LOCUS       sequence-424231-        4448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Priestia megaterium LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4448)
  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 4448)
  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: pCYP59
FEATURES             Location/Qualifiers
     source          1..4448
                     /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(4360..4379)
                     /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 tgtttagtag ggacgtataa
     2101 cccaaatccg gcgggagaga ttttatgaag gaagtgatcg cggtgaaaga aataacccgt
     2161 tttaaaacgc gtactgaaga attttcgccc tacgcttggt gcaaacgtat gcttgaaaac
     2221 gaccctgtta gctatcatga aggcacggat acttggaacg tgtttaaata tgaggatgtg
     2281 aaacgcgtac tgagtgacta taaacatttt agctctgtta gaaaacgtac taccatcagt
     2341 gtcgggacgg actcggaaga aggttccgtg ccagagaaaa tccagattac ggaaagcgac
     2401 ccaccggatc atcgcaaacg ccggagtctg ttggccgcgg ccttcacccc cagatctctg
     2461 cagaactggg aacctcgcat ccaagaaatc gccgatgaac tgattggtca gatggatggc
     2521 ggaacagaga ttgatatagt ggccagcctg gccagccctt taccgattat cgtaatggcc
     2581 gacctgatgg gcgtgccgag caaagaccgc ctgctgttta agaagtgggt ggataccctg
     2641 tttctgccgt ttgatcgcga gaaacaagaa gaggtggata aactgaagca agtcgcggcc
     2701 aaagaatatt atcagtatct ctatccgatc gtggttcaaa agcgactgaa ccctgcagac
     2761 gacatcattt ctgacttgct gaaaagcgaa gttgatggag agatgttcac cgatgacgag
     2821 gtggtgcgca ctacgatgct tatcttaggt gctggtgtgg aaactacgtc acacctcctg
     2881 gcgaacagct tttatagcct gctgtacgac gataaagaag tttaccaaga attgcatgag
     2941 aacttagatc tggtcccgca ggcggtcgag gaaatgttac gcttccgttt caacttgatt
     3001 aagctggacc gtacggtcaa agaagataat gatctcctgg gcgttgagct gaaagaaggt
     3061 gattccgtcg tcgtctggat gagcgccgcg aatatggatg aggagatgtt cgaggacccg
     3121 tttactctga atatccatcg acctaacaac aagaaacacc tgacctttgg caatggtccg
     3181 catttctgtc tgggtgcgcc cttagcaaga ctcgaagcca aaatcgccct gaccgcgttc
     3241 ctcaagaaat ttaaacatat tgaagcagtg cctagctttc agctggaaga gaatttaacc
     3301 gactcggcta cgggccagac gctgacctca ctgcctctga aagcaagtcg tatgtgagga
     3361 tgagctctac aaataagcag aggtggttgt gttgcgaaaa aaaaaaaaaa acaccctaac
     3421 gggtgttttt ttttttttgg tgtccccacg tgtggcgctg gagaccgtcc aatggcggcg
     3481 cgccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     3541 attcagggtg gtgaatatga gcccgaaacg tcgtacccag gcagaacgtg caatggaaac
     3601 ccagggtaaa ctgattgcag cagcactggg tgttctgcgt gaaaaaggtt atgcaggttt
     3661 tcgtattgca gatgttccgg gtgcagccgg tgttagccgt ggtgcacaga gccatcattt
     3721 tccgaccaaa ctggaactgc tgctggcaac ctttgaatgg ctgtatgagc agattaccga
     3781 acgtagccgt gcacgtctgg caaaactgaa accggaagat gatgttattc agcagatgct
     3841 ggatgatgca gcagattttt ttctggatga tgattttagc atcggcctgg atctgattgt
     3901 tgcagcagat cgtgatccgg cactgcgtga aggtattctg cgtaccgttg aacgtaatcg
     3961 ttttgttgtt gaagatatgt ggctgggtgt gctggtgagc cgtggtctga gccgtgatga
     4021 tgccgaagat attctgtggc tgatttttaa cagcgttcgt ggtctgacag ttcgtagcct
     4081 gtggcagaaa gataaagaac gttttgaacg tgtgcgtaat agcaccctgg aaattgcacg
     4141 tgaacgttat gcaaaattca aacgttgata aggatcctaa ttggtaacga atcagacaat
     4201 tgacggctcg agggagtagc atagggtttg cagaatccct gcttcgtcca tttgacaggc
     4261 acattatgca tcgatgataa gctgtcaaac atgagcagat cctctacgcc ggacgcatcg
     4321 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg
     4381 atggggaaga tcgggctcgc cacttcgggc tcatgagcaa atattttatc tgaggtgctt
     4441 cctcgctc
//
