LOCUS       Exported               10533 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pSCGALCaMET15-2, complete sequence.
ACCESSION   LT727324
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10533)
  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  JOURNAL   Unpublished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 10533)
  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 10533)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10533
                     /organism="Yeast expression vector pSCGALCaMET15-2"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4213."
                     /db_xref="taxon:1945499"
     rep_origin      complement(696..1209)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(754..1209)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"
     primer_bind     1697..1713
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1977..3071)
                     /codon_start=1
                     /note="unnamed protein product; LEU2D"
                     /protein_id="SJL88108.1"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     rep_origin      3429..4775
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(3729..4409)
                     /direction=LEFT
                     /label=2mu ori
                     /note="2mu ori"
     misc_feature    4176..4778
                     /label=IR
                     /note="IR"
     regulatory      complement(4491..4595)
                     /regulatory_class="terminator"
                     /note="FLPT"
     regulatory      complement(4781..5051)
                     /regulatory_class="terminator"
                     /note="CYC1T"
     terminator      4784..5031
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    complement(5135..5172)
                     /label=3UTR
                     /note="3UTR"
     CDS             complement(5173..6495)
                     /codon_start=1
                     /note="unnamed protein product; CaMET15"
                     /protein_id="SJL88109.1"
                     /translation="MPSHFDTLQLHAGQPVEKPHQPRAPPIYATTSYVFNDSKHGAQLF
                     GLETPGYIYSRIMNPTNDVFEQRIAALEGGIGALATSSGQSAQFLAIAGLAHAGDNIIS
                     TSYLYGGTYNQFKVAFKRLGIETKFVNGDAAEDFAKLIDDKTKAIYIETIGNPKYNVPD
                     FEKITKLAHEHGIPVVVDNTFGAGGFLVNPIAHGADIVVHSATKWIGGHGTTIAGVIVD
                     SGNFPWTEYPEKYPQFSKPSEGYHGLILNDALGKAAYIGHLRIELLRDLGPALNPFGSF
                     LLLQGLETLSLRVERQSENALKLAQWLEKNPNVESVSYLGLPSHESHELSKKYLNNDAK
                     YFGGALAFTVKDITNTSSDPFNEASPKLVDNLEIASNLANVGDSKTLVIAPWFTTHQQL
                     SDEEKLASGVTKGLIRVSTGTEYIDDIINDFEQAFKKVYNN"
     misc_feature    complement(6496..6511)
                     /label=5UTR
                     /note="5UTR"
     regulatory      complement(6642..7049)
                     /regulatory_class="promoter"
                     /note="GA:CY"
     protein_bind    6894..7011
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             complement(7268..8071)
                     /codon_start=1
                     /note="unnamed protein product; URA3"
                     /protein_id="SJL88110.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(8072..8288)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(8308..8324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8332..8348
                     /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(8356..8386)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(8526..8928)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(8710..9298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9468..10328)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL88111.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(10329..10433)
                     /gene="bla"
                     /label=AmpR promoter