LOCUS       Exported                6054 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pcDNAFLAGhRAIDDMyc, complete sequence.
ACCESSION   LT727264
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6054)
  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  JOURNAL   Unpublished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 6054)
  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 6054)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6054
                     /organism="Mammalian expression vector pcDNAFLAGhRAIDDMyc"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 3842."
                     /db_xref="taxon:1944942"
     regulatory      232..818
                     /regulatory_class="promoter"
                     /note="hCMV"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      863..879
                     /regulatory_class="promoter"
                     /note="T7"
     misc_feature    904..927
                     /label=FLAG
                     /note="FLAG"
     CDS             904..927
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             937..1533
                     /codon_start=1
                     /note="unnamed protein product; hRAIDD"
                     /protein_id="SJL87929.1"
                     /translation="MEARDKQVLRSLRLELGAEVLVEGLVLQYLYQEGILTENHIQEIN
                     AQTTGLRKTMLLLDILPSRGPKAFDTFLDSLQEFPWVREKLKKAREEAMTDLPAGDRLT
                     GIPSHILNSSPSDRQINQLAQRLGPEWEPMVLSLGLSQTDIYRCKANHPHNVQSQVVEA
                     FIRWRQRFGKQATFQSLHNGLRAVEVDPSLLLHMLE"
     misc_feature    1540..1569
                     /label=myc
                     /note="myc"
     CDS             1540..1569
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(1606..1624)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      complement(1608..1624)
                     /regulatory_class="promoter"
                     /note="SP6"
     regulatory      1650..1874
                     /regulatory_class="terminator"
                     /note="polyA"
     polyA_signal    1650..1874
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1920..2348
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      2353..2635
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        2362..2692
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2543..2678
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      2592..2667
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             2759..3553
                     /codon_start=1
                     /note="unnamed protein product; neomycin resistance"
                     /protein_id="SJL87930.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3727..3848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(3727..3801)
                     /regulatory_class="terminator"
                     /note="polyA"
     regulatory      3727..3801
                     /regulatory_class="terminator"
                     /note="polyA"
     primer_bind     complement(3897..3913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3921..3937
                     /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(3945..3975)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3990..4011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4115..4517)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(4299..4887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5058..5918)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL87931.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5919..6023)
                     /gene="bla"
                     /label=AmpR promoter