北京华越洋生物
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首页    载体及质粒    pET-50b(+)
1

pET-50b(+)

pET-50b(+)

pET-50b(+)

 

编号

载体名称

北京华越洋生物VECT4130

pET-50b(+)

 

pET50b载体基本信息

别名:

pET50b, pET 50b

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:

6733 bp

5' 测序引物及序列:  

T7: 5'-TAATACGACTCACTATAGGG-3'

3' 测序引物及序列:  

T7t: 5'-TGCTAGTTATTGCTCAGCGG-3'

载体标签:

N-Nus, N-His, N-HRV 3C, C-S, C-Thrombin

载体抗性:

卡那

备注:

pET47系列载体一样,但是多出了一个NNus标签;载体含有HRV 3C蛋白酶切位点,可以在低温下直接将多余的融合表达标签切除。

稳定性:

瞬时表达

组成型:

组成型

病毒/非病毒:

非病毒

 

pET50b载体质粒图谱和多克隆位点信息


 

pET50b载体简介

 

pET50b载体携带有N端的Nus标签和His纯化标签,紧跟着这些标签的有一个HRV3C蛋白酶切位点。这个HRV3C蛋白酶切位点能够特异性的高效切割 LEVLFQGP”氨基酸序列,酶活力在低温下很强。pET-50b(+)载体上还含有一个可选择的CThrombin蛋白酶切位点和S标签序列。

 

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(Cat. No. 69337-3)的作用下终止蛋白翻译。

 

pET50b载体序列

ORIGIN

        1 ATCCGGATAT AGTTCCTCCT TTCAGCAAAA AACCCCTCAA GACCCGTTTA GAGGCCCCAA

       61 GGGGTTATGC TAGTTATTGC TCAGCGGTGG CAGCAGCCAA CTCAGCTTCC TTTCGGGCTT

      121 TGTTTAGCAG CCTAGGTTAA TTAAGCCTCG AGAGCAGCAG AAGTAGAGCT GTCCATGTGC

      181 TGGCGTTCGA ATTTAGCAGC AGCGGTTTCT TTACTACCGC GTGGCACCAG AGCGAGCTCT

      241 GCGGCCGCAA GCTTGTCGAC GGACGTCGGG CGCGCCAAGG CCTGTACAGA ATTCGGATCC

      301 TGGTACCCGG GTCCCTGAAA GAGGACTTCA AGAGCCGCGG AGTGATGGTG ATGGTGATGA

      361 CCAGAACCAC TAGTCGCTTC GTCACCGAAC CAGCAAATAT TACGGGCAGC CATAATCAGT

      421 GCTCCGGCTT TTTCGTCGGT CAACCCTTCG ATATCAGCCA GATCATCAAT GCCCTGTTCG

      481 GCGAGATCTT CCAGCGTACA AACGCCACGG GCGGCCAGTT TGAATGCCAA ATCACGATCT

      541 ACCCCTTCAA GGTTCAGCAG ATCGTCAGCC GGTTTGTTAT CACCGAGGCT TTCTTCCTGG

      601 GCCTGTGCAA TGGTGGCCAG TGCATTTTTA GCACGCTCGC GCAGTGCTTC AACGGTCGGC

      661 TCATCAAGGC CTTCGATTTC CAACAGCTCT TTCATCGGCA CATAGGCCAA TTCTTCCAGC

      721 GTCGAGAAGC CTTCTTCTAC CAGAACAGTC GCGAAGTCTT CGTCGATGTC GAGATATTTG

      781 GTGAAGGTGT CGATCGCTGC GTGCGCTTCC GCCTGATGCT TAGCTTGCAG GTCGTCAACG

      841 GTCATCACGT TGAGTTCCCA ACCGCTCAGT TGCGAAGCCA GACGCACGTT CTGACCGTTA

      901 CGGCCAATCG CCTGCGCCAG ATTACCGGCT TCAACGGCGA TGTCCATGGT GTGTTTATCT

      961 TCATCCACCA CGATAGAAGC AACGTCTGCC GGTGCCATTG CGTTAATCAC GAACTGCGCC

     1021 GGGTTATCAT CCCACAGGAC GATATCGATA CGCTCGCCAC CCAGTTCAGT AGACACCGCC

     1081 TGAACACGCG CGCCACGCAT ACCTACGCAA GCACCTACCG GATCGATACG TTTATCGTTG

     1141 GTTTTCACCG CGATTTTCGC ACGAGAACCC GGATCGCGAG CCGCTGCTTT AATTTCAATC

     1201 ACTTCTTCGC CGATTTCTGG CACTTCAATA CGGAACAGTT CGATCAGCAT TTCCGGCTTG

     1261 GAACGAGTGA CGAACAGTTG CGCGCCACGC GCTTCCGGGC GAACGGAATA GAGCACGCCA

     1321 CGAACGCGGT CGCCAGGGCG GAAGTTTTCA CGCGGCAGCA TATCTTCGCG CAGGATCACG

     1381 GCTTCAGCGT TGTTGCCCAG ATCCAGAGAG ATGTTGTCGC GGTTTACTTT TTTCACCACG

     1441 CCGGTGATGA TTTCACCTTC GTGTTCACGG AACTGATCAA CCACCATCGC ACGTTCGGCT

     1501 TCACGCACTT TCTGCACGAT AACCTGTTTT GCCGTCTGGG TAGTGATACG GTCAAAGGTA

     1561 ACAGACTCAA TCTGATCTTC AACGTAATCG CCCAGGTTCA GGCTTTCATC TTCATAACGT

     1621 GCGGCTTCAA GGGTGATTTC CTTGGTCGGC TGGGTGACTT CATCAACAAC TAACCAGCGA

     1681 CGGAAAGTGT CAAAATCACC GCTTTTGCGA TCGATCTGTA CGCGGACGTC GATCTCTTGT

     1741 TCATATTTTT TCTTTGTTGC TGTCGCCAGC GCGCTTTCCA ATGCTTCGAA AATCTTCTCG

     1801 CGAGGTAGCG CCTTTTCATT GGATACGGCT TCAACTACAG CCAAAATTTC TTTGTTCATA

     1861 CTAGAATGGT GATGGTGGTG ATGGCTGCTG CCCATATGTA TATCTCCTTC TTAAAGTTAA

     1921 ACAAAATTAT TTCTAGAGGG GAATTGTTAT CCGCTCACAA TTCCCCTATA GTGAGTCGTA

     1981 TTAATTTCGC GGGATCGAGA TCGATCTCGA TCCTCTACGC CGGACGCATC GTGGCCGGCA

     2041 TCACCGGCGC CACAGGTGCG GTTGCTGGCG CCTATATCGC CGACATCACC GATGGGGAAG

     2101 ATCGGGCTCG CCACTTCGGG CTCATGAGCG CTTGTTTCGG CGTGGGTATG GTGGCAGGCC

     2161 CCGTGGCCGG GGGACTGTTG GGCGCCATCT CCTTGCATGC ACCATTCCTT GCGGCGGCGG

     2221 TGCTCAACGG CCTCAACCTA CTACTGGGCT GCTTCCTAAT GCAGGAGTCG CATAAGGGAG

     2281 AGCGTCGAGA TCCCGGACAC CATCGAATGG CGCAAAACCT TTCGCGGTAT GGCATGATAG

     2341 CGCCCGGAAG AGAGTCAATT CAGGGTGGTG AATGTGAAAC CAGTAACGTT ATACGATGTC

     2401 GCAGAGTATG CCGGTGTCTC TTATCAGACC GTTTCCCGCG TGGTGAACCA GGCCAGCCAC

     2461 GTTTCTGCGA AAACGCGGGA AAAAGTGGAA GCGGCGATGG CGGAGCTGAA TTACATTCCC

     2521 AACCGCGTGG CACAACAACT GGCGGGCAAA CAGTCGTTGC TGATTGGCGT TGCCACCTCC

     2581 AGTCTGGCCC TGCACGCGCC GTCGCAAATT GTCGCGGCGA TTAAATCTCG CGCCGATCAA

     2641 CTGGGTGCCA GCGTGGTGGT GTCGATGGTA GAACGAAGCG GCGTCGAAGC CTGTAAAGCG

     2701 GCGGTGCACA ATCTTCTCGC GCAACGCGTC AGTGGGCTGA TCATTAACTA TCCGCTGGAT

     2761 GACCAGGATG CCATTGCTGT GGAAGCTGCC TGCACTAATG TTCCGGCGTT ATTTCTTGAT

     2821 GTCTCTGACC AGACACCCAT CAACAGTATT ATTTTCTCCC ATGAAGACGG TACGCGACTG

     2881 GGCGTGGAGC ATCTGGTCGC ATTGGGTCAC CAGCAAATCG CGCTGTTAGC GGGCCCATTA

     2941 AGTTCTGTCT CGGCGCGTCT GCGTCTGGCT GGCTGGCATA AATATCTCAC TCGCAATCAA

     3001 ATTCAGCCGA TAGCGGAACG GGAAGGCGAC TGGAGTGCCA TGTCCGGTTT TCAACAAACC

     3061 ATGCAAATGC TGAATGAGGG CATCGTTCCC ACTGCGATGC TGGTTGCCAA CGATCAGATG

     3121 GCGCTGGGCG CAATGCGCGC CATTACCGAG TCCGGGCTGC GCGTTGGTGC GGACATCTCG

     3181 GTAGTGGGAT ACGACGATAC CGAAGACAGC TCATGTTATA TCCCGCCGTT AACCACCATC

     3241 AAACAGGATT TTCGCCTGCT GGGGCAAACC AGCGTGGACC GCTTGCTGCA ACTCTCTCAG

     3301 GGCCAGGCGG TGAAGGGCAA TCAGCTGTTG CCCGTCTCAC TGGTGAAAAG AAAAACCACC

     3361 CTGGCGCCCA ATACGCAAAC CGCCTCTCCC CGCGCGTTGG CCGATTCATT AATGCAGCTG

     3421 GCACGACAGG TTTCCCGACT GGAAAGCGGG CAGTGAGCGC AACGCAATTA ATGTAAGTTA

     3481 GCTCACTCAT TAGGCACCGG GATCTCGACC GATGCCCTTG AGAGCCTTCA ACCCAGTCAG

     3541 CTCCTTCCGG TGGGCGCGGG GCATGACTAG CATGATCGTG CTCCTGTCGT TGAGGACCCG

     3601 GCTAGGCTGG CGGGGTTGCC TTACTGGTTA GCAGAATGAA TCACCGATAC GCGAGCGAAC

     3661 GTGAAGCGAC TGCTGCTGCA AAACGTCTGC GACCTGAGCA ACAACATGAA TGGTCTTCGG

     3721 TTTCCGTGTT TCGTAAAGTC TGGAAACGCG GAAGTCAGCG CCCTGCACCA TTATGTTCCG

     3781 GATCTGCATC GCAGGATGCT GCTGGCTACC CTGTGGAACA CCTACATCTG TATTAACGAA

     3841 GCGCTGGCAT TGACCCTGAG TGATTTTTCT CTGGTCCCGC CGCATCCATA CCGCCAGTTG

     3901 TTTACCCTCA CAACGTTCCA GTAACCGGGC ATGTTCATCA TCAGTAACCC GTATCGTGAG

     3961 CATCCTCTCT CGTTTCATCG GTATCATTAC CCCCATGAAC AGAAATCCCC CTTACACGGA

     4021 GGCATCAGTG ACCAAACAGG AAAAAACCGC CCTTAACATG GCCCGCTTTA TCAGAAGCCA

     4081 GACATTAACG CTTCTGGAGA AACTCAACGA GCTGGACGCG GATGAACAGG CAGACATCTG

     4141 TGAATCGCTT CACGACCACG CTGATGAGCT TTACCGCAGC TGCCTCGCGC GTTTCGGTGA

     4201 TGACGGTGAA AACCTCTGAC ACATGCAGCT CCCGGAGACG GTCACAGCTT GTCTGTAAGC

     4261 GGATGCCGGG AGCAGACAAG CCCGTCAGGG CGCGTCAGCG GGTGTTGGCG GGTGTCGGGG

     4321 CGCAGCCATG ACCCAGTCAC GTAGCGATAG CGGAGTGTAT ACTGGCTTAA CTATGCGGCA

     4381 TCAGAGCAGA TTGTACTGAG AGTGCACCAT ATATGCGGTG TGAAATACCG CACAGATGCG

     4441 TAAGGAGAAA ATACCGCATC AGGCGCTCTT CCGCTTCCTC GCTCACTGAC TCGCTGCGCT

     4501 CGGTCGTTCG GCTGCGGCGA GCGGTATCAG CTCACTCAAA GGCGGTAATA CGGTTATCCA

     4561 CAGAATCAGG GGATAACGCA GGAAAGAACA TGTGAGCAAA AGGCCAGCAA AAGGCCAGGA

     4621 ACCGTAAAAA GGCCGCGTTG CTGGCGTTTT TCCATAGGCT CCGCCCCCCT GACGAGCATC

     4681 ACAAAAATCG ACGCTCAAGT CAGAGGTGGC GAAACCCGAC AGGACTATAA AGATACCAGG

     4741 CGTTTCCCCC TGGAAGCTCC CTCGTGCGCT CTCCTGTTCC GACCCTGCCG CTTACCGGAT

     4801 ACCTGTCCGC CTTTCTCCCT TCGGGAAGCG TGGCGCTTTC TCATAGCTCA CGCTGTAGGT

     4861 ATCTCAGTTC GGTGTAGGTC GTTCGCTCCA AGCTGGGCTG TGTGCACGAA CCCCCCGTTC

     4921 AGCCCGACCG CTGCGCCTTA TCCGGTAACT ATCGTCTTGA GTCCAACCCG GTAAGACACG

     4981 ACTTATCGCC ACTGGCAGCA GCCACTGGTA ACAGGATTAG CAGAGCGAGG TATGTAGGCG

     5041 GTGCTACAGA GTTCTTGAAG TGGTGGCCTA ACTACGGCTA CACTAGAAGG ACAGTATTTG

     5101 GTATCTGCGC TCTGCTGAAG CCAGTTACCT TCGGAAAAAG AGTTGGTAGC TCTTGATCCG

     5161 GCAAACAAAC CACCGCTGGT AGCGGTGGTT TTTTTGTTTG CAAGCAGCAG ATTACGCGCA

     5221 GAAAAAAAGG ATCTCAAGAA GATCCTTTGA TCTTTTCTAC GGGGTCTGAC GCTCAGTGGA

     5281 ACGAAAACTC ACGTTAAGGG ATTTTGGTCA TGAACAATAA AACTGTCTGC TTACATAAAC

     5341 AGTAATACAA GGGGTGTTAT GAGCCATATT CAACGGGAAA CGTCTTGCTC TAGGCCGCGA

     5401 TTAAATTCCA ACATGGATGC TGATTTATAT GGGTATAAAT GGGCTCGCGA TAATGTCGGG

     5461 CAATCAGGTG CGACAATCTA TCGATTGTAT GGGAAGCCCG ATGCGCCAGA GTTGTTTCTG

     5521 AAACATGGCA AAGGTAGCGT TGCCAATGAT GTTACAGATG AGATGGTCAG ACTAAACTGG

     5581 CTGACGGAAT TTATGCCTCT TCCGACCATC AAGCATTTTA TCCGTACTCC TGATGATGCA

     5641 TGGTTACTCA CCACTGCGAT CCCCGGCAAA ACAGCATTCC AGGTATTAGA AGAATATCCT

     5701 GATTCAGGTG AAAATATTGT TGATGCGCTG GCAGTGTTCC TGCGCCGGTT GCATTCGATT

     5761 CCTGTTTGTA ATTGTCCTTT TAACAGTGAT CGCGTATTTC GTCTCGCTCA GGCGCAATCA

     5821 CGAATGAATA ACGGTTTGGT TGATGCGAGT GATTTTGATG ACGAGCGTAA TGGCTGGCCT

     5881 GTTGAACAAG TCTGGAAAGA AATGCATAAA CTTTTGCCAT TCTCACCGGA TTCAGTCGTC

     5941 ACTCATGGTG ATTTCTCACT TGATAACCTT ATTTTTGACG AGGGGAAATT AATAGGTTGT

     6001 ATTGATGTTG GACGAGTCGG AATCGCAGAC CGATACCAGG ATCTTGCCAT CCTATGGAAC

     6061 TGCCTCGGTG AGTTTTCTCC TTCATTACAG AAACGGCTTT TTCAAAAATA TGGTATTGAT

     6121 AATCCTGATA TGAATAAATT GCAGTTTCAT TTGATGCTCG ATGAGTTTTT CTAAGAATTA

     6181 ATTCATGAGC GGATACATAT TTGAATGTAT TTAGAAAAAT AAACAAATAG GGGTTCCGCG

     6241 CACATTTCCC CGAAAAGTGC CACCTGAAAT TGTAAACGTT AATATTTTGT TAAAATTCGC

     6301 GTTAAATTTT TGTTAAATCA GCTCATTTTT TAACCAATAG GCCGAAATCG GCAAAATCCC

     6361 TTATAAATCA AAAGAATAGA CCGAGATAGG GTTGAGTGTT GTTCCAGTTT GGAACAAGAG

     6421 TCCACTATTA AAGAACGTGG ACTCCAACGT CAAAGGGCGA AAAACCGTCT ATCAGGGCGA

     6481 TGGCCCACTA CGTGAACCAT CACCCTAATC AAGTTTTTTG GGGTCGAGGT GCCGTAAAGC

     6541 ACTAAATCGG AACCCTAAAG GGAGCCCCCG ATTTAGAGCT TGACGGGGAA AGCCGGCGAA

     6601 CGTGGCGAGA AAGGAAGGGA AGAAAGCGAA AGGAGCGGGC GCTAGGGCGC TGGCAAGTGT

     6661 AGCGGTCACG CTGCGCGTAA CCACCACACC CGCCGCGCTT AATGCGCCGC TACAGGGCGC

     6721 GTCCCATTCG CCA

//

 

pET-50b(+)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(+)