北京华越洋生物
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首页    载体及质粒    pET-44a(+)
1

pET-44a(+)

pET-44a(+)

pET-44a(+)

 

编号

载体名称

北京华越洋生物VECT5250

pET-44a(+)

 

pET44a载体基本信息

别名:

pET44a, pET 44a

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:

7311 bp

5' 测序引物序列:  

T7: 5'-TAATACGACTCACTATAGGG-3'

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:

氨苄

备注:

载体同时含有N端及CHis标签; 含有NThrombinEK蛋白酶酶切位点; pET44a,b,c 载体的差异主要是在多克隆位点.

稳定性:

瞬时表达

组成型:

组成型

病毒/非病毒:

非病毒

 

pET44a载体质粒图谱和多克隆位点信息

 

pET44a载体简介

pET44系列载体是设计用来克隆和高水平表达含有495个氨基酸的Nus标签的蛋白表达载体。与pET-43.1系列载体相比,pET44系列载体额外编码一个N端的His蛋白纯化标签。pET42a载体的单一的多克隆位点见上面的环状质粒图谱。

注意:载体序列是以pBR322质粒的编码规矩进行编码的,所以T7蛋白表达区在质粒图谱上面 是反向的。T7 RNA聚合酶启动的克隆和表达区域在质粒图谱中也被标注了出来。质粒的F1复制子是被定向的,所以在T7噬菌体聚合酶的作用下,包含有蛋白编码序列的病毒 粒子能够产生,并启动蛋白表达,同时蛋白表达将被T7终止子序列的作用下终止蛋白翻译。对pET44a载体的单链测序可以使用COLIDOWN primer. 把表达的基因插入到载体的PshAI或者SmaI限制性内切酶位点时,可以通过EK蛋白酶或者Thrombin蛋白酶将融合表达的Nus标签完全切除。

 

pET44a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 CTCCCGGACT CTTGTCGTCG TCATCAATCG TACCAGAACC CGCGCTGCCC

  721 CGGGGGACCA GACCAGTTGG CGGTGGCGAG TCCATGTGCT GGCGTTCAAA TTTCGCAGCA

  781 GCGGTTTCTT TACCCGCGGA GTGATGGTGA TGGTGATGAC CAGAACCACT AGTCGCTTCG

  841 TCACCGAACC AGCAAATATT ACGGGCAGCC ATAATCAGTG CTCCGGCTTT TTCGTCGGTC

  901 AACCCTTCGA TATCAGCCAG ATCATCAATG CCCTGTTCGG CGAGATCTTC CAGCGTACAA

  961 ACGCCACGGG CGGCCAGTTT GAATGCCAAA TCACGATCTA CCCCTTCAAG GTTCAGCAGA

 1021 TCGTCAGCCG GTTTGTTATC ACCGAGGCTT TCTTCCTGGG CCTGTGCAAT GGTGGCCAGT

 1081 GCATTTTTAG CACGCTCGCG CAGTGCTTCA ACGGTCGGCT CATCAAGGCC TTCGATTTCC

 1141 AACAGCTCTT TCATCGGCAC ATAGGCCAAT TCTTCCAGCG TCGAGAAGCC TTCTTCTACC

 1201 AGAACAGTCG CGAAGTCTTC GTCGATGTCG AGATATTTGG TGAAGGTGTC GATCGCTGCG

 1261 TGCGCTTCCG CCTGATGCTT AGCTTGCAGG TCGTCAACGG TCATCACGTT GAGTTCCCAA

 1321 CCGCTCAGTT GCGAAGCCAG ACGCACGTTC TGACCGTTAC GGCCAATCGC CTGCGCCAGA

 1381 TTACCGGCTT CAACGGCGAT GTCCATGGTG TGTTTATCTT CATCCACCAC GATAGAAGCA

 1441 ACGTCTGCCG GTGCCATTGC GTTAATCACG AACTGCGCCG GGTTATCATC CCACAGGACG

 1501 ATATCGATAC GCTCGCCACC CAGTTCAGTA GACACCGCCT GAACACGCGC GCCACGCATA

 1561 CCTACGCAAG CACCTACCGG ATCGATACGT TTATCGTTGG TTTTCACCGC GATTTTCGCA

 1621 CGAGAACCCG GATCGCGAGC CGCTGCTTTA ATTTCAATCA CTTCTTCGCC GATTTCTGGC

 1681 ACTTCAATAC GGAACAGTTC GATCAGCATT TCCGGCTTGG AACGAGTGAC GAACAGTTGC

 1741 GCGCCACGCG CTTCCGGGCG AACGGAATAG AGCACGCCAC GAACGCGGTC GCCAGGGCGG

 1801 AAGTTTTCAC GCGGCAGCAT ATCTTCGCGC AGGATCACGG CTTCAGCGTT GTTGCCCAGA

 1861 TCCAGAGAGA TGTTGTCGCG GTTTACTTTT TTCACCACGC CGGTGATGAT TTCACCTTCG

 1921 TGTTCACGGA ACTGATCAAC CACCATCGCA CGTTCGGCTT CACGCACTTT CTGCACGATA

 1981 ACCTGTTTTG CCGTCTGGGT AGTGATACGG TCAAAGGTAA CAGACTCAAT CTGATCTTCA

 2041 ACGTAATCGC CCAGGTTCAG GCTTTCATCT TCATAACGTG CGGCTTCAAG GGTGATTTCC

 2101 TTGGTCGGCT GGGTGACTTC ATCAACAACT AACCAGCGAC GGAAAGTGTC AAAATCACCG

 2161 CTTTTGCGAT CGATCTGTAC GCGGACGTCG ATCTCTTGTT CATATTTTTT CTTTGTTGCT

 2221 GTCGCCAGCG CGCTTTCCAA TGCTTCGAAA ATCTTCTCGC GAGGTAGCGC CTTTTCATTG

 2281 GATACGGCTT CAACTACAGC CAAAATTTCT TTGTTCATAC TAGAATGGTG ATGGTGGTGA

 2341 TGGCTGCTGC CCATATGTAT ATCTCCTTCT TAAAGTTAAA CAAAATTATT TCTAGAGGGG

 2401 AATTGTTATC CGCTCACAAT TCCCCTATAG TGAGTCGTAT TAATTTCGCG GGATCGAGAT

 2461 CGATCTCGAT CCTCTACGCC GGACGCATCG TGGCCGGCAT CACCGGCGCC ACAGGTGCGG

 2521 TTGCTGGCGC CTATATCGCC GACATCACCG ATGGGGAAGA TCGGGCTCGC CACTTCGGGC

 2581 TCATGAGCGC TTGTTTCGGC GTGGGTATGG TGGCAGGCCC CGTGGCCGGG GGACTGTTGG

 2641 GCGCCATCTC CTTGCATGCA CCATTCCTTG CGGCGGCGGT GCTCAACGGC CTCAACCTAC

 2701 TACTGGGCTG CTTCCTAATG CAGGAGTCGC ATAAGGGAGA GCGTCGAGAT CCCGGACACC

 2761 ATCGAATGGC GCAAAACCTT TCGCGGTATG GCATGATAGC GCCCGGAAGA GAGTCAATTC

 2821 AGGGTGGTGA ATGTGAAACC AGTAACGTTA TACGATGTCG CAGAGTATGC CGGTGTCTCT

 2881 TATCAGACCG TTTCCCGCGT GGTGAACCAG GCCAGCCACG TTTCTGCGAA AACGCGGGAA

 2941 AAAGTGGAAG CGGCGATGGC GGAGCTGAAT TACATTCCCA ACCGCGTGGC ACAACAACTG

 3001 GCGGGCAAAC AGTCGTTGCT GATTGGCGTT GCCACCTCCA GTCTGGCCCT GCACGCGCCG

 3061 TCGCAAATTG TCGCGGCGAT TAAATCTCGC GCCGATCAAC TGGGTGCCAG CGTGGTGGTG

 3121 TCGATGGTAG AACGAAGCGG CGTCGAAGCC TGTAAAGCGG CGGTGCACAA TCTTCTCGCG

 3181 CAACGCGTCA GTGGGCTGAT CATTAACTAT CCGCTGGATG ACCAGGATGC CATTGCTGTG

 3241 GAAGCTGCCT GCACTAATGT TCCGGCGTTA TTTCTTGATG TCTCTGACCA GACACCCATC

 3301 AACAGTATTA TTTTCTCCCA TGAAGACGGT ACGCGACTGG GCGTGGAGCA TCTGGTCGCA

 3361 TTGGGTCACC AGCAAATCGC GCTGTTAGCG GGCCCATTAA GTTCTGTCTC GGCGCGTCTG

 3421 CGTCTGGCTG GCTGGCATAA ATATCTCACT CGCAATCAAA TTCAGCCGAT AGCGGAACGG

 3481 GAAGGCGACT GGAGTGCCAT GTCCGGTTTT CAACAAACCA TGCAAATGCT GAATGAGGGC

 3541 ATCGTTCCCA CTGCGATGCT GGTTGCCAAC GATCAGATGG CGCTGGGCGC AATGCGCGCC

 3601 ATTACCGAGT CCGGGCTGCG CGTTGGTGCG GACATCTCGG TAGTGGGATA CGACGATACC

 3661 GAAGACAGCT CATGTTATAT CCCGCCGTTA ACCACCATCA AACAGGATTT TCGCCTGCTG

 3721 GGGCAAACCA GCGTGGACCG CTTGCTGCAA CTCTCTCAGG GCCAGGCGGT GAAGGGCAAT

 3781 CAGCTGTTGC CCGTCTCACT GGTGAAAAGA AAAACCACCC TGGCGCCCAA TACGCAAACC

 3841 GCCTCTCCCC GCGCGTTGGC CGATTCATTA ATGCAGCTGG CACGACAGGT TTCCCGACTG

 3901 GAAAGCGGGC AGTGAGCGCA ACGCAATTAA TGTAAGTTAG CTCACTCATT AGGCACCGGG

 3961 ATCTCGACCG ATGCCCTTGA GAGCCTTCAA CCCAGTCAGC TCCTTCCGGT GGGCGCGGGG

 4021 CATGACTAGC ATGATCGTGC TCCTGTCGTT GAGGACCCGG CTAGGCTGGC GGGGTTGCCT

 4081 TACTGGTTAG CAGAATGAAT CACCGATACG CGAGCGAACG TGAAGCGACT GCTGCTGCAA

 4141 AACGTCTGCG ACCTGAGCAA CAACATGAAT GGTCTTCGGT TTCCGTGTTT CGTAAAGTCT

 4201 GGAAACGCGG AAGTCAGCGC CCTGCACCAT TATGTTCCGG ATCTGCATCG CAGGATGCTG

 4261 CTGGCTACCC TGTGGAACAC CTACATCTGT ATTAACGAAG CGCTGGCATT GACCCTGAGT

 4321 GATTTTTCTC TGGTCCCGCC GCATCCATAC CGCCAGTTGT TTACCCTCAC AACGTTCCAG

 4381 TAACCGGGCA TGTTCATCAT CAGTAACCCG TATCGTGAGC ATCCTCTCTC GTTTCATCGG

 4441 TATCATTACC CCCATGAACA GAAATCCCCC TTACACGGAG GCATCAGTGA CCAAACAGGA

 4501 AAAAACCGCC CTTAACATGG CCCGCTTTAT CAGAAGCCAG ACATTAACGC TTCTGGAGAA

 4561 ACTCAACGAG CTGGACGCGG ATGAACAGGC AGACATCTGT GAATCGCTTC ACGACCACGC

 4621 TGATGAGCTT TACCGCAGCT GCCTCGCGCG TTTCGGTGAT GACGGTGAAA ACCTCTGACA

 4681 CATGCAGCTC CCGGAGACGG TCACAGCTTG TCTGTAAGCG GATGCCGGGA GCAGACAAGC

 4741 CCGTCAGGGC GCGTCAGCGG GTGTTGGCGG GTGTCGGGGC GCAGCCATGA CCCAGTCACG

 4801 TAGCGATAGC GGAGTGTATA CTGGCTTAAC TATGCGGCAT CAGAGCAGAT TGTACTGAGA

 4861 GTGCACCATA TATGCGGTGT GAAATACCGC ACAGATGCGT AAGGAGAAAA TACCGCATCA

 4921 GGCGCTCTTC CGCTTCCTCG CTCACTGACT CGCTGCGCTC GGTCGTTCGG CTGCGGCGAG

 4981 CGGTATCAGC TCACTCAAAG GCGGTAATAC GGTTATCCAC AGAATCAGGG GATAACGCAG

 5041 GAAAGAACAT GTGAGCAAAA GGCCAGCAAA AGGCCAGGAA CCGTAAAAAG GCCGCGTTGC

 5101 TGGCGTTTTT CCATAGGCTC CGCCCCCCTG ACGAGCATCA CAAAAATCGA CGCTCAAGTC

 5161 AGAGGTGGCG AAACCCGACA GGACTATAAA GATACCAGGC GTTTCCCCCT GGAAGCTCCC

 5221 TCGTGCGCTC TCCTGTTCCG ACCCTGCCGC TTACCGGATA CCTGTCCGCC TTTCTCCCTT

 5281 CGGGAAGCGT GGCGCTTTCT CATAGCTCAC GCTGTAGGTA TCTCAGTTCG GTGTAGGTCG

 5341 TTCGCTCCAA GCTGGGCTGT GTGCACGAAC CCCCCGTTCA GCCCGACCGC TGCGCCTTAT

 5401 CCGGTAACTA TCGTCTTGAG TCCAACCCGG TAAGACACGA CTTATCGCCA CTGGCAGCAG

 5461 CCACTGGTAA CAGGATTAGC AGAGCGAGGT ATGTAGGCGG TGCTACAGAG TTCTTGAAGT

 5521 GGTGGCCTAA CTACGGCTAC ACTAGAAGGA CAGTATTTGG TATCTGCGCT CTGCTGAAGC

 5581 CAGTTACCTT CGGAAAAAGA GTTGGTAGCT CTTGATCCGG CAAACAAACC ACCGCTGGTA

 5641 GCGGTGGTTT TTTTGTTTGC AAGCAGCAGA TTACGCGCAG AAAAAAAGGA TCTCAAGAAG

 5701 ATCCTTTGAT CTTTTCTACG GGGTCTGACG CTCAGTGGAA CGAAAACTCA CGTTAAGGGA

 5761 TTTTGGTCAT GACCTATTTG TTTATTTTTC TAAATACATT CAAATATGTA TCCGCTCATG

 5821 AGACAATAAC CCTGATAAAT GCTTCAATCA TGATTGAAAA AGGAAGAGTA TGAGTATTCA

 5881 ACATTTCCGT GTCGCCCTTA TTCCCTTTTT TGCGGCATTT TGCCTTCCTG TTTTTGCTCA

 5941 CCCAGAAACG CTGGTGAAAG TAAAAGATGC TGAAGATCAG TTGGGTGCAC GAGTGGGTTA

 6001 CATCGAACTG GATCTCAACA GCGGTAAGAT CCTTGAGAGT TTTCGCCCCG AAGAACGTTT

 6061 TCCAATGATG AGCACTTTTA AAGTTCTGCT ATGTGGCGCG GTATTATCCC GTATTGACGC

 6121 CGGGCAAGAG CAACTCGGTC GCCGCATACA CTATTCTCAG AATGACTTGG TTGAGTACTC

 6181 ACCAGTCACA GAAAAGCATC TTACGGATGG CATGACAGTA AGAGAATTAT GCAGTGCTGC

 6241 CATAACCATG AGTGATAACA CTGCGGCCAA CTTACTTCTG ACAACGATCG GAGGACCGAA

 6301 GGAGCTAACC GCTTTTTTGC ACAACATGGG GGATCATGTA ACTCGCCTTG ATCGTTGGGA

 6361 ACCGGAGCTG AATGAAGCCA TACCAAACGA CGAGCGTGAC ACCACGATGC CTGTAGCAAT

 6421 GGCAACAACG TTGCGCAAAC TATTAACTGG CGAACTACTT ACTCTAGCTT CCCGGCAACA

 6481 ATTAATAGAC TGGATGGAGG CGGATAAAGT TGCAGGACCA CTTCTGCGCT CGGCCCTTCC

 6541 GGCTGGCTGG TTTATTGCTG ATAAATCTGG AGCCGGTGAG CGTGGGTCTC GCGGTATCAT

 6601 TGCAGCACTG GGGCCAGATG GTAAGCCCTC CCGTATCGTA GTTATCTACA CGACGGGGAG

 6661 TCAGGCAACT ATGGATGAAC GAAATAGACA GATCGCTGAG ATAGGTGCCT CACTGATTAA

 6721 GCATTGGTAA CTGTCAGACC AAGTTTACTC ATATATACTT TAGATTGATT TAAAACTTCA

 6781 TTTTTAATTT AAAAGGATCT AGGTGAAGAT CCTTTTTGAT AATCTCATGC CATCGTGCCG

 6841 CCAGAAATTG TAAACGTTAA TATTTTGTTA AAATTCGCGT TAAATTTTTG TTAAATCAGC

 6901 TCATTTTTTA ACCAATAGGC CGAAATCGGC AAAATCCCTT ATAAATCAAA AGAATAGACC

 6961 GAGATAGGGT TGAGTGTTGT TCCAGTTTGG AACAAGAGTC CACTATTAAA GAACGTGGAC

 7021 TCCAACGTCA AAGGGCGAAA AACCGTCTAT CAGGGCGATG GCCCACTACG TGAACCATCA

 7081 CCCTAATCAA GTTTTTTGGG GTCGAGGTGC CGTAAAGCAC TAAATCGGAA CCCTAAAGGG

 7141 AGCCCCCGAT TTAGAGCTTG ACGGGGAAAG CCGGCGAACG TGGCGAGAAA GGAAGGGAAG

 7201 AAAGCGAAAG GAGCGGGCGC TAGGGCGCTG GCAAGTGTAG CGGTCACGCT GCGCGTAACC

 7261 ACCACACCCG CCGCGCTTAA TGCGCCGCTA CAGGGCGCGT CCCATTCGCC 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(+)