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首页    载体及质粒    pET-32a(+)
1

pET-32a(+)

pET-32a(+)

pET-32a(+)

 

编号

名称

北京华越洋VECT-230

pET-32a(+)

 

pet32a载体基本信息

别名:

pET32a, pet 32a

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:

5900bp

5' 测序引物:

T7   5'-TAATACGACTCACTATAGGG-3'

3' 测序引物:  

T7t: 5'-GCTAGTTATTGCTCAGCGG-3'

载体标签:

thioredoxin (N); His (中间和C)

载体抗性:

氨苄

备注:

pET32a载体用于表达可溶性,具有活性功能的蛋白;pET-32a(+)载体具有NThrombin(凝血酶)酶切位点,N端肠激酶酶切位点。pET32啊,pET32bpET32c载体的差异主要是在于多克隆位点,其他地方完全一致。

稳定性:

瞬时表达

组成型:

组成型

病毒/非病毒:

非病毒

 

pet32a载体质粒图谱和多克隆位点信息

 

pet32a载体简介

 

pET32系列载体是设计用来克隆构建和高水平融合表达表达带有109个氨基酸Trx标签的蛋白质序列。将基因插入到载体的多克隆位点,能够获得融合表达蛋白质。同时融合蛋白包含有His标签和S标签,这些可以用于蛋白表达检测和纯化过程。

 

载体的多克隆位点见上面的图谱。需要注意的是图谱中的核苷酸编码顺序是以pBR322载体的复制方向为正方向,所以T7表达区域在图谱中是处于反向的位置。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。

 

pet32a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 tccgatatca gccatggcct tgtcgtcgtc gtcggtaccc

      241 agatctgggc tgtccatgtg ctggcgttcg aatttagcag cagcggtttc tttcatacca

      301 gaaccgcgtg gcaccagacc agaagaatga tgatgatgat ggtgcatatg gccagaacca

      361 gaaccggcca ggttagcgtc gaggaactct ttcaactgac ctttagacag tgcacccact

      421 ttggttgccg ccacttcacc gtttttgaac agcagcagag tcgggatacc acggatgcca

      481 tatttcggcg cagtgccagg gttttgatcg atgttcagtt ttgcaacggt cagtttgccc

      541 tgatattcgt cagcgatttc atccagaatc ggggcgatca ttttgcacgg accgcaccac

      601 tctgcccaga aatcgacgag gatcgccccg tccgctttga gtacatccgt gtcaaaactg

      661 tcgtcagtca ggtgaataat tttatcgctc atatgtatat ctccttctta aagttaaaca

      721 aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg agtcgtatta

      781 atttcgcggg atcgagatcg atctcgatcc tctacgccgg acgcatcgtg gccggcatca

      841 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc

      901 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg

      961 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc

     1021 tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc

     1081 gtcgagatcc cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc

     1141 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca

     1201 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt

     1261 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac

     1321 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt

     1381 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg

     1441 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg

     1501 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac

     1561 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc

     1621 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc

     1681 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt

     1741 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt

     1801 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg

     1861 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg

     1921 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga catctcggta

     1981 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa

     2041 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc

     2101 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg

     2161 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca

     2221 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct

     2281 cactcattag gcaccgggat ctcgaccgat gcccttgaga gccttcaacc cagtcagctc

     2341 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat

     2401 gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg

     2461 ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc

     2521 tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc

     2581 cggcatggcg gccccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg

     2641 ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag

     2701 cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg

     2761 tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg

     2821 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg

     2881 gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc

     2941 ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct

     3001 ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc

     3061 agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt

     3121 aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc

     3181 gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg

     3241 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc

     3301 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc

     3361 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag

     3421 cagattgtac tgagagtgca ccatatatgc ggtgtgaaat accgcacaga tgcgtaagga

     3481 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg

     3541 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat

     3601 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta

     3661 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa

     3721 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc

     3781 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt

     3841 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca

     3901 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg

     3961 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat

     4021 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta

     4081 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct

     4141 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac

     4201 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa

     4261 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa

     4321 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt

     4381 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca

     4441 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca

     4501 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc

     4561 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa

     4621 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc

     4681 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca

     4741 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat

     4801 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag

     4861 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac

     4921 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt

     4981 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt

     5041 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc

     5101 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat

     5161 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca

     5221 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga

     5281 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg

     5341 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg

     5401 ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat attttgttaa

     5461 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca

     5521 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga

     5581 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc

     5641 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc

     5701 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc

     5761 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg

     5821 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac

     5881 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(+)