LOCUS       Exported                6928 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pWCD2336, complete sequence.
ACCESSION   KR232304
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6928)
  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 6928)
  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 6928)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6928
                     /organism="Expression vector pWCD2336"
                     /mol_type="other DNA"
                     /db_xref="taxon:1655392"
     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..1990
                     /codon_start=1
                     /product="norcoclaurine synthase 1"
                     /label=norcoclaurine synthase 1
                     /note="derived from Tinospora cordifolia"
                     /protein_id="AKH61504.1"
                     /translation="MRKELRHELKVPASADDVWEVYSSPDLPKLIVQLLPSVFEKIEVV
                     EGDGGVGTVLHITFPPGSIPISYKEKFVTVDDYKRLKEVRQIEGGYLDMGCTFYMDSFH
                     ILEECHDSCVIVSKTEYEVPQELAANVEPYISIDSLAGMATAISNYVVDKNKNKECEPE
                     GCCDDERDHCSEKEGRHEQSSDESESDCESESDCDGS"
     regulatory      1997..2221
                     /regulatory_class="terminator"
     misc_feature    2369..2374
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    2376..2379
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     regulatory      2420..3067
                     /regulatory_class="promoter"
     promoter        2660..3067
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3068..4162
                     /codon_start=1
                     /product="LEU2p auxotrophy marker"
                     /label=LEU2p auxotrophy marker
                     /protein_id="AKH61502.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     regulatory      4163..4639
                     /regulatory_class="terminator"
     misc_feature    4644..5153
                     /label=similar to LEU2 3
                     /note="similar to LEU2 3"
     regulatory      complement(5166..5192)
                     /regulatory_class="terminator"
     CDS             complement(5193..6008)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AKH61503.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     regulatory      complement(6009..6156)
                     /regulatory_class="promoter"
     rep_origin      complement(6157..6920)
                     /direction=LEFT
     rep_origin      6233..6821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"