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pBad/His A

pBad/His A

pBad/His A

 

编号

载体名称

北京华越洋生物VECT4070

pBad/His   A

 

pBadHis A载体基本信息

载体名称:

pBAD/His A

质粒类型:

大肠杆菌表达载体;诱导表达载体

高拷贝/低拷贝:

低拷贝

克隆方法:

限制性内切酶;多克隆位点

启动子:

araBAD

载体大小:

4102 bp

5' 测序引物及序列:

pBAD Forward: 5′-ATGCCATAGCATTTTTATCC-3′

3' 测序引物及序列:

pBAD Reverse 5′-GATTTAATCTGTATCAGG-3′

载体标签:

6x His TagN-端),Xpress Epitope TagN-端), EK 切割位点

载体抗性:

氨苄青霉素(Ampicillin

克隆菌株:

TOP10

表达菌株:

推荐LMG194

备注:

pBAD/His A载体是阿拉伯糖调控载体;
  在无葡萄糖的培养基中,阿拉伯糖正向调控目的基因   的表达;
  通过调节阿拉伯糖的浓度水平来优化目的蛋白的可溶性表达;
  pBAD/His A,B,C之间仅阅读框不同。

稳定性:

稳表达

组成型/诱导型:

诱导型(阿拉伯糖)

病毒/非病毒:

非病毒

 

pBadHis A载体质粒图谱和多克隆位点信息






 

pBadHis A载体简介

pBAD/HisPBAD/Myc-His载体质粒是衍生于pBR322载体。载体设计用来在大肠杆菌中进行可调节,剂量依赖性的表达和纯化重组目的蛋白。使用大肠杆菌araBAD启动子(pBAD)增强了大肠杆菌重组蛋白可溶性表达的水平。pBAD/HispBAD/Myc His载体上的调节蛋白AraC能够调控pBad启动子。

 

pBAD/His A,B,C 载体介绍

The pBAD/His Kit provides all of the necessary reagents to express your protein in a tightly regulated fashion. The vector pBAD/His allows you to express your protein with an N-terminal tag. The vector provides:

Þ      The araBAD promoter for tightly regulated expression

Þ      Translation initiation signals optimized for E. coliexpression

Þ      N-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin and detection with an Anti-

 

HisG Antibody

Þ      N-terminal Xpress epitope for detection and analysis with an Anti-Xpress Antibody

Þ      Enterokinase cleavage site for removing the N-terminal tag following purification

 

Three vectors are provided (A, B, and C). Each has the N-terminal tag in a different reading frame relative to the multiple cloning site to simplify in-frame cloning of your gene.

 

The pBAD/His and pBAD/Myc-His plasmids are pBR322-derived expression vectors designed for regulated, dose-dependent recombinant protein expression and purification in E. coli. Optimum levels of soluble, recombinant protein are possible using the araBAD promoter (PBAD) from E. coli. The regulatory protein, AraC, is provided on the pBAD/His and pBAD/Myc-His vectors allowing regulation of PBAD.

 

L-阿拉伯糖调控表达

In the presence of L-arabinose, expression from PBAD is turned on while the absence of L-arabinose produces very low levels of transcription from PBAD (Lee, 1980; Lee et al., 1987). Uninduced levels are repressed even further by growth in the presence of glucose. Glucose reduces the levels of 3,5-cyclic AMP, thus lowering expression of the catabolite-repressed PBAD promoter (Miyada et al., 1984). By varying the concentration of L-arabinose, protein expression levels can be optimized to ensure maximum expression of soluble protein. In addition, the tight regulation of PBAD by AraC is useful for expression of potentially toxic or essential genes (Carson et al., 1991; Dalbey and Wickner, 1985; Guzman et al., 1992; Kuhn and Wickner, 1985; Russell et al., 1989; San Millan et al., 1989). For more information on the mechanism of expression and repression of the ara regulon, refer to Schleif, 1992.

 

 

pBadHis A载体序列                    

ORIGIN

    1 AAGAAACCAA TTGTCCATAT TGCATCAGAC ATTGCCGTCA CTGCGTCTTT TACTGGCTCT

   61 TCTCGCTAAC CAAACCGGTA ACCCCGCTTA TTAAAAGCAT TCTGTAACAA AGCGGGACCA

  121 AAGCCATGAC AAAAACGCGT AACAAAAGTG TCTATAATCA CGGCAGAAAA GTCCACATTG

  181 ATTATTTGCA CGGCGTCACA CTTTGCTATG CCATAGCATT TTTATCCATA AGATTAGCGG

  241 ATCCTACCTG ACGCTTTTTA TCGCAACTCT CTACTGTTTC TCCATACCCG TTTTTTGGGC

  301 TAACAGGAGG AATTAACCAT GGGGGGTTCT CATCATCATC ATCATCATGG TATGGCTAGC

  361 ATGACTGGTG GACAGCAAAT GGGTCGGGAT CTGTACGACG ATGACGATAA GGATCGATGG

  421 GGATCCGAGC TCGAGATCTG CAGCTGGTAC CATATGGGAA TTCGAAGCTT GGCTGTTTTG

  481 GCGGATGAGA GAAGATTTTC AGCCTGATAC AGATTAAATC AGAACGCAGA AGCGGTCTGA

  541 TAAAACAGAA TTTGCCTGGC GGCAGTAGCG CGGTGGTCCC ACCTGACCCC ATGCCGAACT

  601 CAGAAGTGAA ACGCCGTAGC GCCGATGGTA GTGTGGGGTC TCCCCATGCG AGAGTAGGGA

  661 ACTGCCAGGC ATCAAATAAA ACGAAAGGCT CAGTCGAAAG ACTGGGCCTT TCGTTTTATC

  721 TGTTGTTTGT CGGTGAACGC TCTCCTGAGT AGGACAAATC CGCCGGGAGC GGATTTGAAC

  781 GTTGCGAAGC AACGGCCCGG AGGGTGGCGG GCAGGACGCC CGCCATAAAC TGCCAGGCAT

  841 CAAATTAAGC AGAAGGCCAT CCTGACGGAT GGCCTTTTTG CGTTTCTACA AACTCTTTTG

  901 TTTATTTTTC TAAATACATT CAAATATGTA TCCGCTCATG AGACAATAAC CCTGATAAAT

  961 GCTTCAATAA TATTGAAAAA GGAAGAGTAT GAGTATTCAA CATTTCCGTG TCGCCCTTAT

 1021 TCCCTTTTTT GCGGCATTTT GCCTTCCTGT TTTTGCTCAC CCAGAAACGC TGGTGAAAGT

 1081 AAAAGATGCT GAAGATCAGT TGGGTGCACG AGTGGGTTAC ATCGAACTGG ATCTCAACAG

 1141 CGGTAAGATC CTTGAGAGTT TTCGCCCCGA AGAACGTTTT CCAATGATGA GCACTTTTAA

 1201 AGTTCTGCTA TGTGGCGCGG TATTATCCCG TGTTGACGCC GGGCAAGAGC AACTCGGTCG

 1261 CCGCATACAC TATTCTCAGA ATGACTTGGT TGAGTACTCA CCAGTCACAG AAAAGCATCT

 1321 TACGGATGGC ATGACAGTAA GAGAATTATG CAGTGCTGCC ATAACCATGA GTGATAACAC

 1381 TGCGGCCAAC TTACTTCTGA CAACGATCGG AGGACCGAAG GAGCTAACCG CTTTTTTGCA

 1441 CAACATGGGG GATCATGTAA CTCGCCTTGA TCGTTGGGAA CCGGAGCTGA ATGAAGCCAT

 1501 ACCAAACGAC GAGCGTGACA CCACGATGCC TGTAGCAATG GCAACAACGT TGCGCAAACT

 1561 ATTAACTGGC GAACTACTTA CTCTAGCTTC CCGGCAACAA TTAATAGACT GGATGGAGGC

 1621 GGATAAAGTT GCAGGACCAC TTCTGCGCTC GGCCCTTCCG GCTGGCTGGT TTATTGCTGA

 1681 TAAATCTGGA GCCGGTGAGC GTGGGTCTCG CGGTATCATT GCAGCACTGG GGCCAGATGG

 1741 TAAGCCCTCC CGTATCGTAG TTATCTACAC GACGGGGAGT CAGGCAACTA TGGATGAACG

 1801 AAATAGACAG ATCGCTGAGA TAGGTGCCTC ACTGATTAAG CATTGGTAAC TGTCAGACCA

 1861 AGTTTACTCA TATATACTTT AGATTGATTT AAAACTTCAT TTTTAATTTA AAAGGATCTA

 1921 GGTGAAGATC CTTTTTGATA ATCTCATGAC CAAAATCCCT TAACGTGAGT TTTCGTTCCA

 1981 CTGAGCGTCA GACCCCGTAG AAAAGATCAA AGGATCTTCT TGAGATCCTT TTTTTCTGCG

 2041 CGTAATCTGC TGCTTGCAAA CAAAAAAACC ACCGCTACCA GCGGTGGTTT GTTTGCCGGA

 2101 TCAAGAGCTA CCAACTCTTT TTCCGAAGGT AACTGGCTTC AGCAGAGCGC AGATACCAAA

 2161 TACTGTCCTT CTAGTGTAGC CGTAGTTAGG CCACCACTTC AAGAACTCTG TAGCACCGCC

 2221 TACATACCTC GCTCTGCTAA TCCTGTTACC AGTGGCTGCT GCCAGTGGCG ATAAGTCGTG

 2281 TCTTACCGGG TTGGACTCAA GACGATAGTT ACCGGATAAG GCGCAGCGGT CGGGCTGAAC

 2341 GGGGGGTTCG TGCACACAGC CCAGCTTGGA GCGAACGACC TACACCGAAC TGAGATACCT

 2401 ACAGCGTGAG CTATGAGAAA GCGCCACGCT TCCCGAAGGG AGAAAGGCGG ACAGGTATCC

 2461 GGTAAGCGGC AGGGTCGGAA CAGGAGAGCG CACGAGGGAG CTTCCAGGGG GAAACGCCTG

 2521 GTATCTTTAT AGTCCTGTCG GGTTTCGCCA CCTCTGACTT GAGCGTCGAT TTTTGTGATG

 2581 CTCGTCAGGG GGGCGGAGCC TATGGAAAAA CGCCAGCAAC GCGGCCTTTT TACGGTTCCT

 2641 GGCCTTTTGC TGGCCTTTTG CTCACATGTT CTTTCCTGCG TTATCCCCTG ATTCTGTGGA

 2701 TAACCGTATT ACCGCCTTTG AGTGAGCTGA TACCGCTCGC CGCAGCCGAA CGACCGAGCG

 2761 CAGCGAGTCA GTGAGCGAGG AAGCGGAAGA GCGCCTGATG CGGTATTTTC TCCTTACGCA

 2821 TCTGTGCGGT ATTTCACACC GCATATGGTG CACTCTCAGT ACAATCTGCT CTGATGCCGC

 2881 ATAGTTAAGC CAGTATACAC TCCGCTATCG CTACGTGACT GGGTCATGGC TGCGCCCCGA

 2941 CACCCGCCAA CACCCGCTGA CGCGCCCTGA CGGGCTTGTC TGCTCCCGGC ATCCGCTTAC

 3001 AGACAAGCTG TGACCGTCTC CGGGAGCTGC ATGTGTCAGA GGTTTTCACC GTCATCACCG

 3061 AAACGCGCGA GGCAGCAGAT CAATTCGCGC GCGAAGGCGA AGCGGCATGC ATAATGTGCC

 3121 TGTCAAATGG ACGAAGCAGG GATTCTGCAA ACCCTATGCT ACTCCGTCAA GCCGTCAATT

 3181 GTCTGATTCG TTACCAATTA TGACAACTTG ACGGCTACAT CATTCACTTT TTCTTCACAA

 3241 CCGGCACGGA ACTCGCTCGG GCTGGCCCCG GTGCATTTTT TAAATACCCG CGAGAAATAG

 3301 AGTTGATCGT CAAAACCAAC ATTGCGACCG ACGGTGGCGA TAGGCATCCG GGTGGTGCTC

 3361 AAAAGCAGCT TCGCCTGGCT GATACGTTGG TCCTCGCGCC AGCTTAAGAC GCTAATCCCT

 3421 AACTGCTGGC GGAAAAGATG TGACAGACGC GACGGCGACA AGCAAACATG CTGTGCGACG

 3481 CTGGCGATAT CAAAATTGCT GTCTGCCAGG TGATCGCTGA TGTACTGACA AGCCTCGCGT

 3541 ACCCGATTAT CCATCGGTGG ATGGAGCGAC TCGTTAATCG CTTCCATGCG CCGCAGTAAC

 3601 AATTGCTCAA GCAGATTTAT CGCCAGCAGC TCCGAATAGC GCCCTTCCCC TTGCCCGGCG

 3661 TTAATGATTT GCCCAAACAG GTCGCTGAAA TGCGGCTGGT GCGCTTCATC CGGGCGAAAG

 3721 AACCCCGTAT TGGCAAATAT TGACGGCCAG TTAAGCCATT CATGCCAGTA GGCGCGCGGA

 3781 CGAAAGTAAA CCCACTGGTG ATACCATTCG CGAGCCTCCG GATGACGACC GTAGTGATGA

 3841 ATCTCTCCTG GCGGGAACAG CAAAATATCA CCCGGTCGGC AAACAAATTC TCGTCCCTGA

 3901 TTTTTCACCA CCCCCTGACC GCGAATGGTG AGATTGAGAA TATAACCTTT CATTCCCAGC

 3961 GGTCGGTCGA TAAAAAAATC GAGATAACCG TTGGCCTCAA TCGGCGTTAA ACCCGCCACC

 4021 AGATGGGCAT TAAACGAGTA TCCCGGCAGC AGGGGATCAT TTTGCGCTTC AGCCATACTT

 4081 TTCATACTCC CGCCATTCAG AG

//

 

pBadHis A其他大肠杆菌表达载体:

pKD13

PinPoint Xa-1

pKD4

pTf16

pTWIN2

pTYB11

pKJE7

pET-17b

pTrcHis2 A

pBad/gIII B

pGEX-6P-2

pG-KJE8

pET-46 EK/LIC

pMal-p2G

pGEX-2TK

pGEX-4T-3

pET-49b(+)

pEZZ18

pMal-c2X

pET300/NT-DEST

pBAD-TOPO

pMal-p2X

pET-33b(+)

pRSET-CFP

pET-23(+)

pMal-c4X

pET-24c(+)

pTrcHis2 B

pET-11b(+)

pET-41a(+)

pET-24(+)

pET-44c(+)

pQE-82L

pGEX-4T-1

pMal-p4X

pET-43.1c(+)

pQE-9

pALEX a,b,c

pBad/gIII C

pET-27b(+)

pQE-70

pKD20

pET102/D-TOPO

pET-21a(+)

pET-16b

pRSET B

pACYCDuet-1

pET-20b(+)

pET-21d(+)

pCOLADuet-1

pCYB1

pET-14b

pRSET-EmGFP

pET-42b(+)

pGEX-5X-2

pET-26b(+)

pET301/CT-DEST

pGEX-6P-1

pET-28c(+)

pET-50b(+)

pGro7

pRSET A

pET-32a(+)

pMal-c2G

pET-11a(+)

pRSET C

ptdTomato

pQE-30

pET-12b(+)

pET-42c(+)

pET-43.1b(+)

pET-41c(+)

pET-23b(+)

pET-39b(+)

pBV220

pET-41c(+)

pBad/His C

pET-32b(+)

pCold IV

pETDuet-1

pBAD/Thio-TOPO

pET-29c(+)

pE-SUMO

pTYB12

pCold-ProS2

pET-24b(+)

pACYC184

pEGM-7ZF(+)

pCold III

pBAD33

pBad/Myc-His A

pSP73

pCold-GST

pAmCyan

PinPoint Xa-2

pTXB1

pET-52b(+)

pColdS-SUMO

pTWIN1

pBad/gIII A

pET-23d(+)

pCold II

pET-24a(+)

pkk223-3

pET-29a(+)

pBAD102/D-TOPO

pET-28a(+)

pBAD18

pET-30 EK/LIC

pBad/Myc-His C

pProEX HTc

pET-44 EK/LIC

pET-32 Xa/LIC

pBad/His B

pMal-p2E

pTrcHis B

pET-40b(+)

pET-23a(+)

pQE-60

pGEX-3X

pET-45b(+)

pET-12c(+)

pQE-32

pTrc99a

pMal-c2E

pET-11c(+)

pBad24

pACYC177

pGEX-5X-1

pQE-80L

pET-12a(+)

pMXB10

pGEX-6P-3

pQE-31

pET-23c(+)

pGEX-KG

pTrcHis A

pET-51b(+)

pBad/His A

pTrcHis C

pGEX-2T

pET-47b(+)

pBad/Myc-His B

pET-3a(+)

pEcoli-6xHN-GFPuv

pET-22b(+)

pBAD202/D-TOPO

pGEX-5X-3

pBR322

pET-19b

pCold I

pET-32 EK/LIC

pET-28b(+)

pET-21b(+)

pCold TF

pET-30 Xa/LIC

pGEX-4T-2

pET-25b(+)

pQE-81L

pET-29b(+)

pET-31b(+)

pET-42a(+)

pQE-16

pGEM-T

pET-43.1 EK/LIC

pET-44a(+)

pQE-40

pBlueScript SK(+)

pMAL-c5x

pTrcHis2 C

pET-48b(+)

pET-SUMO

pkk232-8

pET302/NT-His

pET-21b(+)

pCDFDuet-1

pET-5a(+)

pGFPuv

pET-15b

pBluescript II SK(+)

pET-5b(+)

pBad43

pET-21c(+)

pProEX HTb

pSP64

pG-Tf2

pET-24d(+)

pKD46

PinPoint Xa-3

pSE380

pET-30a(+)

pBluescript II KS(-)

pMAL-p5e

pMAL-p5x

pET-43.1a(+)

pEGM-11ZF(+)

pTYB2

pET-30b(+)

pET-37b(+)

pDsRed-Express2

pKD3

pProEX HTa

pET303/CT-His

pBV221

pSUMO

pTYB1

pRSET-BFP

pET-11d(+)

pET-3b(+)

pET-41 EK/LIC

pET-32c(+)

pET-44b(+)

pRSFDuet-1

pET-30c(+)

pET-His

pET-41b(+)