LOCUS       sequence-423283-        4530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Streptomyces platensis LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4530)
  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 4530)
  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: pCYP33
FEATURES             Location/Qualifiers
     source          1..4530
                     /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(4442..4461)
                     /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 tgtttaccga cactaaagta
     2101 agggggaaca acatgaccac ccgcccgacc gcggccagcc gtgggaataa atatcgtttc
     2161 gctttccgta ctctgtcgtt cctggacgat cgcaaggatg accgctccgg tgtggcagag
     2221 ttaagcggcc cgccgggtcg tgcactcctt gtatggaagc cggagataat aggtcaggtg
     2281 tttcgtggcg atcgtaatat gacattggaa ggctcagata ccctgggtcc gcttgttgga
     2341 gacacgtccc tgttgttcgc aaatgggcca cgtcatgcgg cctaccgtca ggtgattggc
     2401 ccacgccttc gtggtagacc gctgcgcgga tatgaagaac tcatcgccga ggctacacgt
     2461 gcagcaatcg atgaattgcg tccaggcacc gtattccaag tgccagactg gacccgccgg
     2521 ctgaccctcc agattgtctc ccagataatt ctgggtcctg ttgaccatgg cctccttcac
     2581 cgcttcacgt cctggattga gggtgtatta gggagccgtg gacgtacgct cgcgtatcga
     2641 tatctgcgct taccgcacgc gctgccaagt ccctggcgta cctttgttcg tcagcgcgag
     2701 agcctggata aggagctgct gtgtccagtt tcggggaaat ctgcgggtgg gggtgccggt
     2761 agcggtacag ccgaaccaag ccctgacacc ttagcggagg tccttcgttc aggggaagaa
     2821 ccgctcggtc cgctggggga tggcgaactg cgtgatcaaa tcgtcagttt actctttgcc
     2881 ggccacgaaa ccaccgcatc cgcgatttcg tgggcactgt tctggctcgc agagcacgat
     2941 gaagtgcgac gtgacatcat agacgaactg aaagcaacta gcagctctgg ggcagctgcg
     3001 gaggacgtac cgctgttaga tgcggcgtgc cgagaagtat tacgaatttc tcctccggca
     3061 gttgtggcag gcaatcgcgt actgaatgaa gatcaagaaa tcgatggtgt accgcatgat
     3121 gcaggtacac gtctgacgcc gtgtatttac ctggcgcatc aacaaccgga tctttatcca
     3181 caaccgcggc gctttgatcc gcatcgcttc ttaggcaaac gtaaatccgc gcaagaatac
     3241 ctgcctttcg gcggcggcac tcgtcgatgc ctgggcgctg atctggctat gctggaaatg
     3301 cgaatggtcg ttgcagcagt gctccgtcgg cgggaactga aatgcgtcaa tccagagaca
     3361 ggtgtgccac agctgcgtgg cccggcgatg ggacctagcg aagatctgcg tatgacagtc
     3421 acaaaatggc cggcgtaata aggatgagct ctacaaataa gcagaggtgg ttgtgttgcg
     3481 aaaaaaaaaa aaacacccta acgggtgttt tttttttttt ggtgtcccca cgtgtggcgc
     3541 tggagaccgt ccaatggcgg cgcgccatcg aatggtgcaa aacctttcgc ggtatggcat
     3601 gatagcgccc ggaagagagt caattcaggg tggtgaatat gagcccgaaa cgtcgtaccc
     3661 aggcagaacg tgcaatggaa acccagggta aactgattgc agcagcactg ggtgttctgc
     3721 gtgaaaaagg ttatgcaggt tttcgtattg cagatgttcc gggtgcagcc ggtgttagcc
     3781 gtggtgcaca gagccatcat tttccgacca aactggaact gctgctggca acctttgaat
     3841 ggctgtatga gcagattacc gaacgtagcc gtgcacgtct ggcaaaactg aaaccggaag
     3901 atgatgttat tcagcagatg ctggatgatg cagcagattt ttttctggat gatgatttta
     3961 gcatcggcct ggatctgatt gttgcagcag atcgtgatcc ggcactgcgt gaaggtattc
     4021 tgcgtaccgt tgaacgtaat cgttttgttg ttgaagatat gtggctgggt gtgctggtga
     4081 gccgtggtct gagccgtgat gatgccgaag atattctgtg gctgattttt aacagcgttc
     4141 gtggtctgac agttcgtagc ctgtggcaga aagataaaga acgttttgaa cgtgtgcgta
     4201 atagcaccct ggaaattgca cgtgaacgtt atgcaaaatt caaacgttga taaggatcct
     4261 aattggtaac gaatcagaca attgacggct cgagggagta gcatagggtt tgcagaatcc
     4321 ctgcttcgtc catttgacag gcacattatg catcgatgat aagctgtcaa acatgagcag
     4381 atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc
     4441 gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc
     4501 aaatatttta tctgaggtgc ttcctcgctc
//
