LOCUS       sequence-423312-        4659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Kitasatospora sp. CB02891 LRD-modifying cytochrome 
            P450 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4659)
  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 4659)
  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: pCYP27
FEATURES             Location/Qualifiers
     source          1..4659
                     /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(4571..4590)
                     /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 tgtttacaac taatattaac
     2101 ttaaggaatt attcaaatga aaggaaattt acttgccatg tcaaccgcta ccagccccag
     2161 cgtaaccggc tcggtgacga cccctcgtac ggccaccaca ccgggaagaa ccccgggact
     2221 ggcaccgggt ggcgtacccg tgctgggaca tttaccaatg atcctccgta gccgttttga
     2281 attcatcgaa accgtacgca acgcaggccc tgtgacccgc gttaaactgg gtcccaaaac
     2341 tgcttacttc gttaacgact acgagctgct cgcccaaatc ttagtttctg acgcagacaa
     2401 gtttgtccgt ggcattcact ttaaaaagat gcgcaacatg gtaggaaatg gggtcgtcac
     2461 gactagcggg gatctgcatc gcagacagcg gcgaattatg ctcccgtcct tcagtcagcg
     2521 gcgcctggct atgcaccttc cggtcatgcg taaaattatg agcgaatttg tggccgccgt
     2581 tccggaacag cagccatacg acttgatggg accggtcatg ggtgtggggt gcgacatcgt
     2641 gacttctact atgctgggag agaaaacgcc tccaaaagtc cttcgtctgg tacgtgaggc
     2701 agttcctgtt tttgtggaga acgcggccat tcaagcagtg gacgtcacgg gaatctacaa
     2761 gcatttgccg accaaatcga atcgcgactt cgaacggctc ctgaatgcgt tcaacgagta
     2821 tatgtattca gtgatcgacg acaaattccg caatggcgct ggggaggaag ctggcctctt
     2881 ggatatgtta atcaatgcaa ccgacccaga aacgggtgaa aagtttgatc gcacggaggt
     2941 acgtgaccag gccgcgacca tcctgcttgc gtcaacagaa accaccgcaa acacaatttc
     3001 atgggcgtgc tacgaactgg caagacaccc aagaatcttt gctgagtgtc gggccgaaat
     3061 cgatgctctg gttaaagatc gcaactggtt ggacatcgag atcggtcgtc acgatctgcc
     3121 ggcactcaaa cgcgtacttt ttgaagcgct gcgcatgtat ccgtcctcct atttattaag
     3181 ccgtcaagct agcgtggaca ccactctcgg tggttatgcg attccgaaag atgcgaccat
     3241 tttgtattcg cattatggtc aacagcgcga tgaacgtaat ttcccacacg gcgatgaatt
     3301 cgatcctgat cggtggctgg acaaggatgg tacggaagtg accgcatcgg catttatgcc
     3361 tttcggtttt ggcgcctatc gttgcttggg cgaaagcgtg gcggtgctgg aggccaccta
     3421 ctgtttagcc atgatggtgc accagtggga ttttgcactg tctgactact ccgaaccgaa
     3481 aatgaatgcg acgattaccc tgtcaccgaa agacctggaa tttttattca ccaaacgagc
     3541 tgaatcaggt gcacgcgacg aataataagg atgagctcta caaataagca gaggtggttg
     3601 tgttgcgaaa aaaaaaaaaa aacaccctaa cgggtgtttt tttttttttg gtgtccccac
     3661 gtgtggcgct ggagaccgtc caatggcggc gcgccatcga atggtgcaaa acctttcgcg
     3721 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatatg agcccgaaac
     3781 gtcgtaccca ggcagaacgt gcaatggaaa cccagggtaa actgattgca gcagcactgg
     3841 gtgttctgcg tgaaaaaggt tatgcaggtt ttcgtattgc agatgttccg ggtgcagccg
     3901 gtgttagccg tggtgcacag agccatcatt ttccgaccaa actggaactg ctgctggcaa
     3961 cctttgaatg gctgtatgag cagattaccg aacgtagccg tgcacgtctg gcaaaactga
     4021 aaccggaaga tgatgttatt cagcagatgc tggatgatgc agcagatttt tttctggatg
     4081 atgattttag catcggcctg gatctgattg ttgcagcaga tcgtgatccg gcactgcgtg
     4141 aaggtattct gcgtaccgtt gaacgtaatc gttttgttgt tgaagatatg tggctgggtg
     4201 tgctggtgag ccgtggtctg agccgtgatg atgccgaaga tattctgtgg ctgattttta
     4261 acagcgttcg tggtctgaca gttcgtagcc tgtggcagaa agataaagaa cgttttgaac
     4321 gtgtgcgtaa tagcaccctg gaaattgcac gtgaacgtta tgcaaaattc aaacgttgat
     4381 aaggatccta attggtaacg aatcagacaa ttgacggctc gagggagtag catagggttt
     4441 gcagaatccc tgcttcgtcc atttgacagg cacattatgc atcgatgata agctgtcaaa
     4501 catgagcaga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg
     4561 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg
     4621 ctcatgagca aatattttat ctgaggtgct tcctcgctc
//
