LOCUS       sequence-424230-        4648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Kitasatospora griseola LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4648)
  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 4648)
  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: pCYP60
FEATURES             Location/Qualifiers
     source          1..4648
                     /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(4560..4579)
                     /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 tgtttaatat taagcgagcc
     2101 tatttatgaa agggaatctg ccagcgatga gcaccgccac gtcatcatca gtgaccggta
     2161 gtgtcaccac gccccgaact gcgaccacac cgggtcgccg tcctggtttg gcgccaggtg
     2221 gcgttccagt gttaggccac ttgccgatga ttctccgctc acgcttcgaa ttcattgaaa
     2281 cggtccgtaa cgcgggtccg gtgacgcggg tcaaattggg tccaaaaact gcttatttcg
     2341 tgaacgacta tgaacttctg gcacagatcc tggtttccga cgcagataaa tttgtccggg
     2401 gtattcactt taagaaaatg cgcaatatgg tgggtaatgg tgtggttacc acatcaggtg
     2461 atctgcaccg gcgacagcgc cggataatgc tgccgagctt cagccaacgt cgtttagcaa
     2521 tgcatttgcc cgtcatgcgc aaaatcatgt ctgagtttgt ggcaagtgtg ccagagcgcc
     2581 ggccttatga cctcatggga cccgtgatgg gcgtgggctg cgatatcgtg actagcacca
     2641 tgctgggcga gaaaacaccg ccggaagttc tgcgtcttgt tcgggaggct gtgcctgttt
     2701 ttgtcgaaaa tgcggccatc caagccgtcg atgttaccgg gatttataag catctgccta
     2761 ctaagtcgaa ccgggatttt gagcgcctgc ttaacgcatt taatgaatac atgtactcgg
     2821 tgattgatga caagtttaga aatggtgctg gtgaggaagc gggcctgctg gacatgttaa
     2881 tcaacgcgac tgatcctgaa accggagaaa aattcgaccg taccgaagtg cgcgatcaag
     2941 ctgccactat attgcttgct agcactgaaa ccacagcgaa cactatctcg tgggcttgtt
     3001 atgaattggc acgtcatccc cgtatttttg ctgagtgccg cgccgagatc gatgcccttg
     3061 taaaagatcg ggactggctt gacatcgaaa ttggcagaca cgatctgcca gccttgaaac
     3121 gcgtattatt cgaggccttg cgcatgtatc cgtccagtta tctgcttagc cgtcaggcct
     3181 cagtggatac cactctgggc ggttacgcaa tcccgaagga cgcggccatt ttgtattctc
     3241 attacggcca gcaaagagat gaacgtaatt tcccccatgg cgatgagttt gatccggatc
     3301 ggtggctcga taaagatggt gcggaagtta cggcttctgc gtttatgcct tttgggtttg
     3361 gcgcgtacag atgtctgggc gagagtgtag ccgtgctgga ggcgacatac tgtctggcca
     3421 tgatggtgca tcagtgggac ttcgcgctgt cagattattc tgaacctaaa atgaatgcca
     3481 ctatcactct gtcgccgaaa gatctggaat tcctgtttac gaaacgtacc gaaagtggcg
     3541 cgcatgatga ataataagga tgagctctac aaataagcag aggtggttgt gttgcgaaaa
     3601 aaaaaaaaaa acaccctaac gggtgttttt ttttttttgg tgtccccacg tgtggcgctg
     3661 gagaccgtcc aatggcggcg cgccatcgaa tggtgcaaaa cctttcgcgg tatggcatga
     3721 tagcgcccgg aagagagtca attcagggtg gtgaatatga gcccgaaacg tcgtacccag
     3781 gcagaacgtg caatggaaac ccagggtaaa ctgattgcag cagcactggg tgttctgcgt
     3841 gaaaaaggtt atgcaggttt tcgtattgca gatgttccgg gtgcagccgg tgttagccgt
     3901 ggtgcacaga gccatcattt tccgaccaaa ctggaactgc tgctggcaac ctttgaatgg
     3961 ctgtatgagc agattaccga acgtagccgt gcacgtctgg caaaactgaa accggaagat
     4021 gatgttattc agcagatgct ggatgatgca gcagattttt ttctggatga tgattttagc
     4081 atcggcctgg atctgattgt tgcagcagat cgtgatccgg cactgcgtga aggtattctg
     4141 cgtaccgttg aacgtaatcg ttttgttgtt gaagatatgt ggctgggtgt gctggtgagc
     4201 cgtggtctga gccgtgatga tgccgaagat attctgtggc tgatttttaa cagcgttcgt
     4261 ggtctgacag ttcgtagcct gtggcagaaa gataaagaac gttttgaacg tgtgcgtaat
     4321 agcaccctgg aaattgcacg tgaacgttat gcaaaattca aacgttgata aggatcctaa
     4381 ttggtaacga atcagacaat tgacggctcg agggagtagc atagggtttg cagaatccct
     4441 gcttcgtcca tttgacaggc acattatgca tcgatgataa gctgtcaaac atgagcagat
     4501 cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc
     4561 ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcaa
     4621 atattttatc tgaggtgctt cctcgctc
//
