4T-­‐1pGEX4T1载体基本信息别名:pGEX-­‐4T-­‐1,pGEX4T1,pGEX4T1质粒类型:大肠杆菌蛋白表达载体表达水平:高拷贝启动子:Tac克隆方法:多克隆位点，限制性内切酶载体大小:4969bp5'测序引物及序列:pGEX5':GGGCTGGCAAGCCACGTTTGGTG3'测序引物及序列:pGEX3':CCGGGAGCTGCATGTGTCAGAGG载体标签:N-­‐GST载体抗性:Ampicillin氨苄备注:复制子是pMB1，限制性内切酶酶切位点有BamHI,EcoRISmaI,SalI,XhoI,NotI等，可转化进大肠杆菌中高拷贝表达。pGEX-­‐4T-­‐1载体是一种常用的原核表达载体，该载体多克隆位点区域含有多个常用的内切酶位点序列，便于不同基因的克隆；且全序列中含有tac启动子、GST标签序列，是一种高效的蛋白表达载体。Tac启动子在没有诱导物存在的情况下，质粒上携带的lacIq基因产物可以有效抑制tac启动子的转录。可在BL21、Rosstea等表达菌株中高表达融合有GST标签的蛋白，且用凝血酶可将目的蛋白的GST标签切掉，便于下游蛋白纯化。稳定性:瞬时表达Transient组成型:诱导表达病毒/非病毒:非病毒pGEX4T1载体质粒图谱和多克隆位点信息pGEX4T1载体简介TherearearangeofGluthationeSepharose™prepackedcolumnandbulkmediaproductsavailabletopurifyGSTFusionproteins.Formanualpurificationofsamplevolumesupto600µluseGSTSpinTrap™microspincolumnsorGSTMultiTrap™4B96-­‐wellplates.Forsamplevolumesbetween600µland10mluseGSTGraviTrap™gravityflowcolumn.Wheresamplevolumesareabove10ml,useLabMate™reservoirtogetherwithGSTGraviTrap.Allformatsdescribedcanbeusedforpreparationofsamplesinparallel.InadditionGSTHiTrap™1and5mlcolumnsandGSTHiPrep™FF16/10columnareavailableforpurificationinachromatographysystemsuchastheÄKTA™designsystem.Alternatively,GluthationeSepharosebulkmediaareavailablefrom10mlupto500ml.AGSTBulkKitisalsoavailablecombining10mlGluthationeSepharose4Bbulkmediumand5emptycolumnswithrequiredbuffers.pGEX系列载体：pGEX-­‐1λTEcoRI/BAPpGEX-­‐2TpGEX-­‐2TKpGEX-­‐20TpGEX-­‐3XpGEX-­‐4T-­‐1pGEX-­‐4T-­‐2pGEX-­‐4T-­‐3pGEX-­‐5X-­‐1pGEX-­‐5X-­‐2pGEX-­‐5X-­‐3pGEX-­‐6P-­‐1pGEX-­‐6P-­‐2pGEX-­‐6P-­‐3pGEX-­‐KGpGEX4T1载体序列ORIGIN1ACGTTATCGACTGCACGGTGCACCAATGCTTCTGGCGTCAGGCAGCCATCGGAAGCTGTG61GTATGGCTGTGCAGGTCGTAAATCACTGCATAATTCGTGTCGCTCAAGGCGCACTCCCGT121TCTGGATAATGTTTTTTGCGCCGACATCATAACGGTTCTGGCAAATATTCTGAAATGAGC181TGTTGACAATTAATCATCGGCTCGTATAATGTGTGGAATTGTGAGCGGATAACAATTTCA241CACAGGAAACAGTATTCATGTCCCCTATACTAGGTTATTGGAAAATTAAGGGCCTTGTGC301AACCCACTCGACTTCTTTTGGAATATCTTGAAGAAAAATATGAAGAGCATTTGTATGAGC361GCGATGAAGGTGATAAATGGCGAAACAAAAAGTTTGAATTGGGTTTGGAGTTTCCCAATC421TTCCTTATTATATTGATGGTGATGTTAAATTAACACAGTCTATGGCCATCATACGTTATA481TAGCTGACAAGCACAACATGTTGGGTGGTTGTCCAAAAGAGCGTGCAGAGATTTCAATGC541TTGAAGGAGCGGTTTTGGATATTAGATACGGTGTTTCGAGAATTGCATATAGTAAAGACT601TTGAAACTCTCAAAGTTGATTTTCTTAGCAAGCTACCTGAAATGCTGAAAATGTTCGAAG661ATCGTTTATGTCATAAAACATATTTAAATGGTGATCATGTAACCCATCCTGACTTCATGT721TGTATGACGCTCTTGATGTTGTTTTATACATGGACCCAATGTGCCTGGATGCGTTCCCAA781AATTAGTTTGTTTTAAAAAACGTATTGAAGCTATCCCACAAATTGATAAGTACTTGAAAT841CCAGCAAGTATATAGCATGGCCTTTGCAGGGCTGGCAAGCCACGTTTGGTGGTGGCGACC901ATCCTCCAAAATCGGATCTGGTTCCGCGTGGATCCCCGGAATTCCCGGGTCGACTCGAGC961GGCCGCATCGTGACTGACTGACGATCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACC1021TCTGACACATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCA1081GACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCGCAGCCATGACCC1141AGTCACGTAGCGATAGCGGAGTGTATAATTCTTGAAGACGAAAGGGCCTCGTGATACGCC1201TATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTC1261GGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATC1321CGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGA1381GTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTT1441TTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAG1501TGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAG1561AACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTG1621TTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTG1681AGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCA1741GTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAG1801GACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATC1861GTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTG1921CAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCC1981GGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGG2041CCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCG2101GTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGA2161CGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCAC2221TGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAA2281AACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCA2341AAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAG2401GATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCAC2461CGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAA2521CTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCC2581ACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAG2641TGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTAC2701CGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGC2761GAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTC2821CCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCA2881CGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACC2941TCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACG3001CCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCT3061TTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATA3121CCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGC3181GCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATAAATTCCG3241ACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTC3301AATTCAGGGTGGTGAATGTGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTG3361TCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGC3421GGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAAC3481AACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACG3541CGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGG3601TGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTC3661TCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTG3721CTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACAC3781CCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGG3841TCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGC3901GTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGG3961AACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATG4021AGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGC4081GCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACG4141ATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCC4201TGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGG4261GCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGC4321AAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCC4381GACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCA4441CCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAA4501CAATTTCACACAGGAAACAGCTATGACCATGATTACGGATTCACTGGCCGTCGTTTTACA4561ACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCC4621TTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCG4681CAGCCTGAATGGCGAATGGCGCTTTGCCTGGTTTCCGGCACCAGAAGCGGTGCCGGAAAG4741CTGGCTGGAGTGCGATCTTCCTGAGGCCGATACTGTCGTCGTCCCCTCAAACTGGCAGAT4801GCACGGTTACGATGCGCCCATCTACACCAACGTAACCTATCCCATTACGGTCAATCCGCC4861GTTTGTTCCCACGGAGAATCCGACGGGTTGTTACTCGCTCACATTTAATGTTGATGAAAG4921CTGGCTACAGGAAGGCCAGACGCGAATTATTTTTGATGGCGTTGGAATT//其他大肠杆菌表达载体：pBV221ptdTomatopET-52b(+)pAmCyanpDsRed-Express2pBV220pCold-GSTpColdS-SUMOpColdTFpColdIVpColdIIIpColdIIpColdIpE-SUMOpCold-ProS2pBAD102/D-TOPOpBAD202/D-TOPOpACYC184pBAD/Thio-TOPOpBad/Myc-HisCpBad/Myc-HisBpBad/Myc-HisApBad/HisCpBad/HisBpBad/HisApBAD-TOPOpET-23b(+)pET-23a(+)pET-23c(+)pET-23(+)pET-12b(+)pET-12c(+)pET-12a(+)pET-11b(+)pET-11a(+)pET-11c(+)pBad24pQE-82LpQE-81LpQE-80LpQE-32pQE-9pQE-16pQE-31pQE-60pQE-70pQE-40pET-51b(+)pET-50b(+)pET-49b(+)pET-48b(+)pET-47b(+)pET-26b(+)pET-32a(+)pET-21b(+)pET-22b(+)pET-14bpET-16bpET-15bpET-19bpET-20b(+)pET-21d(+)pET-21c(+)pET-21b(+)pET-21a(+)pET-24a(+)pET-24d(+)pET-25b(+)pET-27b(+)pET-28a(+)pET-30a(+)pET-42a(+)pET-43.1c(+)pET-43.1b(+)pET-43.1a(+)pET-44a(+)pET-44c(+)pET-46EK/LICpET-37b(+)pTrcHis2CpTrcHis2BpTrcHis2ApET303/CT-HispET302/NT-HispRSET-CFPpRSET-EmGFPpRSET-BFPpGFPuvpET300/NT-DESTpET301/CT-DESTpGEM-TpBad43pGEX-4T-3pGEX-5X-2pBlueScriptSK(+)pG-Tf2pG-KJE8pGro7pET-SUMOpSE380pET-17bpET102/D-TOPOpCDFDuet-1pMAL-p5xpTf16pET-28c(+)pBluescriptIISK(+)pET-30b(+)pSUMOpProEXHTcpProEXHTbpProEXHTapKD3pKD13pKD46pTYB1pTYB2pTWIN2pBluescriptIIKS(-)pTYB12pMAL-p5epACYCDuet-1pEGM-11ZF(+)pEGM-7ZF(+)PinPointXa-3PinPointXa-2PinPointXa-1pSP73pSP64pTWIN1pTYB11pTXB1pET-5b(+)pBad/gIIICpBad/gIIIBpBad/gIIIApET-5a(+)pMal-p4XpMal-p2Gpkk223-3pkk232-8pCYB1pEZZ18pBAD18pMAL-c5xpMal-p2EpMal-p2XpET-44EK/LICpET-43.1EK/LICpET-41EK/LICpMal-c4XpTrcHisBpET-31b(+)pET-3b(+)pET-41a(+)pGEX-3XpGEX-4T-2pETDuet-1pGEX-4T-1pTrc99apET-28b(+)pET-HispALEXa,b,cpACYC177pBR322pKD4pKD20pMXB10pEcoli-6xHN-GFPuvpKJE7pRSETBpGEX-KGpGEX-2TpRSFDuet-1pCOLADuet-1pTrcHisCpTrcHisApET-41b(+)pET-42b(+)pET-3a(+)pGEX-6P-3pGEX-6P-2pGEX-6P-1pGEX-5X-3pGEX-5X-1pGEX-2TKpRSETApMal-c2GpMal-c2EpMal-c2XpRSETCpQE-30pET-45b(+)pET-44b(+)pET-42c(+)pET-41c(+)pET-40b(+)pET-33b(+)pET-39b(+)pET-32EK/LICpET-32Xa/LICpET-32c(+)pET-32b(+)pET-30Xa/LICpET-30EK/LICpET-30c(+)pET-29c(+)pET-29b(+)pET-29a(+)pET-24c(+)pET-24b(+)pET-24(+)pET-23d(+)pET-11d(+)pBad33