LOCUS       sequence-423391-        4495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Rhizobium mesoamericanum LRD-modifying cytochrome P450
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4495)
  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 4495)
  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: pCYP11
FEATURES             Location/Qualifiers
     source          1..4495
                     /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(4407..4426)
                     /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 tgtttaggca aaaaaaagac
     2101 aaataaggag gaccgaaatg cgtgaaggcc atgatgtcac tcgtgcgaag caaggactga
     2161 tctttaagga aaaaccttca cttggaaagc gcaatacgat tatgcccgag cattccttac
     2221 cgacgctgcc gatgtggcga gtagatcaca tcgaaccctc accggaaatg ctcgcactgc
     2281 gtggtgaggg tccaattcat cgtgtgcggt ttcctagcgg tcatgaaggc tggtgggtta
     2341 cgggttatga cgaagcaaaa gcagttttgt cagacgctgc atttcgaccg gcgggtacgc
     2401 cgacagcctt cacgccggac agtgtaatgt taggcagccc aggttggctg gtgtctcatg
     2461 agggtgatga acacagccgg ctgcgcgcaa ttattgcccc ggcattctct gaccttcgta
     2521 cggatctgct ggctcaacaa gtggaaagta ttgccgaaca gctgtttgca accctttcga
     2581 tccagccaca gccggcggat ctgcgtaaat atctgagttt tcccctgcca gctatggtga
     2641 taagcgccct gatgggcgtt cgctacgaag atcatgcatt ctttgcaggg ctcagcgacg
     2701 aagttatgac gcaccagcac gaaacgggcc cgagaagcgc ttcccggctg gcttgggaag
     2761 aattacgtgt ttacatacgg gatcgcatgc gcgagaaacg ccgtgatccg ggggataatc
     2821 tgttaacgga tcttctgctg gcggtggatc aaggtaaagc gagcgaagaa gaggctatcg
     2881 gcctggcggc tggtatgctg gtggccggcc atgaaagcac ggcagcacaa atagaatgcg
     2941 gcctgttagc gatgttccgc cattcccagc aacgcgaacg cttgatcgcc gatccgagtc
     3001 tgctcgataa agccgttgaa gagattctta gaatgtaccc accgggagcg ggctgggacg
     3061 ggataatgcg ctatccacgt tgcgacgtta ccatcgccgg ggtacacatt cctgcggaat
     3121 caaaagtgct ggtcggcctt ccagcgacca gcttcgatct gcgccacttc gacaaaccgg
     3181 aggttttcga tattgcacga gagggaaatc cgcacctggc gttctcttat ggcccgcact
     3241 actgcatcgg gggagcgctc gcacgtctgg aactgaaggc cgtttttggc tccatcttcc
     3301 aacgctttcc ggggctgcgt ctggcggtgg aaccggaaga actggagtta cgtaaagaaa
     3361 tcatcaccgg gggttttgag gagatgccgg tgctgtggta ataaggatga gctctacaaa
     3421 taagcagagg tggttgtgtt gcgaaaaaaa aaaaaaaaca ccctaacggg tgtttttttt
     3481 tttttggtgt ccccacgtgt ggcgctggag accgtccaat ggcggcgcgc catcgaatgg
     3541 tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     3601 aatatgagcc cgaaacgtcg tacccaggca gaacgtgcaa tggaaaccca gggtaaactg
     3661 attgcagcag cactgggtgt tctgcgtgaa aaaggttatg caggttttcg tattgcagat
     3721 gttccgggtg cagccggtgt tagccgtggt gcacagagcc atcattttcc gaccaaactg
     3781 gaactgctgc tggcaacctt tgaatggctg tatgagcaga ttaccgaacg tagccgtgca
     3841 cgtctggcaa aactgaaacc ggaagatgat gttattcagc agatgctgga tgatgcagca
     3901 gatttttttc tggatgatga ttttagcatc ggcctggatc tgattgttgc agcagatcgt
     3961 gatccggcac tgcgtgaagg tattctgcgt accgttgaac gtaatcgttt tgttgttgaa
     4021 gatatgtggc tgggtgtgct ggtgagccgt ggtctgagcc gtgatgatgc cgaagatatt
     4081 ctgtggctga tttttaacag cgttcgtggt ctgacagttc gtagcctgtg gcagaaagat
     4141 aaagaacgtt ttgaacgtgt gcgtaatagc accctggaaa ttgcacgtga acgttatgca
     4201 aaattcaaac gttgataagg atcctaattg gtaacgaatc agacaattga cggctcgagg
     4261 gagtagcata gggtttgcag aatccctgct tcgtccattt gacaggcaca ttatgcatcg
     4321 atgataagct gtcaaacatg agcagatcct ctacgccgga cgcatcgtgg ccggcatcac
     4381 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     4441 ggctcgccac ttcgggctca tgagcaaata ttttatctga ggtgcttcct cgctc
//
