LOCUS       Exported                7628 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pEF1crmA-T291R, complete sequence.
ACCESSION   LT727085
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7628)
  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  JOURNAL   Unpublished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 7628)
  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 7628)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7628
                     /organism="Mammalian expression vector pEF1crmA-T291R"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5104."
                     /db_xref="taxon:1944974"
     promoter        473..1651
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     regulatory      508..1651
                     /regulatory_class="promoter"
                     /note="hEF1alpha promoter"
     misc_feature    704..1642
                     /label=intron
                     /note="intron"
     intron          704..1642
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     promoter        1668..1686
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1668..1684
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    1754..2028
                     /label=5 UTR of crmA
                     /note="5 UTR of crmA"
     CDS             2029..3054
                     /codon_start=1
                     /note="unnamed protein product; crmA-T291R"
                     /protein_id="SJL87388.1"
                     /translation="MDIFREIASSMKGENVFISPPSISSVLTILYYGANGSTAEQLSKY
                     VEKEADKNKDDISFKSMNKVYGRYSAVFKDSFLRKIGDNFQTVDFTDCRTVDAINKCVD
                     IFTEGKINPLLDEPLSPDTCLLAISAVYFKAKWLMPFEKEFTSDYPFYVSPTEMVDVSM
                     MSMYGEAFNHASVKESFGNFSIIELPYVGDTSMVVILPDNIDGLESIEQNLTDTNFKKW
                     CDSMDAMFIDVHIPKFKVTGSYNLVDALVKLGLTEVFGSTGDYSNMCNSDVSVDAMIHK
                     TYIDVNEEYREAAAATCALVADCASTVTNEFCADHPFIYVIRHVDGKILFVGRYCSPTT
                     N"
     misc_feature    3055..3200
                     /label=3 UTR of crmA
                     /note="3 UTR of crmA"
     misc_feature    3259..3300
                     /label=V5 epitope
                     /note="V5 epitope"
     CDS             3259..3300
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3310..3327
                     /label=His-tag
                     /note="His-tag"
     CDS             3310..3327
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      3356..3580
                     /regulatory_class="terminator"
                     /note="bovine growth hormone polyA"
     polyA_signal    3356..3580
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3626..4054
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      4059..4340
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        4068..4397
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4248..4383
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4297..4372
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             4464..5258
                     /codon_start=1
                     /note="unnamed protein product; neomycin"
                     /protein_id="SJL87389.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5434..5555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(5434..5508)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      5434..5508
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     primer_bind     complement(5604..5620)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5628..5644
                     /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(5652..5682)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5697..5718
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5822..6213)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(5995..6583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6754..7614)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin"
                     /protein_id="SJL87390.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7615..91)
                     /gene="bla"
                     /label=AmpR promoter