LOCUS       sequence-424233-        4498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Rhizobium meliloti LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4498)
  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 4498)
  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: pCYP56
FEATURES             Location/Qualifiers
     source          1..4498
                     /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(4410..4429)
                     /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 tgtttagggg cgggagggag
     2101 aaaatagaag actggggagg tattttatgc gcgtaggcaa cgatcactgc gatgccattg
     2161 gagtctctgt tgcgccaagt caattagata acctgtcctc ggctatactg cagcagggtg
     2221 gcatggcccg tgtgagtctg cccggtgatg tcgtgacgtg ggctgcaggg ggccatcaga
     2281 cactccgacg aatgttatcc gatcaacggt tcaaccgaga ctggcgtcag tggcgcgcct
     2341 tacaggacgg cgaaatccca gaggaccatc ccctgatcgg catgtgtaaa gtggacaaca
     2401 tggtcactgc acacggtgcg gatcaccgtc gcctgcgcgg attgttaagc cgctcatttg
     2461 cgcccgcgcg cattgctttg ttggctcctc ggattgaaca atgtgcggat cgtctgctgg
     2521 ccgaaatggc gcagcgtggt ggctcggccg atctgatgtg tgagtttgcg gttccgctgc
     2581 caaccaatgt gattgcggaa ctgtttggcc tgccggatga acaacgcgaa gaaattgtgg
     2641 cccttaccta ttccctggca aacaccagcg ccacggccgc cgaagtccga cagactcgcc
     2701 agcgattacc cgaattcttt cgacggctga ttgccttcaa acgtggtcag cttggcgatg
     2761 atctcgcaag cgcactgatc gttgcgcgcg ataatggtga actggtgtca gataccgaac
     2821 tgattgatat gctgtttatg gtactgagcg cggggtttgt taccaccacc ggtgttattg
     2881 gcaacggggt gctggccctg ctgacccatc cgcagcaact gcatctggtg cggtccggcc
     2941 aggtaccttg gtcgcaagct attgaggaaa tccttcgttg gggcagttct gtagctaatt
     3001 taccgtttcg gtacgcaact gaagatgttg aaatcgatgg ttgtacggtc cctagagggg
     3061 acgctgttct catggcattt cacgccgcta accgcgacga aaaggcgttt gggcccggag
     3121 ccgatcgttt cgacgttacg cgtcgtcata acccacatct gtcctttggc aaagggccac
     3181 atgtgtgcct tggcgcggct ctggcacgct tagaattgcg ttgcgctttt acggccctgt
     3241 ttgggcggtt ggaagatttg gcactcgcga tcgcggctga agatgtcgtc tatatgccgt
     3301 cctacgttat ccgttgccct cagcgcttgc cggtgacgtt tcgcccgagt atcgtcccgc
     3361 cgattgaaac gaaagcgctg ggacgcacac gtggggcggc ataataagga tgagctctac
     3421 aaataagcag aggtggttgt gttgcgaaaa aaaaaaaaaa acaccctaac gggtgttttt
     3481 ttttttttgg tgtccccacg tgtggcgctg gagaccgtcc aatggcggcg cgccatcgaa
     3541 tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     3601 gtgaatatga gcccgaaacg tcgtacccag gcagaacgtg caatggaaac ccagggtaaa
     3661 ctgattgcag cagcactggg tgttctgcgt gaaaaaggtt atgcaggttt tcgtattgca
     3721 gatgttccgg gtgcagccgg tgttagccgt ggtgcacaga gccatcattt tccgaccaaa
     3781 ctggaactgc tgctggcaac ctttgaatgg ctgtatgagc agattaccga acgtagccgt
     3841 gcacgtctgg caaaactgaa accggaagat gatgttattc agcagatgct ggatgatgca
     3901 gcagattttt ttctggatga tgattttagc atcggcctgg atctgattgt tgcagcagat
     3961 cgtgatccgg cactgcgtga aggtattctg cgtaccgttg aacgtaatcg ttttgttgtt
     4021 gaagatatgt ggctgggtgt gctggtgagc cgtggtctga gccgtgatga tgccgaagat
     4081 attctgtggc tgatttttaa cagcgttcgt ggtctgacag ttcgtagcct gtggcagaaa
     4141 gataaagaac gttttgaacg tgtgcgtaat agcaccctgg aaattgcacg tgaacgttat
     4201 gcaaaattca aacgttgata aggatcctaa ttggtaacga atcagacaat tgacggctcg
     4261 agggagtagc atagggtttg cagaatccct gcttcgtcca tttgacaggc acattatgca
     4321 tcgatgataa gctgtcaaac atgagcagat cctctacgcc ggacgcatcg tggccggcat
     4381 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
     4441 tcgggctcgc cacttcgggc tcatgagcaa atattttatc tgaggtgctt cctcgctc
//
