LOCUS       Exported                8285 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast two-hybrid vector pDG2, complete sequence.
ACCESSION   AY647982
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8285)
  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 8285)
  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 8285)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8285
                     /organism="Yeast two-hybrid vector pDG2"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:284153"
     CDS             434..2059
                     /codon_start=1
                     /product="GDBD-H3-Gcn5(F221A)-HA fusion protein"
                     /label=GDBD-H3-Gcn5
                     /note="derived from Saccharomyces cerevisiae; contains Gal4
                     DNA binding domain, histone H3, Gcn5 F221A mutant allele 
                     and a trimeric HA epitope under the control of yeast ADH1 
                     promoter and ADH1 terminator"
                     /protein_id="AAT97344.1"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSNYLFDDEDTPPNPKKE
                     IEFQLTTMFMARTKQTARKSTGGKAPRKQLASKAARKSAPSTGGVKKPHRYKPGTVALR
                     EIRRFQKSTEPGSPILGYWKGRRDHPPKSDLIEGRGDPEVKRVKLENNVEEIQPEQAET
                     NKQEGTDKENKGKFEKETERIGGSEVVTDVEKGIVKFEFDGVEYTFKERPSVVEENEGK
                     IEFRVVNNDNTKENMMVLTGLKNIFQKQLPKMPKEYIARLVYDRSHLSMAVIRKPLTVV
                     GGITYRPFDKREFAEIVFCAISSTEQVRGYGAHLMNHLKDYVRNTSNIKYFLTYADNYA
                     IGYAKKQGFTKEITLDKSIWMGYIKDYEGGTLMQCNMAIPGGGRIFYPYDVPDYAGYPY
                     DVPDYAGSYPYDVPDYAAQCGRSS"
     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    953..1129
                     /label=histone H3
                     /note="histone H3"
     CDS             1193..1204
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    1208..1912
                     /note="Gcn5 catalytic domain; F221A mutation"
     misc_feature    1945..2035
                     /label=trimeric HA epitope tag
                     /note="trimeric HA epitope tag"
     CDS             1946..2035
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      2474..2661
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    3493..4655
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      3818..4655
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(4494..5168)
                     /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(5169..5270)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     promoter        5376..5480
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5481..6341
                     /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      6512..7100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"