LOCUS       sequence-423393-        4414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Saccharopolyspora shandongensis LRD-modifying 
            cytochrome P450 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4414)
  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 4414)
  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: pCYP44
FEATURES             Location/Qualifiers
     source          1..4414
                     /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(4326..4345)
                     /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 gaacagctaa
     2101 ggaggatatt tatgactggt gctacccagg aggactcatt caccgaacac tattggcgcg
     2161 atcctcatgc cgccctggcg ggcacccgcg aaacctgtcc cgttcgcgaa gttgatttcg
     2221 aaggtggtcg cacttggctg gtcacgcgct atgacgacgt gcgtgccggg cttgctgacg
     2281 agagactggc taaagattgg cgttcgtccc tgcctgcaga agaacgggcg tccgcgcctc
     2341 cgggtttacc ggctccgatg agccacatgc ttacctctct ggacccacca gaacatacac
     2401 gcttacgtag cttggtgaca caggcgttta cggcccgtcg catcgccaca ttaagaccgc
     2461 gcgtggagag cgtcgccgaa gaactgctgg cagatctccc ggaagatgaa cccatcgatc
     2521 ttgtggcccg ctatgcgatt gtgctgccga tggtggtgat ttctgaactt ttgggtgtac
     2581 cggaaattga ccgtgatgaa ttcgcacgtt tgtcaactgt actgattgat gaatgcccgg
     2641 aaccggaact ggaacgcgca tcgacccaaa tggcgactta cctcgaaggt ttggttgctg
     2701 cgaaacgtgc cgcacccgat gacgctctgt taagcgccct catttcggcg tccgatgaag
     2761 gtgatcgctt gagcgattta gaaatcgttg caatggcaat gcttctgctg atggctggcc
     2821 acgaaaccac agcagtgttc attaccaact ctgtccgggc tttagtcgca gatgattcag
     2881 cacgcgaacg cctggcacgc cctgaaagcg tgccctccct gattgaagaa ttactgcgtt
     2941 ggctcagccc agcgctgaat gcatctcttc gctttgctac ggaggacctt gaaattggag
     3001 gcgtgaccat tccgcgtggt gcaagtgtca cgttaagcac aggagccgcg aaccgtgatc
     3061 cggctcgatt tggcgatcca gaagtcttag atccggaccg taacaccgct ggccacctgg
     3121 cgttcgggta tggcattcac cgatgtttag gggcagcgct ggttcgtctg gaaggcgaag
     3181 ttggattggc ggcgctgttc aatcggtttc cgcgccttcg tgttgtcgct gatccggcgg
     3241 atttaacgta ccgtcgcagc gtgcacgtgc atgctccgag aacactgccc gtggtactgg
     3301 ggccgcgtgc gggtggctaa taaggatgag ctctacaaat aagcagaggt ggttgtgttg
     3361 cgaaaaaaaa aaaaaaacac cctaacgggt gttttttttt ttttggtgtc cccacgtgtg
     3421 gcgctggaga ccgtccaatg gcggcgcgcc atcgaatggt gcaaaacctt tcgcggtatg
     3481 gcatgatagc gcccggaaga gagtcaattc agggtggtga atatgagccc gaaacgtcgt
     3541 acccaggcag aacgtgcaat ggaaacccag ggtaaactga ttgcagcagc actgggtgtt
     3601 ctgcgtgaaa aaggttatgc aggttttcgt attgcagatg ttccgggtgc agccggtgtt
     3661 agccgtggtg cacagagcca tcattttccg accaaactgg aactgctgct ggcaaccttt
     3721 gaatggctgt atgagcagat taccgaacgt agccgtgcac gtctggcaaa actgaaaccg
     3781 gaagatgatg ttattcagca gatgctggat gatgcagcag atttttttct ggatgatgat
     3841 tttagcatcg gcctggatct gattgttgca gcagatcgtg atccggcact gcgtgaaggt
     3901 attctgcgta ccgttgaacg taatcgtttt gttgttgaag atatgtggct gggtgtgctg
     3961 gtgagccgtg gtctgagccg tgatgatgcc gaagatattc tgtggctgat ttttaacagc
     4021 gttcgtggtc tgacagttcg tagcctgtgg cagaaagata aagaacgttt tgaacgtgtg
     4081 cgtaatagca ccctggaaat tgcacgtgaa cgttatgcaa aattcaaacg ttgataagga
     4141 tcctaattgg taacgaatca gacaattgac ggctcgaggg agtagcatag ggtttgcaga
     4201 atccctgctt cgtccatttg acaggcacat tatgcatcga tgataagctg tcaaacatga
     4261 gcagatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
     4321 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat
     4381 gagcaaatat tttatctgag gtgcttcctc gctc
//
