LOCUS       sequence-203966-        4080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid contains human codon optimized sequence of mitoLbNOX 
            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 4080)
  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 4080)
  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-mitoLbNOX
FEATURES             Location/Qualifiers
     source          1..4080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(16..38)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     138..157
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     351..373
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     365..382
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     366..382
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1843..1866
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1891..1907)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1891..1907)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1904..1926)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1915..1931
                     /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(1939..1945,1946..1963,1964..1969))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1984..2005
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2122..2139)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2293..2881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2373..2392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3052..3867)
                     /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(3052..3867)
                     /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     3775..3794
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3868..3972)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4040..4058
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
       61 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
      121 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
      181 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
      241 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
      301 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      361 acgttgtaaa acgacggcca gtgaagcggc cgcatgctcg ctacaagggt ctttagcctc
      421 gtcggaaaga gagctatcag cacctccgtc tgcgtgagag ctcataaggt caccgtggtc
      481 ggatgcaccc atgccggcac cttcgccatc aagcaaatcc tcgctgagca ccctgacgcc
      541 gaggtcaccg tctacgagag gaacgatgtg atctccttcc tgtcctgtgg catcgccctc
      601 tacctgggcg gaaaagtggc cgatccccaa ggcctcttct acagctcccc tgaagaactg
      661 cagaagctgg gcgctaatgt gcagatgaac cacaacgtgc tggccatcga ccctgaccaa
      721 aagaccgtca cagtcgagga cctcaccaat cacgcccaga ccaccgagtc ctacgacaaa
      781 ctggtgatga cctccggaag ctggcctatc gtgcccaaaa tccccggcat cgacagcgat
      841 agggtgaagc tctgcaagaa ttgggcccac gcccaggctc tgattgagga cgccaaggag
      901 gccaagagga tcaccgtcat cggcgccgga tacatcggag ccgaactggc cgaggcctac
      961 tccacaacag gccacgacgt caccctgatt gacgccatgg ctagggtcat gcccaagtac
     1021 ttcgatgccg acttcaccga cgtcatcgaa caggactaca gggaccatgg cgtgcaactc
     1081 gctctgggcg agacagtgga gagcttcacc gacagcgcca ccggcctcac aatcaagaca
     1141 gacaagaact cctatgagac cgacctggcc atcctctgca ttggctttag gcccaacaca
     1201 gacctgctga aaggcaaagt ggacatggcc cctaacggcg ccatcattac cgacgactac
     1261 atgaggtcca gcaaccctga tattttcgct gctggcgact ccgccgccgt ccattacaac
     1321 cccacacacc aaaacgccta cattcccctc gctaccaacg ccgtcaggca gggaatcctc
     1381 gtcggaaaga acctcgtcaa gcccacagtg aagtacatgg gaacccagtc cagctccgga
     1441 ctggccctct atgacaggac aattgtctcc acaggcctca cactggccgc cgccaagcaa
     1501 caaggcctca atgccgagca ggtcatcgtg gaggacaact ataggcccga gttcatgcct
     1561 tccaccgagc ccgtcctcat gagcctggtc ttcgaccccg atacacacag aatcctggga
     1621 ggcgccctga tgtccaaata cgacgtgtcc cagagcgcta ataccctgtc cgtctgcatc
     1681 cagaacgaga acaccatcga tgacctggcc atggtggaca tgctgttcca gcccaatttc
     1741 gacaggccct tcaactacct gaacattctc gcccaggctg cccaagctaa agtggcccaa
     1801 tccgtcaacg ctggtggatc tggtggatct ggtggatcta tggattacaa ggatgacgat
     1861 gacaagtaaa cgcgtcttgg ctcgagcatg gtcatagctg tttcctgtgt gaaattgtta
     1921 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1981 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2041 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2101 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2161 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2221 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2281 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2341 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2401 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2461 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2521 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2581 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2641 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2701 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2761 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2821 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2881 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2941 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3001 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttagaaaaa
     3061 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     3121 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     3181 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     3241 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     3301 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     3361 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     3421 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     3481 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     3541 tacctggaat gctgttttcc cagggatcgc agtggtgagt aaccatgcat catcaggagt
     3601 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     3661 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     3721 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     3781 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca
     3841 agacgtttcc cgttgaatat ggctcatact cttccttttt caatattatt gaagcattta
     3901 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3961 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4021 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
//
