| 出品公司: | EMD Biosciences(Novagen) |
|---|---|
| 别名: | pET32c, pet 32c |
| 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: | 5901bp |
| 5' 测序引物: | T7 |
| 5' 测序引物序列: | 5'-TAATACGACTCACTATAGGG-3' |
| 载体标签: | thioredoxin (N端); His (中间和C端) |
| 载体抗性: | Ampicillin |
| 表达宿主菌: | Origami2(DE3) 、 Origami(DE3)、OrigamiB(DE3)、BL21-TrxB(DE3) |
| 备注: |
Production of soluble, active target proteins; N-term thrombin cleavage site;
Nterm enterokinase cleavage site; a,b,c vary by MCS |
| 产品目录号: | 69017-3 |
| 稳定性: | 瞬时表达 Transient |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
| 表达宿主菌2: | BL21(DE3) 、BLR(DE3)、Rosetta(DE3)、OrigamiB(DE3) |
| pET表达手册： |
pET表达手册英文版 pET表达手册中文版 |
| pET载体汇总: |
pET-3a pET-11a pET-12a pET-14b pET-15b pET-16b pET-17b pET-19b pET-20b pET-21a pET-22b pET-23a pET-24a pET-25b pET-26b pET-27b pET-28a pET-29a pET-30a pET-31b pET-32a pET-33b pET-34b pET-35b pET-37b pET-39b pET-40b pET-41a pET-42a pET-43.1a pET-44a pET-45b pET-47b pET-48b pET-49b pET-50b pET-51b pET-52b pETDuet-1 pACYCDuet-1 pCDFDuet-1 pRSFDuet-1 pColaDuet-1 |
买家导航
pET32c载体基本信息
pET32c载体图谱质粒图谱和多克隆位点信息
pET32c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。
pET32c载体序列
LOCUS pET-32c(+) 5901 bp DNA circular 11-SEP-2008
SOURCE
ORGANISM
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
FEATURES Location/Qualifiers
promoter complement(765..781)
/vntifkey="30"
/label=T7\promoter
misc_signal complement(367..693)
/vntifkey="87"
/label=trx\tag
protein_bind complement(328..345)
/vntifkey="31"
/label=His\tag
misc_feature complement(250..294)
/vntifkey="21"
/label=S\tag
protein_bind complement(140..157)
/vntifkey="31"
/label=His\tag
terminator complement(26..72)
/vntifkey="43"
/label=T7\terminator
CDS 1172..2251
/vntifkey="4"
/label=lac\I
rep_origin complement(3685..3685)
/vntifkey="33"
/label=ColE1\pBR322
CDS complement(4446..5303)
/vntifkey="4"
/label=bla\(amp)\sequence
rep_origin complement(5435..5890)
/vntifkey="33"
/label=f1\origin
CDS complement(738..762)
/vntifkey="4"
/label=lac\operator
protein_bind complement(301..318)
/vntifkey="31"
/label=thrombin
protein_bind complement(220..234)
/vntifkey="31"
/label=enterokinase
BASE COUNT 1392 a 1565 c 1559 g 1385 t
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 tccacagata tcccatggcc ttgtcgtcgt cgtcggtacc
241 cagatctggg ctgtccatgt gctggcgttc gaatttagca gcagcggttt ctttcatacc
301 agaaccgcgt ggcaccagac cagaagaatg atgatgatga tggtgcatat ggccagaacc
361 agaaccggcc aggttagcgt cgaggaactc tttcaactga cctttagaca gtgcacccac
421 tttggttgcc gccacttcac cgtttttgaa cagcagcaga gtcgggatac cacggatgcc
481 atatttcggc gcagtgccag ggttttgatc gatgttcagt tttgcaacgg tcagtttgcc
541 ctgatattcg tcagcgattt catccagaat cggggcgatc attttgcacg gaccgcacca
601 ctctgcccag aaatcgacga ggatcgcccc gtccgctttg agtacatccg tgtcaaaact
661 gtcgtcagtc aggtgaataa ttttatcgct catatgtata tctccttctt aaagttaaac
721 aaaattattt ctagagggga attgttatcc gctcacaatt cccctatagt gagtcgtatt
781 aatttcgcgg gatcgagatc gatctcgatc ctctacgccg gacgcatcgt ggccggcatc
841 accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat
901 cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc
961 gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg
1021 ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag
1081 cgtcgagatc ccggacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
1141 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
1201 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
1261 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
1321 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
1381 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
1441 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
1501 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
1561 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
1621 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
1681 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
1741 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
1801 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
1861 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
1921 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt
1981 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
2041 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
2101 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
2161 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
2221 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
2281 tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct
2341 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
2401 tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc
2461 gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg
2521 ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg
2581 ccggcatggc ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag
2641 gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa
2701 gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc
2761 gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct
2821 gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct
2881 ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac
2941 cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc
3001 tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat
3061 cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat
3121 taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat
3181 cgcttcacga ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg
3241 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
3301 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
3361 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
3421 gcagattgta ctgagagtgc accatatatg cggtgtgaaa taccgcacag atgcgtaagg
3481 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
3541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
3601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
3661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
3721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
3781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
3841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
3901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
3961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
4021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
4081 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
4141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
4201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
4261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
4321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
4381 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
4441 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
4501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
4561 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
4621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
4681 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
4741 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
4801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
4861 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
4921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
4981 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
5041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
5101 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
5161 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
5221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
5281 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
5341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
5401 gttccgcgca catttccccg aaaagtgcca cctgaaattg taaacgttaa tattttgtta
5461 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
5521 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
5581 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
5641 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
5701 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
5761 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
5821 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
5881 cagggcgcgt cccattcgcc a
//
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pET32c载体质粒应用举例
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