pET-15b载体质粒图谱
/pET-15b Map.png)
pET-15b载体载体序列
LOCUS Exported 5708 bp ds-DNA circular SYN 01-12-2013
DEFINITION Bacterial vector for inducible expression of N-terminally
6xHis-tagged proteins.
ACCESSION .
VERSION .
KEYWORDS pET-15b
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5708)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
JOURNAL Exported 2017/2/28 from SnapGene 2.3.2
http://www.snapgene.com
COMMENT
FEATURES Location/Qualifiers
source 1..5708
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
terminator 213..260
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
misc_feature 319..335
/note="MCS"
/note="multiple cloning site"
CDS complement(336..353)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(363..380)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(390..392)
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
RBS 400..404
/note="ribosome binding site"
protein_bind 436..460
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(461..479)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 788..865
/gene="lacI"
/note="lacI promoter"
/note="
"
CDS 866..1948
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
CDS 3260..3451
/codon_start=1
/gene="rop"
/product="Rop protein"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(3881..4469)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4640..5500)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5501..5605)
/gene="bla"
/note="AmpR promoter"
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
//
pET-15b载体载体特征
一级属性 |
二级属性 |
属性值 |
起始位置 |
常规 | 长度 | 5708 | |
常规 | 描述 | Bacterial vector for inducible expression of N-terminally 6xHis-tagged proteins. | |
常规 | 是否环形 | 1 | |
常规 | 作者 | Novagen (EMD Millipore) | |
常规 | 注释 | | |
来源 | 实验宿主 | Escherichia coli | 1..5708 |
来源 | 分子类型 | other DNA | 1..5708 |
来源 | 种属 | synthetic DNA construct | 1..5708 |
终止子 | 备注 | T7 terminator,transcription terminator for bacteriophage T7 RNA polymerase | 213..260 |
其它特征区 | 备注 | MCS,multiple cloning site | 319..335 |
编码区 | 翻译起始位点 | 1 | 336..353 |
编码区 | 备注 | thrombin site | 336..353 |
编码区 | 产物 | thrombin recognition and cleavage site | 336..353 |
编码区 | 翻译序列 | LVPRGS | 336..353 |
编码区 | 翻译起始位点 | 1 | 363..380 |
编码区 | 备注 | 6xHis | 363..380 |
编码区 | 产物 | 6xHis affinity tag | 363..380 |
编码区 | 翻译序列 | HHHHHH | 363..380 |
编码区 | 翻译起始位点 | 1 | 390..392 |
编码区 | 备注 | ATG | 390..392 |
编码区 | 产物 | start codon | 390..392 |
编码区 | 翻译序列 | M | 390..392 |
核糖体结合位点 | 备注 | ribosome binding site | 400..404 |
蛋白结合位点 | 嵌合范围 | lac repressor encoded by lacI | 436..460 |
蛋白结合位点 | 备注 | lac operator,The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG). | 436..460 |
启动子 | 备注 | T7 promoter,promoter for bacteriophage T7 RNA polymerase | 461..479 |
启动子 | 基因 | lacI | 788..865 |
启动子 | 备注 | lacI promoter, | 788..865 |
编码区 | 翻译起始位点 | 1 | 866..1948 |
编码区 | 基因 | lacI | 866..1948 |
编码区 | 备注 | lacI,The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG). | 866..1948 |
编码区 | 产物 | lac repressor | 866..1948 |
编码区 | 翻译序列 | MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ | 866..1948 |
编码区 | 翻译起始位点 | 1 | 3260..3451 |
编码区 | 基因 | rop | 3260..3451 |
编码区 | 备注 | rop | 3260..3451 |
编码区 | 产物 | Rop protein | 3260..3451 |
编码区 | 翻译序列 | MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL | 3260..3451 |
复制起点 | 方向 | LEFT | 3881..4469 |
复制起点 | 备注 | ori,high-copy-number ColE1/pMB1/pBR322/pUC origin of replication | 3881..4469 |
编码区 | 翻译起始位点 | 1 | 4640..5500 |
编码区 | 基因 | bla | 4640..5500 |
编码区 | 备注 | AmpR,confers resistance to ampicillin, carbenicillin, and related antibiotics | 4640..5500 |
编码区 | 产物 | beta-lactamase | 4640..5500 |
编码区 | 翻译序列 | MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW | 4640..5500 |
启动子 | 基因 | bla | 5501..5605 |
启动子 | 备注 | AmpR promoter | 5501..5605 |
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