LOCUS       sequence-423267-        4394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Saccharopolyspora antimicrobica LRD-modifying 
            cytochrome P450 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4394)
  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 4394)
  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: pCYP35
FEATURES             Location/Qualifiers
     source          1..4394
                     /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             2104..2112
                     /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(4306..4325)
                     /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 tgtttaggtc ccgagagaga
     2101 ttataaggag gttctatgac tgcagcagag cagcagttct gtccgtttga cgtgggcgag
     2161 gagtttcgtc gcctccgcga ggacgacccg attagccgaa tcgagctgcc cagcggtact
     2221 ccggcgtggt tcgtcgcacg ccacgccgac gcaagacatg tgttgaccga tgaacgttta
     2281 tcttcagtag atgcaagtcg tctgatgcgt ccaggtgccg gggatgatca ggtagcgccg
     2341 cctggtccgg gtgtgctgct gtttcacgat ccgccggaac acacgcgcct ccgacgcatg
     2401 ctggcaggca attttacggt tagacgcatg aaccaattac gtcctatggt ggagtcaatc
     2461 gtcaacgcac agttagacga aatggcgcgc gcgggcagct cgacggactt tgtgcgttcc
     2521 ttcgcgctcc cggtgccctc cctggtaatt tgtgaattgc tcggggttcc gtatgctgat
     2581 cgcgctgaat tccaacggcg taccgcggct caggtggact tagctatggg tgaagaacac
     2641 gcggcggcgc tctgggctga atccaccgaa tacatggccc gtctggttgt agataaacgc
     2701 gccaacccga ccgacgacct tctgggccgc ttggttgccg aacatggagc agaactgtct
     2761 gatgcggagc tggccggaat tggtgttttc ttattgggcg ctgggtatga aacgacagca
     2821 aatatgctgg ggctcggtac actgttatta ctgagacacc cggagcagct cgcacgcctg
     2881 cgcgactcta cgggggatgg cacagcggtc gatcgtgcgg tggaggaact tctgcgtatt
     2941 ctgaccgtgg ttaacaacgc agcaatgcgc caggcaaagg aagatgttac cgtcggcgga
     3001 cagttaattc gcgccgggga atttgttctg gtgtccctgg gctccgctaa ccgcgatgac
     3061 gcagtgcacg aacgccccga cgaatttgac attggccgtg caccggtggg ccacatggcc
     3121 ttcggtcatg gcattcatca ctgtcttgga gccccgctgg cgcgcctcga gatgcaggtc
     3181 gcttttccgg ccctgctcaa acgctttccg gcgctgcggc tggacgttcc gttcgacgag
     3241 attgaatatc tgccgcgcag cttggtccat ggcgtacgtg cgctgccggt ggcttggtaa
     3301 taaggatgag ctctacaaat aagcagaggt ggttgtgttg cgaaaaaaaa aaaaaaacac
     3361 cctaacgggt gttttttttt ttttggtgtc cccacgtgtg gcgctggaga ccgtccaatg
     3421 gcggcgcgcc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     3481 gagtcaattc agggtggtga atatgagccc gaaacgtcgt acccaggcag aacgtgcaat
     3541 ggaaacccag ggtaaactga ttgcagcagc actgggtgtt ctgcgtgaaa aaggttatgc
     3601 aggttttcgt attgcagatg ttccgggtgc agccggtgtt agccgtggtg cacagagcca
     3661 tcattttccg accaaactgg aactgctgct ggcaaccttt gaatggctgt atgagcagat
     3721 taccgaacgt agccgtgcac gtctggcaaa actgaaaccg gaagatgatg ttattcagca
     3781 gatgctggat gatgcagcag atttttttct ggatgatgat tttagcatcg gcctggatct
     3841 gattgttgca gcagatcgtg atccggcact gcgtgaaggt attctgcgta ccgttgaacg
     3901 taatcgtttt gttgttgaag atatgtggct gggtgtgctg gtgagccgtg gtctgagccg
     3961 tgatgatgcc gaagatattc tgtggctgat ttttaacagc gttcgtggtc tgacagttcg
     4021 tagcctgtgg cagaaagata aagaacgttt tgaacgtgtg cgtaatagca ccctggaaat
     4081 tgcacgtgaa cgttatgcaa aattcaaacg ttgataagga tcctaattgg taacgaatca
     4141 gacaattgac ggctcgaggg agtagcatag ggtttgcaga atccctgctt cgtccatttg
     4201 acaggcacat tatgcatcga tgataagctg tcaaacatga gcagatcctc tacgccggac
     4261 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     4321 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcaaatat tttatctgag
     4381 gtgcttcctc gctc
//
