LOCUS       sequence-423281-        4458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Streptomyces argenteolus LRD-modifying cytochrome P450
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4458)
  AUTHORS   Lin GM, Voigt CA
  TITLE     Design of a redox-proficient Escherichia coli for screening 
            terpenoids and modifying cytochrome P450s
  JOURNAL   Nature Catalysis. 6, 1016–1029 (2023). 
            https://doi.org/10.1038/s41929-023-01049-5
REFERENCE   2  (bases 1 to 4458)
  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: "Nature 
            Catalysis. 6, 1016–1029 (2023). 
            https://doi.org/10.1038/s41929-023-01049-5"
COMMENT     Sequence Label: pCYP53
FEATURES             Location/Qualifiers
     source          1..4458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(296..841)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(376..395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(930..1745)
                     /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(930..1745)
                     /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     1653..1672
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1737..1756
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1922..1949
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     protein_bind    1980..2007
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     primer_bind     complement(4370..4389)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 actgactcgc tgcacgaggc agacctcagc gctagcggag tgtatactgg cttactatgt
       61 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
      121 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
      181 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
      241 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
      301 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
      361 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
      421 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
      481 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
      541 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      601 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
      661 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
      721 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
      781 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
      841 agaagatcat cttattaagg ggtctgacgc tcagtggaac gaaaaatcaa tctaaagtat
      901 atatgagtaa acttggtctg acagttacct tagaaaaact catcgagcat caaatgaaac
      961 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     1021 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     1081 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     1141 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
     1201 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     1261 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     1321 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     1381 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     1441 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     1501 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     1561 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     1621 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     1681 tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg ttgaatatgg
     1741 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     1801 atatttttat cttgtgcaat gtacatcaga gattttgaga cacaaccaat tattgaaggc
     1861 ctccctaacg gggggccttt ttttgtttct ggtctcccaa gcgcttaacg atcgttggct
     1921 gaacaaacag acaatctggt ctgtttgtat tatggaaaat ttttctgtat aatagattca
     1981 acaaacagac aatctggtct gtttgtatta tcagcggtca acgcatgtgc tttgcgttct
     2041 gatgagacag tgatgtcgaa accgcctcta caaataattt tgtttagagt acataaggag
     2101 ggatctaaat gaatcgtcgt actatgcgca ccgacattag cgatccctac gatctgagcc
     2161 acctctggcc agaatatacc gaactgcgca aaaatgcccc agtgcatcca attacctcta
     2221 ttaacggaca acgcttgtgg cttgtgtcag gtcacgacgc agtgcgcctt gtggaaacgg
     2281 acccgcgctt ctctatgaac cttgcgaacg ccgttccaga gctggccgat tcgttccacg
     2341 ggagtatccc acgttgggcc cgcccgctcg cagaaacctt gattacaagc gatccgccac
     2401 agcatggtcg tctgcggcgc ctcgtgcagc ctgccttttc accgagaatt ttaaatgcca
     2461 cacgaggctc gattgaacaa atcgtagatg gattactgga ccgtgcacgt gctgccggct
     2521 cgctggaatt tgtggacgaa tttgcttttc cgatgacaat cgcggtggtc agtcgcttct
     2581 taggctttcc gtacgaggac cgtgatcgcg tgcgccaatg gacccaggtg ttagtgcaga
     2641 acttcgctgg tcaggacgtg aaagcgattg aacaggcagc ttgtgagttt attgattacc
     2701 tgatcgattt atttcctcgg aaacgtgcgc atccggggga tgacgtaata agcgccctga
     2761 ttcacgcgcg tgaccaggag gaagtgttaa ctgaggatga gctgctggca atggtgttta
     2821 atattgtggg cgggggcttt gaaacaacca tcgccctgct gacgcatggt actaaggcgc
     2881 tgctggatca cccggagcag ctggaactgt tgcgacggcg tccggacctg acaggctctg
     2941 cagtggaaga actgttacgt tttagcgcag gcggggttgg cccgattcgc tacccacgtg
     3001 aagatgtggt cattgctggt gtgaccattc cgcgcggggc gcctgtaatg cccctgttct
     3061 cctctgcaaa ctttgacccg cgccgctttg atgaacctca tcgcctggat atcacccgca
     3121 gatctaatca acatttttct tttggccacg gcgcgcacta ctgtcttggc gcaccgcttg
     3181 cgcgtatcga agcggaagtg gcgttcccga gtctggtgcg ggcactgcct ggcttgcgcc
     3241 cttcatcaac cattacgcca agcctggtgt ggcaagaaaa caccccgttc gtacggcgca
     3301 ttacgcacat gcatcttgcg tttgatacgg acacggccct ttcaccagaa gtcactagct
     3361 cttaataagg atgagctcta caaataagca gaggtggttg tgttgcgaaa aaaaaaaaaa
     3421 aacaccctaa cgggtgtttt ttttttttgg tgtccccacg tgtggcgctg gagaccgtcc
     3481 aatggcggcg cgccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg
     3541 aagagagtca attcagggtg gtgaatatga gcccgaaacg tcgtacccag gcagaacgtg
     3601 caatggaaac ccagggtaaa ctgattgcag cagcactggg tgttctgcgt gaaaaaggtt
     3661 atgcaggttt tcgtattgca gatgttccgg gtgcagccgg tgttagccgt ggtgcacaga
     3721 gccatcattt tccgaccaaa ctggaactgc tgctggcaac ctttgaatgg ctgtatgagc
     3781 agattaccga acgtagccgt gcacgtctgg caaaactgaa accggaagat gatgttattc
     3841 agcagatgct ggatgatgca gcagattttt ttctggatga tgattttagc atcggcctgg
     3901 atctgattgt tgcagcagat cgtgatccgg cactgcgtga aggtattctg cgtaccgttg
     3961 aacgtaatcg ttttgttgtt gaagatatgt ggctgggtgt gctggtgagc cgtggtctga
     4021 gccgtgatga tgccgaagat attctgtggc tgatttttaa cagcgttcgt ggtctgacag
     4081 ttcgtagcct gtggcagaaa gataaagaac gttttgaacg tgtgcgtaat agcaccctgg
     4141 aaattgcacg tgaacgttat gcaaaattca aacgttgata aggatcctaa ttggtaacga
     4201 atcagacaat tgacggctcg agggagtagc atagggtttg cagaatccct gcttcgtcca
     4261 tttgacaggc acattatgca tcgatgataa gctgtcaaac atgagcagat cctctacgcc
     4321 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     4381 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcaa atattttatc
     4441 tgaggtgctt cctcgctc
//
