质粒名称: | pPM_Pveg-sfgfp |
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出品公司: | Addgene |
目录编号: | 121503 |
存储实验室: | Robert Shields |
载体骨架基本信息 |
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载体名称: | pMZ |
完整质粒大小: | 7666bp |
载体类型: | 细菌细胞表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 未知 |
插入基因信息 |
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插入基因名称: | Superfolder GFP/sfgfp |
物种来源: | Synthetic |
插入大小: | 717bp |
GenBank编号 | MK301203 |
启动子: | Pveg |
标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII(N端) |
克隆方法: | 限制性内切酶 |
5’克隆位点: | SacI(未破坏) |
3’克隆位点: | SphI(未破坏) |
5’测序引物 | mtlA: CAGTCTTAGTCAGGCTTTG |
3’测序引物 | phnA: GATTCCATTCATAAGCACAT |
文献引用方法 |
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材料和方法部分: |
pPM_Pveg-sfgfp was a gift from Robert Shields (Addgene plasmid # 121503 ; http://n2t.net/addgene:121503 ; RRID:Addgene_121503)
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参考文献部分: | Fluorescence tools adapted for real-time monitoring of the behaviors of Streptococcus species. Shields RC, Kaspar JR, Lee K, Underhill SAM, Burne RA. Appl Environ Microbiol. 2019 May 17. pii: AEM.00620-19. doi: 10.1128/AEM.00620-19. 10.1128/AEM.00620-19 PubMed 31101614 |
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LOCUS pPM_Pveg-sfgfp 7666 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Integration vector that expresses gfp in S. mutans. ACCESSION . VERSION . KEYWORDS pPM_Pveg-sfgfp SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7666) AUTHORS Shields RC, Kaspar JR, Lee K, Underhill SAM, Burne RA TITLE Fluorescence tools adapted for real-time monitoring of the behaviors of Streptococcus species. JOURNAL Appl Environ Microbiol. 2019 May 17. pii: AEM.00620-19. doi: 10.1128/AEM.00620-19. PUBMED 31101614 REFERENCE 2 (bases 1 to 7666) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..7666 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..17 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 596..1312 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /note="mammalian codon-optimized" /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" CDS complement(1766..2560) /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" primer_bind complement(4107..4123) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4107..4123) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4120..4142) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4131..4147 /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)." promoter complement(4155..4185) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4200..4221 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4458..4475) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4592..5137) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(4672..4691) /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7077..7096 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(7163..7265) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" primer_bind 7652..8 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7666..17 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" ORIGIN 1 gtaaaacgac ggccagtgaa ttctagaagg cgaatatttt taacacgtgt tccttgtttt 61 aaatctttag gagccccctt aactttaaga tccttaataa gagtgactgt atcaccatca 121 gctaattgtt tgccattcgc atcaataaca acaagtcctt cttctttctc aacttctgca 181 ggattccatt cataagcaca ttctggacaa accaatagaa taccatcttc atagacatac 241 tctgactgac atttaggaca attaggtaaa gacatataaa ctccttttat tttgattacc 301 attaacagtt tacctgagtt tttatcagtt ggcaagtctt atcacaacaa aaagctagct 361 gcttgtactg actctatgag ttaagtatac tgtctaactt atagaaaacg cagaacaaga 421 gctagttctt ttaattataa gagtttccct taatctaaga ttcaacactg gatctccaac 481 gcgttgggag ctctagatcg aattccttat taacgttgat ataatttaaa ttttatttga 541 caaaaatggg ctcgtgttgt acaataaatg tgattaacta ataaggagga caaacatgtc 601 aaaaggagaa gagctgttca caggtgttgt gccgattctc gttgagcttg acggagatgt 661 aaacggacac aaattctctg ttcgcggtga aggtgaagga gatgcaacaa acggcaagct 721 gacattgaag tttatttgca caactggaaa gctgccggtt ccttggccga cacttgtaac 781 gacgctgact tacggcgttc aatgcttctc tcgttatcca gaccacatga aacgccatga 841 tttcttcaaa tctgcaatgc ctgaaggcta cgttcaagag cgtacgatca gcttcaaaga 901 tgacggaacg tacaaaacaa gagcagaagt gaagtttgaa ggtgacacac ttgtgaaccg 961 cattgaattg aaaggcattg atttcaaaga agatggaaac atccttggac acaaacttga 1021 atacaacttc aacagccaca acgtatacat cactgctgac aaacaaaaaa acggcatcaa 1081 agcaaacttc aaaatccgtc ataacgtaga ggacggttct gttcagcttg ctgatcatta 1141 tcagcaaaat acaccgatcg gtgacggccc ggttcttctt cctgataacc attatttatc 1201 aactcaaagc gtattatcaa aagacccaaa tgaaaagcgt gaccacatgg tgctgcttga 1261 atttgtgaca gctgctggta tcactcacgg catggatgag ctttataagt aagcatgcgc 1321 cggtgattga ttgagcaagc tttatgcttg taaaccgttt tgtgaaaaaa tttttaaaat 1381 aaaaaagggg acctctaggg tccccaatta attagtaata taatctatta aaggtcattc 1441 aaaaggtcat ccaccggatc aattcccctg ctcgcgcagg ctgggtgcca agctctcggg 1501 taacatcaag gcccgatcct tggagccctt gccctcccgc acgatgatcg tgccgtgatc 1561 gaaatccaga tccttgaccc gcagttgcaa accctcactg atccggacga tacaaattcc 1621 tcgtaggcgc tcgggacccc tatctagcga acttttagaa aagatataaa acatcagagt 1681 atggacagtt gcggatgtac ttcagaaaag attagatgtc taaaaagctt gtagttaaag 1741 ctttttagac atctaaatct aggtactaaa acaattcatc cagtaaaata taatatttta 1801 ttttctccca atcaggcttg atccccagta agtcaaaaaa tagctcgaca tactgttctt 1861 ccccgatatc ctccctgatc gaccggacgc agaaggcaat gtcataccac ttgtccgccc 1921 tgccgcttct cccaagatca ataaagccac ttactttgcc atctttcaca aagatgttgc 1981 tgtctcccag gtcgccgtgg gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa 2041 aatcatacag ctcgcgcgga tctttaaatg gagtgtcttc ttcccagttt tcgcaatcca 2101 catcggccag atcgttattc agtaagtaat ccaattcggc taagcggctg tctaagctat 2161 tcgtataggg acaatccgat atgtcgatgg agtgaaagag cctgatgcac tccgcataca 2221 gctcgataat cttttcaggg ctttgttcat cttcatactc ttccgagcaa aggacgccat 2281 cggcctcact catgagcaga ttgctccagc catcatgccg ttcaaagtgc aggacctttg 2341 gaacaggcag ctttccttcc agccatagca tcatgtcctt ttcccgttcc acatcatagg 2401 tggtcccttt ataccggctg tccgtcattt ttaaatatag gttttcattt tctcccacca 2461 gcttatatac cttagcagga gacattcctt ccgtatcttt tacgcagcgg tatttttcga 2521 tcagtttttt caattccggt gatattctca ttttagccat ttattatttc cttcctcttt 2581 tctacagtat ttaaagatac cccaagaagc taattataac aagacgaact ccaattcact 2641 gttccttgca ttctaaaacc ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa 2701 gttggcgtat aacatagtat cgacggagcc gattttgaaa ccacaattat gatagaattt 2761 acaagctata aggttattgt cctgggtttc aagcattagt ccatgcaagt ttttatgctt 2821 tgcccattct atagatatat tgataagcgc gctgcctatg ccttgccccc tgaaatcctt 2881 acatacggcg atatcttcta tataaaagat atattatctt atcagtattg tcaatatatt 2941 caaggcaatc tgcctcctca tcctcttcat cctcttcgtc ttggtagctt tttaaatatg 3001 gcgcttcata gagtaattct gtaaaggtcc aattctcgtt ttcatacctc ggtataatct 3061 tacctatcac ctcaaatggt tcgctgggtt tatcgtccgc gcagatcagt tggaagaatt 3121 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataatgtc taacaattcg 3181 ttcaagccga cgccgcttcg cggcgcggct taactcaagc gttagatgca ctaagcacat 3241 aattgctcac agccaaacta tcaggtcaag tctgctttta ttatttttaa gcgtgcataa 3301 taagccctac acaaattggg agatatatca tgaaaggctg gctttttctt gttatcgcaa 3361 tagttggcga agtaatcgca acatccgcat taaaatctag cgagggcttt actaagctga 3421 tccggtggat gaccttttga atgaccttta atagattata ttactaatta attggggacc 3481 ctagaggtcc ccttttttat tttaaaaatt ttttcacaaa acggtttaca agcataaagc 3541 ttgctcaatc aatcaccgga cgagcttgct caatcaatca ccggatcgat cctgttgcaa 3601 taaacagaga tgctgcaacc ccccatgcaa taaaggcacc aatattaggc ataaccatgt 3661 ttgataatga agttccgagt ttttgtaccc gaaccttcaa tgaagatttt cgttccatca 3721 gaaatcccct cctttttatt ctaaaactac tgtaacattt ttcattttct aataaaactc 3781 aaattttggc acaaaagttt tgccaaaatt tgttatccta agcggaagaa taggatttga 3841 caaagcctga ctaagactga cttttttatc actaatattt ttctcatgtt aaaaacttgg 3901 atatataact tttttaacag tttccttagg taatagagac gaaaaaaagc agcagcattc 3961 ctcaagtttc cttttaaact tcaaactgaa tgtctcaact ctttcatata tttcctctga 4021 tggtcaacat tcgttttatc acagcgaaag tggaatttgt tcttttatgt gactttacgg 4081 ttgcttttag gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 4141 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 4201 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 4261 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 4321 gggcgctagc ggagtgtata tactggctta ctatgttggc actgatgagg gtgtcagtga 4381 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac 4441 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga 4501 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac 4561 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc 4621 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc 4681 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg 4741 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg 4801 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc 4861 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg 4921 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg 4981 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa 5041 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc 5101 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat 5161 aaaatatttc taggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat 5221 cgcagtgatc cgcgatacaa attccccgta ggcgctaggg acctctttag ctccttggaa 5281 gctgtcagta gtatacctaa taatttatct acattccctt tagtaacgtg taactttcca 5341 aatttacaaa agcgactcat agaattattt cctcccgtta aataatagat aactattaaa 5401 aatagacaat acttgctcat aagtaacggt acttaaattg tttactttgg cgtgtttcat 5461 tgcttgatga aactgatttt tagtaaacag ttgacgatat tctcgattga cccattttga 5521 aacaaagtac gtatatagct tccaatattt atctggaaca tctgtggtat ggcgggtaag 5581 ttttattaag acactgttta cttttggttt aggatgaaag cattccgctg gcagcttaag 5641 caattgctga atcgagactt gagtgtgcaa gagcaaccct agtgttcggt gaatatccaa 5701 ggtacgcttg tagaatcctt cttcaacaat cagatagatg tcagacgcat ggctttcaaa 5761 aaccactttt ttaataattt gtgtgcttaa atggtaagga atactcccaa caattttata 5821 cctctgtttg ttagggaatt gaaactgtag aatatcttgg tgaattaaag tgacacgagt 5881 attcagtttt aatttttctg acgataagtt gaatagatga ctgtctaatt caatagacgt 5941 tacctgttta cttattttag ccagtttcgt cgttaaatgc cctttacctg ttccaatttc 6001 gtaaacggta tcggtttctt ttaaattcaa ttgttttatt atttggttga gtactttttc 6061 actcgttaaa aagttttgag aatattttat atttttgttc atgtaatcac tccttcttaa 6121 ttacaaattt ttagcatcta atttaacttc aattcctatt atacaaaatt ttaagatact 6181 gcactatcaa cacactctta agtttgcttc taagtcttat ttccataact tcttttacgt 6241 ttccgccatt ctttgctgtt tcgattttta tgatatggtg caagtcagca cgaacacgaa 6301 ccgtcttatc tcccattata tctttttttg cactgattgg tgtatcattt cgtttttctt 6361 tttgtgcgcc taaatttccc acaatcactc acttctttct atttcttctt attcttattt 6421 tatcatcaac aatcacaaat cacttgtgat ttgtgataag tgatttgtga ttaatatata 6481 aaagccctct ttaaagggct tttatgttta ttttgagaaa gatataaaat caatatatcc 6541 cttttctccg atttttacaa cggcattgta ggactttcta tctttcgttt tgattccttt 6601 taccagggtt tcttttccct ctagtaattc ttttacattt gttttggtga gttttttctt 6661 tctaaaatgt tcagctaaag taaacttaca ttcaggataa tttgaacaac cataaaacga 6721 tttttttaat acaatattgt tgccacactt aggacatttt cctacaatac ttttttctgc 6781 ttctttttct ttctgttcct ggtaatcaga aaaatttagt ttttctatat cgttaggtac 6841 agcttccagt aaatgaacaa tgaatttttt gatattcgta ataaagttct cttgattgcc 6901 ttctctttta ccgatttttt ttaaatacgt ttcccattta gccgtcattt cagcactcgt 6961 taaaaggtgc tgactttcaa ctgcctggca caataatttt cctttttcag ttacaacaag 7021 ctcggatcaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa 7081 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat 7141 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa 7201 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc 7261 gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac 7321 caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg 7381 cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca 7441 gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac 7501 ggcaaaagca ccgccggaca tcagcgccat tcgccattca ggctgcgcaa ctgttgggaa 7561 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca 7621 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt //
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