LOCUS       Exported                6750 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pEF1mBID, complete sequence.
ACCESSION   LT727184
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6750)
  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  JOURNAL   Unpublished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 6750)
  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 6750)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6750
                     /organism="Mammalian expression vector pEF1mBID"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4325."
                     /db_xref="taxon:1944982"
     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="hEF1 promoter"
     misc_feature    704..1642
                     /label=intron hEF1 promoter
                     /note="intron hEF1 promoter"
     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"
     CDS             1754..2341
                     /codon_start=1
                     /note="unnamed protein product; mBID"
                     /protein_id="SJL87697.1"
                     /translation="MDSEVSNGSGLGAKHITDLLVFGFLQSSGCTRQELEVLGRELPVQ
                     AYWEADLEDELQTDGSQASRSFNQGRIEPDSESQEEIIHNIARHLAQIGDEMDHNIQPT
                     LVRQLAAQFMNGSLSEEDKRNCLAKALDEVKTAFPRDMENDKAMLIMTMLLAKKVASHA
                     PSLLRDVFHTTVNFINQNLFSYVRNLVRNEMD"
     misc_feature    2381..2422
                     /label=V5 epitope
                     /note="V5 epitope"
     CDS             2381..2422
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2432..2449
                     /label=His-tag
                     /note="His-tag"
     CDS             2432..2449
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      2478..2702
                     /regulatory_class="terminator"
                     /note="BGH polyA"
     polyA_signal    2478..2702
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2748..3176
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      3181..3462
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        3190..3519
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3370..3505
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      3419..3494
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             3586..4380
                     /codon_start=1
                     /note="unnamed protein product; neomycin"
                     /protein_id="SJL87698.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4556..4677
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(4556..4630)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      4556..4630
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     primer_bind     complement(4726..4742)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4750..4766
                     /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(4769..4852)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(4774..4804)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4819..4840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4944..5335)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(5117..5705)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5876..6736)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin"
                     /protein_id="SJL87699.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6737..91)
                     /gene="bla"
                     /label=AmpR promoter