LOCUS       sequence-423396-        4561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Mesorhizobium sp. LSJC277A00 LRD-modifying cytochrome 
            P450 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4561)
  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 4561)
  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: pCYP09
FEATURES             Location/Qualifiers
     source          1..4561
                     /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             2097..2105
                     /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(4473..4492)
                     /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 tgtttagagc aagctttaag
     2101 gaggtttata atgctctcca actgtcgtcg accttgccgt gcccccatcc gttccgcgaa
     2161 ccaacaagtg acgaagatgg acgtgcagga aacgacagcc gcatgccgag atgcgtttgc
     2221 tgaactggca tcccccgcat gtatacaaga tccttatccg ttcatgcgct ggctccgtga
     2281 acatgacccg gttcaccgag ccgcctcggg cctgtttctg ttgagccgcc atgcggatat
     2341 ttactgggcc ttaaaggcta cgggcgacgc attccgtggt ccggcaccgg gtgaattagc
     2401 gcgttacttc ccgcgggctg caacgtccct gtctttaagc ctgcttgcct ccaccctggc
     2461 tatgaaagaa cctccgactc atacccgtct gcgtcggctg atctcgcgcg actttacaat
     2521 cggccagatc gataagctgc gccctacgat ggcgcgtatc gtcgcagcgc ggttggacgg
     2581 tatggcgcca gcactggaac gtggtgaagc ggtagatctg caccaggaat tcgcgctggc
     2641 cctgccgatg cttgtgttcg gggaactgtt cggcatgcct ccggatgata ttctggcctt
     2701 agtcaccggt attagcgcga tattagaagg gctgtctcca catgcctcag atccacagct
     2761 ggcagctggt gacgaggcaa gcgcgcgtgt tcaggcgtat ttcggtgatt tgattcagcg
     2821 taagcgtact gattcacggc acgatatagt atcgatgctg gtgggcgccc atgacgacga
     2881 tgcagaaacc ttaagcgatg ctgaactgat tagtatgctg tgggggatgt tgctgggcgg
     2941 tttcgcgact actgctgcag cgattggcca tgcggttctg gctatgctcg cctatcccga
     3001 acagcgccat tggttgaagg gtgatgcggt cggtgtgaaa gccttcgtcg aagaggccct
     3061 gcgctacgat gccccggcca tgtttagctc tatcccccgt attgcacagc gggatatcga
     3121 aatgggcggt gtggtgattc ctaaaaacgc agacgtacgc gtccttattg cagcaggtaa
     3181 ccgcgaccca gatgctttcg ccgacccgga tcgtttcgac ccggtgcgct tctgtggtac
     3241 atcacccgga atgtcgatcg atgggaaaat tatgctcacc tttggtcacg gcatccattt
     3301 ttgtcttggc gcgcaattag cccgcgtcca actcgctgaa tcgctgccgc aaatccaagc
     3361 acgcttccca acattggctc tggctgaaca gccgacccga gagccgtcag catttcttcg
     3421 caccttccga gcgctgcctg tgcgcctgca cggtcaggaa ggttaataag gatgagctct
     3481 acaaataagc agaggtggtt gtgttgcgaa aaaaaaaaaa aaacacccta acgggtgttt
     3541 tttttttttt tggtgtcccc acgtgtggcg ctggagaccg tccaatggcg gcgcgccatc
     3601 gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     3661 gtggtgaata tgagcccgaa acgtcgtacc caggcagaac gtgcaatgga aacccagggt
     3721 aaactgattg cagcagcact gggtgttctg cgtgaaaaag gttatgcagg ttttcgtatt
     3781 gcagatgttc cgggtgcagc cggtgttagc cgtggtgcac agagccatca ttttccgacc
     3841 aaactggaac tgctgctggc aacctttgaa tggctgtatg agcagattac cgaacgtagc
     3901 cgtgcacgtc tggcaaaact gaaaccggaa gatgatgtta ttcagcagat gctggatgat
     3961 gcagcagatt tttttctgga tgatgatttt agcatcggcc tggatctgat tgttgcagca
     4021 gatcgtgatc cggcactgcg tgaaggtatt ctgcgtaccg ttgaacgtaa tcgttttgtt
     4081 gttgaagata tgtggctggg tgtgctggtg agccgtggtc tgagccgtga tgatgccgaa
     4141 gatattctgt ggctgatttt taacagcgtt cgtggtctga cagttcgtag cctgtggcag
     4201 aaagataaag aacgttttga acgtgtgcgt aatagcaccc tggaaattgc acgtgaacgt
     4261 tatgcaaaat tcaaacgttg ataaggatcc taattggtaa cgaatcagac aattgacggc
     4321 tcgagggagt agcatagggt ttgcagaatc cctgcttcgt ccatttgaca ggcacattat
     4381 gcatcgatga taagctgtca aacatgagca gatcctctac gccggacgca tcgtggccgg
     4441 catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga
     4501 agatcgggct cgccacttcg ggctcatgag caaatatttt atctgaggtg cttcctcgct
     4561 c
//
