LOCUS       sequence-423270-        4449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Pseudonocardia sp. Ae717_Ps2 LRD-modifying cytochrome 
            P450 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4449)
  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 4449)
  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: pCYP43
FEATURES             Location/Qualifiers
     source          1..4449
                     /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(4361..4380)
                     /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 tgtttagcga tactgaagga
     2101 taacggagga gcaattatgt ctgaaacaac aaccacgagc gccggtagcg aggaattcct
     2161 ggaaacctac tggcacgatc cccacacggc tttagctggg gcgcgccgcg attgtcctgt
     2221 gcgtgaagtg actttcgaaa ccggtccgac atggctggta acccgttacg cggatgtgcg
     2281 tgctggcctt gcagatgagc gactggcaaa agactggcgc gccagcttac ctccagctga
     2341 gcgagcagcg gcccctccgg gcttaccggc acctatgagc cacatgctta cgagtctgga
     2401 cccgccagaa cacactcgcc tgcgtcggtt ggtcacccag gcctttaccg cgcgtcgtat
     2461 cgctgccctg cgtcctcgcg tcgagaaatt ggcacatgat ctgttggacg ctctgccggt
     2521 cgatgaaccg gtggatttgg tcggaagata cgcgattccg ttacctatgg aagtcatctg
     2581 cgaattaatt ggcgtaccgg agcttgatcg agaggatttc gctcattggt ctacggtgct
     2641 tattgacgag tgtccggaac ctgaattggt gcgtgcaagc aatcagatgg cagattacct
     2701 gggtggtctg gtagccgctc gtcggcgcga tccggacgaa gctctgttga gcggcttgat
     2761 tgcggcctct gaagatggtg aacagctgtc cgacatcgaa atcgttgcca tggcgatgtt
     2821 attgctgatg gccggccacg aaaccaccgc tgtattcgtg actaactcag ttcgagtgct
     2881 cctgacggaa ccggatgcac gtcgtcgtct tgcgagccct gagactgccc cagcgctggt
     2941 ggaagaactg ctgagatggg catctccggc tctgaatgcg agcctgcgct ttgcgacgga
     3001 agatatggaa attggtggaa ccgcgatacc gcgtggcgcg agcgttacac tgtcaaccgg
     3061 tgcggccaat cgtgatccat cgcgattcga cgacccaacc cgcctcgacc ccgatcgaga
     3121 taccgctggc catgtcgcct tcggtcacgg aatacatcgc tgtttaggag cggcgctggt
     3181 tcgccttgaa gcagacgtag ccttgaccgc cctgttcggg cgcttccccc aactgcgagc
     3241 cactgccgcg gaagatgaac tgacctttcg gcgttcggtg cacgtccatg cgccgcgtac
     3301 cttcccagta ctgctggggc caactccgga tcaggaacag gagcgtagag cgtaataagg
     3361 atgagctcta caaataagca gaggtggttg tgttgcgaaa aaaaaaaaaa aacaccctaa
     3421 cgggtgtttt tttttttttg gtgtccccac gtgtggcgct ggagaccgtc caatggcggc
     3481 gcgccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     3541 aattcagggt ggtgaatatg agcccgaaac gtcgtaccca ggcagaacgt gcaatggaaa
     3601 cccagggtaa actgattgca gcagcactgg gtgttctgcg tgaaaaaggt tatgcaggtt
     3661 ttcgtattgc agatgttccg ggtgcagccg gtgttagccg tggtgcacag agccatcatt
     3721 ttccgaccaa actggaactg ctgctggcaa cctttgaatg gctgtatgag cagattaccg
     3781 aacgtagccg tgcacgtctg gcaaaactga aaccggaaga tgatgttatt cagcagatgc
     3841 tggatgatgc agcagatttt tttctggatg atgattttag catcggcctg gatctgattg
     3901 ttgcagcaga tcgtgatccg gcactgcgtg aaggtattct gcgtaccgtt gaacgtaatc
     3961 gttttgttgt tgaagatatg tggctgggtg tgctggtgag ccgtggtctg agccgtgatg
     4021 atgccgaaga tattctgtgg ctgattttta acagcgttcg tggtctgaca gttcgtagcc
     4081 tgtggcagaa agataaagaa cgttttgaac gtgtgcgtaa tagcaccctg gaaattgcac
     4141 gtgaacgtta tgcaaaattc aaacgttgat aaggatccta attggtaacg aatcagacaa
     4201 ttgacggctc gagggagtag catagggttt gcagaatccc tgcttcgtcc atttgacagg
     4261 cacattatgc atcgatgata agctgtcaaa catgagcaga tcctctacgc cggacgcatc
     4321 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     4381 gatggggaag atcgggctcg ccacttcggg ctcatgagca aatattttat ctgaggtgct
     4441 tcctcgctc
//
