pETDuet™-1载体质粒图谱
/pETDuet-1 Map.png)
pETDuet™-1载体载体序列
LOCUS Exported 5420 bp ds-DNA circular SYN 30-5-2013
DEFINITION Bacterial vector for the co-expression of two genes.
ACCESSION .
VERSION .
KEYWORDS pETDuet-1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5420)
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..5420
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
protein_bind 3..27
/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)."
RBS 58..63
/note="ribosome binding site"
misc_feature 69..168
/note="MCS-1"
/note="multiple cloning site #1"
CDS 71..73
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 83..100
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
promoter 214..232
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 233..257
/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)."
RBS 286..291
/note="ribosome binding site"
misc_feature 297..438
/note="MCS-2"
/note="multiple cloning site #2"
CDS 300..302
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 366..410
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
terminator 462..509
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 546..1001
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(1116..1976)
/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"
promoter complement(1977..2069)
/gene="bla"
/note="AmpR promoter"
rep_origin 2150..2738
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3168..3359)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3931..5013)
/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"
promoter complement(5014..5091)
/gene="lacI"
/note="lacI promoter"
/note="
"
promoter 5404..2
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
301 tggcagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt
361 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg
421 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
481 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
541 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
601 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
661 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
721 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
781 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
841 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
901 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
961 atttaacgcg aattttaaca aaatattaac gtttacaatt tctggcggca cgatggcatg
1021 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
1081 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
1141 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
1201 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
1261 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
1321 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
1381 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
1441 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
1501 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
1561 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
1621 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
1681 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
1741 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
1801 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
1861 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
1921 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
1981 ttcctttttc aatcatgatt gaagcattta tcagggttat tgtctcatga gcggatacat
2041 atttgaatgt atttagaaaa ataaacaaat aggtcatgac caaaatccct taacgtgagt
2101 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
2161 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
2221 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
2281 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
2341 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
2401 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
2461 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
2521 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
2581 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
2641 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
2701 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
2761 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
2821 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
2881 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
2941 tccttacgca tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg
3001 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
3061 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
3121 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
3181 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
3241 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
3301 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
3361 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
3421 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
3481 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
3541 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
3601 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
3661 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
3721 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
3781 cctagccggg tcctcaacga caggagcacg atcatgctag tcatgccccg cgcccaccgg
3841 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
3901 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
3961 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
4021 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
4081 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
4141 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
4201 cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
4261 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
4321 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
4381 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
4441 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
4501 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
4561 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
4621 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
4681 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
4741 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
4801 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
4861 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
4921 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
4981 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
5041 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
5101 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
5161 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
5221 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
5281 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
5341 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatcgatctc gatcccgcga
5401 aattaatacg actcactata
//
pETDuet™-1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 | 5420 | |
常规 | 描述 | Bacterial vector for the co-expression of two genes. | |
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..5420 |
来源 | 分子类型 | other DNA | 1..5420 |
来源 | 种属 | synthetic DNA construct | 1..5420 |
蛋白结合位点 | 嵌合范围 | lac repressor encoded by lacI | 3..27 |
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). | 3..27 |
核糖体结合位点 | 备注 | ribosome binding site | 58..63 |
其它特征区 | 备注 | MCS-1,multiple cloning site #1 | 69..168 |
编码区 | 翻译起始位点 | 1 | 71..73 |
编码区 | 备注 | ATG | 71..73 |
编码区 | 产物 | start codon | 71..73 |
编码区 | 翻译序列 | M | 71..73 |
编码区 | 翻译起始位点 | 1 | 83..100 |
编码区 | 备注 | 6xHis | 83..100 |
编码区 | 产物 | 6xHis affinity tag | 83..100 |
编码区 | 翻译序列 | HHHHHH | 83..100 |
启动子 | 备注 | T7 promoter,promoter for bacteriophage T7 RNA polymerase | 214..232 |
蛋白结合位点 | 嵌合范围 | lac repressor encoded by lacI | 233..257 |
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). | 233..257 |
核糖体结合位点 | 备注 | ribosome binding site | 286..291 |
其它特征区 | 备注 | MCS-2,multiple cloning site #2 | 297..438 |
编码区 | 翻译起始位点 | 1 | 300..302 |
编码区 | 备注 | ATG | 300..302 |
编码区 | 产物 | start codon | 300..302 |
编码区 | 翻译序列 | M | 300..302 |
编码区 | 翻译起始位点 | 1 | 366..410 |
编码区 | 备注 | S-Tag | 366..410 |
编码区 | 产物 | affinity and epitope tag derived from pancreatic ribonuclease A | 366..410 |
编码区 | 翻译序列 | KETAAAKFERQHMDS | 366..410 |
终止子 | 备注 | T7 terminator,transcription terminator for bacteriophage T7 RNA polymerase | 462..509 |
复制起点 | 方向 | RIGHT | 546..1001 |
复制起点 | 备注 | f1 ori,f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis | 546..1001 |
编码区 | 翻译起始位点 | 1 | 1116..1976 |
编码区 | 基因 | bla | 1116..1976 |
编码区 | 备注 | AmpR,confers resistance to ampicillin, carbenicillin, and related antibiotics | 1116..1976 |
编码区 | 产物 | beta-lactamase | 1116..1976 |
编码区 | 翻译序列 | MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW | 1116..1976 |
启动子 | 基因 | bla | 1977..2069 |
启动子 | 备注 | AmpR promoter | 1977..2069 |
复制起点 | 方向 | RIGHT | 2150..2738 |
复制起点 | 备注 | ori,high-copy-number ColE1/pMB1/pBR322/pUC origin of replication | 2150..2738 |
编码区 | 翻译起始位点 | 1 | 3168..3359 |
编码区 | 基因 | rop | 3168..3359 |
编码区 | 备注 | rop | 3168..3359 |
编码区 | 产物 | Rop protein, which maintains plasmids at low copy number | 3168..3359 |
编码区 | 翻译序列 | MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL | 3168..3359 |
编码区 | 翻译起始位点 | 1 | 3931..5013 |
编码区 | 基因 | lacI | 3931..5013 |
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). | 3931..5013 |
编码区 | 产物 | lac repressor | 3931..5013 |
编码区 | 翻译序列 | MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ | 3931..5013 |
启动子 | 基因 | lacI | 5014..5091 |
启动子 | 备注 | lacI promoter, | 5014..5091 |
启动子 | 备注 | T7 promoter,promoter for bacteriophage T7 RNA polymerase | 2..5404 |
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