LOCUS       Exported                9637 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast two-hybrid vector pGADCg, complete sequence.
ACCESSION   FJ696408
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9637)
  AUTHORS   Stellberger T, Uetz P.
  TITLE     Yeast two-hybrid vectors with C-terminal fusion of Gal4 activation 
            and DNA-binding domains
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9637)
  AUTHORS   Stellberger T, Uetz P.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-FEB-2009) Institut fuer Toxikologie und Genetik, 
            Forschungszentrum Karlsruhe, Hermann-von-Helmholtz-Platz 1, 
            Eggenstein-Leopoldshafen, Baden-Wuerttemberg 76344, Germany
REFERENCE   3  (bases 1 to 9637)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9637
                     /organism="Yeast two-hybrid vector pGADCg"
                     /mol_type="other DNA"
                     /db_xref="taxon:620855"
     regulatory      7..1479
                     /regulatory_class="promoter"
                     /note="yADH1"
     promoter        771..1476
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     misc_feature    1501..1554
                     /label=SV40_NLS
                     /note="SV40_NLS"
     CDS             1522..1542
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    1561..1685
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    1561..1680
                     /note="attR1; binding site"
     promoter        1710..1740
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1794..2453
                     /codon_start=1
                     /product="CamR"
                     /label=CamR
                     /note="chloramphenicol resistance"
                     /protein_id="ACN64936.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2795..3100
                     /codon_start=1
                     /product="CcdB"
                     /label=CcdB
                     /note="F plasmid CcdB protein"
                     /protein_id="ACN64937.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3141..3265)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    3141..3241
                     /note="attR2; binding site"
     CDS             3279..3620
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     misc_feature    3282..3620
                     /label=GAL4 activation domain
                     /note="GAL4 activation domain"
     regulatory      3928..4254
                     /regulatory_class="terminator"
                     /note="yADH2_terminator"
     terminator      4065..4252
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(4369..5463)
                     /codon_start=1
                     /product="Leu2"
                     /label=Leu2
                     /note="beta-isopropylmalate dehydrogenase (IMDH); catalyzes
                     the third step in the leucine biosynthesis pathway"
                     /protein_id="ACN64938.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(5464..5869)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(5911..5927)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5935..5951
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5959..5989)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6004..6025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(6080..6113)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(6191..6224)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      6449..7068
                     /label=pBR322_origin
                     /note="pBR322_origin"
     rep_origin      complement(6464..7052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7223..8083)
                     /codon_start=1
                     /product="AmpC"
                     /label=AmpC
                     /note="beta lactamase"
                     /protein_id="ACN64935.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8084..8188)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8470..9634
                     /label=2micron2_origin
                     /note="2micron2_origin"