LOCUS       sequence-423397-        4498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Acidovorax avenae subsp. Avenae 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: pCYP10
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 tgtttagagc tactacttaa
     2101 gggggaaatt gatgactatt caggaagagg caattttaac ttgtcgtgat gccttcgccg
     2161 aactggccag cccggcatgc ctgcatgatc catatccgtt tatgcatcaa ctgcgcaccc
     2221 acgatccggt gcatcgggcc gcgagtggtc tgttcttgct gtctcgtcac gccgatatct
     2281 tttgggttct gaaggcgacc ggtgatgctt ttcgcggacc ggctccgggt gagcttgccc
     2341 gttactatcc gagagcggcg actagtctgt cgcttcaact cttggcttca acactggcaa
     2401 tgaaagaacc tccaacgcat actcgattgc gccgtctgat ctcacgcgac tttaccctgc
     2461 gtcaaatcga tgtccttcgt ccagacattg ttcgcattat cgaagaacgt ttggatggca
     2521 tggcgccggc cttggaacgt ggtgaggccg tcgacttaca tcgggagttc tcacttgcgg
     2581 tgagtatgct gggttttgct cgtctgttcg gtatgcctcg agatgatatg tctcggatag
     2641 ccgccggtat cggtgcaatc ctcgaaggcc tcagtccgcg agcaagcgat gcgcaacttg
     2701 ccgcagcgga tgccgcaagc gcgggtgcac aggcatactt tgaggaactg attgcgtgta
     2761 aacgtgcagc gccgcaggac gacctggtca gcttattggt aggtgcgcac gacggcgacg
     2821 ctgatacgct gtctgatcct gagctcacct cgatgctctg gggtatgtta ctgggcggat
     2881 ttgctaccac ggcggcgacc atggatcatg ccgcgctggc tatgataagc caacctggtg
     2941 cgcagcgttg gctgcaaggt gatgccgccg gtgttaaggc tttcgttgaa gaagtcctcc
     3001 gtcaccaagc gccggccatg ttcagctcaa ttccgcgcat tgcgcaacgc gatatcgaac
     3061 tgggcggggt tgtaattcca aaggatagcg atgttcgcgt gttgatagcg gccggaaacc
     3121 gtgatccgga cgcctttccg gacccggata gatttgatcc ggcccgcttt ctgggcacct
     3181 cgccgggctt tagcactgat ggccaagtga tgctctcttt cgggcatgga atccacttct
     3241 gcgttggcgc acagctggca cgtgtggaag tggcggaggg attgtcacgg ttgcacgctc
     3301 gttttccggc actgagcctg gctggaactc cagttcgcga gccaagcgca tttctgcgca
     3361 cgtttcgcgc gctcccggtg cgcttacaga ctccgggcgg ttaataagga 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
//
