LOCUS       Exported                9038 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pRM2ScMET15, complete sequence.
ACCESSION   LT727226
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9038)
  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  JOURNAL   Unpublished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 9038)
  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 9038)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9038
                     /organism="Yeast expression vector pRM2ScMET15"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4490."
                     /db_xref="taxon:1945496"
     rep_origin      complement(63..763)
                     /direction=LEFT
                     /label=ARS2
                     /note="ARS2"
     CDS             complement(1662..2474)
                     /codon_start=1
                     /note="unnamed protein product; URA3"
                     /protein_id="SJL87813.1"
                     /translation="MTVNTKTYSERAETHASPVAQRLFRLMELKKTNLCASIDVDTTKE
                     FLELIDKLGPYVCLIKTHIDIINDFSYESTIEPLLELSRKHQFMIFEDRKFADIGNTVK
                     KQYIGGVYKISSWADITNAHGVTGNGVVEGLKQGAKETTTNQEPRGLLMLAELSSVGSL
                     AYGEYSQKTVEIAKSDKEFVIGFIAQRDMGGQEEGFDWLIMTPGVGLDDKGDGLGQQYR
                     TVDEVVSTGTDIIIVGRGLFGKGRDPDIEGKRYRNAGWNAYLKKTGQL"
     promoter        complement(3202..3306)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3578..3594
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      3603..4179
                     /regulatory_class="promoter"
                     /note="ScMET"
     promoter        3829..4179
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             4180..5514
                     /codon_start=1
                     /note="unnamed protein product; ScMET15"
                     /protein_id="SJL87814.1"
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     misc_feature    5516..6097
                     /label=3 UTR of ScMET15
                     /note="3 UTR of ScMET15"
     rep_origin      6103..6783
                     /label=ARS3
                     /note="ARS3"
     primer_bind     complement(6817..6833)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6841..6857
                     /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(6860..6943)
                     /regulatory_class="promoter"
                     /note="lac"
     promoter        complement(6865..6895)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6910..6931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7035..7437)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(7219..7807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7978..8838)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL87815.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8839..8943)
                     /gene="bla"
                     /label=AmpR promoter