LOCUS       Exported               10609 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pEX2kanMX4, complete sequence.
ACCESSION   LT727065
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10609)
  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  JOURNAL   Unpublished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 10609)
  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 10609)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10609
                     /organism="Yeast expression vector pEX2kanMX4"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5010."
                     /db_xref="taxon:1945471"
     rep_origin      3..840
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     rep_origin      94..838
                     /label=ARS1
                     /note="ARS1"
     regulatory      complement(839..1109)
                     /regulatory_class="terminator"
                     /note="S. cerevisiae CYC1 terminator"
     terminator      842..1089
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     regulatory      complement(1229..2210)
                     /regulatory_class="promoter"
                     /note="S. cerevisiae CYC1 promoter"
     promoter        3632..3847
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3848..4651
                     /codon_start=1
                     /note="unnamed protein product; S. cerevisiae URA3"
                     /protein_id="SJL87328.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      5577..6919
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     regulatory      5757..5861
                     /regulatory_class="terminator"
                     /note="FLP terminator"
     protein_bind    5909..5956
                     /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)."
     rep_origin      5939..6919
                     /label=2mu ori
                     /note="2mu ori"
     misc_feature    7061..7201
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7203..7605)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(7387..7975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8146..9006)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL87329.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9007..9111)
                     /gene="bla"
                     /label=AmpR promoter
     gene            complement(9145..10501)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     regulatory      complement(9145..9342)
                     /regulatory_class="terminator"
                     /note="E. gossypii TEF1 terminator"
     terminator      9145..9342
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(9348..10157)
                     /codon_start=1
                     /note="unnamed protein product; Tn903 neomycin resistance"
                     /protein_id="SJL87330.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     regulatory      complement(10158..10501)
                     /regulatory_class="promoter"
                     /note="E. gossypii TEF1 promoter"
     promoter        complement(10158..10501)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"