LOCUS       Exported                8171 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pWCD2111, complete sequence.
ACCESSION   KR232318
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8171)
  AUTHORS   DeLoache WC, Russ ZN, Narcross L, Gonzales AM, Martin VJ, Dueber JE.
  TITLE     An enzyme-coupled biosensor enables (S)-reticuline production in 
            yeast from glucose
  JOURNAL   Nat. Chem. Biol. 11 (7), 465-471 (2015)
  PUBMED    25984720
REFERENCE   2  (bases 1 to 8171)
  AUTHORS   DeLoache WC, Russ ZN, Narcross L, Gonzales AM, Martin VJJ., Dueber 
            JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-APR-2015) Department of Bioengineering, University of 
            California, Berkeley, 2151 Berkeley Way Room 542, Berkeley, CA 
            94704, USA
REFERENCE   3  (bases 1 to 8171)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8171
                     /organism="Expression vector pWCD2111"
                     /mol_type="other DNA"
                     /db_xref="taxon:1655378"
     misc_feature    5..524
                     /label=similar to URA3 5
                     /note="similar to URA3 5"
     regulatory      573..1252
                     /regulatory_class="promoter"
     promoter        586..1252
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase"
     CDS             1259..2755
                     /codon_start=1
                     /product="CYP76AD3/P450 tyrosinase fusion protein"
                     /label=CYP76AD3/P450 tyrosinase fusion protein
                     /protein_id="AKH61558.1"
                     /translation="MDFLTLVMILSIIFFFYNLLKMKFTTHSDAQLPPGPKPMPIFGNI
                     FELGEKPHRSFANLAKTHGPLMSLRLGSVTTIVVSSAEVAKEMFLKNDQSLADRSVPNS
                     VTAGDHHKLTMSWLPVSPKWKNFRKITAVHLLSPQRLDACHALRHAKVKQLYEYVQECA
                     LKGEAVDIGKAAFTTSLNLLSNLFFSVELANHTSNTSQEFKQLIWDIMEDIGKPNYADY
                     FPLLKYVDPSGIRRRLAANFDKLIDVFQSFISKRLSSAYSSATSLDDVLDVLLKLLKEK
                     ELNMGEINHLLVDIFDAGTDTTSNTLEWAMAELMRNPIMMKRAQNEIALVLGKDNATIQ
                     ESDIANMPYLQAIIKETLRLHPPTVFLLPRKAITNVKLYGYIVPKNAQILVNLWAIGRD
                     PKVWKNPNEFLPDRFLNSDIDVKGRDFGLLPFGAGRRICPGMNLAYRMLTLMLATLLQS
                     FDWKLPHRNSPLDLDMDEKFGIALQKTKPLEIIPLIKYGS"
     misc_feature    2177..2179
                     /label=F307L mutation
                     /note="F307L mutation"
     regulatory      2762..2985
                     /regulatory_class="terminator"
     regulatory      3019..3718
                     /regulatory_class="promoter"
     CDS             3725..4534
                     /codon_start=1
                     /product="DOPA 4,5-dioxygenase"
                     /label=DOPA 4,5-dioxygenase
                     /note="MjDOD (DOPA 4,5-dioxygenase) derived from Mirabilis 
                     jalapa"
                     /protein_id="AKH61561.1"
                     /translation="MKGTYYINHGDPLMYLKKHIKLRQFLEGWQENVVIEKPKSILIIS
                     AHWDTNVPTVNFVEHCDTIHDFDDYPDPLYQIQYRAPGAPNLAKKVEELLKESGMECEI
                     DTKRGLDHAAWFPLMFMYPEANIPICELSVQPSKDGIHHYNVGKALSPLLQQGVLIIGS
                     GGTVHPSDDTPHCPNGVAPWAIEFDNWLEDALLSGRYEDVNNFKKLAPNWEISHPGQEH
                     LYPLHVALGAAGKNPKTQLIHRSWAANGVFGYSTYNFTPTTQKTDGS"
     regulatory      4541..4765
                     /regulatory_class="terminator"
     terminator      4541..4728
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     regulatory      4814..4990
                     /regulatory_class="promoter"
     CDS             4991..5794
                     /codon_start=1
                     /product="URA3p uracil auxotrophy marker"
                     /label=URA3p uracil auxotrophy marker
                     /protein_id="AKH61560.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     regulatory      5795..5872
                     /regulatory_class="terminator"
     misc_feature    5877..6396
                     /label=similar to URA3 3
                     /note="similar to URA3 3"
     regulatory      complement(6409..6435)
                     /regulatory_class="terminator"
     CDS             complement(6436..7251)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AKH61559.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     regulatory      complement(7252..7399)
                     /regulatory_class="promoter"
     rep_origin      complement(7400..8163)
                     /direction=LEFT
     rep_origin      7476..8064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"