LOCUS       sequence-175126-        4047 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 4047)
  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 4047)
  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-TPNOX
FEATURES             Location/Qualifiers
     source          1..4047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1381..1404
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1448..1464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1448..1464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1461..1483)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1472..1488
                     /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(1496..1502,1503..1520,1521..1526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1541..1562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1679..1696)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1850..2438)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1930..1949)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2609..3424)
                     /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(2609..3424)
                     /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     3332..3351
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3425..3529)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3597..3615
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3653..3675)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3775..3794
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(join(3874..4047,1..1452))
                     /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
                     GVRVLSEDLLDGEGAGMGASDHGDLHAAASQYASIHWPSFYNVVTGKTLALPNLIALQH
                     IPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     3988..4010
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4002..4019
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4003..4019
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atgaaggtca ccgtggtcgg atgcacccat gccggcacct tcgccatcaa gcaaatcctc
       61 gctgagcacc ctgacgccga ggtcaccgtc tacgagagga acgatgtgat ctccttcctg
      121 tcctgtggca tcgccctcta cctgggcgga aaagtggccg atccccaagg cctcttctac
      181 agctcccctg aagaactgca gaagctgggc gctaatgtgc agatgaacca caacgtgctg
      241 gccatcgacc ctgaccaaaa gaccgtcaca gtcgaggacc tcaccaatca cgcccagacc
      301 accgagtcct acgacaaact ggtgatgacc tccggaagct ggcctatcgt gcccaaaatc
      361 cccggcatcg acagcgatag ggtgaagctc tgcaagaatt gggcccacgc ccaggctctg
      421 attgaggacg ccaaggaggc caagaggatc accgtcatcg gcgccggata catcgccgcc
      481 gaactggccg aggcctactc cacaacaggc cacgacgtca ccctgattgc cagaagcgct
      541 agggtcatga ggaagtactt cgatgccgac ttcaccgacg tcatcgaaca ggactacagg
      601 gaccatggcg tgcaactcgc tctgggcgag acagtggaga gcttcaccga cagcgccacc
      661 ggcctcacaa tcaagacaga caagaactcc tatgagaccg acctggccat cctctgcatt
      721 ggctttaggc ccaacacaga cctgctgaaa ggcaaagtgg acatggcccc taacggcgcc
      781 atcattaccg acgactacat gaggtccagc aaccctgata ttttcgctgc tggcgactcc
      841 gccgccgtcc attacaaccc cacacaccaa aacgcctaca ttcccctcgc taccaacgcc
      901 gtcaggcagg gaatcctcgt cggaaagaac ctcgtcaagc ccacagtgaa gtacatggga
      961 acccagtcca gctccggact ggccctctat gacaggacaa ttgtctccac aggcctcaca
     1021 ctggccgccg ccaagcaaca aggcctcaat gccgagcagg tcatcgtgga ggacaactat
     1081 aggcccgagt tcatgccttc caccgagccc gtcctcatga gcctggtctt cgaccccgat
     1141 acacacagaa tcctgggagg cgccctgatg tccaaatacg acgtgtccca gagcgctaat
     1201 accctgtccg tctgcatcca gaacgagaac accatcgatg acctggccat ggtggacatg
     1261 ctgttccagc ccaatttcga caggcccttc aactacctga acattctcgc ccaggctgcc
     1321 caagctaaag tggcccaatc cgtcaacgct ggtggatctg gtggatctgg tggatctatg
     1381 gattacaagg atgacgatga caagtaaacg cgtatcccaa tggcgcgccg agcttggctc
     1441 gagcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1501 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1561 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1621 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1681 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1741 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1801 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1861 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1921 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1981 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2041 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2101 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2161 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2221 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2281 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2341 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2401 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2461 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2521 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2581 gtatatatga gtaaacttgg tctgacagtt agaaaaactc atcgagcatc aaatgaaact
     2641 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     2701 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     2761 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     2821 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca
     2881 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     2941 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     3001 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     3061 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccag
     3121 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     3181 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     3241 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     3301 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     3361 cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc
     3421 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3481 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3541 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     3601 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     3661 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     3721 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     3781 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     3841 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     3901 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     3961 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     4021 aattgacgcg tattgggatg cggccgc
//
