LOCUS       sequence-423600-        4562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Streptomyces clavuligerus LRD-modifying cytochrome 
            P450 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4562)
  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 4562)
  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: pCYP63
FEATURES             Location/Qualifiers
     source          1..4562
                     /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)"
     RBS             2100..2108
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(4474..4493)
                     /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 tgtttaggca ctgtacacat
     2101 aaggaggtat cttttatgac cggcggcacc atcgcgcgcg ttccgggagc attgccgtta
     2161 ataggtcaca cgggcgcgtt cctgaaatcc ccgctggaat ttctgaaagg tctgtccgcg
     2221 catggcgaat tggttgaact gcgcttcggc agaaaacgag ttttagttgt atgcgaccct
     2281 gaccttacgc aggagatgtt acgccgtgat cgggtgtatg ataaaggggg ccccatttac
     2341 gatcgcatgc gcgaggcggc cggaaatggc ttggcaactt gcccgagtcg ggaccaccga
     2401 cggcagcgcc ggactatcca accggcgttt catcatagtc gcctgccggg ttatgcgcgc
     2461 accatgactg aagtaatcga tgaggcagtt cgcggttggg acggcccgga ggtagacctc
     2521 tgtcacgaaa cgcgcagcat tgcgagcagc gtcctgctga caactctgtt tggtaccgcg
     2581 tttgctccgg aggcccgtga cgcgttagcg cgcgatctgg aaaccatcct gcgaggcggc
     2641 cttcgccgcg ccgtactgcc tgaagctctg tcggccctgc ctacccccgc gaacttacgt
     2701 tatcagcgtg cgcgccgccg tgccagaggt atgatggctg acctcatccg tgcagtacgc
     2761 gcaggcgacg gatctggtga ttcactgttc gccaccctgg taaatcaaca cggcgcggga
     2821 acgggcgacc tgccaatgac agatgaggaa ttagttgatc aggctattac catgtacgtc
     2881 gcgggtacgg aaacaaccgc aggagtggta tgttgggcac tgcatctggc agcacagcat
     2941 ccgcaagccg gccgtcgcct ggctgatgaa tgccgtagcg tgctcggtca tcgcgcggca
     3001 acctgggagg atcttcctca tctgccgtac actcgtcaga cgctcttaga agcgctgcgc
     3061 ctcaaccctc cagggtggct tatcacccgg ctgaccacga cggatgatgc acgattaggc
     3121 gaccatgcgg taccacgggg tagtaccatc gtctacagcg catacctgct ctcccggctg
     3181 ccggcgcgat acgaagaacc tgaacatttt gcaccggagc gttgggcgcc gggtaccatg
     3241 ggtgcaaaag ttccagacgg agtattcggt ggtggtgcgc gtaaatgcat cggagatgaa
     3301 tatgcgctga ttgaaggtgt cctgatgctc gcaacaatca ccgcccgttg ggacgttcgt
     3361 actgccccgg atgctccaaa gcgcccgcat ctgcccgcgc ttacgctgaa tccgacccgt
     3421 ttgatggcga ctgttacccc agcagcggcg ccagatggct cgcgctaata aggatgagct
     3481 ctacaaataa gcagaggtgg ttgtgttgcg aaaaaaaaaa aaaaacaccc taacgggtgt
     3541 tttttttttt ttggtgtccc cacgtgtggc gctggagacc gtccaatggc ggcgcgccat
     3601 cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     3661 ggtggtgaat atgagcccga aacgtcgtac ccaggcagaa cgtgcaatgg aaacccaggg
     3721 taaactgatt gcagcagcac tgggtgttct gcgtgaaaaa ggttatgcag gttttcgtat
     3781 tgcagatgtt ccgggtgcag ccggtgttag ccgtggtgca cagagccatc attttccgac
     3841 caaactggaa ctgctgctgg caacctttga atggctgtat gagcagatta ccgaacgtag
     3901 ccgtgcacgt ctggcaaaac tgaaaccgga agatgatgtt attcagcaga tgctggatga
     3961 tgcagcagat ttttttctgg atgatgattt tagcatcggc ctggatctga ttgttgcagc
     4021 agatcgtgat ccggcactgc gtgaaggtat tctgcgtacc gttgaacgta atcgttttgt
     4081 tgttgaagat atgtggctgg gtgtgctggt gagccgtggt ctgagccgtg atgatgccga
     4141 agatattctg tggctgattt ttaacagcgt tcgtggtctg acagttcgta gcctgtggca
     4201 gaaagataaa gaacgttttg aacgtgtgcg taatagcacc ctggaaattg cacgtgaacg
     4261 ttatgcaaaa ttcaaacgtt gataaggatc ctaattggta acgaatcaga caattgacgg
     4321 ctcgagggag tagcataggg tttgcagaat ccctgcttcg tccatttgac aggcacatta
     4381 tgcatcgatg ataagctgtc aaacatgagc agatcctcta cgccggacgc atcgtggccg
     4441 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
     4501 aagatcgggc tcgccacttc gggctcatga gcaaatattt tatctgaggt gcttcctcgc
     4561 tc
//
