LOCUS       sequence-175127-        4116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid contains human codon optimized sequence of 
            Lactobacillus brevis mutant of a water-forming oxidase that can be 
            used to increase NADP+/NADPH ratio in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4116)
  AUTHORS   Cracan V, Titov DV, Shen H, Grabarek Z, Mootha VK
  TITLE     A genetically encoded tool for manipulation of NADP+/NADPH in living
            cells.
  JOURNAL   Nat Chem Biol. 2017 Aug 7. doi: 10.1038/nchembio.2454.
  PUBMED    28805804
REFERENCE   2  (bases 1 to 4116)
  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"; journalName: "Nat Chem 
            Biol. 2017 Aug 7. doi: 10.1038/nchembio.2454."
COMMENT     Sequence Label: pUC57-mitoTPNOX
FEATURES             Location/Qualifiers
     source          1..4116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1450..1473
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1517..1533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1517..1533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1530..1552)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1541..1557
                     /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(1565..1571,1572..1589,1590..1595))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1610..1631
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1748..1765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1919..2507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1999..2018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2678..3493)
                     /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(2678..3493)
                     /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     3401..3420
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3494..3598)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3666..3684
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3722..3744)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3844..3863
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(join(3943..4116,1..1521))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMLEPSSARHWDTRLLVIVILVIHRSTRSTRSTSVDGLGHFSLG
                     SLGENVQVVEGPVEIGLEQHVHHGQVIDGVLVLDADGQGISALGHVVFGHQGASQDSVC
                     IGVEDQAHEDGLGGRHELGPIVVLHDDLLGIEALLLGGGQCEACGDNCPVIEGQSGAGL
                     GSHVLHCGLDEVLSDEDSLPDGVGSEGNVGVLVCGVVMDGGGVASSENIRVAGPHVVVG
                     NDGAVRGHVHFAFQQVCVGPKANAEDGQVGLIGVLVCLDCEAGGAVGEALHCLAQSELH
                     AMVPVVLFDDVGEVGIEVLPHDPSASGNQGDVVACCGVGLGQFGGDVSGADDGDPLGLL
                     GVLNQSLGVGPILAELHPIAVDAGDFGHDRPASGGHHQFVVGLGGLGVIGEVLDCDGLL
                     VRVDGQHVVVHLHISAQLLQFFRGAVEEALGIGHFSAQVEGDATGQEGDHIVPLVDGDL
                     GVRVLSEDLLDGEGAGMGASDHGDLMSSHADGGADSSLSDEAKDPCSEHAAASQYASIH
                     WPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     4057..4079
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4071..4088
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4072..4088
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atgctcgcta caagggtctt tagcctcgtc ggaaagagag ctatcagcac ctccgtctgc
       61 gtgagagctc ataaggtcac cgtggtcgga tgcacccatg ccggcacctt cgccatcaag
      121 caaatcctcg ctgagcaccc tgacgccgag gtcaccgtct acgagaggaa cgatgtgatc
      181 tccttcctgt cctgtggcat cgccctctac ctgggcggaa aagtggccga tccccaaggc
      241 ctcttctaca gctcccctga agaactgcag aagctgggcg ctaatgtgca gatgaaccac
      301 aacgtgctgg ccatcgaccc tgaccaaaag accgtcacag tcgaggacct caccaatcac
      361 gcccagacca ccgagtccta cgacaaactg gtgatgacct ccggaagctg gcctatcgtg
      421 cccaaaatcc ccggcatcga cagcgatagg gtgaagctct gcaagaattg ggcccacgcc
      481 caggctctga ttgaggacgc caaggaggcc aagaggatca ccgtcatcgg cgccggatac
      541 atcgccgccg aactggccga ggcctactcc acaacaggcc acgacgtcac cctgattgcc
      601 agaagcgcta gggtcatgag gaagtacttc gatgccgact tcaccgacgt catcgaacag
      661 gactacaggg accatggcgt gcaactcgct ctgggcgaga cagtggagag cttcaccgac
      721 agcgccaccg gcctcacaat caagacagac aagaactcct atgagaccga cctggccatc
      781 ctctgcattg gctttaggcc caacacagac ctgctgaaag gcaaagtgga catggcccct
      841 aacggcgcca tcattaccga cgactacatg aggtccagca accctgatat tttcgctgct
      901 ggcgactccg ccgccgtcca ttacaacccc acacaccaaa acgcctacat tcccctcgct
      961 accaacgccg tcaggcaggg aatcctcgtc ggaaagaacc tcgtcaagcc cacagtgaag
     1021 tacatgggaa cccagtccag ctccggactg gccctctatg acaggacaat tgtctccaca
     1081 ggcctcacac tggccgccgc caagcaacaa ggcctcaatg ccgagcaggt catcgtggag
     1141 gacaactata ggcccgagtt catgccttcc accgagcccg tcctcatgag cctggtcttc
     1201 gaccccgata cacacagaat cctgggaggc gccctgatgt ccaaatacga cgtgtcccag
     1261 agcgctaata ccctgtccgt ctgcatccag aacgagaaca ccatcgatga cctggccatg
     1321 gtggacatgc tgttccagcc caatttcgac aggcccttca actacctgaa cattctcgcc
     1381 caggctgccc aagctaaagt ggcccaatcc gtcaacgctg gtggatctgg tggatctggt
     1441 ggatctatgg attacaagga tgacgatgac aagtaaacgc gtatcccaat ggcgcgccga
     1501 gcttggctcg agcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1561 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1621 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1681 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1741 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1801 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1861 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1921 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1981 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2041 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2101 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2161 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2221 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2281 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2341 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2401 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2461 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2521 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2581 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2641 caatctaaag tatatatgag taaacttggt ctgacagtta gaaaaactca tcgagcatca
     2701 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     2761 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     2821 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     2881 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     2941 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     3001 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     3061 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     3121 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     3181 ttttcccagg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     3241 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     3301 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     3361 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     3421 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     3481 gaatatggct catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3541 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3601 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     3661 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     3721 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     3781 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     3841 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     3901 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     3961 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     4021 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     4081 cggccagtga attgacgcgt attgggatgc ggccgc
//
