LOCUS       sequence-423264-        4428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Saccharopolyspora elongata LRD-modifying cytochrome 
            P450 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4428)
  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 4428)
  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: pCYP36
FEATURES             Location/Qualifiers
     source          1..4428
                     /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(4340..4359)
                     /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 tgtttatggc agcttattac
     2101 tctaaggggg acattaatga ccaatgtgca acacgatgcg ccagcgtttc cgatggcccg
     2161 tccgcctggt tgccccttag atcctccggc ggaatatggc cgcatgcgcg cggaagctcc
     2221 ggttactccg gctattttac ccgatggcag tcgcacctgg ctgatctcac ggtatgcgga
     2281 cgtacgtgcg gtgttgcgcg atccgcgggt gtctgccgac ctgcggaaac ccggattccc
     2341 gattatcagc gcggccgaac gtaccctggt tgacgttggc tttcatcccg ggtttatacg
     2401 caccgatcca cccgagcatg gacagctgcg tcgcatggtc agtgcggatt ttacgatccg
     2461 tcgggttaaa gcgctccgac cggccatcca acgtttggtc gatgaacgcg tagatcgtat
     2521 tctggacggt ccgaaacctg ttgatctggt agctgaactg gcgttaccgg tcccgtccac
     2581 ggtaatctgt tggttattag gtgtcccagt ggcagacatt ggtaaattca atgaatggac
     2641 tcgcgcagtc gtggacgccg atacgacgcc ggaacaaacc caggaggcaa acgctgccat
     2701 cctgggttat ttcgacgagt taattgcgat caaacaacgc gaaccagccg atgatattgt
     2761 atcgcgtttg gtccgtcaac acgaagcggg aaagctgacg cgcccggagg tcttaacaac
     2821 ttccatgtta ctgttggtag ctggccatga aaccaccgcc aatatgattt ccctgggtgt
     2881 actgaccctt ctgcagcatc cggagcaagc cgccgcattg cgcgagaacg cgggactcgc
     2941 ctctcgggct gttgaagaac tgctgcgcta tctttcaatc agcgaatttg ccaccactcg
     3001 cgtcgcaaca gaggacctgg aagtcggtgg ccggctgatc cgcgcgggtg atggcatcgt
     3061 agccctgacg ccctctgcca accgcgacgc gacagcgttt ccagaccctg acactgtaga
     3121 tattcaccgt aagagccgtc agcatgttgc attcggcttt gggccgcatc agtgcgtagg
     3181 ccagtcgttg gccagagcgg aactgcagat cgtctttccg accctgctgc gtcgtattcc
     3241 tggtttgcga ttagccgttc ctgcggaaga actgcggtat aaactgaacg aaggtatctt
     3301 cggcgtcctg agcttacctg tagcctggga ttaataagga tgagctctac aaataagcag
     3361 aggtggttgt gttgcgaaaa aaaaaaaaaa acaccctaac gggtgttttt ttttttttgg
     3421 tgtccccacg tgtggcgctg gagaccgtcc aatggcggcg cgccatcgaa tggtgcaaaa
     3481 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatatga
     3541 gcccgaaacg tcgtacccag gcagaacgtg caatggaaac ccagggtaaa ctgattgcag
     3601 cagcactggg tgttctgcgt gaaaaaggtt atgcaggttt tcgtattgca gatgttccgg
     3661 gtgcagccgg tgttagccgt ggtgcacaga gccatcattt tccgaccaaa ctggaactgc
     3721 tgctggcaac ctttgaatgg ctgtatgagc agattaccga acgtagccgt gcacgtctgg
     3781 caaaactgaa accggaagat gatgttattc agcagatgct ggatgatgca gcagattttt
     3841 ttctggatga tgattttagc atcggcctgg atctgattgt tgcagcagat cgtgatccgg
     3901 cactgcgtga aggtattctg cgtaccgttg aacgtaatcg ttttgttgtt gaagatatgt
     3961 ggctgggtgt gctggtgagc cgtggtctga gccgtgatga tgccgaagat attctgtggc
     4021 tgatttttaa cagcgttcgt ggtctgacag ttcgtagcct gtggcagaaa gataaagaac
     4081 gttttgaacg tgtgcgtaat agcaccctgg aaattgcacg tgaacgttat gcaaaattca
     4141 aacgttgata aggatcctaa ttggtaacga atcagacaat tgacggctcg agggagtagc
     4201 atagggtttg cagaatccct gcttcgtcca tttgacaggc acattatgca tcgatgataa
     4261 gctgtcaaac atgagcagat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
     4321 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
     4381 cacttcgggc tcatgagcaa atattttatc tgaggtgctt cctcgctc
//
