LOCUS       Exported                9011 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast two-hybrid vector pGBKCg, complete sequence.
ACCESSION   FJ696409
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9011)
  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  JOURNAL   Unpublished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 9011)
  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 9011)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9011
                     /organism="Yeast two-hybrid vector pGBKCg"
                     /mol_type="other DNA"
                     /db_xref="taxon:620856"
     promoter        30..734
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     regulatory      338..734
                     /regulatory_class="promoter"
                     /note="yADH1*_promoter"
     protein_bind    769..893
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    769..888
                     /note="attR1; binding site"
     promoter        918..948
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1002..1661
                     /codon_start=1
                     /product="CamR"
                     /label=CamR
                     /note="chloramphenicol resistance"
                     /protein_id="ACN64939.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2003..2308
                     /codon_start=1
                     /product="CcdB"
                     /label=CcdB
                     /note="F plasmid CcdB protein"
                     /protein_id="ACN64940.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2349..2473)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2349..2449
                     /note="attR2; binding site"
     misc_feature    2487..2927
                     /label=GAL4 DNA binding domain
                     /note="GAL4 DNA binding domain"
     CDS             2487..2927
                     /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"
     promoter        2938..2956
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2976..3005
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      3093..3140
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      3143..3310
                     /regulatory_class="terminator"
                     /note="yADH1_terminator"
     primer_bind     complement(3330..3346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3354..3370
                     /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(3378..3408)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      3717..4336
                     /label=pBR322_origin
                     /note="pBR322_origin"
     rep_origin      complement(3732..4320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    4649..4696
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(4929..5723)
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    complement(4929..5711)
                     /note="NeoR/KanR variant; kanamycin resistance"
     promoter        complement(5724..5828)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5855..7197
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      5855..6826
                     /label=2micron2_origin
                     /note="2micron2_origin"
     protein_bind    complement(6818..6865)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        7456..7737
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             7738..8412
                     /codon_start=1
                     /product="TRP1"
                     /label=TRP1
                     /protein_id="ACN64941.1"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(8515..8970)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"