LOCUS       Exported                5611 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCR3Flag-E8-FLIP, complete sequence.
ACCESSION   LT727397
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5611)
  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  JOURNAL   Unpublished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 5611)
  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 5611)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5611
                     /organism="Mammalian expression vector pCR3Flag-E8-FLIP"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 3813."
                     /db_xref="taxon:1944958"
     regulatory      10..593
                     /regulatory_class="promoter"
                     /note="hCMV-IE promoter and enhancer"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      638..654
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    705..728
                     /label=FLAG epitope
                     /note="FLAG epitope"
     CDS             705..728
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             735..1250
                     /codon_start=1
                     /note="unnamed protein product; E8-FLIP"
                     /protein_id="SJL88342.1"
                     /translation="MSHYSMIDTYFSLDEDETETYLYLCRDLLKNKGEFQCTRDAFKFL
                     SDYACLSAANQMELLFRVGRLDLIRRIFGQTWTPDSCPRYYMPICSPFRCLMALVNDFL
                     SDKEVEEMYFLCAPRLESHLEPGSKKSFLRLASLLEDLELLGGDKLTFLRHLLTTIGRA
                     DLVKNLQV"
     promoter        complement(1326..1344)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      complement(1328..1344)
                     /regulatory_class="promoter"
                     /note="SP6 promoter"
     regulatory      1370..1594
                     /regulatory_class="terminator"
                     /note="BGH polyA"
     polyA_signal    1370..1594
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1618..1944)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(1726..2314)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    2643..2690
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      complement(2662..2731)
                     /regulatory_class="terminator"
                     /note="HSV-TK polyA"
     CDS             complement(2922..3710)
                     /codon_start=1
                     /note="unnamed protein product; Tn5 neomycin resistance"
                     /protein_id="SJL88343.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDQEHQGL
                     APAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALA
                     TRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(3745..4102)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3759..3894
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(3770..3845)
                     /direction=LEFT
                     /label=SV40 ori
                     /note="SV40 ori"
     regulatory      complement(3802..4083)
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     CDS             complement(4162..5022)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL88344.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5023..5127)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5154..5609
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 ori"