LOCUS       sequence-423269-        4602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Streptomyces sp. HB202 LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4602)
  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 4602)
  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: pCYP25
FEATURES             Location/Qualifiers
     source          1..4602
                     /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             2103..2111
                     /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(4514..4533)
                     /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 accgaggaca
     2101 gataaggagg tatattatgc cggaactgcc ggctattcga aaagcaccgc gtagtttgcc
     2161 gttagccggc cacgccgtgc cgctgctcgt tcagcccctg gaatttctga aaagcctgcc
     2221 ggcgctcggt ggattagtgc gggttgggat gggtccggat gacgcttatg tgccatgcga
     2281 tccagctctg tttcgtcaaa tccttaaaga tacgcgcctc tacgataaag gcggtctgtt
     2341 ctatgatcgc ggccgtgagg ccgtagggaa cggcttggtt acttcccgtt gggcagacca
     2401 tcggcgccag cgtcccctga tgcaaccgtc ctttaatcac cgtcacctgg gtcactatgc
     2461 cgaactgatg gccgatgaaa cggaagtctt aatgcgctcc tggcgtagcg gcgaaaccgt
     2521 ggatgtagat aaagcgatgg ctcggctgac cctgcgcatt acgacgcgga gcttgtttag
     2581 cgtgccagcg gatcatgcat tggttgccca ggtcgagaag tggctgccta ttctgatgga
     2641 tggattcttt cgcagaatgt ttgttccggc tcgcgtgctg tctctggtgc ctacccgtgt
     2701 caatcggctg tacccgagag cgatagccga gatgagaaaa ctcactgaag aaattattga
     2761 cgaggtacgt cacaagaaag acgaagatcc tggccttctg gcctctctgc ttgatgcccg
     2821 agatgaaaca acaggtgccc ctctggatac ccaggaaatc tttgaccaag tgctgatttt
     2881 actcatcgcc ggttctgaaa ccacggcaac tgccttggcc tttacctttc acttgctcgg
     2941 cgcgcatcca ggcatcggcg cgcgtttacg cgaagaagtt gacggcgcgc ttggcggtcg
     3001 tacgcccaga tttgaggacc tgtctgccct gccatatacc agacaagtgt taatggagtc
     3061 cctgcgtttg tatccgcccg cctggatgtt tacccgctct acaaccacgg cctgcgagct
     3121 gggcggccat gccttccccg aaggtacaac cttcctgtta tctccgtaca tcttacagca
     3181 cgatcctgaa ctgtttccac gccccgaacg tttcgatccg gatcggtggg ctcccggggc
     3241 aatgagcgat gtagcgcgcc gtagtgttgt gccgttcggt gctggcggtc gcaagtgcat
     3301 cggcgaccag tttgccttat ctgaggccct gttagccata gcggttattg caggacgctg
     3361 ggacttgacg ccggtgggcg atcgtccagt tcgcccgatt gcacgtgcca cgttgaaaac
     3421 tggcccatta cccatgcgcc ttcgcgaacg tccgcatgag ccggtacgca atgctacaac
     3481 aacggatctg cgtgatcacc aacgctaata aggatgagct ctacaaataa gcagaggtgg
     3541 ttgtgttgcg aaaaaaaaaa aaaaacaccc taacgggtgt tttttttttt ttggtgtccc
     3601 cacgtgtggc gctggagacc gtccaatggc ggcgcgccat cgaatggtgc aaaacctttc
     3661 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat atgagcccga
     3721 aacgtcgtac ccaggcagaa cgtgcaatgg aaacccaggg taaactgatt gcagcagcac
     3781 tgggtgttct gcgtgaaaaa ggttatgcag gttttcgtat tgcagatgtt ccgggtgcag
     3841 ccggtgttag ccgtggtgca cagagccatc attttccgac caaactggaa ctgctgctgg
     3901 caacctttga atggctgtat gagcagatta ccgaacgtag ccgtgcacgt ctggcaaaac
     3961 tgaaaccgga agatgatgtt attcagcaga tgctggatga tgcagcagat ttttttctgg
     4021 atgatgattt tagcatcggc ctggatctga ttgttgcagc agatcgtgat ccggcactgc
     4081 gtgaaggtat tctgcgtacc gttgaacgta atcgttttgt tgttgaagat atgtggctgg
     4141 gtgtgctggt gagccgtggt ctgagccgtg atgatgccga agatattctg tggctgattt
     4201 ttaacagcgt tcgtggtctg acagttcgta gcctgtggca gaaagataaa gaacgttttg
     4261 aacgtgtgcg taatagcacc ctggaaattg cacgtgaacg ttatgcaaaa ttcaaacgtt
     4321 gataaggatc ctaattggta acgaatcaga caattgacgg ctcgagggag tagcataggg
     4381 tttgcagaat ccctgcttcg tccatttgac aggcacatta tgcatcgatg ataagctgtc
     4441 aaacatgagc agatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
     4501 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
     4561 gggctcatga gcaaatattt tatctgaggt gcttcctcgc tc
//
