LOCUS       sequence-423280-        4654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Actinomadura sp. KC06 LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4654)
  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 4654)
  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: pCYP17
FEATURES             Location/Qualifiers
     source          1..4654
                     /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(4566..4585)
                     /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 tgtttaggta cacattaaag
     2101 tctaaggaag gtattttatg ccaactcagc tcgacctgcc cgagtcaccg caacgtagtc
     2161 gcgcgggtgg gttgcttgga gctggattta cagaaggctc acgtatgtcc gttgacgcac
     2221 gtattcgcac cgtccataaa gcgcagacgc tgagcatcag ccatctgcct ggccttctgc
     2281 gtgatccgct gcgtgaattt gaggaaatgg gacgcgaagc agacggtcga atcgttcgag
     2341 ctaaccttgg gccctttcgt ccttacctga taacccaccc tgatcactta cagcacgtgc
     2401 ttcgggataa tgtggataac tatcggagag atggcatgat gtggaaaccg ctttccagac
     2461 tggtaggtga aactagcgat gctgatccga gttggccggt caagcgccgc gtctaccagg
     2521 acttgttatc cggaccgtcc actgctctgg tggcagacaa aatggttgcg actatctcgg
     2581 atgccgtggg ggaactgggt gagcgtgcag accctgaacg tccggtggac gcatacgagg
     2641 aaatgacccg catcgtgtac cgaacgatta ttagagtgtt gattggtgac cgtatctcag
     2701 tcagccaaat cgacgagctt ggccaggtta tggttacggc cactacaagc agttttcgat
     2761 cacgtatgct gatgccattt gtgccgctca gtgtgccgct gccgggcgac cgcaccttta
     2821 atcgttctgt ccaggcggta gacgacctga ttttcccgat cgtgcgtgag gcacgacgca
     2881 atggagctgg tggtcgcgac gttgtgagtt tgctgttaaa cgcgagagga gaagatggaa
     2941 aaggcattga tgaccgtgca gtgcgcgatg gcataaccgg cttgttcgta gccgccaccg
     3001 acaccactgt tagcgtatta accttcctct gggccgccct cgatctgaat ccggaagtcc
     3061 gggcacgcct gtacgatgaa atggaccgcg tcgtcggaga gggcccaccg agttttgagc
     3121 acgtggcaga tctgacctac accaaaatgg tgctccatga agcgttgcgt ttgtactctg
     3181 tcgcttggat cgcgccgcgt gttgtggaag ctgatgacct gatcgacggg gttcgtatca
     3241 aagcgggtgc tatagttgcg gtaagtccgt ttctgaccca ccgcctgcca gggatttggc
     3301 cccagccgta tgtgttcgat ccggaacgct ttgcaaccgg tcgtagccgc catcgcttcg
     3361 cattcatggc atttggaggt ggcccgaatc agtgcgcggg tcgtgtcttt ttcttcattg
     3421 aagcaccact cattttggcg accctgctgt ccaaatatcg ccctattctg cattcgccac
     3481 acccagtcga accgcgcgtg gatataactc tgaagccgcg tgatcgtgtg aaaatcacct
     3541 tagaaccggt gcgacgttaa taaggatgag ctctacaaat aagcagaggt ggttgtgttg
     3601 cgaaaaaaaa aaaaaaacac cctaacgggt gttttttttt ttttggtgtc cccacgtgtg
     3661 gcgctggaga ccgtccaatg gcggcgcgcc atcgaatggt gcaaaacctt tcgcggtatg
     3721 gcatgatagc gcccggaaga gagtcaattc agggtggtga atatgagccc gaaacgtcgt
     3781 acccaggcag aacgtgcaat ggaaacccag ggtaaactga ttgcagcagc actgggtgtt
     3841 ctgcgtgaaa aaggttatgc aggttttcgt attgcagatg ttccgggtgc agccggtgtt
     3901 agccgtggtg cacagagcca tcattttccg accaaactgg aactgctgct ggcaaccttt
     3961 gaatggctgt atgagcagat taccgaacgt agccgtgcac gtctggcaaa actgaaaccg
     4021 gaagatgatg ttattcagca gatgctggat gatgcagcag atttttttct ggatgatgat
     4081 tttagcatcg gcctggatct gattgttgca gcagatcgtg atccggcact gcgtgaaggt
     4141 attctgcgta ccgttgaacg taatcgtttt gttgttgaag atatgtggct gggtgtgctg
     4201 gtgagccgtg gtctgagccg tgatgatgcc gaagatattc tgtggctgat ttttaacagc
     4261 gttcgtggtc tgacagttcg tagcctgtgg cagaaagata aagaacgttt tgaacgtgtg
     4321 cgtaatagca ccctggaaat tgcacgtgaa cgttatgcaa aattcaaacg ttgataagga
     4381 tcctaattgg taacgaatca gacaattgac ggctcgaggg agtagcatag ggtttgcaga
     4441 atccctgctt cgtccatttg acaggcacat tatgcatcga tgataagctg tcaaacatga
     4501 gcagatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
     4561 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat
     4621 gagcaaatat tttatctgag gtgcttcctc gctc
//
