北京华越洋生物
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首页    载体及质粒    pET-30 Xa/LIC
1

pET-30 Xa/LIC

pET-30 Xa/LIC

pET-30 Xa/LIC

 

编号

名称

北京华越洋VECT-660

pET-30 Xa/LIC

 

pET30 Xa/LIC载体基本信息

别名:

pET30 Xa/LIC, pet 30a Xa/LIC

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:

5448 bp

5' 测序引物:

T7

5' 测序引物序列:  

5'-TAATACGACTCACTATAGGG-3'

载体标签:

N-His, N-S, N-Xa Factor, C-His

载体抗性:

Kanamycin

备注:

For directional cloning of PCR-amplified   DNA; ligation independent cloning; Factor Xa 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

 

pET30 Xa/LIC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

lacI

328

1934

T7_transl_en_RBS

398

382

lacO

443

416

T7_promoter

461

443

tet (300 - 563)

497

760

pBRrevBam_primer

568

549

tet (576 - 851)

1993

2268

ROP

2743

2934

pGEX_3_primer

2950

2928

pBR322_origin

3968

3349

KanR2

4074

4889

 

ORF

Start

End

ORF frame 2

353

1102

ORF frame 3

975

1934

ORF frame 3

4074

4889

 

Enzyme Name

Cut

XhoI

158

NotI

166

EagI

166

HindIII

173

SalI

179

SacI

190

EcoRI

192

BamHI

198

EcoRV

206

NcoI

211

MscI

216

AgeI

257

KpnI

264

BglII

267

BstBI

294

NdeI

372

XbaI

410

BclI

1216

ApaI

1413

HpaI

1708

FspI

2284

NruI

4162

SmaI

4379

XmaI

4377

 

pET30 Xa/LIC载体简介

The pET-30 Xa/LIC vector is designed for rapid, directional cloning of PCR-amplified DNA for high-level expression of polypeptides containing N-terminal HisTag® and STag sequences for detection and purification. Using specifically designed primers for amplification and the pET-30Xa/LIC Vector Kit, inserts can be efficiently cloned without the need for restriction digestion or ligation, and the resulting fusion proteins can be cleaved with Factor Xa to precisely remove all vector-encoded amino acids. 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 polymeraseis 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.

 

pET30 Xa/LIC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 TCCGATATCG CCATGGCCAG AGGAGAGTTA GAGCGACCCT
  241 CAATACCGGA GCCACCACCG GTACCCAGAT CTGGGCTGTC CATGTGCTGG CGTTCGAATT
  301 TAGCAGCAGC GGTTTCTTTC ATACCAGAAC CGCGTGGCAC CAGACCAGAA GAATGATGAT
  361 GATGATGGTG CATATGTATA TCTCCTTCTT AAAGTTAAAC AAAATTATTT CTAGAGGGGA
  421 ATTGTTATCC GCTCACAATT CCCCTATAGT GAGTCGTATT AATTTCGCGG GATCGAGATC
  481 GATCTCGATC CTCTACGCCG GACGCATCGT GGCCGGCATC ACCGGCGCCA CAGGTGCGGT
  541 TGCTGGCGCC TATATCGCCG ACATCACCGA TGGGGAAGAT CGGGCTCGCC ACTTCGGGCT
  601 CATGAGCGCT TGTTTCGGCG TGGGTATGGT GGCAGGCCCC GTGGCCGGGG GACTGTTGGG
  661 CGCCATCTCC TTGCATGCAC CATTCCTTGC GGCGGCGGTG CTCAACGGCC TCAACCTACT
  721 ACTGGGCTGC TTCCTAATGC AGGAGTCGCA TAAGGGAGAG CGTCGAGATC CCGGACACCA
  781 TCGAATGGCG CAAAACCTTT CGCGGTATGG CATGATAGCG CCCGGAAGAG AGTCAATTCA
  841 GGGTGGTGAA TGTGAAACCA GTAACGTTAT ACGATGTCGC AGAGTATGCC GGTGTCTCTT
  901 ATCAGACCGT TTCCCGCGTG GTGAACCAGG CCAGCCACGT TTCTGCGAAA ACGCGGGAAA
  961 AAGTGGAAGC GGCGATGGCG GAGCTGAATT ACATTCCCAA CCGCGTGGCA CAACAACTGG
 1021 CGGGCAAACA GTCGTTGCTG ATTGGCGTTG CCACCTCCAG TCTGGCCCTG CACGCGCCGT
 1081 CGCAAATTGT CGCGGCGATT AAATCTCGCG CCGATCAACT GGGTGCCAGC GTGGTGGTGT
 1141 CGATGGTAGA ACGAAGCGGC GTCGAAGCCT GTAAAGCGGC GGTGCACAAT CTTCTCGCGC
 1201 AACGCGTCAG TGGGCTGATC ATTAACTATC CGCTGGATGA CCAGGATGCC ATTGCTGTGG
 1261 AAGCTGCCTG CACTAATGTT CCGGCGTTAT TTCTTGATGT CTCTGACCAG ACACCCATCA
 1321 ACAGTATTAT TTTCTCCCAT GAAGACGGTA CGCGACTGGG CGTGGAGCAT CTGGTCGCAT
 1381 TGGGTCACCA GCAAATCGCG CTGTTAGCGG GCCCATTAAG TTCTGTCTCG GCGCGTCTGC
 1441 GTCTGGCTGG CTGGCATAAA TATCTCACTC GCAATCAAAT TCAGCCGATA GCGGAACGGG
 1501 AAGGCGACTG GAGTGCCATG TCCGGTTTTC AACAAACCAT GCAAATGCTG AATGAGGGCA
 1561 TCGTTCCCAC TGCGATGCTG GTTGCCAACG ATCAGATGGC GCTGGGCGCA ATGCGCGCCA
 1621 TTACCGAGTC CGGGCTGCGC GTTGGTGCGG ACATCTCGGT AGTGGGATAC GACGATACCG
 1681 AAGACAGCTC ATGTTATATC CCGCCGTTAA CCACCATCAA ACAGGATTTT CGCCTGCTGG
 1741 GGCAAACCAG CGTGGACCGC TTGCTGCAAC TCTCTCAGGG CCAGGCGGTG AAGGGCAATC
 1801 AGCTGTTGCC CGTCTCACTG GTGAAAAGAA AAACCACCCT GGCGCCCAAT ACGCAAACCG
 1861 CCTCTCCCCG CGCGTTGGCC GATTCATTAA TGCAGCTGGC ACGACAGGTT TCCCGACTGG
 1921 AAAGCGGGCA GTGAGCGCAA CGCAATTAAT GTAAGTTAGC TCACTCATTA GGCACCGGGA
 1981 TCTCGACCGA TGCCCTTGAG AGCCTTCAAC CCAGTCAGCT CCTTCCGGTG GGCGCGGGGC
 2041 ATGACTATCG TCGCCGCACT TATGACTGTC TTCTTTATCA TGCAACTCGT AGGACAGGTG
 2101 CCGGCAGCGC TCTGGGTCAT TTTCGGCGAG GACCGCTTTC GCTGGAGCGC GACGATGATC
 2161 GGCCTGTCGC TTGCGGTATT CGGAATCTTG CACGCCCTCG CTCAAGCCTT CGTCACTGGT
 2221 CCCGCCACCA AACGTTTCGG CGAGAAGCAG GCCATTATCG CCGGCATGGC GGCCCCACGG
 2281 GTGCGCATGA TCGTGCTCCT GTCGTTGAGG ACCCGGCTAG GCTGGCGGGG TTGCCTTACT
 2341 GGTTAGCAGA ATGAATCACC GATACGCGAG CGAACGTGAA GCGACTGCTG CTGCAAAACG
 2401 TCTGCGACCT GAGCAACAAC ATGAATGGTC TTCGGTTTCC GTGTTTCGTA AAGTCTGGAA
 2461 ACGCGGAAGT CAGCGCCCTG CACCATTATG TTCCGGATCT GCATCGCAGG ATGCTGCTGG
 2521 CTACCCTGTG GAACACCTAC ATCTGTATTA ACGAAGCGCT GGCATTGACC CTGAGTGATT
 2581 TTTCTCTGGT CCCGCCGCAT CCATACCGCC AGTTGTTTAC CCTCACAACG TTCCAGTAAC
 2641 CGGGCATGTT CATCATCAGT AACCCGTATC GTGAGCATCC TCTCTCGTTT CATCGGTATC
 2701 ATTACCCCCA TGAACAGAAA TCCCCCTTAC ACGGAGGCAT CAGTGACCAA ACAGGAAAAA
 2761 ACCGCCCTTA ACATGGCCCG CTTTATCAGA AGCCAGACAT TAACGCTTCT GGAGAAACTC
 2821 AACGAGCTGG ACGCGGATGA ACAGGCAGAC ATCTGTGAAT CGCTTCACGA CCACGCTGAT
 2881 GAGCTTTACC GCAGCTGCCT CGCGCGTTTC GGTGATGACG GTGAAAACCT CTGACACATG
 2941 CAGCTCCCGG AGACGGTCAC AGCTTGTCTG TAAGCGGATG CCGGGAGCAG ACAAGCCCGT
 3001 CAGGGCGCGT CAGCGGGTGT TGGCGGGTGT CGGGGCGCAG CCATGACCCA GTCACGTAGC
 3061 GATAGCGGAG TGTATACTGG CTTAACTATG CGGCATCAGA GCAGATTGTA CTGAGAGTGC
 3121 ACCATATATG CGGTGTGAAA TACCGCACAG ATGCGTAAGG AGAAAATACC GCATCAGGCG
 3181 CTCTTCCGCT TCCTCGCTCA CTGACTCGCT GCGCTCGGTC GTTCGGCTGC GGCGAGCGGT
 3241 ATCAGCTCAC TCAAAGGCGG TAATACGGTT ATCCACAGAA TCAGGGGATA ACGCAGGAAA
 3301 GAACATGTGA GCAAAAGGCC AGCAAAAGGC CAGGAACCGT AAAAAGGCCG CGTTGCTGGC
 3361 GTTTTTCCAT AGGCTCCGCC CCCCTGACGA GCATCACAAA AATCGACGCT CAAGTCAGAG
 3421 GTGGCGAAAC CCGACAGGAC TATAAAGATA CCAGGCGTTT CCCCCTGGAA GCTCCCTCGT
 3481 GCGCTCTCCT GTTCCGACCC TGCCGCTTAC CGGATACCTG TCCGCCTTTC TCCCTTCGGG
 3541 AAGCGTGGCG CTTTCTCATA GCTCACGCTG TAGGTATCTC AGTTCGGTGT AGGTCGTTCG
 3601 CTCCAAGCTG GGCTGTGTGC ACGAACCCCC CGTTCAGCCC GACCGCTGCG CCTTATCCGG
 3661 TAACTATCGT CTTGAGTCCA ACCCGGTAAG ACACGACTTA TCGCCACTGG CAGCAGCCAC
 3721 TGGTAACAGG ATTAGCAGAG CGAGGTATGT AGGCGGTGCT ACAGAGTTCT TGAAGTGGTG
 3781 GCCTAACTAC GGCTACACTA GAAGGACAGT ATTTGGTATC TGCGCTCTGC TGAAGCCAGT
 3841 TACCTTCGGA AAAAGAGTTG GTAGCTCTTG ATCCGGCAAA CAAACCACCG CTGGTAGCGG
 3901 TGGTTTTTTT GTTTGCAAGC AGCAGATTAC GCGCAGAAAA AAAGGATCTC AAGAAGATCC
 3961 TTTGATCTTT TCTACGGGGT CTGACGCTCA GTGGAACGAA AACTCACGTT AAGGGATTTT
 4021 GGTCATGAAC AATAAAACTG TCTGCTTACA TAAACAGTAA TACAAGGGGT GTTATGAGCC
 4081 ATATTCAACG GGAAACGTCT TGCTCTAGGC CGCGATTAAA TTCCAACATG GATGCTGATT
 4141 TATATGGGTA TAAATGGGCT CGCGATAATG TCGGGCAATC AGGTGCGACA ATCTATCGAT
 4201 TGTATGGGAA GCCCGATGCG CCAGAGTTGT TTCTGAAACA TGGCAAAGGT AGCGTTGCCA
 4261 ATGATGTTAC AGATGAGATG GTCAGACTAA ACTGGCTGAC GGAATTTATG CCTCTTCCGA
 4321 CCATCAAGCA TTTTATCCGT ACTCCTGATG ATGCATGGTT ACTCACCACT GCGATCCCCG
 4381 GGAAAACAGC ATTCCAGGTA TTAGAAGAAT ATCCTGATTC AGGTGAAAAT ATTGTTGATG
 4441 CGCTGGCAGT GTTCCTGCGC CGGTTGCATT CGATTCCTGT TTGTAATTGT CCTTTTAACA
 4501 GCGATCGCGT ATTTCGTCTC GCTCAGGCGC AATCACGAAT GAATAACGGT TTGGTTGATG
 4561 CGAGTGATTT TGATGACGAG CGTAATGGCT GGCCTGTTGA ACAAGTCTGG AAAGAAATGC
 4621 ATAAACTTTT GCCATTCTCA CCGGATTCAG TCGTCACTCA TGGTGATTTC TCACTTGATA
 4681 ACCTTATTTT TGACGAGGGG AAATTAATAG GTTGTATTGA TGTTGGACGA GTCGGAATCG
 4741 CAGACCGATA CCAGGATCTT GCCATCCTAT GGAACTGCCT CGGTGAGTTT TCTCCTTCAT
 4801 TACAGAAACG GCTTTTTCAA AAATATGGTA TTGATAATCC TGATATGAAT AAATTGCAGT
 4861 TTCATTTGAT GCTCGATGAG TTTTTCTAAG AATTAATTCA TGAGCGGATA CATATTTGAA
 4921 TGTATTTAGA AAAATAAACA AATAGGGGTT CCGCGCACAT TTCCCCGAAA AGTGCCACCT
 4981 GAAATTGTAA ACGTTAATAT TTTGTTAAAA TTCGCGTTAA ATTTTTGTTA AATCAGCTCA
 5041 TTTTTTAACC AATAGGCCGA AATCGGCAAA ATCCCTTATA AATCAAAAGA ATAGACCGAG
 5101 ATAGGGTTGA GTGTTGTTCC AGTTTGGAAC AAGAGTCCAC TATTAAAGAA CGTGGACTCC
 5161 AACGTCAAAG GGCGAAAAAC CGTCTATCAG GGCGATGGCC CACTACGTGA ACCATCACCC
 5221 TAATCAAGTT TTTTGGGGTC GAGGTGCCGT AAAGCACTAA ATCGGAACCC TAAAGGGAGC
 5281 CCCCGATTTA GAGCTTGACG GGGAAAGCCG GCGAACGTGG CGAGAAAGGA AGGGAAGAAA
 5341 GCGAAAGGAG CGGGCGCTAG GGCGCTGGCA AGTGTAGCGG TCACGCTGCG CGTAACCACC
 5401 ACACCCGCCG CGCTTAATGC GCCGCTACAG GGCGCGTCCC ATTCGCCA
//

 

 

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(+)