| 出品公司: | EMD Biosciences(Novagen) |
|---|---|
| 别名: | pET-12b, pet 12b |
| 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: | 4673bp |
| 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: |
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-12b(+)载体 pet12b质粒基本信息
pET-12b(+)质粒图谱 pet12b载体图谱和pET-12b(+)载体序列 pet12b质粒序列多克隆位点信息
| Feature Name | Start | End | |
|---|---|---|---|
| tet (w/ gaps) | 86 | 1586 | |
| T7_terminator | 507 | 379 | |
| T7_Terminal_primer | 447 | 465 | |
| T7_transl_en_RBS | 605 | 589 | |
| T7_promoter | 660 | 642 | |
| pBRrevBam_primer | 763 | 744 | |
| ROP | 2225 | 2416 | |
| pGEX_3_primer | 2432 | 2410 | |
| Ampicillin | 4465 | 3605 | |
| AmpR_promoter | 4535 | 4507 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 2 | 797 | 1384 | |
| ORF frame 1 | 1391 | 906 | |
| ORF frame 3 | 969 | 1586 | |
| ORF frame 2 | 4465 | 3605 |
| Enzyme Name | Cut |
|---|---|
| ClaI | 24 |
| HindIII | 29 |
| NheI | 229 |
| BamHI | 510 |
| SalI | 516 |
| NdeI | 579 |
| XbaI | 617 |
| BglII | 675 |
| EagI | 1249 |
| MscI | 1756 |
| PvuII | 2376 |
| PstI | 3923 |
| AatII | 4600 |
| EcoRI | 4671 |
pET-12b(+)质粒 pet12b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。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-12b(+)质粒序列 pet12b载体序列
LOCUS pET12b 4673 bp DNA circular SYN
DEFINITION pET12b
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..4673
/organism="pET12b"
/mol_type="other DNA"
gene 86..1586
/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(589..605)
/label="T7_transl_en_RBS"
promoter complement(642..660)
/label="T7_promoter"
misc_feature complement(744..763)
/label="pBRrevBam_primer"
CDS 797..1384
/label="ORF frame 2"
/translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
LMQESHKGERRSTDALESLQPSQLLPVGAGHDYRRRTYDCLLYHATRRTGAGSALGHF
RRGPLSLERDDDRPVACGIRNLARPRSSLRHWSRHQTFRREAGHYRRHGGRRAGLRLA
GVRDARLDGLPHYDSSRFRRHRDARVAGHAVQAGR*"
CDS complement(906..1391)
/label="ORF frame 1"
/translation="MVVIYLPGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQ
RVGRHAGDNGLLLAETFGGGTSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPER
CRHLSYELHDKEDSHKCGDDSHAPRPPEGADWVEGSQGHRSIDALPYATPALGSSPVV
G*"
CDS 969..1586
/label="ORF frame 3"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
misc_feature 2225..2416
/label="ROP"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
misc_feature complement(2410..2432)
/label="pGEX_3_primer"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
gene complement(3605..4465)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
CDS complement(3605..4465)
/label="ORF frame 2"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(4507..4535)
/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 GATCCGTCGA CGCAAAAGAG CTGATCGCGA
541 TAGGGGTTGT CAGGACTATT CCTAGGAGTT TCGCCCGCAT ATGTATATCT CCTTCTTAAA
601 GTTAAACAAA ATTATTTCTA GAGGGAAACC GTTGTGGTCT CCCTATAGTG AGTCGTATTA
661 ATTTCGCGGG ATCGAGATCT CGATCCTCTA CGCCGGACGC ATCGTGGCCG GCATCACCGG
721 CGCCACAGGT GCGGTTGCTG GCGCCTATAT CGCCGACATC ACCGATGGGG AAGATCGGGC
781 TCGCCACTTC GGGCTCATGA GCGCTTGTTT CGGCGTGGGT ATGGTGGCAG GCCCCGTGGC
841 CGGGGGACTG TTGGGCGCCA TCTCCTTGCA TGCACCATTC CTTGCGGCGG CGGTGCTCAA
901 CGGCCTCAAC CTACTACTGG GCTGCTTCCT AATGCAGGAG TCGCATAAGG GAGAGCGTCG
961 ATCGACCGAT GCCCTTGAGA GCCTTCAACC CAGTCAGCTC CTTCCGGTGG GCGCGGGGCA
1021 TGACTATCGT CGCCGCACTT ATGACTGTCT TCTTTATCAT GCAACTCGTA GGACAGGTGC
1081 CGGCAGCGCT CTGGGTCATT TTCGGCGAGG ACCGCTTTCG CTGGAGCGCG ACGATGATCG
1141 GCCTGTCGCT TGCGGTATTC GGAATCTTGC ACGCCCTCGC TCAAGCCTTC GTCACTGGTC
1201 CCGCCACCAA ACGTTTCGGC GAGAAGCAGG CCATTATCGC CGGCATGGCG GCCGACGCGC
1261 TGGGCTACGT CTTGCTGGCG TTCGCGACGC GAGGCTGGAT GGCCTTCCCC ATTATGATTC
1321 TTCTCGCTTC CGGCGGCATC GGGATGCCCG CGTTGCAGGC CATGCTGTCC AGGCAGGTAG
1381 ATGACGACCA TCAGGGACAG CTTCAAGGAT CGCTCGCGGC TCTTACCAGC CTAACTTCGA
1441 TCACTGGACC GCTGATCGTC ACGGCGATTT ATGCCGCCTC GGCGAGCACA TGGAACGGGT
1501 TGGCATGGAT TGTAGGCGCC GCCCTATACC TTGTCTGCCT CCCCGCGTTG CGTCGCGGTG
1561 CATGGAGCCG GGCCACCTCG ACCTGAATGG AAGCCGGCGG CACCTCGCTA ACGGATTCAC
1621 CACTCCAAGA ATTGGAGCCA ATCAATTCTT GCGGAGAACT GTGAATGCGC AAACCAACCC
1681 TTGGCAGAAC ATATCCATCG CGTCCGCCAT CTCCAGCAGC CGCACGCGGC GCATCTCGGG
1741 CAGCGTTGGG TCCTGGCCAC GGGTGCGCAT GATCGTGCTC CTGTCGTTGA GGACCCGGCT
1801 AGGCTGGCGG GGTTGCCTTA CTGGTTAGCA GAATGAATCA CCGATACGCG AGCGAACGTG
1861 AAGCGACTGC TGCTGCAAAA CGTCTGCGAC CTGAGCAACA ACATGAATGG TCTTCGGTTT
1921 CCGTGTTTCG TAAAGTCTGG AAACGCGGAA GTCAGCGCCC TGCACCATTA TGTTCCGGAT
1981 CTGCATCGCA GGATGCTGCT GGCTACCCTG TGGAACACCT ACATCTGTAT TAACGAAGCG
2041 CTGGCATTGA CCCTGAGTGA TTTTTCTCTG GTCCCGCCGC ATCCATACCG CCAGTTGTTT
2101 ACCCTCACAA CGTTCCAGTA ACCGGGCATG TTCATCATCA GTAACCCGTA TCGTGAGCAT
2161 CCTCTCTCGT TTCATCGGTA TCATTACCCC CATGAACAGA AATCCCCCTT ACACGGAGGC
2221 ATCAGTGACC AAACAGGAAA AAACCGCCCT TAACATGGCC CGCTTTATCA GAAGCCAGAC
2281 ATTAACGCTT CTGGAGAAAC TCAACGAGCT GGACGCGGAT GAACAGGCAG ACATCTGTGA
2341 ATCGCTTCAC GACCACGCTG ATGAGCTTTA CCGCAGCTGC CTCGCGCGTT TCGGTGATGA
2401 CGGTGAAAAC CTCTGACACA TGCAGCTCCC GGAGACGGTC ACAGCTTGTC TGTAAGCGGA
2461 TGCCGGGAGC AGACAAGCCC GTCAGGGCGC GTCAGCGGGT GTTGGCGGGT GTCGGGGCGC
2521 AGCCATGACC CAGTCACGTA GCGATAGCGG AGTGTATACT GGCTTAACTA TGCGGCATCA
2581 GAGCAGATTG TACTGAGAGT GCACCATATA TGCGGTGTGA AATACCGCAC AGATGCGTAA
2641 GGAGAAAATA CCGCATCAGG CGCTCTTCCG CTTCCTCGCT CACTGACTCG CTGCGCTCGG
2701 TCGTTCGGCT GCGGCGAGCG GTATCAGCTC ACTCAAAGGC GGTAATACGG TTATCCACAG
2761 AATCAGGGGA TAACGCAGGA AAGAACATGT GAGCAAAAGG CCAGCAAAAG GCCAGGAACC
2821 GTAAAAAGGC CGCGTTGCTG GCGTTTTTCC ATAGGCTCCG CCCCCCTGAC GAGCATCACA
2881 AAAATCGACG CTCAAGTCAG AGGTGGCGAA ACCCGACAGG ACTATAAAGA TACCAGGCGT
2941 TTCCCCCTGG AAGCTCCCTC GTGCGCTCTC CTGTTCCGAC CCTGCCGCTT ACCGGATACC
3001 TGTCCGCCTT TCTCCCTTCG GGAAGCGTGG CGCTTTCTCA TAGCTCACGC TGTAGGTATC
3061 TCAGTTCGGT GTAGGTCGTT CGCTCCAAGC TGGGCTGTGT GCACGAACCC CCCGTTCAGC
3121 CCGACCGCTG CGCCTTATCC GGTAACTATC GTCTTGAGTC CAACCCGGTA AGACACGACT
3181 TATCGCCACT GGCAGCAGCC ACTGGTAACA GGATTAGCAG AGCGAGGTAT GTAGGCGGTG
3241 CTACAGAGTT CTTGAAGTGG TGGCCTAACT ACGGCTACAC TAGAAGGACA GTATTTGGTA
3301 TCTGCGCTCT GCTGAAGCCA GTTACCTTCG GAAAAAGAGT TGGTAGCTCT TGATCCGGCA
3361 AACAAACCAC CGCTGGTAGC GGTGGTTTTT TTGTTTGCAA GCAGCAGATT ACGCGCAGAA
3421 AAAAAGGATC TCAAGAAGAT CCTTTGATCT TTTCTACGGG GTCTGACGCT CAGTGGAACG
3481 AAAACTCACG TTAAGGGATT TTGGTCATGA GATTATCAAA AAGGATCTTC ACCTAGATCC
3541 TTTTAAATTA AAAATGAAGT TTTAAATCAA TCTAAAGTAT ATATGAGTAA ACTTGGTCTG
3601 ACAGTTACCA ATGCTTAATC AGTGAGGCAC CTATCTCAGC GATCTGTCTA TTTCGTTCAT
3661 CCATAGTTGC CTGACTCCCC GTCGTGTAGA TAACTACGAT ACGGGAGGGC TTACCATCTG
3721 GCCCCAGTGC TGCAATGATA CCGCGAGACC CACGCTCACC GGCTCCAGAT TTATCAGCAA
3781 TAAACCAGCC AGCCGGAAGG GCCGAGCGCA GAAGTGGTCC TGCAACTTTA TCCGCCTCCA
3841 TCCAGTCTAT TAATTGTTGC CGGGAAGCTA GAGTAAGTAG TTCGCCAGTT AATAGTTTGC
3901 GCAACGTTGT TGCCATTGCT GCAGGCATCG TGGTGTCACG CTCGTCGTTT GGTATGGCTT
3961 CATTCAGCTC CGGTTCCCAA CGATCAAGGC GAGTTACATG ATCCCCCATG TTGTGCAAAA
4021 AAGCGGTTAG CTCCTTCGGT CCTCCGATCG TTGTCAGAAG TAAGTTGGCC GCAGTGTTAT
4081 CACTCATGGT TATGGCAGCA CTGCATAATT CTCTTACTGT CATGCCATCC GTAAGATGCT
4141 TTTCTGTGAC TGGTGAGTAC TCAACCAAGT CATTCTGAGA ATAGTGTATG CGGCGACCGA
4201 GTTGCTCTTG CCCGGCGTCA ACACGGGATA ATACCGCGCC ACATAGCAGA ACTTTAAAAG
4261 TGCTCATCAT TGGAAAACGT TCTTCGGGGC GAAAACTCTC AAGGATCTTA CCGCTGTTGA
4321 GATCCAGTTC GATGTAACCC ACTCGTGCAC CCAACTGATC TTCAGCATCT TTTACTTTCA
4381 CCAGCGTTTC TGGGTGAGCA AAAACAGGAA GGCAAAATGC CGCAAAAAAG GGAATAAGGG
4441 CGACACGGAA ATGTTGAATA CTCATACTCT TCCTTTTTCA ATATTATTGA AGCATTTATC
4501 AGGGTTATTG TCTCATGAGC GGATACATAT TTGAATGTAT TTAGAAAAAT AAACAAATAG
4561 GGGTTCCGCG CACATTTCCC CGAAAAGTGC CACCTGACGT CTAAGAAACC ATTATTATCA
4621 TGACATTAAC CTATAAAAAT AGGCGTATCA CGAGGCCCTT TCGTCTTCAA GAA
//
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pET-12b(+)质粒 pet12b载体应用举例
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