| 出品公司: | |
|---|---|
| 别名: | pET15b, pet 15b |
| 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: | 5708bp |
| 5' 测序引物: | T7 |
| 5' 测序引物序列: | 5'-TAATACGACTCACTATAGGG-3' |
| 载体标签: | N-His, N-thrombin |
| 载体抗性: | Ampicillin |
| 备注: |
Hosts: E.coli. Related vectors: pBR322.
|
| 产品目录号: | ZY69661-3 |
| 稳定性: | 瞬时表达 Transient |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pET-15b
- 库存：
20
- 供应商：
泽叶生物
pet15b载体基本信息
pet15b载体质粒图谱和多克隆位点信息
| Feature Name | Start | End | |
|---|---|---|---|
| T7_terminator | 316 | 188 | |
| T7_Terminal_primer | 256 | 274 | |
| T7_transl_en_RBS | 416 | 400 | |
| lacO | 461 | 434 | |
| T7_promoter | 479 | 461 | |
| pBRrevBam_primer | 582 | 563 | |
| lacI | 857 | 1948 | |
| tet (576 - 1190) | 2007 | 2621 | |
| ROP | 3260 | 3451 | |
| pGEX_3_primer | 3467 | 3445 | |
| Ampicillin | 5500 | 4640 | |
| AmpR_promoter | 5570 | 5542 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 1 | 616 | 1116 | |
| ORF frame 2 | 989 | 1948 | |
| ORF frame 1 | 2426 | 1731 | |
| ORF frame 3 | 2004 | 2621 | |
| ORF frame 2 | 5500 | 4640 |
| Enzyme Name | Cut |
|---|---|
| ClaI | 24 |
| HindIII | 29 |
| BamHI | 319 |
| XhoI | 324 |
| NdeI | 331 |
| NcoI | 389 |
| XbaI | 428 |
| BglII | 494 |
| BclI | 1230 |
| ApaI | 1427 |
| HpaI | 1722 |
| EagI | 2284 |
| NruI | 2319 |
| MscI | 2791 |
| PstI | 4958 |
| AatII | 5635 |
| EcoRI | 5706 |
pet15b载体简介
pet15b载体序列
LOCUS pET-15b 5708 bp DNA circular SYN
DEFINITION pET-15b
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..5708
/organism="pET-15b"
/mol_type="other DNA"
terminator complement(188..316)
/label="T7_terminator"
misc_feature 256..274
/label="T7_Terminal_primer"
misc_feature complement(400..416)
/label="T7_transl_en_RBS"
misc_feature complement(434..461)
/label="lacO"
promoter complement(461..479)
/label="T7_promoter"
misc_feature complement(563..582)
/label="pBRrevBam_primer"
CDS 616..1116
/label="ORF frame 1"
/translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
ANCRGD*"
misc_feature 857..1948
/label="lacI"
/translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
ANCRGD*"
CDS 989..1948
/label="ORF frame 2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS complement(1731..2426)
/label="ORF frame 1"
/translation="MVVIYLPGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQ
RVGRHAGDNGLLLAETFGGGTSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPER
CRHLSYELHDKEDSHKCGDDSHAPRPPEGADWVEGSQGHRSRSRCLMSELTYINCVAL
TARFPVGKPVVPAALMNRPTRGERRFAYWAPGWFFFSPVRRATADCPSPPGPERVAAS
GPRWFAPAGENPV*"
CDS 2004..2621
/label="ORF frame 3"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
gene 2007..2621
/label="tet (576 - 1190)"
/gene="tet (576 - 1190)"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
misc_feature 3260..3451
/label="ROP"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
misc_feature complement(3445..3467)
/label="pGEX_3_primer"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
gene complement(4640..5500)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
CDS complement(4640..5500)
/label="ORF frame 2"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(5542..5570)
/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 cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
301 cgggctttgt tagcagccgg atcctcgagc atatggctgc cgcgcggcac caggccgctg
361 ctgtgatgat gatgatgatg gctgctgccc atggtatatc tccttcttaa agttaaacaa
421 aattatttct agaggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa
481 tttcgcggga tcgagatctc gatcctctac gccggacgca tcgtggccgg catcaccggc
541 gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct
601 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc
661 gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac
721 ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga
781 gatcccggac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga
841 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
901 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
961 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
1021 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
1081 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
1141 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
1201 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
1261 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
1321 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
1381 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
1441 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
1501 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
1561 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
1621 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
1681 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga
1741 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
1801 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
1861 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
1921 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc
1981 attaggcacc gggatctcga ccgatgccct tgagagcctt caacccagtc agctccttcc
2041 ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac
2101 tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga
2161 gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag
2221 ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca
2281 tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct
2341 tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc
2401 tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc gcggctctta
2461 ccagcctaac ttcgatcact ggaccgctga tcgtcacggc gatttatgcc gcctcggcga
2521 gcacatggaa cgggttggca tggattgtag gcgccgccct ataccttgtc tgcctccccg
2581 cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg aatggaagcc ggcggcacct
2641 cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa
2701 tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac
2761 gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc
2821 gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat
2881 acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg
2941 aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac
3001 cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc
3061 tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca
3121 taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac
3181 ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc
3241 cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt
3301 tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca
3361 ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc
3421 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
3481 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
3541 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
3601 aactatgcgg catcagagca gattgtactg agagtgcacc atatatgcgg tgtgaaatac
3661 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
3721 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
3781 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
3841 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
3901 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
3961 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
4021 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
4081 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
4141 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
4201 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
4261 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
4321 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
4381 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
4441 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
4501 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
4561 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
4621 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
4681 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
4741 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
4801 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
4861 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
4921 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
4981 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
5041 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
5101 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
5161 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
5221 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
5281 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
5341 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
5401 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
5461 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
5521 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
5581 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
5641 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
5701 ttcaagaa
DEFINITION pET-15b
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..5708
/organism="pET-15b"
/mol_type="other DNA"
terminator complement(188..316)
/label="T7_terminator"
misc_feature 256..274
/label="T7_Terminal_primer"
misc_feature complement(400..416)
/label="T7_transl_en_RBS"
misc_feature complement(434..461)
/label="lacO"
promoter complement(461..479)
/label="T7_promoter"
misc_feature complement(563..582)
/label="pBRrevBam_primer"
CDS 616..1116
/label="ORF frame 1"
/translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
ANCRGD*"
misc_feature 857..1948
/label="lacI"
/translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
ANCRGD*"
CDS 989..1948
/label="ORF frame 2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS complement(1731..2426)
/label="ORF frame 1"
/translation="MVVIYLPGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQ
RVGRHAGDNGLLLAETFGGGTSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPER
CRHLSYELHDKEDSHKCGDDSHAPRPPEGADWVEGSQGHRSRSRCLMSELTYINCVAL
TARFPVGKPVVPAALMNRPTRGERRFAYWAPGWFFFSPVRRATADCPSPPGPERVAAS
GPRWFAPAGENPV*"
CDS 2004..2621
/label="ORF frame 3"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
gene 2007..2621
/label="tet (576 - 1190)"
/gene="tet (576 - 1190)"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
misc_feature 3260..3451
/label="ROP"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
misc_feature complement(3445..3467)
/label="pGEX_3_primer"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
gene complement(4640..5500)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVI
FGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVL
LAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITG
PLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
CDS complement(4640..5500)
/label="ORF frame 2"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(5542..5570)
/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 cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
301 cgggctttgt tagcagccgg atcctcgagc atatggctgc cgcgcggcac caggccgctg
361 ctgtgatgat gatgatgatg gctgctgccc atggtatatc tccttcttaa agttaaacaa
421 aattatttct agaggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa
481 tttcgcggga tcgagatctc gatcctctac gccggacgca tcgtggccgg catcaccggc
541 gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct
601 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc
661 gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac
721 ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga
781 gatcccggac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga
841 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
901 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
961 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
1021 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
1081 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
1141 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
1201 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
1261 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
1321 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
1381 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
1441 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
1501 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
1561 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
1621 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
1681 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga
1741 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
1801 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
1861 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
1921 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc
1981 attaggcacc gggatctcga ccgatgccct tgagagcctt caacccagtc agctccttcc
2041 ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac
2101 tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga
2161 gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag
2221 ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca
2281 tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct
2341 tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc
2401 tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc gcggctctta
2461 ccagcctaac ttcgatcact ggaccgctga tcgtcacggc gatttatgcc gcctcggcga
2521 gcacatggaa cgggttggca tggattgtag gcgccgccct ataccttgtc tgcctccccg
2581 cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg aatggaagcc ggcggcacct
2641 cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa
2701 tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac
2761 gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc
2821 gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat
2881 acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg
2941 aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac
3001 cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc
3061 tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca
3121 taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac
3181 ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc
3241 cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt
3301 tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca
3361 ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc
3421 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
3481 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
3541 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
3601 aactatgcgg catcagagca gattgtactg agagtgcacc atatatgcgg tgtgaaatac
3661 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
3721 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
3781 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
3841 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
3901 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
3961 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
4021 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
4081 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
4141 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
4201 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
4261 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
4321 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
4381 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
4441 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
4501 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
4561 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
4621 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
4681 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
4741 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
4801 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
4861 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
4921 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
4981 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
5041 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
5101 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
5161 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
5221 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
5281 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
5341 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
5401 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
5461 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
5521 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
5581 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
5641 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
5701 ttcaagaa
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文献和实验相关实验
表达载体的构建是生物技术和所需蛋白质产生的基本工具，本文总结了pET-32α（+）载体技术的构建，该技术通常用于研究实验室。该方法包括获得用于构建载体的外源DNA片段，将DNA片段亚克隆到pET-32α（+）表达载体中，在大肠杆菌 BL21（DE3）中进行蛋白质表达以及在大肠杆菌中在天然条件下进行蛋白质纯化。裂解液。介绍作为分子生物学实验室中非常有用的实践，重组DNA技术（或基因克隆）是指将DNA片段从一种生物体转移到自我复制的遗传元件如表达载体。然后插入的DNA可以在外源宿主细胞中繁殖
相关专题 1)噬菌体DE3溶源化的菌株如BL21(DE3)是最常用的表达菌株，噬菌体DE3是λ噬菌体的衍生株，构建好的表达载体 可以直接转入表达菌株中，诱导蛋白的表达方式与lac启动子一样都是IPTG诱导。 2)表达菌株BL21(DE3)上带有lacI基因，T7 RNA 聚合酶编码基因以及lacUV5启动子，lacUV5启动子可以启动T7 RNA 聚合酶的表达; 3)表达质粒 如pET-28质粒带有T7启动子和编码阻遏
（ DE3 ）表达的活性蛋白比其它宿主菌高 10 倍以上。 Origami 宿主菌与氨苄抗性质粒相容，可用于 pET-32 载体，硫氧还蛋白标签能够进一步增强在胞浆内形成二硫键。 TrxB 和 gor 突变可分别用卡那霉素和四环素选择，因此该菌株建议用于带氨苄抗性标记 bla 的 pET 质粒。 Origami B 宿主菌来源于 BL21 lacZY 突变株，还带有与原始 Origami 菌株相同的 TrxB / gor 突变。 Origami B 菌株集 BL21 、 Tuner
技术资料暂无技术资料 索取技术资料
pET-15b载体
¥1100
