LOCUS       sequence-424229-        4461 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 4461)
  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 4461)
  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: pCYP61
FEATURES             Location/Qualifiers
     source          1..4461
                     /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(4373..4392)
                     /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 tgtttaagag cttcaataag
     2101 ttaaggagag ttttatatgg cggatctgga aacaaaattt ccgcgttacg aacttattag
     2161 ttctggcaag tatgtggacc gtatcccgga attacacgag ctgcgagaga agagtccgat
     2221 tgcatgggta ccggttatgg acgctgcgtt cctcacgcgc cacgcggata tcgttcgcgt
     2281 tttgaaagat caccgcatgg cccccgctaa cctgacccaa gggattcggc tgctgagtcc
     2341 ggaacagcag gaagaattgg aacccctgag ttcagctgtg aagaaatgga tgggacatac
     2401 cgttcctgcc gaccatcagc gcttcattgg tctgcttaaa cgctatttta ccccggcgat
     2461 gattgaccgt atgcgtccgc gtgttcgaca gctcagtcat gagctcctgg atgcagttga
     2521 accggcaggc cgcatggata tagtatcaga tatcgcgtat ccgctgccgg cgtgcgtgat
     2581 cgctgaaatg ttaggagttc cgatggataa ccgggctcag ttgctggcct ggagcgcgga
     2641 catcggcgcc attgcggaga tcgtcagcta cgaccgcctg atggaatgcc agcgtagtct
     2701 gctggccatg caggactttg tgctcgaagt tgtgaaagaa cgtcgtgccg aaccgaaaga
     2761 tgatctgatc tcaatgttcg tcgcggcgga gcgtgagggc ctcgtcagtg aagccgaaat
     2821 tctgtcgaat tgtgtcatgt tgttgtttag cggacatgag acaaccggag gtttgatcac
     2881 cagtggctta gttcagctgt ttgatcatcc tgatcaactg gaactgctga aatctgatcc
     2941 tgatttgatg ccaggcgctg tcgaggaaat gctgcgtctg gccggtcctg cgagcgttat
     3001 ttcacgtgtt agtactgaac ctgtggaagt agcaggctca agcatccgtt ctggcgctgc
     3061 tgtaccgcct ggtgcggatg gtggtaaacg tgatcctcgt gtctttgagg accccgatcg
     3121 tctggacgct acccgccgcc cgaacgatca cttggcgttt gcgaccggta tgttttactg
     3181 ccttggggcc gctcttgcac gtatggaagc cgatgaattt ttccgcatcc ttctggatcg
     3241 tttcccggat gtgaatccgg gctatgaaac accggactgg cagcctgtgc tgttgatttc
     3301 tcggcggtta aaaaccttac cggtgaattt acgaggggtg ggcggctctg gggcgggaga
     3361 tgagtaataa ggatgagctc tacaaataag cagaggtggt tgtgttgcga aaaaaaaaaa
     3421 aaaacaccct aacgggtgtt tttttttttt tggtgtcccc acgtgtggcg ctggagaccg
     3481 tccaatggcg gcgcgccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc
     3541 cggaagagag tcaattcagg gtggtgaata tgagcccgaa acgtcgtacc caggcagaac
     3601 gtgcaatgga aacccagggt aaactgattg cagcagcact gggtgttctg cgtgaaaaag
     3661 gttatgcagg ttttcgtatt gcagatgttc cgggtgcagc cggtgttagc cgtggtgcac
     3721 agagccatca ttttccgacc aaactggaac tgctgctggc aacctttgaa tggctgtatg
     3781 agcagattac cgaacgtagc cgtgcacgtc tggcaaaact gaaaccggaa gatgatgtta
     3841 ttcagcagat gctggatgat gcagcagatt tttttctgga tgatgatttt agcatcggcc
     3901 tggatctgat tgttgcagca gatcgtgatc cggcactgcg tgaaggtatt ctgcgtaccg
     3961 ttgaacgtaa tcgttttgtt gttgaagata tgtggctggg tgtgctggtg agccgtggtc
     4021 tgagccgtga tgatgccgaa gatattctgt ggctgatttt taacagcgtt cgtggtctga
     4081 cagttcgtag cctgtggcag aaagataaag aacgttttga acgtgtgcgt aatagcaccc
     4141 tggaaattgc acgtgaacgt tatgcaaaat tcaaacgttg ataaggatcc taattggtaa
     4201 cgaatcagac aattgacggc tcgagggagt agcatagggt ttgcagaatc cctgcttcgt
     4261 ccatttgaca ggcacattat gcatcgatga taagctgtca aacatgagca gatcctctac
     4321 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
     4381 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag caaatatttt
     4441 atctgaggtg cttcctcgct c
//
