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1

pET-28c(+)

pET-28c(+)

pET-28c(+)

 

编号

名称

北京华越洋VECT-360

pET-28c(+)

 

pET28c载体基本信息

别名:

pET28c, pet 28c

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:

5367 bp

5' 测序引物及序列:

T7: 5'-TAATACGACTCACTATAGGG-3'

3' 测序引物及序列:  

T7t: 5'-GCTAGTTATTGCTCAGCGG-3'

载体标签:

N-His, N-Thrombin, N-T7, C-His

载体抗性:

Kanamycin

备注:

Nterm thrombin 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

 

pET28c载体质粒图谱和多克隆位点信息


pET28c载体简介

pET-28a-c(+)载体带有一个N端的His/Thrombin/T7蛋白标签,同时含有一个可以选择的CHis标签。pET28c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。

 

The pET-28a-c(+) vectors carry an N-terminal His•Tag®/ thrombin/ T7•Tag® configuration plus an optional C-terminal His•Tag sequence. 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 circular 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 (Cat. No. 69337-3).

 

pET28c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 tccgacccat ttgctgtcca ccagtcatgc tagccatatg

  241 gctgccgcgc ggcaccaggc cgctgctgtg atgatgatga tgatggctgc tgcccatggt

  301 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca

  361 attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg

  421 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga

  481 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt

  541 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc

  601 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca

  661 ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc

  721 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag

  781 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg

  841 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg

  901 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga

  961 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta

 1021 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg

 1081 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca

 1141 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc

 1201 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg

 1261 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc

 1321 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat

 1381 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt

 1441 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg

 1501 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg

 1561 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc

 1621 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct

 1681 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg

 1741 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg

 1801 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac

 1861 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga

 1921 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt

 1981 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt

 2041 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc

 2101 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc

 2161 gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg

 2221 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg

 2281 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca

 2341 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc

 2401 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca

 2461 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc

 2521 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta

 2581 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat

 2641 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc

 2701 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa

 2761 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc

 2821 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca

 2881 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt

 2941 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc

 3001 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat

 3061 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac

 3121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt

 3181 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca

 3241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc

 3301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact

 3361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct

 3421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag

 3481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca

 3541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa

 3601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc

 3661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag

 3721 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg

 3781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca

 3841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc

 3901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgaaca ataaaactgt

 3961 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt

 4021 gctctaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc

 4081 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc

 4141 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg

 4201 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta

 4261 ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat

 4321 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc

 4381 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg

 4441 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc

 4501 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac

 4561 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga

 4621 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg

 4681 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa

 4741 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt

 4801 ttttctaaga attaattcat gagcggatac atatttgaat gtatttagaa aaataaacaa

 4861 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaattgtaaa cgttaatatt

 4921 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa

 4981 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca

 5041 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc

 5101 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg

 5161 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg

 5221 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg

 5281 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg

 5341 ccgctacagg gcgcgtccca ttcgcca

//

 

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(+)