LOCUS       Exported                9267 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEG(KG), complete sequence.
ACCESSION   FJ526991
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9267)
  AUTHORS   Pierce BD, Wendland B.
  TITLE     Sequence of the yeast protein expression plasmid pEG(KT)
  JOURNAL   Yeast 26 (6), 349-353 (2009)
  PUBMED    19350530
REFERENCE   2  (bases 1 to 9267)
  AUTHORS   Pierce BD, Wendland B.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-DEC-2008) Biology, The Johns Hopkins University, 3400 
            N. Charles St., Mudd Hall, Baltimore, MD 21218, USA
REFERENCE   3  (bases 1 to 9267)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9267
                     /organism="Cloning vector pEG(KG)"
                     /mol_type="other DNA"
                     /db_xref="taxon:596394"
     CDS             1..18
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     rep_origin      327..1309
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             1676..2770
                     /codon_start=1
                     /gene="LEU2"
                     /allele="leu2-d"
                     /product="beta-isopropylmalate dehydrogenase"
                     /EC_number="1.1.1.85"
                     /label=LEU2
                     /note="Leu2; 3-isopropylmalate dehydrogenase; enzyme in 
                     leucine biosynthetic pathway"
                     /protein_id="ACM86781.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     gene            1676..2770
                     /gene="LEU2"
                     /allele="leu2-d"
                     /label=LEU2
     primer_bind     complement(3021..3037)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3477..3990
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4735..4839
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4840..5700
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /EC_number="3.5.2.6"
                     /label=bla
                     /note="resistance to ampicillin"
                     /protein_id="ACM86782.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4840..5700
                     /gene="bla"
                     /label=bla
     rep_origin      5871..6459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6746..6767
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6782..6812
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6820..6836
                     /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)."
     primer_bind     6844..6860
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6880..7091
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             7092..7895
                     /codon_start=1
                     /gene="URA3"
                     /allele="URA3"
                     /product="orotidine-5-phosphate decarboxylase"
                     /EC_number="4.1.1.23"
                     /label=URA3
                     /note="Ura3; enzyme in uracil biosynthetic pathway"
                     /protein_id="ACM86783.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            7092..7895
                     /gene="URA3"
                     /allele="URA3"
                     /label=URA3
     regulatory      7974..8339
                     /regulatory_class="promoter"
                     /note="GAL1/10 intergenic region; galactose-regulated 
                     promoter"
     protein_bind    8152..8269
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     regulatory      8344..8589
                     /regulatory_class="promoter"
                     /note="CYC1"
     CDS             8608..93
                     /codon_start=1
                     /gene="GST"
                     /product="glutathione-S-transferase"
                     /EC_number="2.5.1.18"
                     /label=GST
                     /note="with C-terminal linker containing a 
                     thrombin-cleavage site followed by a glycine kinker"
                     /protein_id="ACM86784.1"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLV
                     PRGSPGISGGGGGILDSMGRLELKLNSS"
     gene            8608..93
                     /gene="GST"
                     /label=GST
     CDS             8608..9261
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"