pET-32a质粒

启动子：	T7/lac
复制子：	ColE1 ori，F1 ori
终止子：	T7 terminator
质粒分类：	大肠系列质粒；大肠表达质粒；pET系列质粒
质粒大小：	5900bp
质粒标签：	N-Trx，N-6×His，N-thrombin，N-S，N-enterokinase，C-6×His
原核抗性：	氨苄青霉素Amp（100μg/ml）
克隆菌株：	DH5α等大肠杆菌
培养条件：	37℃，有氧，LB
表达宿主：	BL21(DE3)等大肠杆菌
诱导方式：	IPTG或乳糖及其类似物
5'测序引物：	T7（TAATACGACTCACTATAGGG）
3'测序引物：	T7-ter（TGCTAGTTATTGCTCAGCGG）

质粒简介

The pET-32 series is designed for cloning and high-level expression of peptide sequences fused with the 109aa Trx•Tag™ thioredoxin protein (1). Cloning sites are available for producing fusion proteins also containing cleavable His•Tag® and S•Tag™ sequences for detection and purification. 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.

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