LOCUS       Exported                8578 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pGBKT7-Cezanne(17-443), complete sequence.
ACCESSION   LT726928
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8578)
  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  JOURNAL   Unpublished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 8578)
  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 8578)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8578
                     /organism="Yeast expression vector pGBKT7-Cezanne(17-443)"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5572."
                     /db_xref="taxon:1945484"
     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="SJL86978.1"
                     /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"
     CDS             1305..2588
                     /codon_start=1
                     /note="unnamed protein product; Cezanne(17-444)"
                     /protein_id="SJL86979.1"
                     /translation="TLDMDAVLSDFVRSTGAEPGLARDLLEGKNWDVNAALSDFEQLRQ
                     VHAGNLPPSFSEGSGGSRTPEKGFSDREPTRPPRPILQRQDDIVQEKRLSRGISHASSS
                     IVSLARSHVSSNGGGGGSNEHPLEMPICAFQLPDLTVYNEDFRSFIERDLIEQSMLVAL
                     EQAGRLNWWVSVDPTSQRLLPLATTGDGNCLLHAASLGMWGFHDRDLMLRKALYALMEK
                     GVEKEALKRRWRWQQTQQNKESGLVYTEDEWQKEWNELIKLASSEPRMHLGTNGANCGG
                     VESSEEPVYESLEEFHVFVLAHVLRRPIVVVADTMLRDSGGEAFAPIPFGGIYLPLEVP
                     ASQCHRSPLVLAYDQAHFSALVSMEQKENTKEQAVIPLTDSEYKLLPLHFAVDPGKGWE
                     WGKDDSDNVRLASVILSLEVKLHLLHSYM"
     regulatory      2612..2659
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      2612..2659
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2686..2873
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     regulatory      2686..2787
                     /regulatory_class="terminator"
                     /note="ADH1 terminator"
     primer_bind     complement(2897..2913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2921..2937
                     /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(2940..3023)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(2945..2975)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(3115..3517)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(3299..3887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    4216..4263
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      complement(4235..4304)
                     /regulatory_class="terminator"
                     /note="HSV-TK polyA"
     CDS             complement(4496..5290)
                     /codon_start=1
                     /note="unnamed protein product; Tn5 neomyciin"
                     /protein_id="SJL86980.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5291..5395)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    complement(5291..5325)
                     /label=5 UTR of amp, incl. RBS
                     /note="5 UTR of amp, incl. RBS"
     regulatory      complement(5326..5421)
                     /regulatory_class="promoter"
                     /note="ampicillin promoter"
     rep_origin      5422..6764
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(5422..6402)
                     /direction=LEFT
                     /label=2mu ori
                     /note="2mu ori"
     protein_bind    complement(6385..6432)
                     /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(6480..6584)
                     /regulatory_class="terminator"
                     /note="FLP terminator"
     promoter        7023..7304
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             7305..7979
                     /codon_start=1
                     /note="unnamed protein product; TRP1"
                     /protein_id="SJL86981.1"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(8082..8537)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"