LOCUS       sequence-423274-        4412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Herbihabitans rhizosphaerae LRD-modifying cytochrome 
            P450 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4412)
  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 4412)
  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: pCYP52
FEATURES             Location/Qualifiers
     source          1..4412
                     /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(4324..4343)
                     /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 tgtttagcga cttaccgttc
     2101 ataagcgagg tttaaatgaa tacccggaac acaacgccgg ctgacgttcc tgtacgcgat
     2161 ttcaaccacc acgcggaccc ggatctgcgg atcgatccct tcgcaacctt tgaccgcttc
     2221 cgagatgaac gcgtgttttg gacgcccgaa ttagatggct tttgggtcct tacgcgctac
     2281 gacgatattc gtgccgtatt gcgcgacacg gaacgcttct ctaacgagag tacagcaatc
     2341 ccgccgattg attggaatgg tcagcgcctt attcccgtgg aattggaccc tccggaccac
     2401 ggcaaatacc ggaccctgct gaccagatac ttgaacgggc cggcgaacgc tgtacttgcc
     2461 actgcggtgg agaaagaatg tttacgtctg gttgaacgcc tggcccctgc tggagaatgc
     2521 gaactcgttg gcgacttcgc ccgcaagctg cagaacgcat tactcggcgc tatgttcgac
     2581 gaaccgggcg aagaggacga tgtgcctgac ttcggtcggc tgtccgacga cttggttcag
     2641 gccagtgaac ctgcgcggcg aagtcgtgca attcgtgaca ttatggctta ccttcgtcgt
     2701 gcaatcgcag gacatcagac tcgccgaacc gattctacta gcctcatcgc gtccttgtcg
     2761 ggagctgaaa ttgacggtag accgctgtca gaatcggaga ctcttaacat ggcatttctg
     2821 atgacgatgg cagctatcga taccctgacg tcctcgatga gtttcggctt tcggtatctg
     2881 gcagaacacc cggcccacca gcgtcgctta gcggaagatc ctggagcagc ggcggctgcc
     2941 gccgacgaac tgcttcgtct gcatagtgtc gtaagtatcg cgcgcattgt agcgcgcgac
     3001 actgaattcg caggagttcg tatgcgcgcc ggtgaacgtg tgatgctgtc actgagctta
     3061 gcagggagag atccgcatca gtatccggaa gccacgactg cagattttga tcgaccgaat
     3121 agagcctccc acttagcatt tggttttgga gctcatcgct gcgcgggcgc ccgcattgca
     3181 acaggtggaa tgattgctgc cttacgcgaa tggcacgcgg caattcccga atatagcgtg
     3241 agacttggga ccgaactggt taccgggggt ggggctgcag cgtcactgga atctttgccg
     3301 ttagtctgga aaacctaata aggatgagct ctacaaataa gcagaggtgg ttgtgttgcg
     3361 aaaaaaaaaa aaaaacaccc taacgggtgt tttttttttt ttggtgtccc cacgtgtggc
     3421 gctggagacc gtccaatggc ggcgcgccat cgaatggtgc aaaacctttc gcggtatggc
     3481 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat atgagcccga aacgtcgtac
     3541 ccaggcagaa cgtgcaatgg aaacccaggg taaactgatt gcagcagcac tgggtgttct
     3601 gcgtgaaaaa ggttatgcag gttttcgtat tgcagatgtt ccgggtgcag ccggtgttag
     3661 ccgtggtgca cagagccatc attttccgac caaactggaa ctgctgctgg caacctttga
     3721 atggctgtat gagcagatta ccgaacgtag ccgtgcacgt ctggcaaaac tgaaaccgga
     3781 agatgatgtt attcagcaga tgctggatga tgcagcagat ttttttctgg atgatgattt
     3841 tagcatcggc ctggatctga ttgttgcagc agatcgtgat ccggcactgc gtgaaggtat
     3901 tctgcgtacc gttgaacgta atcgttttgt tgttgaagat atgtggctgg gtgtgctggt
     3961 gagccgtggt ctgagccgtg atgatgccga agatattctg tggctgattt ttaacagcgt
     4021 tcgtggtctg acagttcgta gcctgtggca gaaagataaa gaacgttttg aacgtgtgcg
     4081 taatagcacc ctggaaattg cacgtgaacg ttatgcaaaa ttcaaacgtt gataaggatc
     4141 ctaattggta acgaatcaga caattgacgg ctcgagggag tagcataggg tttgcagaat
     4201 ccctgcttcg tccatttgac aggcacatta tgcatcgatg ataagctgtc aaacatgagc
     4261 agatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
     4321 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
     4381 gcaaatattt tatctgaggt gcttcctcgc tc
//
