LOCUS       sequence-423395-        4582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Thermomonospora curvata LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4582)
  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 4582)
  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: pCYP20
FEATURES             Location/Qualifiers
     source          1..4582
                     /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             2092..2100
                     /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(4494..4513)
                     /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 tgtttactaa ataaggaggt
     2101 ttcctatgag caccacgttg ggggcgcact cggccgcatc tactgcatct ctggaacgtg
     2161 tacctttgcg ccgtttaata tctctctacc gtcaaggtgg tgccttgcat gccctggaat
     2221 ctatcggccg catggcggag ggcgcaatcg cgcgcatcga cttaggattg tttcgtccat
     2281 atcttatcac ggacccagcc cacgtgcaac atgtactgcg ggatcgtgcc gataactatc
     2341 agcggggcgt ggttatgtgg aaacctgtat cccgtttagt cggcaatgga ataagcagcg
     2401 agggcccgct gtggcgaagc tcgcgccaag tccttgctcg cgttttcagt gcgcaatata
     2461 tctcacattt cacggaaaga atggccgaag ccatcaacgc ggctgcggat gatcttctgg
     2521 cgcgagcggc cgatcagccg gtggacgtac gtcgtgaaat gaatcgtctt atgcacagag
     2581 ccatcagtct ggttttcttc gacgaccgac tgtccgttgc cgaaattgac cgtctgggcg
     2641 ccgcagtcag cgctgcatcc tcatcattca tgtggcgcat ggctatgccc tttgtaccaa
     2701 acgccgttcc tatgccgggt gatcgcactt ttcgtcgggc cacccgcacc gttgatgaaa
     2761 ttttacgtcc tctggttgct gagggccgta aacgtccgcc gggtggccat gatgtagtga
     2821 gcaagctgat agaggcccgc gaagaagatg gtaccccgtt ctctgaccaa cgtattcgcg
     2881 atgatgttgt tgcgttgttc gtgtctggaa ccgagtctac cgcggatggg ctgacctggc
     2941 tgtgggtggc attacaccgc tacccggaaa ttgccgaacg tgtgtatgcg gaaattcgca
     3001 gcgaagttgg tgatgggccg gcgcgtccag agcatattcg tcggttgccc tatacccaaa
     3061 tgtttctcca agaagtctta cgtatgcatg cggtcggctg ggctgttcca cgcgtggcag
     3121 tggatgatga tgtgatcgcc ggtgtgccta ttaagaaagg tgcgacggtt gtgctgtccc
     3181 catacctgac ccatcgcgat cgccgcatct gggagcgccc tgaggaattt gaccccgaac
     3241 ggttcacacc tgaacgtatt aaagcacgta ttcagcgtca tggccgcttc gcctactttc
     3301 cgttcggagg tggggatcat gtttgccttg gtgaacagtt atttaaaatt gagacaacgc
     3361 tgataatggc gtccctgctg agtcgcgcgc gccctgtgtt gcgtaatgac gggccaatcg
     3421 aaagtcagct gacgctgacg gtacgcccta gccgccccgt acaaatgaca ctgcaggccc
     3481 atcgctaata aggatgagct ctacaaataa gcagaggtgg ttgtgttgcg aaaaaaaaaa
     3541 aaaaacaccc taacgggtgt tttttttttt ttggtgtccc cacgtgtggc gctggagacc
     3601 gtccaatggc ggcgcgccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc
     3661 ccggaagaga gtcaattcag ggtggtgaat atgagcccga aacgtcgtac ccaggcagaa
     3721 cgtgcaatgg aaacccaggg taaactgatt gcagcagcac tgggtgttct gcgtgaaaaa
     3781 ggttatgcag gttttcgtat tgcagatgtt ccgggtgcag ccggtgttag ccgtggtgca
     3841 cagagccatc attttccgac caaactggaa ctgctgctgg caacctttga atggctgtat
     3901 gagcagatta ccgaacgtag ccgtgcacgt ctggcaaaac tgaaaccgga agatgatgtt
     3961 attcagcaga tgctggatga tgcagcagat ttttttctgg atgatgattt tagcatcggc
     4021 ctggatctga ttgttgcagc agatcgtgat ccggcactgc gtgaaggtat tctgcgtacc
     4081 gttgaacgta atcgttttgt tgttgaagat atgtggctgg gtgtgctggt gagccgtggt
     4141 ctgagccgtg atgatgccga agatattctg tggctgattt ttaacagcgt tcgtggtctg
     4201 acagttcgta gcctgtggca gaaagataaa gaacgttttg aacgtgtgcg taatagcacc
     4261 ctggaaattg cacgtgaacg ttatgcaaaa ttcaaacgtt gataaggatc ctaattggta
     4321 acgaatcaga caattgacgg ctcgagggag tagcataggg tttgcagaat ccctgcttcg
     4381 tccatttgac aggcacatta tgcatcgatg ataagctgtc aaacatgagc agatcctcta
     4441 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     4501 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcaaatattt
     4561 tatctgaggt gcttcctcgc tc
//
