LOCUS       Exported                6435 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pWCD1610, complete sequence.
ACCESSION   KR232334
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6435)
  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 6435)
  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 6435)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6435
                     /organism="Expression vector pWCD1610"
                     /mol_type="other DNA"
                     /db_xref="taxon:1655362"
     misc_feature    6..11
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    13..16
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    17..541
                     /label=similar to URA3 5
                     /note="similar to URA3 5"
     regulatory      complement(542..619)
                     /regulatory_class="terminator"
     CDS             complement(620..1423)
                     /codon_start=1
                     /product="URA3p uracil auxotrophy marker"
                     /label=URA3p uracil auxotrophy marker
                     /protein_id="AKH61615.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     regulatory      complement(1424..1600)
                     /regulatory_class="promoter"
     regulatory      1607..2306
                     /regulatory_class="promoter"
     CDS             2313..3821
                     /codon_start=1
                     /product="mKate2 red fluorescent protein/DOPA 
                     4,5-dioxygenase fusion protein"
                     /label=mKate2 red fluorescent protein/DOPA 4,5-dioxyge
                     /note="mKate2 red fluorescent protein fused to MjDOD (DOPA 
                     4,5-dioxygenase) derived from Mirabilis jalapa"
                     /protein_id="AKH61613.1"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHRGSKGTYYINHGDPLMYLKKHIKLRQFLEGWQENVVIEKPKSILIISAHW
                     DTNVPTVNFVEHCDTIHDFDDYPDPLYQIQYRAPGAPNLAKKVEELLKESGMECEIDTK
                     RGLDHAAWFPLMFMYPEANIPICELSVQPSKDGIHHYNVGKALSPLLQQGVLIIGSGGT
                     VHPSDDTPHCPNGVAPWAIEFDNWLEDALLSGRYEDVNNFKKLAPNWEISHPGQEHLYP
                     LHVALGAAGKNPKTQLIHRSWAANGVFGYSTYNFTPTTQKTDGS"
     CDS             2313..3008
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     misc_feature    2316..3008
                     /label=mKate2 red fluorescent protein
                     /note="mKate2 red fluorescent protein"
     misc_feature    3015..3812
                     /note="MjDOD (DOPA 4,5-dioxygenase) derived from Mirabilis 
                     jalapa"
     regulatory      3828..4052
                     /regulatory_class="terminator"
     terminator      3828..4015
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    4061..4540
                     /label=similar to URA3 3
                     /note="similar to URA3 3"
     misc_feature    complement(4542..4545)
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     misc_feature    complement(4547..4552)
                     /label=BsmBI recognition site
                     /note="BsmBI recognition site"
     rep_origin      4572..5335
     rep_origin      complement(4671..5259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      5336..5432
                     /regulatory_class="terminator"
     CDS             complement(5430..6290)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AKH61614.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     regulatory      complement(6291..6421)
                     /regulatory_class="promoter"
     promoter        complement(6291..6395)
                     /gene="bla"
                     /label=AmpR promoter