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基本信息

載體類型: 

大腸桿菌蛋白表達(dá)載體 

載體抗性: 

Amp 

載體標(biāo)簽: 

N-GST 

表達(dá)水平: 

高拷貝 

載體大小: 

4969 bp 

啟動(dòng)子: 

Tac 

克隆方法: 

多克隆位點(diǎn),限制性內(nèi)切酶 

5' 測(cè)序引物: 

pGEX5':  5’-GGGCTGGCAAGCCACGTTTGGTG-3’ 

3' 測(cè)序引物 

pGEX3': 5’-CCGGGAGCTGCATGTGTCAGAGG-3’ 

備注:

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質(zhì)粒圖譜和多克隆位點(diǎn)信息

注意:

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載體描述

pGEX-4T-1載體是一種常用的原核表達(dá)載體,具有Amp抗性,質(zhì)粒大小為4.9 kb。該載體多克隆位點(diǎn)區(qū)域含有多個(gè)常用的內(nèi)切酶位點(diǎn)序列,便于不同基因的克??;且全序列中含有tac啟動(dòng)子、GST標(biāo)簽序列,是一種高效的蛋白表達(dá)載體。Tac啟動(dòng)子在沒有誘導(dǎo)物存在的情況下,質(zhì)粒上攜帶的lacIq基因產(chǎn)物可以有效抑制tac啟動(dòng)子的轉(zhuǎn)錄??稍贐L21、Rosstea等表達(dá)菌株中高表達(dá)融合有GST標(biāo)簽的蛋白,且用凝血酶可將目的蛋白的GST標(biāo)簽切掉,便于下游蛋白純化。

載體序列

ORIGIN

        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg

       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt

      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc

      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca

      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc

      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc

      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc

      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata

      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc

      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact

      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag

      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt

      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa

      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat

      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc

      901 atcctccaaa atcggatctg gttccgcgtg gatccccgga attcccgggt cgactcgagc

      961 ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc

     1021 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca

     1081 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc

     1141 agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc

     1201 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc

     1261 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc

     1321 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga

     1381 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt

     1441 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag

     1501 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag

     1561 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg

     1621 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg

     1681 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca

     1741 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag

     1801 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc

     1861 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg

     1921 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc

     1981 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg

     2041 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg

     2101 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga

     2161 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac

     2221 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa

     2281 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca

     2341 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag

     2401 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac

     2461 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa

     2521 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc

     2581 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag

     2641 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac

     2701 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc

     2761 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc

     2821 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca

     2881 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc

     2941 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg

     3001 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct

     3061 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata

     3121 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc

     3181 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg

     3241 acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc

     3301 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg

     3361 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc

     3421 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac

     3481 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg

     3541 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg

     3601 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc

     3661 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg

     3721 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac

     3781 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg

     3841 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc

     3901 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg

     3961 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg

     4021 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc

     4081 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg

     4141 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc

     4201 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg

     4261 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc

     4321 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc

     4381 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca

     4441 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa

     4501 caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca

     4561 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc

     4621 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg

     4681 cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag

     4741 ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat

     4801 gcacggttac gatgcgccca tctacaccaa cgtaacctat cccattacgg tcaatccgcc

     4861 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag

     4921 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatt

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