LOCUS       sequence-422120-        6358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Priestia megaterium LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6358)
  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 6358)
  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: pCYP64
FEATURES             Location/Qualifiers
     source          1..6358
                     /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(6270..6289)
                     /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 tgtttaatct ccgctctatc
     2101 cccaaggacc tatatatgac aattaaagaa atgcctcagc caaaaacgtt tggagagctt
     2161 aaaaatttac cgttattaaa cacagataaa ccggttcaag ctttgatgaa aattgcggat
     2221 gaattaggag aaatctttaa attcgaggcg cctggtcgtg taacgcgcta cttatcaagt
     2281 cagcgtctaa ttaaagaagc atgcgatgaa tcacgctttg ataaaaactt aagtcaagcg
     2341 cttaaattcg cgcgtgattt tggcggagac gggttagtta caagctggac gcatgaaaaa
     2401 aattggaaaa aagcgcataa tatcttactt ccaagcttta gtcagcaggc aatgaaaggc
     2461 tatcatgcga tgatggtcga tatcgccgtg cagcttgttc aaaagtggga gcgtctaaat
     2521 gcagatgagc atattgaagt atcggaagac atgacacgtt taacgcttga tacaattggt
     2581 ctttgcggct ttaactatcg ctttaacagc ttttaccgag atcagcctca tccatttatt
     2641 ataagtgcgg tccgtgcact ggatgaagta atgaacaagc tgcagcgagc aaatccagac
     2701 gacccagctt atgatgaaaa caagcgccag tgtcaagaag atatcaaggt gatgaacgac
     2761 ctagtagata aaattattgc agatcgcaaa gcaaggggtg aacaaagcga tgatttatta
     2821 acgcagatgc taaacggaaa agatccagaa acgggtgagc cgcttgatga cgggaacatt
     2881 agctatcaaa ttattacatt cttaattgcg ggacacgaaa caacaagtgg tcttttatca
     2941 tttgcgctgt atttcttagt gaaaaatcca catgtattac aaaaagtagc agaagaagca
     3001 gcacgagttc tagtagatcc tgttccaagc tacaaacaag tcaaacagct taaatatgtc
     3061 ggcatggtct taaacgaagc gctgcgctta tggccaactg cgcctgcgtt ttccctatat
     3121 gcaaaagaag atacggtgct tggaggagaa tatcctttag aaaaaggcga cgaagtaatg
     3181 gttctgattc ctcagcttca ccgtgataaa acaatttggg gagacgatgt ggaggagttc
     3241 cgtccagagc gttttgaaaa tccaagtgcg attccgcagc atgcgtttaa accgtttgga
     3301 aacggtcagc gtgcgtgtat cggtcagcag ttcgctcttc atgaagcaac gctggtactt
     3361 ggtatgatgc taaaacactt tgactttgaa gatcatacaa actacgagct cgatattaaa
     3421 gaaactttaa cgttaaaacc tgaaggcttt gtggtaaaag caaaatcgaa aaaaattccg
     3481 cttggcggta ttccttcacc tagcactgaa cagtctgcta aaaaagtacg caaaaaggca
     3541 gaaaacgctc ataatacgcc gctgcttgtg ctatacggtt caaatatggg aacagctgaa
     3601 ggaacggcgc gtgatttagc agatattgca atgagcaaag gatttgcacc gcaggtcgca
     3661 acgcttgatt cacacgccgg aaatcttccg cgcgaaggag ctgtattaat tgtaacggcg
     3721 tcttataacg gtcatccgcc tgataacgca aagcaatttg tcgactggtt agaccaagcg
     3781 tctgctgatg aagtaaaagg cgttcgctac tccgtatttg gatgcggcga taaaaactgg
     3841 gctactacgt atcaaaaagt gcctgctttt atcgatgaaa cgcttgccgc taaaggggca
     3901 gaaaacatcg ctgaccgcgg tgaagcagat gcaagcgacg actttgaagg cacatatgaa
     3961 gaatggcgtg aacatatgtg gagtgacgta gcagcctact ttaacctcga cattgaaaac
     4021 agtgaagata ataaatctac tctttcactt caatttgtcg acagcgccgc ggatatgccg
     4081 cttgcgaaaa tgcacggtgc gttttcaacg aacgtcgtag caagcaaaga acttcaacag
     4141 ccaggcagtg cacgaagcac gcgacatctt gaaattgaac ttccaaaaga agcttcttat
     4201 caagaaggag atcatttagg tgttattcct cgcaactatg aaggaatagt aaaccgtgta
     4261 acagcaaggt tcggcctaga tgcatcacag caaatccgtc tggaagcaga agaagaaaaa
     4321 ttagctcatt tgccactcgc taaaacagta tccgtagaag agcttctgca atacgtggag
     4381 cttcaagatc ctgttacgcg cacgcagctt cgcgcaatgg ctgctaaaac ggtctgcccg
     4441 ccgcataaag tagagcttga agccttgctt gaaaagcaag cctacaaaga acaagtgctg
     4501 gcaaaacgtt taacaatgct tgaactgctt gaaaaatacc cggcgtgtga aatgaaattc
     4561 agcgaattta tcgcccttct gccaagcata cgcccgcgct attactcgat ttcttcatca
     4621 cctcgtgtcg atgaaaaaca agcaagcatc acggtcagcg ttgtctcagg agaagcgtgg
     4681 agcggatatg gagaatataa aggaattgcg tcgaactatc ttgccgagct gcaagaagga
     4741 gatacgatta cgtgctttat ttccacaccg cagtcagaat ttacgctgcc aaaagaccct
     4801 gaaacgccgc ttatcatggt cggaccggga acaggcgtcg cgccgtttag aggctttgtg
     4861 caggcgcgca aacagctaaa agaacaagga cagtcacttg gagaagcaca tttatacttc
     4921 ggctgccgtt cacctcatga agactatctg tatcaagaag agcttgaaaa cgcccaaagc
     4981 gaaggcatca ttacgcttca taccgctttt tctcgcatgc caaatcagcc gaaaacatac
     5041 gttcagcacg taatggaaca agacggcaag aaattgattg aacttcttga tcaaggagcg
     5101 cacttctata tttgcggaga cggaagccaa atggcacctg ccgttgaagc aacgcttatg
     5161 aaaagctatg ctgacgttca ccaagtgagt gaagcagacg ctcgcttatg gctgcagcag
     5221 ctagaagaaa aaggccgata cgcaaaagac gtgtgggctg ggtaataagg atgagctcta
     5281 caaataagca gaggtggttg tgttgcgaaa aaaaaaaaaa acaccctaac gggtgttttt
     5341 ttttttttgg tgtccccacg tgtggcgctg gagaccgtcc aatggcggcg cgccatcgaa
     5401 tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     5461 gtgaatatga gcccgaaacg tcgtacccag gcagaacgtg caatggaaac ccagggtaaa
     5521 ctgattgcag cagcactggg tgttctgcgt gaaaaaggtt atgcaggttt tcgtattgca
     5581 gatgttccgg gtgcagccgg tgttagccgt ggtgcacaga gccatcattt tccgaccaaa
     5641 ctggaactgc tgctggcaac ctttgaatgg ctgtatgagc agattaccga acgtagccgt
     5701 gcacgtctgg caaaactgaa accggaagat gatgttattc agcagatgct ggatgatgca
     5761 gcagattttt ttctggatga tgattttagc atcggcctgg atctgattgt tgcagcagat
     5821 cgtgatccgg cactgcgtga aggtattctg cgtaccgttg aacgtaatcg ttttgttgtt
     5881 gaagatatgt ggctgggtgt gctggtgagc cgtggtctga gccgtgatga tgccgaagat
     5941 attctgtggc tgatttttaa cagcgttcgt ggtctgacag ttcgtagcct gtggcagaaa
     6001 gataaagaac gttttgaacg tgtgcgtaat agcaccctgg aaattgcacg tgaacgttat
     6061 gcaaaattca aacgttgata aggatcctaa ttggtaacga atcagacaat tgacggctcg
     6121 agggagtagc atagggtttg cagaatccct gcttcgtcca tttgacaggc acattatgca
     6181 tcgatgataa gctgtcaaac atgagcagat cctctacgcc ggacgcatcg tggccggcat
     6241 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
     6301 tcgggctcgc cacttcgggc tcatgagcaa atattttatc tgaggtgctt cctcgctc
//
