LOCUS       sequence-423389-        4415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Streptomyces olivaceus LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4415)
  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 4415)
  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: pCYP47
FEATURES             Location/Qualifiers
     source          1..4415
                     /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(4327..4346)
                     /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 tgtttagagc gacctagcga
     2101 acaagggagg taatttatga cagcgacgga acagggaacc cggttagcag accaaagcgc
     2161 tgacgtcgcg acaatgctgg catggttctg gagaatgcgt gaatccgagc cagttagtca
     2221 tgatgaagaa gccggggttt ggcacgtgtt tcgtcatgct gatgtagagc gcgtcctggc
     2281 agatcctggg acatttagct ctgatttctc gagcttcatg ccggcgcagg aggatgtgga
     2341 tcgctttgtg aaaggaaacc tgctccgtaa agatccaccc caacatcgca aactgcgcac
     2401 gttagttagc aaagccttta cccctgcagt agttgcgcgc ctggctcccc ggattgaggt
     2461 cattgcggac gaacttctcg acgccaaagc gggcgccgat cagcttgaac tggtcgcgga
     2521 tctggcgtca ccgctgccgg tcaccgttat tgcagaattg cttggtatac ctgctcagga
     2581 ccggccgatg tttggtcgat gggccgacgc gctcgtggcg ccggagtcac agacctcttt
     2641 cattccgaat gaggaccgta caaaagcaat ggccgtcgtc atgcgtgaaa tgaatgcgta
     2701 ctgtcttgaa catattcgta gacgtcgtgc cgcgcctggg caggacttag ttagcaaact
     2761 ggtagctgcc gaagttgacg ggcagagatt ggatgacgat gaaatcacgg gctttgtggg
     2821 gcttttactg ctggcaggcc atattacaac gaccgcactg ttaactaatg tagtcctttg
     2881 cctggacaag tttcctgatg ttgcggccgt gctgcgtgca gatccgggtg cgttaccagg
     2941 cgctatcgag gaagttctgc gcttccgcac accgttagcc cctagtatgc gtcgcaccac
     3001 tcgcgaagta cgcgtggggg aacacacggt gccagccggc cgtatcgttc tggcctggtt
     3061 agcgagcgcg aaccgtgatg aacgtcagtt taccgcaccg gacacgtttg atattcgccg
     3121 cgctccgaat ccccatctga gtctgggtca tggtatccat ttctgccttg gtgcaccact
     3181 ggcccgactg gaagtgcgca tcgccgttga aaagatgctg ggccgttggt ccggcatgac
     3241 ggttgctgat ggcgtggaat accatgacgc tcgcggaatt gttggcgcca agaaactgcc
     3301 tctgcatctg cgctgggcgt aataaggatg agctctacaa ataagcagag gtggttgtgt
     3361 tgcgaaaaaa aaaaaaaaca ccctaacggg tgtttttttt tttttggtgt ccccacgtgt
     3421 ggcgctggag accgtccaat ggcggcgcgc catcgaatgg tgcaaaacct ttcgcggtat
     3481 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatatgagcc cgaaacgtcg
     3541 tacccaggca gaacgtgcaa tggaaaccca gggtaaactg attgcagcag cactgggtgt
     3601 tctgcgtgaa aaaggttatg caggttttcg tattgcagat gttccgggtg cagccggtgt
     3661 tagccgtggt gcacagagcc atcattttcc gaccaaactg gaactgctgc tggcaacctt
     3721 tgaatggctg tatgagcaga ttaccgaacg tagccgtgca cgtctggcaa aactgaaacc
     3781 ggaagatgat gttattcagc agatgctgga tgatgcagca gatttttttc tggatgatga
     3841 ttttagcatc ggcctggatc tgattgttgc agcagatcgt gatccggcac tgcgtgaagg
     3901 tattctgcgt accgttgaac gtaatcgttt tgttgttgaa gatatgtggc tgggtgtgct
     3961 ggtgagccgt ggtctgagcc gtgatgatgc cgaagatatt ctgtggctga tttttaacag
     4021 cgttcgtggt ctgacagttc gtagcctgtg gcagaaagat aaagaacgtt ttgaacgtgt
     4081 gcgtaatagc accctggaaa ttgcacgtga acgttatgca aaattcaaac gttgataagg
     4141 atcctaattg gtaacgaatc agacaattga cggctcgagg gagtagcata gggtttgcag
     4201 aatccctgct tcgtccattt gacaggcaca ttatgcatcg atgataagct gtcaaacatg
     4261 agcagatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
     4321 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
     4381 tgagcaaata ttttatctga ggtgcttcct cgctc
//
