LOCUS       sequence-258257-        4011 bp    DNA     circular SYN 15-DEC-2025
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    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.aad4017"; journalName: "Science"; date: "2016-04-8-
            8"; volume: "352"; issue: "6282"; pages: "231-5"
COMMENT     Sequence Label: pUC57-LbNOX
FEATURES             Location/Qualifiers
     source          1..4011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(join(1864..1870,1871..1888,1889..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
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     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
//
