LOCUS       Exported                8267 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast two-hybrid vector pMK498-TEV, complete sequence.
ACCESSION   AY647985
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8267)
  AUTHORS   Guo D, Hazbun TR, Xu XJ, Ng SL, Fields S, Kuo MH.
  TITLE     A tethered catalysis, two-hybrid system to identify protein-protein 
            interactions requiring post-translational modifications
  JOURNAL   Nat. Biotechnol. 22 (7), 888-892 (2004)
  PUBMED    15208639
REFERENCE   2  (bases 1 to 8267)
  AUTHORS   Kuo M-H., Xu X-J., Ng S-L.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUN-2004) Biochem. Mol. Biol., Michigan State 
            University, 309 BCH Building, East Lansing, MI 48824, USA
REFERENCE   3  (bases 1 to 8267)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8267
                     /organism="Yeast two-hybrid vector pMK498-TEV"
                     /mol_type="genomic DNA"
                     /label=tethered catalysis bait construct
                     /note="tethered catalysis bait construct"
                     /db_xref="taxon:284156"
     CDS             434..2041
                     /codon_start=1
                     /product="GDBD-TEV-Kin28-HA fusion protein"
                     /label=GDBD-TEV-Kin28-HA fusion protein
                     /note="derived from Saccharomyces cerevisiae"
                     /protein_id="AAT97347.1"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSNYLFDDEDTPPNPKKE
                     IELENLYFQGEFQLTTMAMKVNMEYTKEKKVGEGTYAVVYLGCQHSTGRKIAIKEIKTS
                     EFKDGLDMSAIREVKYLQEMQHPNVIELIDIFMAYDNLNLVLEFLPTDLEVVIKDKSIL
                     FTPADIKAWMLMTLRGVYHCHRNFILHRDLKPNNLLFSPDGQIKVADFGLARAIPAPHE
                     ILTSNVVTRWYRAPELLFGAKHYTSAIDIWSVGVIFAELMLRIPYLPGQNDVDQMEVTF
                     RALGTPTDRDWPEVSSFMTYNKLQIYPPPSRDELRKRFIAASEYALDFMCGMLTMNPQK
                     RWTAVQCLESDYFKELPPPSDPSSIKIRNVMAIPGGGRIFYPYDVPDYAGYPYDVPDYA
                     GSYPYDVPDYAAQCGRSS"
     misc_feature    434..874
                     /label=Gal4 DNA binding domain
                     /note="Gal4 DNA binding domain"
     CDS             434..874
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     misc_feature    932..952
                     /label=TEV protease cleavage sequence
                     /note="TEV protease cleavage sequence"
     CDS             932..952
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    977..1894
                     /label=Kin28 protein kinase
                     /note="Kin28 protein kinase"
     CDS             1928..2017
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      2456..2643
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    3475..4637
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      3800..4637
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(4476..5150)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(5151..5252)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     promoter        5358..5462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5463..6323
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6494..7082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"