LOCUS       Exported                8189 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast two-hybrid vector pDG4, complete sequence.
ACCESSION   AY647984
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8189)
  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 8189)
  AUTHORS   Kuo M-H., Guo D.
  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 8189)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8189
                     /organism="Yeast two-hybrid vector pDG4"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:284155"
     CDS             434..1963
                     /codon_start=1
                     /product="GDBD-H4-Gcn5(F221A)-HA fusion protein"
                     /label=GDBD-H4-Gcn5
                     /note="derived from Saccharomyces cerevisiae; contains Gal4
                     DNA binding domain, histone H4, Gcn5 F221A mutant allele 
                     and a trimeric HA epitope under the control of yeast ADH1 
                     promoter and ADH1 terminator"
                     /protein_id="AAT97346.1"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSNYLFDDEDTPPNPKKE
                     IEFQLTTMSGRGKGGKGLGKGGAKRHRKILRDNIQGISGSPILGYWKGRRDHPPKSDLI
                     EGRGDPEVKRVKLENNVEEIQPEQAETNKQEGTDKENKGKFEKETERIGGSEVVTDVEK
                     GIVKFEFDGVEYTFKERPSVVEENEGKIEFRVVNNDNTKENMMVLTGLKNIFQKQLPKM
                     PKEYIARLVYDRSHLSMAVIRKPLTVVGGITYRPFDKREFAEIVFCAISSTEQVRGYGA
                     HLMNHLKDYVRNTSNIKYFLTYADNYAIGYAKKQGFTKEITLDKSIWMGYIKDYEGGTL
                     MQCSMAIPGGGRIFYPYDVPDYAGYPYDVPDYAGSYPYDVPDYAAQCGRSS"
     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"
     CDS             1097..1108
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1850..1939
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      2378..2565
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    3397..4559
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      3722..4559
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(4398..5072)
                     /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(5073..5174)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     promoter        5280..5384
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5385..6245
                     /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      6416..7004
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"