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1

pET-33b(+)

pET-33b(+)

pET-33b(+)

 

编号

名称

北京华越洋VECT-590

pET-33b(+)

 

pET33b载体基本信息

别名:

pET33b, pET 33b

质粒类型:

大肠杆菌蛋白表达

表达水平:

克隆方法:

多克隆位点,限制性内切酶

载体大小:

5383 bp

5' 测序引物:

T7

5' 测序引物序列:  

T7: 5'-TAATACGACTCACTATAGGG-3'

载体标签:

His (中间和C), N-Thrombin

载体抗性:

Kanamycin

备注:

Production of target proteins suitable for site-specific   32P-labeling; Nterm thrombin cleavage site; Nterm enterokinase cleavage site

稳定性:

瞬时表达 Transient

组成型:

组成型 Constitutive

病毒/非病毒:

非病毒

 

pET33b载体质粒图谱和多克隆位点信息

Feature Name

Start

End

T7_terminator

129

1

T7_Terminal_primer

69

87

T7_leader

238

206

Xpress_fwd_primer

239

221

T7_transl_en_RBS

337

321

lacO

382

355

T7_promoter

400

382

tet (300 - 563)

432

695

pBRrevBam_primer

503

484

lacI

778

1869

tet (576 - 851)

1928

2203

ROP

2678

2869

pGEX_3_primer

2885

2863

pBR322_origin

3903

3284

KanR2

4009

4824

 

ORF

Start

End

ORF frame 3

537

1037

ORF frame 1

910

1869

ORF frame 1

4009

4824

 

Enzyme Name

Cut

XhoI

158

NotI

166

EagI

166

HindIII

173

SalI

179

SacI

190

EcoRI

192

BamHI

198

NheI

230

NdeI

237

NcoI

310

XbaI

349

BglII

415

BclI

1151

ApaI

1348

EcoRV

1587

HpaI

1643

FspI

2219

NruI

4097

ClaI

4131

XmaI

4312

SmaI

4314

 

pET33b载体简介

The pET-33b(+) vector is derived from pET-28b(+) and carries a 15bp sequence encoding the protein kinase A (PKA) site RRASV, located between the thrombin cleavage and Nde I sites (1). Proteins expressed in pET-33b(+) can be easily purified by metal chelation chromatography (via either N- or C-terminal His•Tag® sequences) and efficiently labeled with 32P- or 33P-gATP and the catalytic subunit of cAMP-dependent protein kinase from heart muscle. Labeled proteins can be used as direct probes in protein-protein interaction studies. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/ expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the T7 terminator primer.

 

pET33b载体序列

ORIGIN
    1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
   61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
  121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
  181 cgacggagct cgaattcgga tcccgaccca tttgctgtcc accagtcatg ctagccatat
  241 gaacagatgc acgacggctg ccgcgcggca ccaggccgct gctgtgatga tgatgatgat
  301 ggctgctgcc catggtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
  361 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatct
  421 cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
  481 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
  541 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca
  601 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg
  661 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa
  721 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
  781 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
  841 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
  901 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
  961 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
 1021 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
 1081 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
 1141 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
 1201 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
 1261 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
 1321 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
 1381 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
 1441 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
 1501 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
 1561 gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac
 1621 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
 1681 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
 1741 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
 1801 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
 1861 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg
 1921 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
 1981 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
 2041 agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct
 2101 gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc
 2161 caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccc cacgggtgcg
 2221 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
 2281 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
 2341 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
 2401 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc
 2461 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct
 2521 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc
 2581 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac
 2641 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc
 2701 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
 2761 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
 2821 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
 2881 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
 2941 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
 3001 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
 3061 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
 3121 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
 3181 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
 3241 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
 3301 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
 3361 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
 3421 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
 3481 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
 3541 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
 3601 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
 3661 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
 3721 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
 3781 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
 3841 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
 3901 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
 3961 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
 4021 caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat
 4081 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
 4141 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
 4201 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
 4261 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa
 4321 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
 4381 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
 4441 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
 4501 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
 4561 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
 4621 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
 4681 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
 4741 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
 4801 ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat
 4861 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat
 4921 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
 4981 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
 5041 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
 5101 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
 5161 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
 5221 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
 5281 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
 5341 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg cca
//

 

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(+)