LOCUS       Exported                6910 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pWCD2339, complete sequence.
ACCESSION   KR232301
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6910)
  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 6910)
  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 6910)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6910
                     /organism="Expression vector pWCD2339"
                     /mol_type="other DNA"
                     /db_xref="taxon:1655395"
     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..1381
                     /regulatory_class="promoter"
     promoter        715..1381
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase"
     CDS             1388..1972
                     /codon_start=1
                     /product="norcoclaurine synthase"
                     /label=norcoclaurine synthase
                     /note="derived from Thalictrum flavum"
                     /protein_id="AKH61495.1"
                     /translation="MQKLILTGRPFLHHQGIINQVSTVTKVIHHELEVAASADDIWTVY
                     SWPGLAKHLPDLLPGAFEKLEIIGDGGVGTILDMTFVPGEFPHEYKEKFILVDNEHRLK
                     KVQMIEGGYLDLGVTYYMDTIHVVPTGKDSCVIKSSTEYHVKPEFVKIVEPLITTGPLA
                     AMADAISKLVLEHKSKSNSDEIEAAIITVGS"
     regulatory      1979..2203
                     /regulatory_class="terminator"
     misc_feature    2351..2356
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    2358..2361
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     regulatory      2402..3049
                     /regulatory_class="promoter"
     promoter        2642..3049
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3050..4144
                     /codon_start=1
                     /product="LEU2p auxotrophy marker"
                     /label=LEU2p auxotrophy marker
                     /protein_id="AKH61493.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     regulatory      4145..4621
                     /regulatory_class="terminator"
     misc_feature    4626..5135
                     /label=similar to LEU2 3
                     /note="similar to LEU2 3"
     regulatory      complement(5148..5174)
                     /regulatory_class="terminator"
     CDS             complement(5175..5990)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AKH61494.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     regulatory      complement(5991..6138)
                     /regulatory_class="promoter"
     rep_origin      complement(6139..6902)
                     /direction=LEFT
     rep_origin      6215..6803
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"