LOCUS       sequence-423394-        4469 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses the Saccharomonospora piscinae LRD-modifying cytochrome 
            P450 enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4469)
  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 4469)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pCYP31
FEATURES             Location/Qualifiers
     source          1..4469
                     /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(4381..4400)
                     /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 tgtttaggac tgagtaattt
     2101 aaggagggat tgataatgaa tagaacggag ctgcgcctgg ctctgcgtac cctgccgttc
     2161 ctggacgcac accgtgcaga acccgcgcgg cacgtcgttc tggccgccga tccacctaag
     2221 ctgcttgtgt gggacccggg tactgtggag tggcttttcc gcagtgacgg cgagctggac
     2281 catccaggtt cgagaagcct ccgaccgctt tttggcccgc attcgttgct gtgggttgat
     2341 ggggaacgcc ataccgcata ccggagaacg cttgggccgc cactgcgcgg gcgtagatta
     2401 tcagcctacg aaggactgat cgcggatacg gtgcgcgatg ccgtgtctgc gctgcgtcct
     2461 ggcgataccg tgggactggc tgattggacg cgtgcgctgg cactgcggat catcgcccgc
     2521 atcgtgttag gtgacctgga tacccgaacg gaacgcgtat tagatggctt cactgcgtgg
     2581 atcgatgacg ccctgggctc acgccgtcgc acattggcac accgcttttt aagaggcgga
     2641 ttgccccgtt ccggcgcggg ccttgacgct gaactggtta gctgggcaag agcagagtcc
     2701 cgcccggacc ctccggcatt agccgcctta ctgcgacagg ccgatggacc gctggctgac
     2761 cttccagacg gcgaactgcg tgaccaactg gtttctctgt tattcgcagg ccatgaaaca
     2821 accgcctctg cggctgcctg gacgctgtac cgtctgagcc gtgacgaagc gctccgcgat
     2881 gaagttctcg cagaattgga tgcccctgga gcagctggac gagcagcacc accagcaagt
     2941 gcggttccac ttctgactag cgtcattagc gaaaccctgc gtctggcgcc gcctgtgacg
     3001 gttgcagaaa accggcggtt acgcgcacca gcacgtctgt tgggtcgtaa tctgccggaa
     3061 ggtactaccc tcacgaccag tatctatctg gcccaacgcc agggtgaggc attcgccgat
     3121 ccgctccatt tcgatccaca tcgttttgtt agcagacgtt acgatccggc ccgttacttg
     3181 ccttttgggg gcggctcacg acgatgcctt ggcgcgtccc ttgcgatgtt agagctgcgt
     3241 gtgatcgtgg ctgcggttct gcgtcgtcga gaatggcgct gcctgaaccc gggatctgcc
     3301 agactttctc tgcgcgggca tgccatggcc ccgtcgtcgc gtctgcgtat gagagtgacc
     3361 gcgtgccgtg attaataagg atgagctcta caaataagca gaggtggttg tgttgcgaaa
     3421 aaaaaaaaaa aacaccctaa cgggtgtttt tttttttttg gtgtccccac gtgtggcgct
     3481 ggagaccgtc caatggcggc gcgccatcga atggtgcaaa acctttcgcg gtatggcatg
     3541 atagcgcccg gaagagagtc aattcagggt ggtgaatatg agcccgaaac gtcgtaccca
     3601 ggcagaacgt gcaatggaaa cccagggtaa actgattgca gcagcactgg gtgttctgcg
     3661 tgaaaaaggt tatgcaggtt ttcgtattgc agatgttccg ggtgcagccg gtgttagccg
     3721 tggtgcacag agccatcatt ttccgaccaa actggaactg ctgctggcaa cctttgaatg
     3781 gctgtatgag cagattaccg aacgtagccg tgcacgtctg gcaaaactga aaccggaaga
     3841 tgatgttatt cagcagatgc tggatgatgc agcagatttt tttctggatg atgattttag
     3901 catcggcctg gatctgattg ttgcagcaga tcgtgatccg gcactgcgtg aaggtattct
     3961 gcgtaccgtt gaacgtaatc gttttgttgt tgaagatatg tggctgggtg tgctggtgag
     4021 ccgtggtctg agccgtgatg atgccgaaga tattctgtgg ctgattttta acagcgttcg
     4081 tggtctgaca gttcgtagcc tgtggcagaa agataaagaa cgttttgaac gtgtgcgtaa
     4141 tagcaccctg gaaattgcac gtgaacgtta tgcaaaattc aaacgttgat aaggatccta
     4201 attggtaacg aatcagacaa ttgacggctc gagggagtag catagggttt gcagaatccc
     4261 tgcttcgtcc atttgacagg cacattatgc atcgatgata agctgtcaaa catgagcaga
     4321 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
     4381 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagca
     4441 aatattttat ctgaggtgct tcctcgctc
//
