质粒名称: | pET15b_Bst-LF |
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出品公司: | addgene |
目录编号: | 153313 |
存储实验室: | Drew Endy, Philippa Marrack |
载体骨架基本信息 |
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载体名称: | pET-15b |
完整质粒大小: | 7473bp |
载体类型: | 细菌细胞表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 30 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | Bst Large Fragment |
物种来源: | 其他 |
文献引用方法 |
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材料和方法部分: | pET15b_Bst-LF was a gift from Drew Endy & Philippa Marrack (Addgene plasmid # 153313 ; http://n2t.net/addgene:153313 ; RRID:Addgene_153313) |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pET15b_Bst-LF 7473 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression of the Bst Large Fragment. ACCESSION . VERSION . KEYWORDS pET15b_Bst-LF SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7473) TITLE COVID-19 Diagnostic Toolkit Enzymes REFERENCE 2 (bases 1 to 7473) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..7473 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 14..33 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 117..136 /label=T7 /note="T7 promoter, forward primer" promoter 117..135 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 136..160 /label=lac operator /bound_moiety="lac repressor encoded by 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)." RBS 175..197 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 216..233 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 243..260 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(2090..2108) /label=T7 Term /note="T7 terminator, reverse primer" terminator 2104..2151 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(2326..2354) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" primer_bind complement(2381..2399) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2467..2571 /gene="bla" /label=AmpR promoter CDS 2572..3432 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(2790..2809) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3603..4191 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4092..4111 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4345..4362 /label=L4440 /note="L4440 vector, forward primer" misc_feature 4377..4516 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4442..4461) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4602..4624 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(4618..4809) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(6121..7203) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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" primer_bind 7165..7184 /label=LacI-R /note="LacI, reverse primer" promoter complement(7204..7281) /gene="lacI" /label=lacI promoter ORIGIN 1 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg 61 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat 121 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt 181 taactttaag aaggagatat accatgggca gcagccatca tcatcatcat cacagcagcg 241 gcctggtgcc gcgcggcagc catagccata tgaccgacga aggcgaaaaa ccgctggcgg 301 gtatggactt tgcgatcgct gatagcgtta ccgacgagat gctggcagat aaagcagcgc 361 tagtagttga agttgtgggc gataactacc atcatgcgcc gatcgtgggt atcgctttgg 421 cgaacgagcg cggccgtttt ttcctgcgcc cggaaaccgc actggcggac ccgaagtttc 481 tggcttggct gggcgacgaa acgaaaaaaa agactatgtt cgattctaaa cgtgcagcgg 541 tggctctcaa atggaaaggt atcgaattgc gcggtgttgt gttcgacctg ctgctggctg 601 cgtacctgct cgatcctgct caggctgcgg gtgacgtggc agctgtagcg aaaatgcacc 661 agtacgaagc cgtacgctct gatgaagcgg tttatggtaa aggtgcgaaa cgtaccgtgc 721 cagacgaacc aaccctggcg gaacacctgg ctcgcaaagc agccgctatc tgggccctgg 781 aggagcctct gatggatgaa cttcgccgca acgaacagga ccgtctgctc acagaactgg 841 aacagccgtt ggcgggtatt ctggcaaaca tggagttcac tggtgtaaaa gtggacacta 901 agcgcctgga acaaatgggt gccgaactga ctgagcagct tcaggcagtt gaacgccgca 961 tctacgaact ggcgggtcag gagtttaaca tcaactctcc taaacagctg ggcaccgtgt 1021 tgttcgacaa actgcaactg ccggtgttga agaaaacgaa aaccggctac agcacctccg 1081 ccgatgttct ggaaaaactg gcaccgcatc acgaaatcgt tgaacacatc ctgcactacc 1141 gtcagctggg taaactgcaa tctacctaca tcgagggctt actgaaagta gtgcacccgg 1201 tgaccggtaa agtgcacact atgttcaacc aggctctgac gcagaccggc cgcctgtcct 1261 ccgttgaacc gaacctgcaa aacatcccca tccgtctgga agagggccgt aagatccgtc 1321 aggcattcgt gccgtctgaa ccggactggc tgattttcgc tgctgattac tctcaaatcg 1381 aactgcgtgt tcttgctcac attgcggagg acgataacct gattgaagca ttccgtcgtg 1441 gtctggacat tcacactaaa acggctatgg acattttcca tgtttccgaa gaagatgtga 1501 ccgctaatat gcgtcgtcag gctaaagcgg taaactttgg tatcgtatac ggtatctccg 1561 attacggcct ggcacagaac ctgaacatca cccgcaaaga agcggctgag tttatcgaac 1621 gttatttcgc ctcgttcccc ggtgttaaac agtacatgga taacatcgtt caagaagcta 1681 aacagaaggg ttatgtgacc accctgctgc accgtcgtcg ttacctgccg gacatcacct 1741 cccgtaactt caacgtgcgt tctttcgccg aacgtacggc tatgaatacc cctatccagg 1801 gcagcgcggc ggacatcatc aaaaaagcca tgatcgacct gagcgtgcgc ctgcgcgaag 1861 aacgtttgca ggctcgcctg cttctgcaag tgcacgatga gctgatccta gaagctccaa 1921 aagaggaaat tgaacgtttg tgccgtctgg ttccggaagt tatggaacag gctgtcgcac 1981 tgcgtgttcc gcttaaagtt gattaccatt acggtcccac ttggtacgac gccaagtgag 2041 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 2101 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 2161 ggaactatat ccggatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc 2221 ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac 2281 acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa agcttatcga 2341 tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat acgcctattt 2401 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga 2461 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 2521 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 2581 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 2641 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 2701 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 2761 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac 2821 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 2881 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 2941 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 3001 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 3061 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca 3121 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 3181 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 3241 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 3301 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 3361 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 3421 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 3481 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 3541 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 3601 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 3661 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 3721 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac 3781 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 3841 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 3901 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 3961 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 4021 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 4081 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 4141 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 4201 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 4261 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct 4321 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg 4381 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca 4441 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga 4501 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg 4561 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca 4621 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg 4681 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc 4741 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg 4801 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat 4861 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga 4921 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca 4981 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca 5041 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact 5101 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca 5161 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca 5221 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca 5281 ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga 5341 tatgttctgc caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc 5401 aattcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg 5461 gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct 5521 acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc 5581 agcggtccaa tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc 5641 tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg 5701 ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc 5761 aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa 5821 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc 5881 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc 5941 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg 6001 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag 6061 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc 6121 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 6181 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga 6241 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc 6301 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata 6361 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag 6421 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat 6481 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc 6541 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg 6601 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat 6661 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga 6721 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt 6781 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag 6841 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct 6901 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc 6961 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa 7021 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat 7081 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg 7141 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt 7201 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt 7261 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta 7321 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat 7381 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga 7441 aacaagcgct catgagcccg aagtggcgag ccc //