LOCUS       Exported                8993 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast vector pDEST-GBKT7, complete sequence.
ACCESSION   LT727256
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8993)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8993)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 8993)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8993
                     /organism="Yeast vector pDEST-GBKT7"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4543."
                     /db_xref="taxon:1945506"
     promoter        30..734
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     regulatory      336..739
                     /regulatory_class="promoter"
                     /note="ADH1 promoter"
     CDS             762..1202
                     /codon_start=1
                     /note="unnamed protein product; GALbd"
                     /protein_id="SJL87900.1"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     promoter        1213..1231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1213..1229
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    1251..1280
                     /label=Myc epitope
                     /note="Myc epitope"
     CDS             1251..1280
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      complement(1308..1432)
                     /regulatory_class="attenuator"
                     /note="attR1"
     protein_bind    1308..1432
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1436..1492
                     /regulatory_class="promoter"
                     /note="lacUV5 promoter"
     promoter        1457..1487
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1541..2200
                     /codon_start=1
                     /note="unnamed protein product; chloramphenicol resistance"
                     /protein_id="SJL87901.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2542..2847
                     /codon_start=1
                     /note="unnamed protein product; ccdB"
                     /protein_id="SJL87902.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     regulatory      2888..3012
                     /regulatory_class="attenuator"
                     /note="attR2"
     protein_bind    complement(2888..3012)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      3027..3074
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      3027..3074
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3101..3288
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     regulatory      3101..3202
                     /regulatory_class="terminator"
                     /note="ADH1 terminator"
     primer_bind     complement(3312..3328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3336..3352
                     /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)."
     regulatory      complement(3355..3438)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(3360..3390)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(3530..3932)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(3714..4302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    4631..4678
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      complement(4650..4719)
                     /regulatory_class="terminator"
                     /note="HSV-TK polyA"
     CDS             complement(4911..5705)
                     /codon_start=1
                     /note="unnamed protein product; Tn5 neomycin resistance"
                     /protein_id="SJL87903.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5706..5810)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    complement(5706..5740)
                     /label=5 UTR of amp, incl. RBS
                     /note="5 UTR of amp, incl. RBS"
     regulatory      complement(5741..5836)
                     /regulatory_class="promoter"
                     /note="ampicillin promoter"
     rep_origin      5837..7179
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(5837..6817)
                     /direction=LEFT
                     /label=2mu ori
                     /note="2mu ori"
     protein_bind    complement(6800..6847)
                     /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)."
     regulatory      complement(6895..6999)
                     /regulatory_class="terminator"
                     /note="FLP terminator"
     promoter        7438..7719
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             7720..8394
                     /codon_start=1
                     /note="unnamed protein product; TRP1"
                     /protein_id="SJL87904.1"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(8497..8952)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"