| 出品公司: | EMD Biosciences(Novagen) |
|---|---|
| 别名: | pET-12a, pet 12a |
| 质粒类型: | 大肠杆菌表达载体 |
| 表达水平: | 高 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 4674bp |
| 5' 测序引物: | T7/ lacI |
| 5' 测序引物序列: | 5'-TAATACGACTCACTATAGGG-3' |
| 载体标签: | OmpT |
| 载体抗性: | Ampicillin |
| 克隆菌株: | DH5a |
| 表达菌株: | BL21系列 |
| 备注: |
Hosts: E.coli. Related vectors: pBR322；
pET-12a，b，c仅阅读框（读码框）不同，彼此相差1个bp。 |
| 产品目录号: | -- |
| 稳定性: | 稳表达 |
| 组成型: | 诱导型（IPTG) |
| 病毒/非病毒: | 非病毒 |
| 表达宿主菌: | 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
pET-12a(+)载体 pet12a质粒基本信息
pET-12a(+)质粒图谱 pet12a载体图谱和pET-12a(+)载体序列 pet12a质粒序列多克隆位点信息
| Feature Name | Start | End | |
|---|---|---|---|
| tet (w/ gaps) | 86 | 1587 | |
| T7_terminator | 507 | 379 | |
| T7_Terminal_primer | 447 | 465 | |
| T7_transl_en_RBS | 606 | 590 | |
| T7_promoter | 661 | 643 | |
| pBRrevBam_primer | 764 | 745 | |
| ROP | 2226 | 2417 | |
| pGEX_3_primer | 2433 | 2411 | |
| Ampicillin | 4466 | 3606 | |
| AmpR_promoter | 4536 | 4508 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 3 | 798 | 1385 | |
| ORF frame 1 | 1392 | 907 | |
| ORF frame 1 | 970 | 1587 | |
| ORF frame 2 | 4466 | 3606 |
| Enzyme Name | Cut |
|---|---|
| ClaI | 24 |
| HindIII | 29 |
| NheI | 229 |
| BamHI | 510 |
| SalI | 517 |
| NdeI | 580 |
| XbaI | 618 |
| BglII | 676 |
| EagI | 1250 |
| MscI | 1757 |
| PvuII | 2377 |
| PstI | 3924 |
| AatII | 4601 |
| EcoRI | 4672 |
pET-12a(+)质粒 pet12a载体简介
pET-12a-c载体具有N端ompT标签，用于周质腔蛋白表达。pET12a载体独特的酶切位点在载体图谱中被标注出来。需要注意的是，这些载体序列是依据载体pBR322的方向来进行编号的，所以T7标签表达区域在载体图谱上是反向的。
pET-12a(+)质粒序列 pet12a载体序列
LOCUS pET12a 4674 bp DNA circular SYN
DEFINITION pET12a
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
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
source 1..4674
/organism="pET12a"
/mol_type="other DNA"
gene 86..1587
/label="tet (w/ gaps)"
/gene="tet (w/ gaps)"
terminator complement(379..507)
/label="T7_terminator"
misc_feature 447..465
/label="T7_Terminal_primer"
misc_feature complement(590..606)
/label="T7_transl_en_RBS"
promoter complement(643..661)
/label="T7_promoter"
misc_feature complement(745..764)
/label="pBRrevBam_primer"
CDS 798..1385
/label="ORF frame 3"
/translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
LMQESHKGERRSTDALESLQPSQLLPVGAGHDYRRRTYDCLLYHATRRTGAGSALGHF
RRGPLSLERDDDRPVACGIRNLARPRSSLRHWSRHQTFRREAGHYRRHGGRRAGLRLA
GVRDARLDGLPHYDSSRFRRHRDARVAGHAVQAGR*"
CDS complement(907..1392)
/label="ORF frame 1"
/translation="MVVIYLPGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQ
RVGRHAGDNGLLLAETFGGGTSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPER
CRHLSYELHDKEDSHKCGDDSHAPRPPEGADWVEGSQGHRSIDALPYATPALGSSPVV
G*"
CDS 970..1587
/label="ORF frame 1"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
misc_feature 2226..2417
/label="ROP"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
misc_feature complement(2411..2433)
/label="pGEX_3_primer"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
gene complement(3606..4466)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
CDS complement(3606..4466)
/label="ORF frame 2"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(4508..4536)
/label="AmpR_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
ORIGIN
1 TTCTCATGTT TGACAGCTTA TCATCGATAA GCTTTAATGC GGTAGTTTAT CACAGTTAAA
61 TTGCTAACGC AGTCAGGCAC CGTGTATGAA ATCTAACAAT GCGCTCATCG TCATCCTCGG
121 CACCGTCACC CTGGATGCTG TAGGCATAGG CTTGGTTATG CCGGTACTGC CGGGCCTCTT
181 GCGGGATATC GTCCATTCCG ACAGCATCGC CAGTCACTAT GGCGTGCTGC TAGCGCTATA
241 TGCGTTGATG CAATTTCTAT GCGCACCCGT TCTCGGAGCA CTGTCCGACC GCTTTGGCCG
301 CCGCCCAGTC CTGCTCGCTT CGCTACTTGG AGCCACTATC GACTACGCGA TCATGGCGAC
361 CACACCCGTC CTGTGGATAT CCGGATATAG TTCCTCCTTT CAGCAAAAAA CCCCTCAAGA
421 CCCGTTTAGA GGCCCCAAGG GGTTATGCTA GTTATTGCTC AGCGGTGGCA GCAGCCAACT
481 CAGCTTCCTT TCGGGCTTTG TTAGCAGCCG GATCCCGTCG ACGCAAAAGA GCTGATCGCG
541 ATAGGGGTTG TCAGGACTAT TCCTAGGAGT TTCGCCCGCA TATGTATATC TCCTTCTTAA
601 AGTTAAACAA AATTATTTCT AGAGGGAAAC CGTTGTGGTC TCCCTATAGT GAGTCGTATT
661 AATTTCGCGG GATCGAGATC TCGATCCTCT ACGCCGGACG CATCGTGGCC GGCATCACCG
721 GCGCCACAGG TGCGGTTGCT GGCGCCTATA TCGCCGACAT CACCGATGGG GAAGATCGGG
781 CTCGCCACTT CGGGCTCATG AGCGCTTGTT TCGGCGTGGG TATGGTGGCA GGCCCCGTGG
841 CCGGGGGACT GTTGGGCGCC ATCTCCTTGC ATGCACCATT CCTTGCGGCG GCGGTGCTCA
901 ACGGCCTCAA CCTACTACTG GGCTGCTTCC TAATGCAGGA GTCGCATAAG GGAGAGCGTC
961 GATCGACCGA TGCCCTTGAG AGCCTTCAAC CCAGTCAGCT CCTTCCGGTG GGCGCGGGGC
1021 ATGACTATCG TCGCCGCACT TATGACTGTC TTCTTTATCA TGCAACTCGT AGGACAGGTG
1081 CCGGCAGCGC TCTGGGTCAT TTTCGGCGAG GACCGCTTTC GCTGGAGCGC GACGATGATC
1141 GGCCTGTCGC TTGCGGTATT CGGAATCTTG CACGCCCTCG CTCAAGCCTT CGTCACTGGT
1201 CCCGCCACCA AACGTTTCGG CGAGAAGCAG GCCATTATCG CCGGCATGGC GGCCGACGCG
1261 CTGGGCTACG TCTTGCTGGC GTTCGCGACG CGAGGCTGGA TGGCCTTCCC CATTATGATT
1321 CTTCTCGCTT CCGGCGGCAT CGGGATGCCC GCGTTGCAGG CCATGCTGTC CAGGCAGGTA
1381 GATGACGACC ATCAGGGACA GCTTCAAGGA TCGCTCGCGG CTCTTACCAG CCTAACTTCG
1441 ATCACTGGAC CGCTGATCGT CACGGCGATT TATGCCGCCT CGGCGAGCAC ATGGAACGGG
1501 TTGGCATGGA TTGTAGGCGC CGCCCTATAC CTTGTCTGCC TCCCCGCGTT GCGTCGCGGT
1561 GCATGGAGCC GGGCCACCTC GACCTGAATG GAAGCCGGCG GCACCTCGCT AACGGATTCA
1621 CCACTCCAAG AATTGGAGCC AATCAATTCT TGCGGAGAAC TGTGAATGCG CAAACCAACC
1681 CTTGGCAGAA CATATCCATC GCGTCCGCCA TCTCCAGCAG CCGCACGCGG CGCATCTCGG
1741 GCAGCGTTGG GTCCTGGCCA CGGGTGCGCA TGATCGTGCT CCTGTCGTTG AGGACCCGGC
1801 TAGGCTGGCG GGGTTGCCTT ACTGGTTAGC AGAATGAATC ACCGATACGC GAGCGAACGT
1861 GAAGCGACTG CTGCTGCAAA ACGTCTGCGA CCTGAGCAAC AACATGAATG GTCTTCGGTT
1921 TCCGTGTTTC GTAAAGTCTG GAAACGCGGA AGTCAGCGCC CTGCACCATT ATGTTCCGGA
1981 TCTGCATCGC AGGATGCTGC TGGCTACCCT GTGGAACACC TACATCTGTA TTAACGAAGC
2041 GCTGGCATTG ACCCTGAGTG ATTTTTCTCT GGTCCCGCCG CATCCATACC GCCAGTTGTT
2101 TACCCTCACA ACGTTCCAGT AACCGGGCAT GTTCATCATC AGTAACCCGT ATCGTGAGCA
2161 TCCTCTCTCG TTTCATCGGT ATCATTACCC CCATGAACAG AAATCCCCCT TACACGGAGG
2221 CATCAGTGAC CAAACAGGAA AAAACCGCCC TTAACATGGC CCGCTTTATC AGAAGCCAGA
2281 CATTAACGCT TCTGGAGAAA CTCAACGAGC TGGACGCGGA TGAACAGGCA GACATCTGTG
2341 AATCGCTTCA CGACCACGCT GATGAGCTTT ACCGCAGCTG CCTCGCGCGT TTCGGTGATG
2401 ACGGTGAAAA CCTCTGACAC ATGCAGCTCC CGGAGACGGT CACAGCTTGT CTGTAAGCGG
2461 ATGCCGGGAG CAGACAAGCC CGTCAGGGCG CGTCAGCGGG TGTTGGCGGG TGTCGGGGCG
2521 CAGCCATGAC CCAGTCACGT AGCGATAGCG GAGTGTATAC TGGCTTAACT ATGCGGCATC
2581 AGAGCAGATT GTACTGAGAG TGCACCATAT ATGCGGTGTG AAATACCGCA CAGATGCGTA
2641 AGGAGAAAAT ACCGCATCAG GCGCTCTTCC GCTTCCTCGC TCACTGACTC GCTGCGCTCG
2701 GTCGTTCGGC TGCGGCGAGC GGTATCAGCT CACTCAAAGG CGGTAATACG GTTATCCACA
2761 GAATCAGGGG ATAACGCAGG AAAGAACATG TGAGCAAAAG GCCAGCAAAA GGCCAGGAAC
2821 CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC CATAGGCTCC GCCCCCCTGA CGAGCATCAC
2881 AAAAATCGAC GCTCAAGTCA GAGGTGGCGA AACCCGACAG GACTATAAAG ATACCAGGCG
2941 TTTCCCCCTG GAAGCTCCCT CGTGCGCTCT CCTGTTCCGA CCCTGCCGCT TACCGGATAC
3001 CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG GCGCTTTCTC ATAGCTCACG CTGTAGGTAT
3061 CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG CTGGGCTGTG TGCACGAACC CCCCGTTCAG
3121 CCCGACCGCT GCGCCTTATC CGGTAACTAT CGTCTTGAGT CCAACCCGGT AAGACACGAC
3181 TTATCGCCAC TGGCAGCAGC CACTGGTAAC AGGATTAGCA GAGCGAGGTA TGTAGGCGGT
3241 GCTACAGAGT TCTTGAAGTG GTGGCCTAAC TACGGCTACA CTAGAAGGAC AGTATTTGGT
3301 ATCTGCGCTC TGCTGAAGCC AGTTACCTTC GGAAAAAGAG TTGGTAGCTC TTGATCCGGC
3361 AAACAAACCA CCGCTGGTAG CGGTGGTTTT TTTGTTTGCA AGCAGCAGAT TACGCGCAGA
3421 AAAAAAGGAT CTCAAGAAGA TCCTTTGATC TTTTCTACGG GGTCTGACGC TCAGTGGAAC
3481 GAAAACTCAC GTTAAGGGAT TTTGGTCATG AGATTATCAA AAAGGATCTT CACCTAGATC
3541 CTTTTAAATT AAAAATGAAG TTTTAAATCA ATCTAAAGTA TATATGAGTA AACTTGGTCT
3601 GACAGTTACC AATGCTTAAT CAGTGAGGCA CCTATCTCAG CGATCTGTCT ATTTCGTTCA
3661 TCCATAGTTG CCTGACTCCC CGTCGTGTAG ATAACTACGA TACGGGAGGG CTTACCATCT
3721 GGCCCCAGTG CTGCAATGAT ACCGCGAGAC CCACGCTCAC CGGCTCCAGA TTTATCAGCA
3781 ATAAACCAGC CAGCCGGAAG GGCCGAGCGC AGAAGTGGTC CTGCAACTTT ATCCGCCTCC
3841 ATCCAGTCTA TTAATTGTTG CCGGGAAGCT AGAGTAAGTA GTTCGCCAGT TAATAGTTTG
3901 CGCAACGTTG TTGCCATTGC TGCAGGCATC GTGGTGTCAC GCTCGTCGTT TGGTATGGCT
3961 TCATTCAGCT CCGGTTCCCA ACGATCAAGG CGAGTTACAT GATCCCCCAT GTTGTGCAAA
4021 AAAGCGGTTA GCTCCTTCGG TCCTCCGATC GTTGTCAGAA GTAAGTTGGC CGCAGTGTTA
4081 TCACTCATGG TTATGGCAGC ACTGCATAAT TCTCTTACTG TCATGCCATC CGTAAGATGC
4141 TTTTCTGTGA CTGGTGAGTA CTCAACCAAG TCATTCTGAG AATAGTGTAT GCGGCGACCG
4201 AGTTGCTCTT GCCCGGCGTC AACACGGGAT AATACCGCGC CACATAGCAG AACTTTAAAA
4261 GTGCTCATCA TTGGAAAACG TTCTTCGGGG CGAAAACTCT CAAGGATCTT ACCGCTGTTG
4321 AGATCCAGTT CGATGTAACC CACTCGTGCA CCCAACTGAT CTTCAGCATC TTTTACTTTC
4381 ACCAGCGTTT CTGGGTGAGC AAAAACAGGA AGGCAAAATG CCGCAAAAAA GGGAATAAGG
4441 GCGACACGGA AATGTTGAAT ACTCATACTC TTCCTTTTTC AATATTATTG AAGCATTTAT
4501 CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA TTTAGAAAAA TAAACAAATA
4561 GGGGTTCCGC GCACATTTCC CCGAAAAGTG CCACCTGACG TCTAAGAAAC CATTATTATC
4621 ATGACATTAA CCTATAAAAA TAGGCGTATC ACGAGGCCCT TTCGTCTTCA AGAA
//
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pET-12a(+)质粒 pet12a载体应用举例
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