LOCUS       Exported                5246 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pSV-Sport-Raf-1E, complete sequence.
ACCESSION   LT726992
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5246)
  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  JOURNAL   Unpublished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 5246)
  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 5246)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5246
                     /organism="Mammalian expression vector pSV-Sport-Raf-1E"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5264."
                     /db_xref="taxon:1945160"
     regulatory      1..282
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        10..339
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      190..325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      239..314
                     /label=SV40 ori
                     /note="SV40 ori"
     promoter        345..363
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      345..361
                     /regulatory_class="promoter"
                     /note="SP6 promoter"
     CDS             395..2338
                     /codon_start=1
                     /note="unnamed protein product; hRaf1"
                     /protein_id="SJL87143.1"
                     /translation="MEHIQGAWKTISNGFGFKDAVFDGSSCISPTIVQQFGYQRRASDD
                     GKLTDPSKTSNTIRVFLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLHE
                     HKGKKARLDWNTDAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGF
                     RCQTCGYKFHEHCSTKVPTMCVDWSNIRQLLLFPNSTIGDSGVPALPSLTMRRMRESVS
                     RMPVSSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAI
                     RSHSESASPSALSSSPNNLSPTGWSQPKTPVPAQRERAPVSGTQEKNKIRPRGQRDSSY
                     YWEIEASEVMLSTRIGSGSFGTVYKGKWHGDVAVKILKVVDPTPEQFQAFRNEVAVLRK
                     TRHVNILLFMGYMTKDNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDY
                     LHAKNIIHRDMKSNNIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVI
                     RMQDNNPFSFQSDVYSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSKLYKN
                     CPKAMKRLVADCVKKVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDIN
                     ACTLTTSPRLPVF"
     misc_feature    2348..2386
                     /label=E-tag
                     /note="E-tag"
     promoter        complement(2564..2582)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(2566..2582)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    2671..2710
                     /label=intron of SV40 small t-antigen
                     /note="intron of SV40 small t-antigen"
     regulatory      complement(2947..3043)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      2947..3043
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     polyA_signal    2969..3103
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3156..3558)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     CDS             complement(4089..4949)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin"
                     /protein_id="SJL87144.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4950..5054)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      5145..5202
                     /regulatory_class="promoter"
                     /note="lacUV5 promoter"
     protein_bind    5205..5221
                     /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)."