LOCUS       sequence-423311-        4413 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 4413)
  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 4413)
  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: pCYP13
FEATURES             Location/Qualifiers
     source          1..4413
                     /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(4325..4344)
                     /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 tgtttaggag tcaagaagaa
     2101 aagaaaggag gtaaaaatga gcgaaacacc gctgccgacg ttgcctatgt ggcgcgctca
     2161 ccacattgaa ccgagccctg atatgctggc gattcaggcc cgtggtccta ttcatcgggt
     2221 gcggttcccc agtggccatg aaggctggtg ggttactggc tatgatgaag ccaaagcggt
     2281 ccttaccaat gcggcgtttc gaccagccgg catgccgccg ccagcgttca cgcctgatag
     2341 cgtgattctc ggttccccgg gttggctggt aagccatgag ggtaccgaac atgcgcgtct
     2401 gcgcgcgatt gtggcaccgg ccttctcaaa ccgccgtgtg gaactgttag cgggacaggt
     2461 tgaagcgatt accgagcgcc tgttcgatac tcttgccgcc caaccacagc cggccgattt
     2521 aagacgccat ctttcttttc cgctgccggc gatggtgatt agcgccttga tgggggtgcc
     2581 gtacgaagat cacgcgttct ttgccggcct gagcgatgaa gtgatgacgc atcagcacga
     2641 gtcgggtcca cgctccgcgt cacgacttgc ctgggaagaa ctgcgcggct atatccgcga
     2701 caaaatgcgc gccaagcgtg aagccccagg cgacgatttg ttaacggatc tcctcaccgc
     2761 agtggatcag ggtaaagcta ctgaggaaga agccatcggc cttgcagcgg gtatgcttgt
     2821 ggccggtcat gaatctaccg tggctcaaat cgaattcggg ctgttagcgc tgttccgtca
     2881 ccctcggcaa agagcccgcc tggcagggga cccgagtctg gttgacaatg ccgttgaaga
     2941 gatcctgcgt atgtatccgc cgggtgccgg atgggacgga atcatgcgtt atccgcgtac
     3001 cgacgtgacg attgccgggg tccatatccc tgccgaatcc aaagtgttgg ttggtctgcc
     3061 cgcgacatct ttcgatccta gacatttttc cgatccccaa gcatttgata taggtcgcga
     3121 aggcaagcca catcttgcgt tctcttacgg tccgcattat tgtattggag ttgccctggc
     3181 ccgcttggaa ctgcgtgcgg tgttcggttc tatcttccag cgttttccta gccttcagtt
     3241 agcagtagca cctgaagcat tgcggttacg taaagagatc ataactggtg ggttcgaggc
     3301 ctttccggtg cgttggtaat aaggatgagc tctacaaata agcagaggtg gttgtgttgc
     3361 gaaaaaaaaa aaaaaacacc ctaacgggtg tttttttttt tttggtgtcc ccacgtgtgg
     3421 cgctggagac cgtccaatgg cggcgcgcca tcgaatggtg caaaaccttt cgcggtatgg
     3481 catgatagcg cccggaagag agtcaattca gggtggtgaa tatgagcccg aaacgtcgta
     3541 cccaggcaga acgtgcaatg gaaacccagg gtaaactgat tgcagcagca ctgggtgttc
     3601 tgcgtgaaaa aggttatgca ggttttcgta ttgcagatgt tccgggtgca gccggtgtta
     3661 gccgtggtgc acagagccat cattttccga ccaaactgga actgctgctg gcaacctttg
     3721 aatggctgta tgagcagatt accgaacgta gccgtgcacg tctggcaaaa ctgaaaccgg
     3781 aagatgatgt tattcagcag atgctggatg atgcagcaga tttttttctg gatgatgatt
     3841 ttagcatcgg cctggatctg attgttgcag cagatcgtga tccggcactg cgtgaaggta
     3901 ttctgcgtac cgttgaacgt aatcgttttg ttgttgaaga tatgtggctg ggtgtgctgg
     3961 tgagccgtgg tctgagccgt gatgatgccg aagatattct gtggctgatt tttaacagcg
     4021 ttcgtggtct gacagttcgt agcctgtggc agaaagataa agaacgtttt gaacgtgtgc
     4081 gtaatagcac cctggaaatt gcacgtgaac gttatgcaaa attcaaacgt tgataaggat
     4141 cctaattggt aacgaatcag acaattgacg gctcgaggga gtagcatagg gtttgcagaa
     4201 tccctgcttc gtccatttga caggcacatt atgcatcgat gataagctgt caaacatgag
     4261 cagatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
     4321 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
     4381 agcaaatatt ttatctgagg tgcttcctcg ctc
//
