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首页    载体及质粒    pET-44b(+)

pET-44b(+)

pET-44b(+)

pET-44b(+)

 

编号

名称

北京华越洋VECT-580

pET-44b(+)

 

pET44b载体基本信息

别名:

pET44b, pET 44b

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:

7310 bp

5' 测序引物:

T7

5' 测序引物序列:  

T7: 5'-TAATACGACTCACTATAGGG-3'

3' 测序引物:

ColiDOWN

3' 测序引物序列:  

ColiDOWN: 5'-TTCACTTCTGAGTTCGGCATG-3'

载体标签:

C-HSV, N-His, C-His, N-Thrombin, N-Nus, N-EK

载体抗性:

Ampicillin

备注:

Both Nterm and Cterm His tags; Nterm thrombin cleavage site; Nterm   enterokinase cleavage site; a,b,c vary by MCS.

稳定性:

瞬时表达 Transient

组成型:

组成型 Constitutive

病毒/非病毒:

非病毒

 

pET44b载体质粒图谱和多克隆位点信息


Feature Name

Start

End

T7_Terminal_primer

69

87

rrnB_T2_terminator

168

141

rrnB_T1_terminator

343

300

rrnB_terminator

466

309

EK

694

680

S15

790

746

6xHIS

2341

2324

T7_transl_en_RBS

2378

2362

lacO

2423

2396

T7_promoter

2441

2423

tet (300 - 563)

2477

2740

pBRrevBam_primer

2548

2529

lacI

2823

3914

ROP

4486

4677

pGEX_3_primer

4693

4671

pBR322_origin

5711

5092

AmpR_promoter

5798

5826

Ampicillin

5869

6729

 

ORF

Start

End

ORF frame 2

2353

509

ORF frame 2

803

2131

ORF frame 2

2324

3082

ORF frame 3

2955

3914

ORF frame 1

5869

6729

 

Enzyme Name

Cut

PacI

503

XhoI

530

PmlI

576

NotI

592

EagI

592

HindIII

599

KpnI

624

SalI

626

PstI

639

AscI

641

EcoRI

654

BamHI

660

SacI

672

SmaI

720

XmaI

718

SacII

796

SpeI

827

BglII

942

MscI

1074

StuI

1127

NcoI

1402

NdeI

2352

XbaI

2390

ApaI

3393

HpaI

3688

FspI

6433

 

pET44b载体简介

The pET-44 vectors are designed for cloning and high-level expression of peptide sequences fused with the 495 aa Nus•Tag™ protein. Compared to the pET-43.1 series, the pET-44 vectors encode an additional N-terminal His•Tag. 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 COLIDOWN primer (Cat. No. 70845-3). Vector encoded sequence can be completely removed when cloning into the PshA I or Sma I sites (as shown below) by cleaving the Nus•Tag fusion protein with enterokinase or thrombin, respectively.

 

pET44b载体序列

ORIGIN
    1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
   61 ggggttatgc tagttattgc tcagcggata catatttgaa tgtatttaga aaaataaaca
  121 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
  181 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt
  241 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
  301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
  361 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
  421 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
  481 agcctaggaa cgcccaactt aattaacatt agtggtggtg gtggtggtgc tcgagatcct
  541 ctgggtcttc aggagcgagt tctggctggc ttgcacgtgt atacagctgt gcggccgcaa
  601 gcttcgaacg cgtatcgatg gtaccgtcga cgtcctgcag gcgcgcctgt acagaattcg
  661 gatccacgag ctccggactc ttgtcgtcgt catcaatcgt accagaaccc gcgctgcccc
  721 gggggaccag accagttggc ggtggcgagt ccatgtgctg gcgttcaaat ttcgcagcag
  781 cggtttcttt acccgcggag tgatggtgat ggtgatgacc agaaccacta gtcgcttcgt
  841 caccgaacca gcaaatatta cgggcagcca taatcagtgc tccggctttt tcgtcggtca
  901 acccttcgat atcagccaga tcatcaatgc cctgttcggc gagatcttcc agcgtacaaa
  961 cgccacgggc ggccagtttg aatgccaaat cacgatctac cccttcaagg ttcagcagat
 1021 cgtcagccgg tttgttatca ccgaggcttt cttcctgggc ctgtgcaatg gtggccagtg
 1081 catttttagc acgctcgcgc agtgcttcaa cggtcggctc atcaaggcct tcgatttcca
 1141 acagctcttt catcggcaca taggccaatt cttccagcgt cgagaagcct tcttctacca
 1201 gaacagtcgc gaagtcttcg tcgatgtcga gatatttggt gaaggtgtcg atcgctgcgt
 1261 gcgcttccgc ctgatgctta gcttgcaggt cgtcaacggt catcacgttg agttcccaac
 1321 cgctcagttg cgaagccaga cgcacgttct gaccgttacg gccaatcgcc tgcgccagat
 1381 taccggcttc aacggcgatg tccatggtgt gtttatcttc atccaccacg atagaagcaa
 1441 cgtctgccgg tgccattgcg ttaatcacga actgcgccgg gttatcatcc cacaggacga
 1501 tatcgatacg ctcgccaccc agttcagtag acaccgcctg aacacgcgcg ccacgcatac
 1561 ctacgcaagc acctaccgga tcgatacgtt tatcgttggt tttcaccgcg attttcgcac
 1621 gagaacccgg atcgcgagcc gctgctttaa tttcaatcac ttcttcgccg atttctggca
 1681 cttcaatacg gaacagttcg atcagcattt ccggcttgga acgagtgacg aacagttgcg
 1741 cgccacgcgc ttccgggcga acggaataga gcacgccacg aacgcggtcg ccagggcgga
 1801 agttttcacg cggcagcata tcttcgcgca ggatcacggc ttcagcgttg ttgcccagat
 1861 ccagagagat gttgtcgcgg tttacttttt tcaccacgcc ggtgatgatt tcaccttcgt
 1921 gttcacggaa ctgatcaacc accatcgcac gttcggcttc acgcactttc tgcacgataa
 1981 cctgttttgc cgtctgggta gtgatacggt caaaggtaac agactcaatc tgatcttcaa
 2041 cgtaatcgcc caggttcagg ctttcatctt cataacgtgc ggcttcaagg gtgatttcct
 2101 tggtcggctg ggtgacttca tcaacaacta accagcgacg gaaagtgtca aaatcaccgc
 2161 ttttgcgatc gatctgtacg cggacgtcga tctcttgttc atattttttc tttgttgctg
 2221 tcgccagcgc gctttccaat gcttcgaaaa tcttctcgcg aggtagcgcc ttttcattgg
 2281 atacggcttc aactacagcc aaaatttctt tgttcatact agaatggtga tggtggtgat
 2341 ggctgctgcc catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga
 2401 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc
 2461 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
 2521 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
 2581 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
 2641 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
 2701 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
 2761 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
 2821 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
 2881 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
 2941 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
 3001 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
 3061 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
 3121 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
 3181 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
 3241 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
 3301 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
 3361 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
 3421 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
 3481 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
 3541 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
 3601 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg
 3661 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
 3721 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
 3781 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
 3841 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
 3901 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
 3961 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
 4021 atgactagca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
 4081 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
 4141 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
 4201 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
 4261 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
 4321 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
 4381 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
 4441 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
 4501 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
 4561 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
 4621 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
 4681 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
 4741 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
 4801 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
 4861 tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
 4921 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
 4981 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
 5041 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
 5101 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
 5161 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
 5221 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
 5281 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
 5341 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
 5401 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
 5461 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
 5521 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
 5581 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
 5641 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
 5701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
 5761 tttggtcatg acctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
 5821 gacaataacc ctgataaatg cttcaatcat gattgaaaaa ggaagagtat gagtattcaa
 5881 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
 5941 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
 6001 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
 6061 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
 6121 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
 6181 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
 6241 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
 6301 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
 6361 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
 6421 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
 6481 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
 6541 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
 6601 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
 6661 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
 6721 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
 6781 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgcc atcgtgccgc
 6841 cagaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
 6901 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
 6961 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
 7021 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
 7081 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
 7141 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
 7201 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
 7261 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//

 

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(+)