pET-52b(+)载体质粒图谱
/pET-52b(+) Map.png)
pET-52b(+)载体载体序列
LOCUS Exported 5227 bp ds-DNA circular SYN 03-12-2013
DEFINITION Bacterial vector for expressing proteins with a cleavable N-terminal
Strep-Tag II tag and a cleavable C-terminal 10xHis tag.
ACCESSION .
VERSION .
KEYWORDS pET-52b(+)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5227)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
JOURNAL Exported 2017/2/28 from SnapGene 2.3.2
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5227
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(110..139)
/codon_start=1
/product="10xHis affinity tag"
/note="10xHis"
/translation="HHHHHHHHHH"
CDS complement(146..163)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
misc_feature 167..208
/note="MCS"
/note="multiple cloning site"
CDS complement(209..232)
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C protease"
/note="HRV 3C site"
/translation="LEVLFQGP"
CDS complement(239..262)
/codon_start=1
/product="peptide that binds Strep-Tactin, an engineered
form of streptavidin"
/note="Strep-Tag II"
/translation="WSHPQFEK"
CDS complement(269..271)
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
RBS 279..284
/note="ribosome binding site"
protein_bind 315..339
/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(340..358)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 671..748
/gene="lacI"
/note="lacI promoter"
/note="
"
CDS 749..1831
/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 2403..2594
/codon_start=1
/gene="rop"
/product="Rop protein"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(3024..3612)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 3786..4646
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(4761..5216)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccta ggttaattag tggtggtgat
121 ggtgatgatg gtggtgatga gcggaactac cgcgtggcac cagagcgagc tctgcggccg
181 cgtcgacttg tacaggatcc tggtacccgg gtccctgaaa gaggacttca agtgcaccct
241 tttcgaactg cgggtggctc cagcttgcca tggtatatct ccttcttaaa gttaaacaaa
301 attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag tcgtattaat
361 ttcgcgggat cgagatcgat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc
421 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg
481 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg
541 gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc
601 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt
661 cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
721 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
781 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
841 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
901 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
961 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
1021 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
1081 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
1141 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
1201 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
1261 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
1321 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
1381 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
1441 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
1501 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggaca tctcggtagt
1561 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
1621 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
1681 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
1741 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
1801 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca
1861 ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
1921 tccggtgggc gcggggcatg actagcatga tcgtgctcct gtcgttgagg acccggctag
1981 gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa
2041 gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc
2101 gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct
2161 gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct
2221 ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac
2281 cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc
2341 tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat
2401 cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat
2461 taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat
2521 cgcttcacga ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg
2581 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
2641 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
2701 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
2761 gcagattgta ctgagagtgc accatatatg cggtgtgaaa taccgcacag atgcgtaagg
2821 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
2881 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
2941 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
3001 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
3061 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
3121 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
3181 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
3241 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
3301 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
3361 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
3421 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
3481 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
3541 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
3601 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
3661 aactcacgtt aagggatttt ggtcatgacc tatttgttta tttttctaaa tacattcaaa
3721 tatgtatccg ctcatgagac aataaccctg ataaatgctt caatcatgat tgaaaaagga
3781 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
3841 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
3901 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
3961 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
4021 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
4081 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
4141 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
4201 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
4261 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
4321 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
4381 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
4441 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
4501 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
4561 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
4621 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
4681 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
4741 tcatgccatc gtgccgccag aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
4801 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
4861 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
4921 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
4981 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
5041 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
5101 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
5161 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
5221 ttcgcca
//
pET-52b(+)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 | 5227 | |
常规 | 描述 | Bacterial vector for expressing proteins with a cleavable N-terminal Strep-Tag II tag and a cleavable C-terminal 10xHis tag. | |
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..5227 |
来源 | 分子类型 | other DNA | 1..5227 |
来源 | 种属 | synthetic DNA construct | 1..5227 |
终止子 | 备注 | T7 terminator,transcription terminator for bacteriophage T7 RNA polymerase | 26..73 |
编码区 | 翻译起始位点 | 1 | 110..139 |
编码区 | 备注 | 10xHis | 110..139 |
编码区 | 产物 | 10xHis affinity tag | 110..139 |
编码区 | 翻译序列 | HHHHHHHHHH | 110..139 |
编码区 | 翻译起始位点 | 1 | 146..163 |
编码区 | 备注 | thrombin site | 146..163 |
编码区 | 产物 | thrombin recognition and cleavage site | 146..163 |
编码区 | 翻译序列 | LVPRGS | 146..163 |
其它特征区 | 备注 | MCS,multiple cloning site | 167..208 |
编码区 | 翻译起始位点 | 1 | 209..232 |
编码区 | 备注 | HRV 3C site | 209..232 |
编码区 | 产物 | recognition and cleavage site for human rhinovirus 3C protease | 209..232 |
编码区 | 翻译序列 | LEVLFQGP | 209..232 |
编码区 | 翻译起始位点 | 1 | 239..262 |
编码区 | 备注 | Strep-Tag II | 239..262 |
编码区 | 产物 | peptide that binds Strep-Tactin, an engineered form of streptavidin | 239..262 |
编码区 | 翻译序列 | WSHPQFEK | 239..262 |
编码区 | 翻译起始位点 | 1 | 269..271 |
编码区 | 备注 | ATG | 269..271 |
编码区 | 产物 | start codon | 269..271 |
编码区 | 翻译序列 | M | 269..271 |
核糖体结合位点 | 备注 | ribosome binding site | 279..284 |
蛋白结合位点 | 嵌合范围 | lac repressor encoded by lacI | 315..339 |
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). | 315..339 |
启动子 | 备注 | T7 promoter,promoter for bacteriophage T7 RNA polymerase | 340..358 |
启动子 | 基因 | lacI | 671..748 |
启动子 | 备注 | lacI promoter, | 671..748 |
编码区 | 翻译起始位点 | 1 | 749..1831 |
编码区 | 基因 | lacI | 749..1831 |
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). | 749..1831 |
编码区 | 产物 | lac repressor | 749..1831 |
编码区 | 翻译序列 | MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ | 749..1831 |
编码区 | 翻译起始位点 | 1 | 2403..2594 |
编码区 | 基因 | rop | 2403..2594 |
编码区 | 备注 | rop | 2403..2594 |
编码区 | 产物 | Rop protein | 2403..2594 |
编码区 | 翻译序列 | MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL | 2403..2594 |
复制起点 | 方向 | LEFT | 3024..3612 |
复制起点 | 备注 | ori,high-copy-number ColE1/pMB1/pBR322/pUC origin of replication | 3024..3612 |
编码区 | 翻译起始位点 | 1 | 3786..4646 |
编码区 | 基因 | bla | 3786..4646 |
编码区 | 备注 | AmpR,confers resistance to ampicillin, carbenicillin, and related antibiotics | 3786..4646 |
编码区 | 产物 | beta-lactamase | 3786..4646 |
编码区 | 翻译序列 | MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW | 3786..4646 |
复制起点 | 方向 | LEFT | 4761..5216 |
复制起点 | 备注 | f1 ori,f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis | 4761..5216 |
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