LOCUS       Exported                6449 bp ds-DNA    circular SYN 23-2-2014
DEFINITION  Yeast two-hybrid vector with a GAL1 promoter, for fusing the B42 
            transcriptional activator to a partner protein. Also known as 
            pB42AD.
ACCESSION   U89961.1
VERSION     .
KEYWORDS    pJG4-5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6449)
  AUTHORS   Gyuris J, Golemis E, Chertkov H, Brent R.
  TITLE     Cdi1, a human G1 and S phase protein phosphatase that associates 
            with Cdk2.
  JOURNAL   Cell 1993;75:791-803.
  PUBMED    8242750
REFERENCE   2  (bases 1 to 6449)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6449
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     promoter        81..522
                     /gene="S. cerevisiae GAL1"
                     /note="GAL1 promoter"
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    81..198
                     /bound_moiety="Gal4"
                     /note="UAS"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             534..536
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     CDS             546..566
                     /codon_start=1
                     /locus_tag="
                     "
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /note="SV40 NLS"
                     /note="
                     "
                     /protein_id="
                     "
                     /translation="PKKKRKV"
     CDS             573..809
                     /codon_start=1
                     /product="yeast transcriptional activator cretaed from E. 
                     coli genomic DNA fragments (Ma and Ptashne, 1987)"
                     /note="B42 transcriptional activator"
                     /translation="INKDIEECNAIIEQFIDYLRTGQEMPMEMADQAINVVPGMTPKTI
                     LHAGPPIQPDWLKSNGFHEIEADVNDTSLLLSGD"
     CDS             816..842
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /note="HA"
                     /translation="YPYDVPDYA"
     misc_feature    849..866
                     /note="MCS"
                     /note="multiple cloning site"
     terminator      1004..1191
                     /gene="S. cerevisiae ADH1"
                     /note="ADH1 terminator"
                     /note="transcription terminator for alcohol dehydrogenase 
                     1"
     rep_origin      1713..3055
                     /note="2u ori"
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3416..4090)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /note="TRP1"
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(4091..4192)
                     /gene="S. cerevisiae TRP1"
                     /note="TRP1 promoter"
     promoter        4295..4399
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             4403..5263
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="SIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIE
                     LDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSP
                     VTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWE
                     PELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSAL
                     PAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASL
                     IKHW*L"
     rep_origin      5431..6019
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"