LOCUS       sequence-423271-        4459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Prauserella muralis LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4459)
  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 4459)
  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: pCYP32
FEATURES             Location/Qualifiers
     source          1..4459
                     /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(4371..4390)
                     /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 tgtttatagg ccatttgcac
     2101 gaatttaagg agagtttttt atgaaccgta cggaactgcg tatggcgttg cgtcccctgg
     2161 cctttttgga cgcgcatgcc cactcgcccc atcgttatgt agagctgagt caagaaccag
     2221 ccccgaaact gttggtctgg gacccggaca ctgtagattg gattttccgt tcagaccact
     2281 cgctggggca cccgggttca cgcgcaatgc gtcccgtgct ggggtctaaa agcctgttat
     2341 ggaccgatgg ccctcgctat gcggcgtata gacgtgtgct gggtccgcca ctgggtacac
     2401 ggcgtttggg tgattaccat gatttaatta cgggaattgc gggccgtgca atcgacgcgc
     2461 ttacccctgg cacagtcttt accctggctg attggacgcg ccaggtaact ctgaaagtag
     2521 cggcaaccat tgtattaggt cgtgcggata ccgcggtgct cggacagttt accgcgtgga
     2581 tcgaccgtgc acttgcgtcg gtgccccgca cctttgcata tcgccttttg cgcgggggcc
     2641 ttccgactag ttcccgtgaa ctggatcgtg ctctcgtaac gcaagcgcgc gctggcgtta
     2701 gcggtccaca ggcaacattg gcatctttac ttctcagcga tgactccccg gtcggcgcga
     2761 tcgatgatga tgaactgcgc gatcaaatcg tgtctctgtt attcgccggc catgaaacca
     2821 ccgcgtcggc tacggcgtgg acgctgttct ggctggatag acacgatgat gtgcgtcatg
     2881 acgtgctggc tgagctgagt agcacctctg atagtggtgc agatgcaaaa gccgtgcctt
     2941 tactgcaagc gtgtgtccag gaggccttac gcattacccc tccggtgcca gcagctggga
     3001 atcgtcgact cccgggagat acagaattgc tgggtcgccg cttgccagcc ggctcggtcg
     3061 ttgcgccaag catctactta gcgcatcggc aaccggacag ctttccgggt ccgcgccgtt
     3121 ttgatccggc acgatttctg ggacgccgcc agccgccgca gcactatctg ccgtttggtg
     3181 gcggtgcccg tcactgcctg ggtagccagc tgggtcagct ggaagtgcgc gtgatcgctg
     3241 ccgccctgct gcgtcgacgt tcactgcgtt gtatcgatcc tcacgccggt gtaccggcct
     3301 tacgcggaca tgcaatggcc ccggctcgcg cgttgcggat ggaggtaaca gcatgtcatg
     3361 attaataagg atgagctcta caaataagca gaggtggttg tgttgcgaaa aaaaaaaaaa
     3421 aacaccctaa cgggtgtttt tttttttttg gtgtccccac gtgtggcgct ggagaccgtc
     3481 caatggcggc gcgccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg
     3541 gaagagagtc aattcagggt ggtgaatatg agcccgaaac gtcgtaccca ggcagaacgt
     3601 gcaatggaaa cccagggtaa actgattgca gcagcactgg gtgttctgcg tgaaaaaggt
     3661 tatgcaggtt ttcgtattgc agatgttccg ggtgcagccg gtgttagccg tggtgcacag
     3721 agccatcatt ttccgaccaa actggaactg ctgctggcaa cctttgaatg gctgtatgag
     3781 cagattaccg aacgtagccg tgcacgtctg gcaaaactga aaccggaaga tgatgttatt
     3841 cagcagatgc tggatgatgc agcagatttt tttctggatg atgattttag catcggcctg
     3901 gatctgattg ttgcagcaga tcgtgatccg gcactgcgtg aaggtattct gcgtaccgtt
     3961 gaacgtaatc gttttgttgt tgaagatatg tggctgggtg tgctggtgag ccgtggtctg
     4021 agccgtgatg atgccgaaga tattctgtgg ctgattttta acagcgttcg tggtctgaca
     4081 gttcgtagcc tgtggcagaa agataaagaa cgttttgaac gtgtgcgtaa tagcaccctg
     4141 gaaattgcac gtgaacgtta tgcaaaattc aaacgttgat aaggatccta attggtaacg
     4201 aatcagacaa ttgacggctc gagggagtag catagggttt gcagaatccc tgcttcgtcc
     4261 atttgacagg cacattatgc atcgatgata agctgtcaaa catgagcaga tcctctacgc
     4321 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
     4381 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagca aatattttat
     4441 ctgaggtgct tcctcgctc
//
