LOCUS       Exported                7467 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pWCD2163, complete sequence.
ACCESSION   KR232313
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7467)
  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 7467)
  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 7467)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7467
                     /organism="Expression vector pWCD2163"
                     /mol_type="other DNA"
                     /db_xref="taxon:1655383"
     misc_feature    5..524
                     /label=similar to LEU2 5
                     /note="similar to LEU2 5"
     misc_feature    complement(544..547)
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    complement(549..554)
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     regulatory      702..1401
                     /regulatory_class="promoter"
     CDS             1408..2520
                     /codon_start=1
                     /product="ARO4 protein"
                     /label=ARO4 protein
                     /note="ARO4 derived from Saccharomyces cerevisiae with 
                     K229L feedback resistant mutation"
                     /protein_id="AKH61540.1"
                     /translation="MSESPMFAANGMPKVNQGAEEDVRILGYDPLASPALLQVQIPATP
                     TSLETAKRGRREAIDIITGKDDRVLVIVGPCSIHDLEAAQEYALRLKKLSDELKGDLSI
                     IMRAYLEKPRTTVGWKGLINDPDVNNTFNINKGLQSARQLFVNLTNIGLPIGSEMLDTI
                     SPQYLADLVSFGAIGARTTESQLHRELASGLSFPVGFKNGTDGTLNVAVDACQAAAHSH
                     HFMGVTLHGVAAITTTKGNEHCFVILRGGKKGTNYDAKSVAEAKAQLPAGSNGLMIDYS
                     HGNSNKDFRNQPKVNDVVCEQIANGENAITGVMIESNINEGNQGIPAEGKAGLKYGVSI
                     TDACIGWETTEDVLRKLAAAVRQRREVNKK"
     regulatory      2536..2760
                     /regulatory_class="terminator"
     misc_feature    2908..2913
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    2915..2918
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     regulatory      2959..3606
                     /regulatory_class="promoter"
     promoter        3199..3606
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3607..4701
                     /codon_start=1
                     /product="LEU2p auxotrophy marker"
                     /label=LEU2p auxotrophy marker
                     /protein_id="AKH61541.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     regulatory      4702..5178
                     /regulatory_class="terminator"
     misc_feature    5183..5692
                     /label=similar to LEU2 3
                     /note="similar to LEU2 3"
     regulatory      complement(5705..5731)
                     /regulatory_class="terminator"
     CDS             complement(5732..6547)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AKH61542.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     regulatory      complement(6548..6695)
                     /regulatory_class="promoter"
     rep_origin      complement(6696..7459)
                     /direction=LEFT
     rep_origin      6772..7360
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"