pET-41b(+)
pET-41b(+)
编号 | 名称 |
北京华越洋VECT-540 | pET-41b(+) |
pET41b载体基本信息
别名: | pET41b, pET 41b |
质粒类型: | 大肠杆菌蛋白表达 |
表达水平: | 高 |
克隆方法: | 多克隆位点，限制性内切酶 |
载体大小: | 5932 bp |
5' 测序引物: | T7或pGEX-5‘ |
5' 测序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3'; pGEX-5': 5'-GGGCTGGCAAGCCACGTTTGGTG-3' |
载体标签: | N-GST, N-His, N-Thrombin |
载体抗性: | Kanamycin |
备注: | Encodes GST fusion tag; Nterm thrombin cleavage site; Nterm enterokinase cleavage site; PshAI blunt cloning 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 |
pET41b载体质粒图谱和多克隆位点信息
Feature Name | Start | End |
T7_terminator | 120 | 1 |
T7_Terminal_primer | 69 | 87 |
EK | 278 | 264 |
S15 | 353 | 309 |
6xHIS | 413 | 396 |
GST (variant) | 1094 | 444 |
T7_transl_en_RBS | 1119 | 1103 |
lacO | 1164 | 1137 |
T7_promoter | 1182 | 1164 |
tet (300 - 563) | 1218 | 1481 |
pBRrevBam_primer | 1289 | 1270 |
lacI | 1564 | 2655 |
ROP | 3227 | 3418 |
pGEX_3_primer | 3434 | 3412 |
pBR322_origin | 4452 | 3833 |
KanR2 | 4558 | 5373 |
ORF | Start | End |
ORF frame 3 | 1094 | 147 |
ORF frame 3 | 1095 | 1823 |
ORF frame 1 | 1696 | 2655 |
ORF frame 1 | 4558 | 5373 |
Enzyme Name | Cut |
XhoI | 174 |
NotI | 182 |
EagI | 182 |
HindIII | 189 |
SalI | 195 |
SacI | 206 |
PstI | 214 |
AscI | 216 |
StuI | 226 |
EcoRI | 235 |
BamHI | 241 |
EcoRV | 251 |
NcoI | 254 |
AgeI | 291 |
KpnI | 298 |
BglII | 301 |
SacII | 392 |
SpeI | 423 |
MscI | 886 |
NdeI | 1093 |
XbaI | 1131 |
ApaI | 2134 |
HpaI | 2429 |
NruI | 4646 |
SmaI | 4863 |
XmaI | 4861 |
pET41b载体简介
The pET-41 series is designed for cloning and high-level expression of peptide sequences fused with the 220 aa GST•Tag™ protein. 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 T7 terminator primer. Vector encoded sequence can be completely removed when cloning into the PshAI site and then cleaving the GST fusion protein with Enterokinase.
pET41b载体序列
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
181 gcggccgcaa gcttgtcgac ggagctcgcc tgcaggcgcg ccaaggcctg tacagaattc
241 ggatccccga tatccatggg actcttgtcg tcgtcatcac cggagccacc accggtaccc
301 agatctgggc tgtccatgtg ctggcgttcg aatttagcag cagcggtttc tttcatacca
361 attgcagtac taccgcgtgg caccagaccc gcggagtgat ggtgatggtg atgaccagaa
421 ccactagttg aaccatccga ttttggagga tggtcgccac caccaaacgt ggcttgccag
481 ccctgcaaag gccatgctat atacttgctg gatttcaagt acttatcaat ttgtgggata
541 gcttcaatac gttttttaaa acaaactaat tttgggaacg catccaggca cattgggtcc
601 atgtataaaa caacatcaag agcgtcatac aacatgaagt caggatgggt tacatgatca
661 ccatttaaat atgttttatg acataaacga tcttcgaaca ttttcagcat ttcaggtagc
721 ttgctaagaa aatcaacttt gagagtttca aagtctttac tatatgcaat tctcgaaaca
781 ccgtatctaa tatccaaaac cgctccttca agcattgaaa tctctgcacg ctcttttgga
841 caaccaccca acatgttgtg cttgtcagct atataacgta tgatggccat agactgtgtt
901 aatttaacat caccatcaat ataataagga agattgggaa actccaaacc caattcaaac
961 tttttgtttc gccatttatc accttcatcg cgctcataca aatgctcttc atatttttct
1021 tcaagatatt ccaaaagaag tcgagtgggt tgcacaaggc ccttaatttt ccaataacct
1081 agtatagggg acatatgtat atctccttct taaagttaaa caaaattatt tctagagggg
1141 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat
1201 cgatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
1261 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
1321 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
1381 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
1441 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
1501 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
1561 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
1621 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
1681 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
1741 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
1801 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
1861 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
1921 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
1981 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
2041 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
2101 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
2161 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
2221 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
2281 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
2341 attaccgagt ccgggctgcg cgttggtgcg gacatctcgg tagtgggata cgacgatacc
2401 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
2461 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
2521 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
2581 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
2641 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
2701 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
2761 catgactagc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
2821 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
2881 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
2941 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
3001 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
3061 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
3121 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
3181 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
3241 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
3301 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
3361 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
3421 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
3481 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
3541 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
3601 gtgcaccata tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
3661 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
3721 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
3781 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
3841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
3901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
3961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
4021 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
4081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
4141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
4201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
4261 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
4321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
4381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
4441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
4501 ttttggtcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
4561 agccatattc aacgggaaac gtcttgctct aggccgcgat taaattccaa catggatgct
4621 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
4681 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
4741 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
4801 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
4861 cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
4921 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
4981 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
5041 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
5101 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
5161 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
5221 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
5281 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
5341 cagtttcatt tgatgctcga tgagtttttc taagaattaa ttcatgagcg gatacatatt
5401 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
5461 acctgaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
5521 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
5581 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
5641 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
5701 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
5761 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
5821 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
5881 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc ca
//
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