LOCUS       sequence-423266-        4504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Streptomyces sp. BK447 LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4504)
  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 4504)
  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: pCYP45
FEATURES             Location/Qualifiers
     source          1..4504
                     /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(4416..4435)
                     /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 tgtttatggt ctagcctgtc
     2101 taggaggtca agtaatgagc atcacacccc cggccgtaac cgcggtgccg ttgcatggtc
     2161 ctgagcatgc cgctaaccca cagcgctctt atgaactgtt aagactccag ggccctgttg
     2221 gcctggcgga agtagcgccg ggagtggtgg tccatctggt aaccgactac cgggctgcat
     2281 tagacctgtt acaagacacc gatacctgga ccaaagatac ccgcagttgg ctgccgcagg
     2341 ttcctgaaaa ctctcccatt cgcccattgg tagaatggcg cccaagcctc ttctttgcgg
     2401 atggcgaaga acatgcgcat ttgcgaaaag tgattacaga ttcgttcggc cttcttgatc
     2461 cttacgatgt tcgcgttctg acctttcgcc acgctgatcg cttactgcaa gatttcgcag
     2521 cggacggaac cgttgatctg gttagccagt atgctcagca gctgccgctg atggttttta
     2581 atgcactgtt cggcatgggt gatgagtatg gcccgcagct tgtgcgcgct ttaagcgcaa
     2641 tgatggatac agatcctgaa aagaaagccg aaggcgcaca gtcattcggg gcctatctgc
     2701 agacactgta ccagacaaaa gcggcgcaaa gagggcatga tctgacctcc tggtttatcg
     2761 atcatccgaa tggtttgaac caggaagaag caattaacca gattctgata acactgggcg
     2821 cagggaatga gcctaccgcc aacctgatcg cgaacacttg tgtgcggatg ctctccgatg
     2881 atagatatta tggcactctg acaacggggt cattaccgat tcagcatgcg attgatgatg
     2941 ctttatggaa tgatagcccg ctggccaact actcttttca ttttccaaag gaggacatta
     3001 cattccatgg caaacgtata ccagccggaa gcccggtgat ggttagttat gcggctgcta
     3061 atacgtgtcc tcatgccggg gtcccgagtg acggttatcg gagcggtacc aaagcgcact
     3121 tagcatttgc tgccggcccg catacgtgtc cggcacgcga cttagctctg ttgatcgcga
     3181 cggcggctat cgaacgtctg gcatcatatc tcccagacat tgaactggcg gtaagcccgg
     3241 accagttaac ctaccgggac ggcgcgatcc atcgcacact tacgtcattg ccagcgagat
     3301 ttactccgct caccccggat gcgaacggcg caaccccgtg gagtcgtaat acttcatccc
     3361 ctaaaaccga tactggtccg ccacagcaac aagcgacagc ttcgcgctaa taaggatgag
     3421 ctctacaaat aagcagaggt ggttgtgttg cgaaaaaaaa aaaaaaacac cctaacgggt
     3481 gttttttttt ttttggtgtc cccacgtgtg gcgctggaga ccgtccaatg gcggcgcgcc
     3541 atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     3601 agggtggtga atatgagccc gaaacgtcgt acccaggcag aacgtgcaat ggaaacccag
     3661 ggtaaactga ttgcagcagc actgggtgtt ctgcgtgaaa aaggttatgc aggttttcgt
     3721 attgcagatg ttccgggtgc agccggtgtt agccgtggtg cacagagcca tcattttccg
     3781 accaaactgg aactgctgct ggcaaccttt gaatggctgt atgagcagat taccgaacgt
     3841 agccgtgcac gtctggcaaa actgaaaccg gaagatgatg ttattcagca gatgctggat
     3901 gatgcagcag atttttttct ggatgatgat tttagcatcg gcctggatct gattgttgca
     3961 gcagatcgtg atccggcact gcgtgaaggt attctgcgta ccgttgaacg taatcgtttt
     4021 gttgttgaag atatgtggct gggtgtgctg gtgagccgtg gtctgagccg tgatgatgcc
     4081 gaagatattc tgtggctgat ttttaacagc gttcgtggtc tgacagttcg tagcctgtgg
     4141 cagaaagata aagaacgttt tgaacgtgtg cgtaatagca ccctggaaat tgcacgtgaa
     4201 cgttatgcaa aattcaaacg ttgataagga tcctaattgg taacgaatca gacaattgac
     4261 ggctcgaggg agtagcatag ggtttgcaga atccctgctt cgtccatttg acaggcacat
     4321 tatgcatcga tgataagctg tcaaacatga gcagatcctc tacgccggac gcatcgtggc
     4381 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
     4441 ggaagatcgg gctcgccact tcgggctcat gagcaaatat tttatctgag gtgcttcctc
     4501 gctc
//
