LOCUS       sequence-422115-        4550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Sinorhizobium fredii LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4550)
  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 4550)
  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: pCYP02
FEATURES             Location/Qualifiers
     source          1..4550
                     /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(4462..4481)
                     /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 tgtttaggcc aagaacaccc
     2101 gggaggttat aatgaatgtc ctgctgaatc cgttgaatcg ccgccatcgt ctgcgatatg
     2161 acattccagt aatgcctggt gcgttccccc ttgtcgggca tttgccggcg attgtatgtg
     2221 atctgccacg cttactccgc cgggcagaac gtactctcgg tagtcatttt tggctggatt
     2281 tcggtccggc aggtcatttg atgacctgtg ttgatccgca cgcgttcgcg ctgttgcgcc
     2341 acaaagatgt tagtagcgcg ttgattgagg aaatcgctcc cgaactgctc ggcggcacct
     2401 tggtagcaca agatggcggt gcccatcgcc aggcgcgtga tgcaatcaaa gcagccttcc
     2461 tgccagaagg tttaacccag gcggggattg gcgatctttt tgctccggtg attcgtgcgc
     2521 gtgtgcaagc gtggcgtgat cgtggcgacg taaccatact gcctgaaaca ggcgacctga
     2581 tgcttaaact gatttttacc ctgatgggcg ttcctgcaca ggacctgccg ggttggcacc
     2641 gtaaatatcg tcaacttctg caacttattg tcgcccctag tgtggatctg ccaggattgc
     2701 cacttcgccg tggccgcgca gcacgtgatt ggattgacgc gcaactgcgc caattcgttc
     2761 gcgacgcgcg tgcacatgcc gcccggacag gtctgattaa tgatatggta agcgcgttcg
     2821 atcgctccga tgatgccctg tctgacgatc tgttggtggc taatattcgt ctgctgttgc
     2881 ttgcaggaca tgataccacg gcgagcacaa tggcatggat ggtgattgaa ctggccagac
     2941 aaccaatgtt gtgggatgcg ctggttgaag aagcccagcg tgtgggagcg gttccgaccc
     3001 gtcatgccga tttagaacaa tgtcctgtgg cggaagccct gttccgcgaa acactgcgcg
     3061 ttcacccggc gacgacttta ctgccacgcc gcgcattaca agaactccag ttgggccaac
     3121 gcagaattcc tgcgggcaca catctgtgca taccgttact gcattttagc acctctgctc
     3181 tgttacatga agccccggac cagtttcgcc tggcacgctg gttgcagcga accgagccga
     3241 tacggccggt tgatatgctg cagtttggaa ccggcccgca tgtgtgtatt ggttaccacc
     3301 tggtctggct tgagcttgtc cagttcagta tagccctggc gctgacaatg cataaagctg
     3361 gagtgcgccc gctcctgctt agcggcgttg aaaaaggtcg cagatactac ccgacggcgc
     3421 atccgagcat gacgattcgt atcggatttt cgtaataagg atgagctcta caaataagca
     3481 gaggtggttg tgttgcgaaa aaaaaaaaaa aaacacccta acgggtgttt tttttttttt
     3541 ggtgtcccca cgtgtggcgc tggagaccgt ccaatggcgg cgcgccatcg aatggtgcaa
     3601 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatat
     3661 gagcccgaaa cgtcgtaccc aggcagaacg tgcaatggaa acccagggta aactgattgc
     3721 agcagcactg ggtgttctgc gtgaaaaagg ttatgcaggt tttcgtattg cagatgttcc
     3781 gggtgcagcc ggtgttagcc gtggtgcaca gagccatcat tttccgacca aactggaact
     3841 gctgctggca acctttgaat ggctgtatga gcagattacc gaacgtagcc gtgcacgtct
     3901 ggcaaaactg aaaccggaag atgatgttat tcagcagatg ctggatgatg cagcagattt
     3961 ttttctggat gatgatttta gcatcggcct ggatctgatt gttgcagcag atcgtgatcc
     4021 ggcactgcgt gaaggtattc tgcgtaccgt tgaacgtaat cgttttgttg ttgaagatat
     4081 gtggctgggt gtgctggtga gccgtggtct gagccgtgat gatgccgaag atattctgtg
     4141 gctgattttt aacagcgttc gtggtctgac agttcgtagc ctgtggcaga aagataaaga
     4201 acgttttgaa cgtgtgcgta atagcaccct ggaaattgca cgtgaacgtt atgcaaaatt
     4261 caaacgttga taaggatcct aattggtaac gaatcagaca attgacggct cgagggagta
     4321 gcatagggtt tgcagaatcc ctgcttcgtc catttgacag gcacattatg catcgatgat
     4381 aagctgtcaa acatgagcag atcctctacg ccggacgcat cgtggccggc atcaccggcg
     4441 ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc
     4501 gccacttcgg gctcatgagc aaatatttta tctgaggtgc ttcctcgctc
//
