LOCUS       Exported                6407 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pcDNA3mB7-2, complete sequence.
ACCESSION   LT727375
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6407)
  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  JOURNAL   Unpublished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 6407)
  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 6407)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6407
                     /organism="Mammalian expression vector pcDNA3mB7-2"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 3696."
                     /db_xref="taxon:1944935"
     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"
     CDS             956..1885
                     /codon_start=1
                     /note="unnamed protein product; mB7-2"
                     /protein_id="SJL88271.1"
                     /translation="MDPRCTMGLAILIFVTVLLISDAVSVETQAYFNGTAYLPCPFTKA
                     QNISLSELVVFWQDQQKLVLYEHYLGTEKLDSVNAKYLGRTSFDRNNWTLRLHNVQIKD
                     MGSYDCFIQKKPPTGSIILQQTLTELSVIANFSEPEIKLAQNVTGNSGINLTCTSKQGH
                     PKPKKMYFLITNSTNEYGDNMQISQDNVTELFSISNSLSLSFPDGVWHMTVVCVLETES
                     MKISSKPLNFTQEFPSPQTYWKEITASVTVALLLVMLLIIVCHKKPNQPSRPSNTASKL
                     ERDSNADRETINLKELEPQIASAKPNAE"
     promoter        complement(1959..1977)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      complement(1961..1977)
                     /regulatory_class="promoter"
                     /note="SP6"
     regulatory      2003..2227
                     /regulatory_class="terminator"
                     /note="polyA"
     polyA_signal    2003..2227
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2273..2701
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      2706..2988
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        2715..3045
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2896..3031
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      2945..3020
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             3112..3906
                     /codon_start=1
                     /note="unnamed protein product; neomycin resistance"
                     /protein_id="SJL88272.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4080..4201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(4080..4154)
                     /regulatory_class="terminator"
                     /note="polyA"
     regulatory      4080..4154
                     /regulatory_class="terminator"
                     /note="polyA"
     primer_bind     complement(4250..4266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4274..4290
                     /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(4298..4328)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4343..4364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4468..4870)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(4652..5240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5411..6271)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL88273.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6272..6376)
                     /gene="bla"
                     /label=AmpR promoter