质粒名称: | pUC57-LbNOX |
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出品公司: | Addgene |
目录编号: | 75285 |
存储实验室: | Vamsi Mootha |
载体骨架基本信息 |
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载体名称: | pUC57 |
空载体大小: | 2710 bp |
完整质粒大小: | 4131 bp |
载体抗性: | 卡那霉素, 50 μg/mL |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | LbNOX,H2O-forming NADH oxidase from Lactobacillus brevis |
物种来源: | Lactobacillus brevis |
插入大小: | 1421 bp |
基因突变: | Human codon optimized |
标签/融合蛋白: | 带接头的C端子FLAG标签(插入件上的C端子) |
克隆方法: | 限制性内切酶 |
5’克隆位点: | NotI(未破坏) |
3’克隆位点: | MluI (未破坏) |
5’测序引物 | TGTAAAACGACGGCCAGT |
3’测序引物 | CAGGAAACGCTATGAC |
文献引用方法 |
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材料和方法部分: | pUC57-LbNOX was a gift from Vamsi Mootha (Addgene plasmid # 75285 ; http://n2t.net/addgene:75285 ; RRID:Addgene_75285) |
参考文献部分: | Complementation of mitochondrial electron transport chain by manipulation of the NAD+/NADH ratio. Titov DV, Cracan V, Goodman RP, Peng J, Grabarek Z, Mootha VK. Science. 2016 Apr 8;352(6282):231-5. doi: 10.1126/science.aad4017. Epub 2016 Apr 7. 10.1126/science.aad4017 PubMed 27124460 |
买家导航
pUC57LbNOX载体基本信息
pUC57LbNOX载体图谱质粒图谱和多克隆位点信息
pUC57LbNOX载体简介
pUC57LbNOX载体序列
LOCUS pUC57-LbNOX 4011 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION This plasmid contains human codon optimized sequence of LbNOX which is a water-forming NADH oxidase that can be used to increase NAD+/NADH ratio in mammalian cells.. ACCESSION . VERSION . KEYWORDS pUC57-LbNOX SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4011) AUTHORS Titov DV, Cracan V, Goodman RP, Peng J, Grabarek Z, Mootha VK TITLE Complementation of mitochondrial electron transport chain by manipulation of the NAD+/NADH ratio. JOURNAL Science. 2016 Apr 8;352(6282):231-5. doi: 10.1126/science.aad4017. Epub 2016 Apr 7. PUBMED 27124460 REFERENCE 2 (bases 1 to 4011) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4011 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(10..32) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 132..151 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 345..367 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 359..376 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 360..376 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 1768..1791 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(1816..1832) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1816..1832) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1829..1851) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1840..1856 /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(1864..1894) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1909..1930 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2047..2064) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2218..2806) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2298..2317) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2977..3792) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 3700..3719 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" promoter complement(3793..3897) /gene="bla" /label=AmpR promoter primer_bind 3965..3983 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 61 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 121 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 181 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 241 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 301 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 361 taaaacgacg gccagtgaag cggccgcatg aaggtcaccg tggtcggatg cacccatgcc 421 ggcaccttcg ccatcaagca aatcctcgct gagcaccctg acgccgaggt caccgtctac 481 gagaggaacg atgtgatctc cttcctgtcc tgtggcatcg ccctctacct gggcggaaaa 541 gtggccgatc cccaaggcct cttctacagc tcccctgaag aactgcagaa gctgggcgct 601 aatgtgcaga tgaaccacaa cgtgctggcc atcgaccctg accaaaagac cgtcacagtc 661 gaggacctca ccaatcacgc ccagaccacc gagtcctacg acaaactggt gatgacctcc 721 ggaagctggc ctatcgtgcc caaaatcccc ggcatcgaca gcgatagggt gaagctctgc 781 aagaattggg cccacgccca ggctctgatt gaggacgcca aggaggccaa gaggatcacc 841 gtcatcggcg ccggatacat cggagccgaa ctggccgagg cctactccac aacaggccac 901 gacgtcaccc tgattgacgc catggctagg gtcatgccca agtacttcga tgccgacttc 961 accgacgtca tcgaacagga ctacagggac catggcgtgc aactcgctct gggcgagaca 1021 gtggagagct tcaccgacag cgccaccggc ctcacaatca agacagacaa gaactcctat 1081 gagaccgacc tggccatcct ctgcattggc tttaggccca acacagacct gctgaaaggc 1141 aaagtggaca tggcccctaa cggcgccatc attaccgacg actacatgag gtccagcaac 1201 cctgatattt tcgctgctgg cgactccgcc gccgtccatt acaaccccac acaccaaaac 1261 gcctacattc ccctcgctac caacgccgtc aggcagggaa tcctcgtcgg aaagaacctc 1321 gtcaagccca cagtgaagta catgggaacc cagtccagct ccggactggc cctctatgac 1381 aggacaattg tctccacagg cctcacactg gccgccgcca agcaacaagg cctcaatgcc 1441 gagcaggtca tcgtggagga caactatagg cccgagttca tgccttccac cgagcccgtc 1501 ctcatgagcc tggtcttcga ccccgataca cacagaatcc tgggaggcgc cctgatgtcc 1561 aaatacgacg tgtcccagag cgctaatacc ctgtccgtct gcatccagaa cgagaacacc 1621 atcgatgacc tggccatggt ggacatgctg ttccagccca atttcgacag gcccttcaac 1681 tacctgaaca ttctcgccca ggctgcccaa gctaaagtgg cccaatccgt caacgctggt 1741 ggatctggtg gatctggtgg atctatggat tacaaggatg acgatgacaa gtaaacgcgt 1801 cttggctcga gcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc 1861 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta 1921 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 1981 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 2041 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 2101 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 2161 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 2221 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 2281 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 2341 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 2401 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 2461 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 2521 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 2581 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 2641 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt 2701 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 2761 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 2821 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 2881 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 2941 aatctaaagt atatatgagt aaacttggtc tgacagttag aaaaactcat cgagcatcaa 3001 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt 3061 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg 3121 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat 3181 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag 3241 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc 3301 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg 3361 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc 3421 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt 3481 tttcccaggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt 3541 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac 3601 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc 3661 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc 3721 atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg 3781 aatatggctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 3841 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 3901 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 3961 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg a //
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