pET-41b(+)大肠杆菌表达载体说明

pET-41b(+)

编号 名称

北京华越洋VECT--‐540 pET--‐41b(+)

pET41b载体基本信息

别名: pET41b, p ET 41b

质粒类型: 大肠杆菌蛋白表达

表达水平: 高

克隆方法: 多克隆位点，限制性内切酶 载体大小: 5932 b p

5' 测序引物: T7或pGEX--‐5‘

5' 测序引物序列: T7: 5'--‐TAATACGACTCACTATAGGG--‐3'; pGEX--‐5': 5'--‐GGGCTGGCAAGCCACGTTTGGTG--‐3'

载体标签: N--‐GST, N--‐His, N--‐Thrombin 载体抗性: Kanamycin

备注: Encodes GST fusion tag; Nterm thrombin cleavage site; Nterm enterokinase c leavage s ite; P shAI b lunt c loning s ite; a,b,c v ary b y M CS

稳定性: 瞬时表达 Transient 组成型: 组成型 Constitutive 病毒/非病毒: 非病毒

pET41b载体质粒图谱和多克隆位点信息

Feature N ame Start End T7_terminator 120 1 T7_Terminal_primer 69 87 EK 278 264 S15 353 309 6xHIS 413 396 GST (variant) 1094 444 T7_transl_en_RBS 1119 1103 lacO 1164 1137 T7_promoter 1182 1164 tet (300 --‐ 563) 1218 1481 pBRrevBam_primer 1289 1270 lacI 1564 2655 ROP

3227 3418 pGEX_3_primer 3434 3412 pBR322_origin

4452

3833

Feature N ame Start End

KanR2 4558 5373

ORF Start End

ORF f rame 3 1094 147

ORF f rame 3 1095 1823

ORF f rame 1 1696 2655

ORF f rame 1 4558 5373

Enzyme N ame Cut

XhoI 174

NotI 182

EagI 182

HindIII 189

SalI 195

SacI 206

PstI 214

AscI 216

StuI 226

EcoRI 235

BamHI 241

EcoRV 251

NcoI 254

AgeI 291

KpnI 298

BglII 301

SacII 392

SpeI 423

MscI 886

NdeI 1093

XbaI 1131

ApaI 2134

HpaI 2429

NruI 4646

SmaI 4863

XmaI 4861

pET41b载体简介

The pET--‐41 series is designed for cloning and high--‐level expression of peptide sequences fused with the 220 aa GST?Tag? protein. Unique sites are shown on the circle map. Note that the sequence i s n umbered b y t he p BR322 c onvention, s o t he T7 e xpression r egion i s r eversed o n t he circle m ap. T he c loning/expression r egion o f t he c oding s trand t ranscribed b y T7 R NA p olymerase is s hown b elow. T he f1 o rigin i s o riented s o t hat i nfection w ith h elper p hage w ill p roduce v irions containing s ingle s tranded D NA t hat c orresponds t o t he c oding s trand. T herefore, s ingle s tranded

sequencing should be performed using the T7 terminator primer. Vector encoded sequence can be completely removed when cloning into the PshAI site and then cleaving the GST fusion protein w ith E nterokinase.

pET41b载体序列

ORIGIN

1 ATCCGGATAT AGTTCCTCCT TTCAGCAAAA AACCCCTCAA GACCCGTTTA GAGGCCCCAA 61 GGGGTTATGC TAGTTATTGC TCAGCGGTGG CAGCAGCCAA CTCAGCTTCC TTTCGGGCTT 121 TGTTTAGCAG CCTAGGTATT AATCAATTAG TGGTGGTGGT GGTGGTGGTG GTGCTCGAGT 181 GCGGCCGCAA GCTTGTCGAC GGAGCTCGCC TGCAGGCGCG CCAAGGCCTG TACAGAATTC 241 GGATCCCCGA TATCCATGGG ACTCTTGTCG TCGTCATCAC CGGAGCCACC ACCGGTACCC 301 AGATCTGGGC TGTCCATGTG CTGGCGTTCG AATTTAGCAG CAGCGGTTTC TTTCATACCA 361 ATTGCAGTAC TACCGCGTGG CACCAGACCC GCGGAGTGAT GGTGATGGTG ATGACCAGAA 421 CCACTAGTTG AACCATCCGA TTTTGGAGGA TGGTCGCCAC CACCAAACGT GGCTTGCCAG 481 CCCTGCAAAG GCCATGCTAT ATACTTGCTG GATTTCAAGT ACTTATCAAT TTGTGGGATA 541 GCTTCAATAC GTTTTTTAAA ACAAACTAAT TTTGGGAACG CATCCAGGCA CATTGGGTCC 601 ATGTATAAAA CAACATCAAG AGCGTCATAC AACATGAAGT CAGGATGGGT TACATGATCA 661 CCATTTAAAT ATGTTTTATG ACATAAACGA TCTTCGAACA TTTTCAGCAT TTCAGGTAGC 721 TTGCTAAGAA AATCAACTTT GAGAGTTTCA AAGTCTTTAC TATATGCAAT TCTCGAAACA 781 CCGTATCTAA TATCCAAAAC CGCTCCTTCA AGCATTGAAA TCTCTGCACG CTCTTTTGGA 841 CAACCACCCA ACATGTTGTG CTTGTCAGCT ATATAACGTA TGATGGCCAT AGACTGTGTT 901 AATTTAACAT CACCATCAAT ATAATAAGGA AGATTGGGAA ACTCCAAACC CAATTCAAAC 961 TTTTTGTTTC GCCATTTATC ACCTTCATCG CGCTCATACA AATGCTCTTC ATATTTTTCT 1021 TCAAGATATT CCAAAAGAAG TCGAGTGGGT TGCACAAGGC CCTTAATTTT CCAATAACCT 1081 AGTATAGGGG ACATATGTAT ATCTCCTTCT TAAAGTTAAA CAAAATTATT TCTAGAGGGG 1141 AATTGTTATC CGCTCACAAT TCCCCTATAG TGAGTCGTAT TAATTTCGCG GGATCGAGAT 1201 CGATCTCGAT CCTCTACGCC GGACGCATCG TGGCCGGCAT CACCGGCGCC ACAGGTGCGG 1261 TTGCTGGCGC CTATATCGCC GACATCACCG ATGGGGAAGA TCGGGCTCGC CACTTCGGGC 1321 TCATGAGCGC TTGTTTCGGC GTGGGTATGG TGGCAGGCCC CGTGGCCGGG GGACTGTTGG 1381 GCGCCATCTC CTTGCATGCA CCATTCCTTG CGGCGGCGGT GCTCAACGGC CTCAACCTAC 1441 TACTGGGCTG CTTCCTAATG CAGGAGTCGC ATAAGGGAGA GCGTCGAGAT CCCGGACACC 1501 ATCGAATGGC GCAAAACCTT TCGCGGTATG GCATGATAGC GCCCGGAAGA GAGTCAATTC 1561 AGGGTGGTGA ATGTGAAACC AGTAACGTTA TACGATGTCG CAGAGTATGC CGGTGTCTCT 1621 TATCAGACCG TTTCCCGCGT GGTGAACCAG GCCAGCCACG TTTCTGCGAA AACGCGGGAA 1681 AAAGTGGAAG CGGCGATGGC GGAGCTGAAT TACATTCCCA ACCGCGTGGC ACAACAACTG 1741 GCGGGCAAAC AGTCGTTGCT GATTGGCGTT GCCACCTCCA GTCTGGCCCT GCACGCGCCG 1801 TCGCAAATTG TCGCGGCGAT TAAATCTCGC GCCGATCAAC TGGGTGCCAG CGTGGTGGTG 1861 TCGATGGTAG AACGAAGCGG CGTCGAAGCC TGTAAAGCGG CGGTGCACAA TCTTCTCGCG 1921 CAACGCGTCA GTGGGCTGAT CATTAACTAT CCGCTGGATG ACCAGGATGC CATTGCTGTG 1981 GAAGCTGCCT GCACTAATGT TCCGGCGTTA TTTCTTGATG TCTCTGACCA GACACCCATC 2041 AACAGTATTA TTTTCTCCCA TGAAGACGGT ACGCGACTGG GCGTGGAGCA TCTGGTCGCA 2101 TTGGGTCACC AGCAAATCGC GCTGTTAGCG GGCCCATTAA GTTCTGTCTC GGCGCGTCTG 2161 CGTCTGGCTG GCTGGCATAA ATATCTCACT CGCAATCAAA TTCAGCCGAT AGCGGAACGG 2221 GAAGGCGACT GGAGTGCCAT GTCCGGTTTT CAACAAACCA TGCAAATGCT GAATGAGGGC

2281 ATCGTTCCCA CTGCGATGCT GGTTGCCAAC GATCAGATGG CGCTGGGCGC AATGCGCGCC 2341 ATTACCGAGT CCGGGCTGCG CGTTGGTGCG GACATCTCGG TAGTGGGATA CGACGATACC 2401 GAAGACAGCT CATGTTATAT CCCGCCGTTA ACCACCATCA AACAGGATTT TCGCCTGCTG 2461 GGGCAAACCA GCGTGGACCG CTTGCTGCAA CTCTCTCAGG GCCAGGCGGT GAAGGGCAAT 2521 CAGCTGTTGC CCGTCTCACT GGTGAAAAGA AAAACCACCC TGGCGCCCAA TACGCAAACC 2581 GCCTCTCCCC GCGCGTTGGC CGATTCATTA ATGCAGCTGG CACGACAGGT TTCCCGACTG 2641 GAAAGCGGGC AGTGAGCGCA ACGCAATTAA TGTAAGTTAG CTCACTCATT AGGCACCGGG 2701 ATCTCGACCG ATGCCCTTGA GAGCCTTCAA CCCAGTCAGC TCCTTCCGGT GGGCGCGGGG 2761 CATGACTAGC ATGATCGTGC TCCTGTCGTT GAGGACCCGG CTAGGCTGGC GGGGTTGCCT 2821 TACTGGTTAG CAGAATGAAT CACCGATACG CGAGCGAACG TGAAGCGACT GCTGCTGCAA 2881 AACGTCTGCG ACCTGAGCAA CAACATGAAT GGTCTTCGGT TTCCGTGTTT CGTAAAGTCT 2941 GGAAACGCGG AAGTCAGCGC CCTGCACCAT TATGTTCCGG ATCTGCATCG CAGGATGCTG 3001 CTGGCTACCC TGTGGAACAC CTACATCTGT ATTAACGAAG CGCTGGCATT GACCCTGAGT 3061 GATTTTTCTC TGGTCCCGCC GCATCCATAC CGCCAGTTGT TTACCCTCAC AACGTTCCAG 3121 TAACCGGGCA TGTTCATCAT CAGTAACCCG TATCGTGAGC ATCCTCTCTC GTTTCATCGG 3181 TATCATTACC CCCATGAACA GAAATCCCCC TTACACGGAG GCATCAGTGA CCAAACAGGA 3241 AAAAACCGCC CTTAACATGG CCCGCTTTAT CAGAAGCCAG ACATTAACGC TTCTGGAGAA 3301 ACTCAACGAG CTGGACGCGG ATGAACAGGC AGACATCTGT GAATCGCTTC ACGACCACGC 3361 TGATGAGCTT TACCGCAGCT GCCTCGCGCG TTTCGGTGAT GACGGTGAAA ACCTCTGACA 3421 CATGCAGCTC CCGGAGACGG TCACAGCTTG TCTGTAAGCG GATGCCGGGA GCAGACAAGC 3481 CCGTCAGGGC GCGTCAGCGG GTGTTGGCGG GTGTCGGGGC GCAGCCATGA CCCAGTCACG 3541 TAGCGATAGC GGAGTGTATA CTGGCTTAAC TATGCGGCAT CAGAGCAGAT TGTACTGAGA 3601 GTGCACCATA TATGCGGTGT GAAATACCGC ACAGATGCGT AAGGAGAAAA TACCGCATCA 3661 GGCGCTCTTC CGCTTCCTCG CTCACTGACT CGCTGCGCTC GGTCGTTCGG CTGCGGCGAG 3721 CGGTATCAGC TCACTCAAAG GCGGTAATAC GGTTATCCAC AGAATCAGGG GATAACGCAG 3781 GAAAGAACAT GTGAGCAAAA GGCCAGCAAA AGGCCAGGAA CCGTAAAAAG GCCGCGTTGC 3841 TGGCGTTTTT CCATAGGCTC CGCCCCCCTG ACGAGCATCA CAAAAATCGA CGCTCAAGTC 3901 AGAGGTGGCG AAACCCGACA GGACTATAAA GATACCAGGC GTTTCCCCCT GGAAGCTCCC 3961 TCGTGCGCTC TCCTGTTCCG ACCCTGCCGC TTACCGGATA CCTGTCCGCC TTTCTCCCTT 4021 CGGGAAGCGT GGCGCTTTCT CATAGCTCAC GCTGTAGGTA TCTCAGTTCG GTGTAGGTCG 4081 TTCGCTCCAA GCTGGGCTGT GTGCACGAAC CCCCCGTTCA GCCCGACCGC TGCGCCTTAT 4141 CCGGTAACTA TCGTCTTGAG TCCAACCCGG TAAGACACGA CTTATCGCCA CTGGCAGCAG 4201 CCACTGGTAA CAGGATTAGC AGAGCGAGGT ATGTAGGCGG TGCTACAGAG TTCTTGAAGT 4261 GGTGGCCTAA CTACGGCTAC ACTAGAAGGA CAGTATTTGG TATCTGCGCT CTGCTGAAGC 4321 CAGTTACCTT CGGAAAAAGA GTTGGTAGCT CTTGATCCGG CAAACAAACC ACCGCTGGTA 4381 GCGGTGGTTT TTTTGTTTGC AAGCAGCAGA TTACGCGCAG AAAAAAAGGA TCTCAAGAAG 4441 ATCCTTTGAT CTTTTCTACG GGGTCTGACG CTCAGTGGAA CGAAAACTCA CGTTAAGGGA 4501 TTTTGGTCAT GAACAATAAA ACTGTCTGCT TACATAAACA GTAATACAAG GGGTGTTATG 4561 AGCCATATTC AACGGGAAAC GTCTTGCTCT AGGCCGCGAT TAAATTCCAA CATGGATGCT 4621 GATTTATATG GGTATAAATG GGCTCGCGAT AATGTCGGGC AATCAGGTGC GACAATCTAT 4681 CGATTGTATG GGAAGCCCGA TGCGCCAGAG TTGTTTCTGA AACATGGCAA AGGTAGCGTT 4741 GCCAATGATG TTACAGATGA GATGGTCAGA CTAAACTGGC TGACGGAATT TATGCCTCTT 4801 CCGACCATCA AGCATTTTAT CCGTACTCCT GATGATGCAT GGTTACTCAC CACTGCGATC 4861 CCCGGGAAAA CAGCATTCCA GGTATTAGAA GAATATCCTG ATTCAGGTGA AAATATTGTT

4921 GATGCGCTGG CAGTGTTCCT GCGCCGGTTG CATTCGATTC CTGTTTGTAA TTGTCCTTTT 4981 AACAGCGATC GCGTATTTCG TCTCGCTCAG GCGCAATCAC GAATGAATAA CGGTTTGGTT 5041 GATGCGAGTG ATTTTGATGA CGAGCGTAAT GGCTGGCCTG TTGAACAAGT CTGGAAAGAA 5101 ATGCATAAAC TTTTGCCATT CTCACCGGAT TCAGTCGTCA CTCATGGTGA TTTCTCACTT 5161 GATAACCTTA TTTTTGACGA GGGGAAATTA ATAGGTTGTA TTGATGTTGG ACGAGTCGGA 5221 ATCGCAGACC GATACCAGGA TCTTGCCATC CTATGGAACT GCCTCGGTGA GTTTTCTCCT 5281 TCATTACAGA AACGGCTTTT TCAAAAATAT GGTATTGATA ATCCTGATAT GAATAAATTG 5341 CAGTTTCATT TGATGCTCGA TGAGTTTTTC TAAGAATTAA TTCATGAGCG GATACATATT 5401 TGAATGTATT TAGAAAAATA AACAAATAGG GGTTCCGCGC ACATTTCCCC GAAAAGTGCC 5461 ACCTGAAATT GTAAACGTTA ATATTTTGTT AAAATTCGCG TTAAATTTTT GTTAAATCAG 5521 CTCATTTTTT AACCAATAGG CCGAAATCGG CAAAATCCCT TATAAATCAA AAGAATAGAC 5581 CGAGATAGGG TTGAGTGTTG TTCCAGTTTG GAACAAGAGT CCACTATTAA AGAACGTGGA 5641 CTCCAACGTC AAAGGGCGAA AAACCGTCTA TCAGGGCGAT GGCCCACTAC GTGAACCATC 5701 ACCCTAATCA AGTTTTTTGG GGTCGAGGTG CCGTAAAGCA CTAAATCGGA ACCCTAAAGG 5761 GAGCCCCCGA TTTAGAGCTT GACGGGGAAA GCCGGCGAAC GTGGCGAGAA AGGAAGGGAA 5821 GAAAGCGAAA GGAGCGGGCG CTAGGGCGCT GGCAAGTGTA GCGGTCACGC TGCGCGTAAC 5881 CACCACACCC GCCGCGCTTA ATGCGCCGCT ACAGGGCGCG TCCCATTCGC CA

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其他大肠杆菌表达载体：

pBV221 ptdTomato pET--‐52b(+) pAmCyan

pDsRed--‐Express2 pBV220 pCold--‐GST pColdS--‐SUMO

pCold T F pCold I V pCold I II pCold I I

pCold I pE--‐SUMO pCold--‐ProS2 pBAD102/D--‐TOPO

pBAD202/D--‐TOPO pACYC184 pBAD/Thio--‐TOPO pBad/Myc--‐His C

pBad/Myc--‐His B pBad/Myc--‐His A pBad/His C pBad/His B

pBad/His A pBAD--‐TOPO pET--‐23b(+) pET--‐23a(+)

pET--‐23c(+) pET--‐23(+) pET--‐12b(+) pET--‐12c(+)

pET--‐12a(+) pET--‐11b(+) pET--‐11a(+) pET--‐11c(+)

pBad24 pQE--‐82L pQE--‐81L pQE--‐80L

pQE--‐32 pQE--‐9 pQE--‐16 pQE--‐31

pQE--‐60 pQE--‐70 pQE--‐40 pET--‐51b(+)

pET--‐50b(+) pET--‐49b(+) pET--‐48b(+) pET--‐47b(+)

pET--‐26b(+) pET--‐32a(+) pET--‐21b(+) pET--‐22b(+)

pET--‐14b pET--‐16b pET--‐15b pET--‐19b

pET--‐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--‐46 E K/LIC pET--‐37b(+) pTrcHis2 C

pTrcHis2 B pTrcHis2 A pET303/CT--‐His pET302/NT--‐His

pRSET--‐CFP pRSET--‐EmGFP pRSET--‐BFP pGFPuv

pET300/NT--‐DEST pET301/CT--‐DEST pGEM--‐T pBad43

pGEX--‐4T--‐3 pGEX--‐5X--‐2 pBlueScript S K(+) pG--‐Tf2

pG--‐KJE8 pGro7 pET--‐SUMO pSE380

pET--‐17b pET102/D--‐TOPO pCDFDuet--‐1 pMAL--‐p5x pTf16 pET--‐28c(+) pBluescript I I S K(+) pET--‐30b(+) pSUMO pProEX H Tc pProEX H Tb pProEX H Ta pKD3 pKD13 pKD46 pTYB1 pTYB2 pTWIN2 pBluescript I I K S(--‐) pTYB12 pMAL--‐p5e pACYCDuet--‐1 pEGM--‐11ZF(+) pEGM--‐7ZF(+) PinPoint X a--‐3 PinPoint X a--‐2 PinPoint X a--‐1 pSP73

pSP64 pTWIN1 pTYB11 pTXB1

pET--‐5b(+) pBad/gIII C pBad/gIII B pBad/gIII A pET--‐5a(+) pMal--‐p4X pMal--‐p2G pkk223--‐3 pkk232--‐8 pCYB1 pEZZ18 pBAD18 pMAL--‐c5x pMal--‐p2E pMal--‐p2X pET--‐44 E K/LIC pET--‐43.1 E K/LIC pET--‐41 E K/LIC pMal--‐c4X pTrcHis B pET--‐31b(+) pET--‐3b(+) pET--‐41a(+) pGEX--‐3X pGEX--‐4T--‐2 pETDuet--‐1 pGEX--‐4T--‐1 pTrc99a pET--‐28b(+) pET--‐His pALEX a,b,c pACYC177 pBR322 pKD4 pKD20 pMXB10 pEcoli--‐6xHN--‐GFPuv pKJE7 pRSET B pGEX--‐KG pGEX--‐2T pRSFDuet--‐1 pCOLADuet--‐1 pTrcHis C pTrcHis A pET--‐41b(+) pET--‐42b(+) pET--‐3a(+) pGEX--‐6P--‐3 pGEX--‐6P--‐2 pGEX--‐6P--‐1 pGEX--‐5X--‐3 pGEX--‐5X--‐1 pGEX--‐2TK pRSET A pMal--‐c2G pMal--‐c2E pMal--‐c2X pRSET C pQE--‐30

pET--‐45b(+) pET--‐44b(+) pET--‐42c(+) pET--‐41c(+) pET--‐40b(+) pET--‐33b(+) pET--‐39b(+) pET--‐32 E K/LIC pET--‐32 X a/LIC pET--‐32c(+) pET--‐32b(+) pET--‐30 X a/LIC pET--‐30 E K/LIC pET--‐30c(+) pET--‐29c(+) pET--‐29b(+) pET--‐29a(+) pET--‐24c(+) pET--‐24b(+) pET--‐24(+) pET--‐23d(+) pET--‐11d(+) pBad33