LOCUS       sequence-424244-        4554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Salinispora arenicola LRD-modifying cytochrome P450 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4554)
  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 4554)
  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: pCYP55
FEATURES             Location/Qualifiers
     source          1..4554
                     /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             2100..2108
                     /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(4466..4485)
                     /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 tgtttaccac ctcttctcat
     2101 aaggaggttt tttatgccag cctggaaagc cctgcctgcg gccgttcgtg atactcatcg
     2161 tgcctttgtg gatgtgggta actggtcaga cggtgatgtg gtgcgcgttt cccttggtgt
     2221 cagccgcccg tatctcgtga ccaatccggc gcacgtgcag gaagtgttac atgaacgggc
     2281 ggcaatctat ccgcgcggag atgataccgc actctggcgt agtgtacgca agttggtggg
     2341 tgatggcatc ttggctgaag gcgacgcgtg ggccgctagt agaagagtcc ttgcgccaat
     2401 gtttcgccct gcccgtatca acgccatggt agataccatg gcggatgcta tcgcaggcgc
     2461 tatagatgat ttgcacggcg cggcaacggc aggcacgccg attgacgttg gtcgcgaact
     2521 gagccgcatt gtgtgcagtg ccataatgcg tgtgttcttt gcagaccgta tcacggtacg
     2581 cgacgccctt cggatcatga aagcccagga aaccattgtg actgccatgg cgccgcgcat
     2641 tttagcacct ctggtaccgt ggtggattcc catgccgggg gatcgtagat tccgcgccgc
     2701 ggtgcgttca atcgatgaca ttctgctgcc cgtgcttaga caggcccagc gtcagcccga
     2761 cgatggcgac gatttattat cccgtttggt gcacgcgcgc gcagatgatg gtcgcgccct
     2821 gagcgaaaaa cgtatgcgcg acgatttggt gagcatggtc gcggttacta ccgaaacaag
     2881 caccgtggta ctgacatggt tgtggccttt gctggcgaac catcccgatg ttgcaaatcg
     2941 tctgtatgac gaaattgatc gcgttgttgg cggcggacca gtccgtggtg atcatctggc
     3001 ggaactgacg tatacccgca tggttctgga tgaattgtta cgactgtacc cggccggatg
     3061 gattctccct cgtcgggcag cgaccacgga tattctggga ggggtgcgta taaataaagg
     3121 tgccaccgta atcctgtcgc cctatgtaac acagcgtatg accgcatggt ggggtcccac
     3181 ggcagaagcc ttcgacccgg aaagattcgc ggcgggtcga gaagccgcgg atgggagaca
     3241 tcggtacgct tactatccct tcggcgtcgg catgcaccga tgtctggggg agcatctgtt
     3301 taacctggaa gctatcctca tcgttgccac gcttttgtca cgtttccgct ttgctctgac
     3361 ggatacaagc atgccgggag ttaaagtggc tgcctccacg cggccggctc gaaccgtaga
     3421 ggtgattctg aaacctgtcg cgccggttcc agcgcgttaa taaggatgag ctctacaaat
     3481 aagcagaggt ggttgtgttg cgaaaaaaaa aaaaaaacac cctaacgggt gttttttttt
     3541 ttttggtgtc cccacgtgtg gcgctggaga ccgtccaatg gcggcgcgcc atcgaatggt
     3601 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     3661 atatgagccc gaaacgtcgt acccaggcag aacgtgcaat ggaaacccag ggtaaactga
     3721 ttgcagcagc actgggtgtt ctgcgtgaaa aaggttatgc aggttttcgt attgcagatg
     3781 ttccgggtgc agccggtgtt agccgtggtg cacagagcca tcattttccg accaaactgg
     3841 aactgctgct ggcaaccttt gaatggctgt atgagcagat taccgaacgt agccgtgcac
     3901 gtctggcaaa actgaaaccg gaagatgatg ttattcagca gatgctggat gatgcagcag
     3961 atttttttct ggatgatgat tttagcatcg gcctggatct gattgttgca gcagatcgtg
     4021 atccggcact gcgtgaaggt attctgcgta ccgttgaacg taatcgtttt gttgttgaag
     4081 atatgtggct gggtgtgctg gtgagccgtg gtctgagccg tgatgatgcc gaagatattc
     4141 tgtggctgat ttttaacagc gttcgtggtc tgacagttcg tagcctgtgg cagaaagata
     4201 aagaacgttt tgaacgtgtg cgtaatagca ccctggaaat tgcacgtgaa cgttatgcaa
     4261 aattcaaacg ttgataagga tcctaattgg taacgaatca gacaattgac ggctcgaggg
     4321 agtagcatag ggtttgcaga atccctgctt cgtccatttg acaggcacat tatgcatcga
     4381 tgataagctg tcaaacatga gcagatcctc tacgccggac gcatcgtggc cggcatcacc
     4441 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg
     4501 gctcgccact tcgggctcat gagcaaatat tttatctgag gtgcttcctc gctc
//
