pET-30b(+)载体质粒图谱
/pET-30b(+) Map.png)
pET-30b(+)载体载体序列
LOCUS Exported 5421 bp ds-DNA circular SYN 02-12-2013
DEFINITION Bacterial vector for expression of N-terminally S-tagged proteins
with an enterokinase site. For other reading frames, use pET-30a(+)
or pET-30c(+).
ACCESSION .
VERSION .
KEYWORDS pET-30b(+)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5421)
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..5421
/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(140..157)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
misc_feature 158..216
/note="MCS"
/note="multiple cloning site"
CDS complement(218..232)
/codon_start=1
/product="enterokinase recognition and cleavage site"
/note="enterokinase site"
/translation="DDDDK"
CDS complement(248..292)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
CDS complement(299..316)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(326..343)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(344..346)
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
RBS 355..360
/note="ribosome binding site"
protein_bind 391..415
/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(416..434)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 747..824
/gene="lacI"
/note="lacI promoter"
/note="
"
CDS 825..1907
/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 2716..2907
/codon_start=1
/gene="rop"
/product="Rop protein"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(3337..3925)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 4047..4862
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin complement(4955..5410)
/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 cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tccgatatcg ccatggcctt gtcgtcgtcg tcggtaccca
241 gatctgggct gtccatgtgc tggcgttcga atttagcagc agcggtttct ttcataccag
301 aaccgcgtgg caccagacca gaagaatgat gatgatgatg gtgcatatgt atatctcctt
361 cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca attcccctat
421 agtgagtcgt attaatttcg cgggatcgag atcgatctcg atcctctacg ccggacgcat
481 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
541 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat
601 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct
661 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc
721 gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta
781 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
841 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
901 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
961 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
1021 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
1081 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
1141 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
1201 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
1261 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
1321 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
1381 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
1441 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
1501 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
1561 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
1621 cggacatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
1681 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
1741 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
1801 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
1861 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
1921 aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc
1981 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
2041 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc
2101 gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc
2161 ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag
2221 caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct cctgtcgttg
2281 aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc accgatacgc
2341 gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg
2401 gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt
2461 atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta
2521 ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg catccatacc
2581 gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc agtaacccgt
2641 atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag aaatccccct
2701 tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc ccgctttatc
2761 agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga tgaacaggca
2821 gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg cctcgcgcgt
2881 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
2941 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
3001 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact
3061 atgcggcatc agagcagatt gtactgagag tgcaccatat atgcggtgtg aaataccgca
3121 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
3181 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
3241 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
3301 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
3361 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
3421 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
3481 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
3541 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
3601 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
3661 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
3721 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
3781 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
3841 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
3901 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
3961 tcagtggaac gaaaactcac gttaagggat tttggtcatg aacaataaaa ctgtctgctt
4021 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcta
4081 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
4141 atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt
4201 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
4261 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
4321 atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag
4381 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
4441 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
4501 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
4561 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
4621 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
4681 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
4741 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
4801 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
4861 aagaattaat tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
4921 gttccgcgca catttccccg aaaagtgcca cctgaaattg taaacgttaa tattttgtta
4981 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
5041 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
5101 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
5161 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
5221 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
5281 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
5341 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
5401 cagggcgcgt cccattcgcc a
//
pET-30b(+)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 | 5421 | |
常规 | 描述 | Bacterial vector for expression of N-terminally S-tagged proteins with an enterokinase site. For other reading frames, use pET-30a(+) or pET-30c(+). | |
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..5421 |
来源 | 分子类型 | other DNA | 1..5421 |
来源 | 种属 | synthetic DNA construct | 1..5421 |
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 | 140..157 |
编码区 | 备注 | 6xHis | 140..157 |
编码区 | 产物 | 6xHis affinity tag | 140..157 |
编码区 | 翻译序列 | HHHHHH | 140..157 |
其它特征区 | 备注 | MCS,multiple cloning site | 158..216 |
编码区 | 翻译起始位点 | 1 | 218..232 |
编码区 | 备注 | enterokinase site | 218..232 |
编码区 | 产物 | enterokinase recognition and cleavage site | 218..232 |
编码区 | 翻译序列 | DDDDK | 218..232 |
编码区 | 翻译起始位点 | 1 | 248..292 |
编码区 | 备注 | S-Tag | 248..292 |
编码区 | 产物 | affinity and epitope tag derived from pancreatic ribonuclease A | 248..292 |
编码区 | 翻译序列 | KETAAAKFERQHMDS | 248..292 |
编码区 | 翻译起始位点 | 1 | 299..316 |
编码区 | 备注 | thrombin site | 299..316 |
编码区 | 产物 | thrombin recognition and cleavage site | 299..316 |
编码区 | 翻译序列 | LVPRGS | 299..316 |
编码区 | 翻译起始位点 | 1 | 326..343 |
编码区 | 备注 | 6xHis | 326..343 |
编码区 | 产物 | 6xHis affinity tag | 326..343 |
编码区 | 翻译序列 | HHHHHH | 326..343 |
编码区 | 翻译起始位点 | 1 | 344..346 |
编码区 | 备注 | ATG | 344..346 |
编码区 | 产物 | start codon | 344..346 |
编码区 | 翻译序列 | M | 344..346 |
核糖体结合位点 | 备注 | ribosome binding site | 355..360 |
蛋白结合位点 | 嵌合范围 | lac repressor encoded by lacI | 391..415 |
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). | 391..415 |
启动子 | 备注 | T7 promoter,promoter for bacteriophage T7 RNA polymerase | 416..434 |
启动子 | 基因 | lacI | 747..824 |
启动子 | 备注 | lacI promoter, | 747..824 |
编码区 | 翻译起始位点 | 1 | 825..1907 |
编码区 | 基因 | lacI | 825..1907 |
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). | 825..1907 |
编码区 | 产物 | lac repressor | 825..1907 |
编码区 | 翻译序列 | MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ | 825..1907 |
编码区 | 翻译起始位点 | 1 | 2716..2907 |
编码区 | 基因 | rop | 2716..2907 |
编码区 | 备注 | rop | 2716..2907 |
编码区 | 产物 | Rop protein | 2716..2907 |
编码区 | 翻译序列 | MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL | 2716..2907 |
复制起点 | 方向 | LEFT | 3337..3925 |
复制起点 | 备注 | ori,high-copy-number ColE1/pMB1/pBR322/pUC origin of replication | 3337..3925 |
编码区 | 翻译起始位点 | 1 | 4047..4862 |
编码区 | 基因 | aph(3')-Ia | 4047..4862 |
编码区 | 备注 | KanR,confers resistance to kanamycin | 4047..4862 |
编码区 | 产物 | aminoglycoside phosphotransferase | 4047..4862 |
编码区 | 翻译序列 | MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF | 4047..4862 |
复制起点 | 方向 | LEFT | 4955..5410 |
复制起点 | 备注 | f1 ori,f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis | 4955..5410 |
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