LOCUS       Exported                5496 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCD-Lck-Flag, complete sequence.
ACCESSION   LT727010
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5496)
  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  JOURNAL   Unpublished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 5496)
  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 5496)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5496
                     /organism="Mammalian expression vector pCD-Lck-Flag"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5350."
                     /db_xref="taxon:1944909"
     regulatory      235..818
                     /regulatory_class="promoter"
                     /note="hCMV-IE promoter and enhancer"
     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 promoter"
     CDS             914..952
                     /codon_start=1
                     /note="unnamed protein product; mLck 
                     myristoylation-targeting sequence"
                     /protein_id="SJL87192.1"
                     /translation="MGCVCSSNPEDDW"
     misc_feature    968..991
                     /label=FLAG epitope
                     /note="FLAG epitope"
     CDS             968..991
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1091..1315
                     /regulatory_class="terminator"
                     /note="BGH polyA"
     polyA_signal    1091..1315
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1361..1789
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      1794..2075
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        1803..2132
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1983..2118
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      2032..2107
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             2199..2993
                     /codon_start=1
                     /note="unnamed protein product; Tn5 neomycin resistance"
                     /protein_id="SJL87193.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3169..3290
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(3169..3243)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      3169..3243
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     primer_bind     complement(3339..3355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3363..3379
                     /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(3382..3465)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(3387..3417)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3432..3453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3557..3959)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(3741..4329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4500..5360)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL87194.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5361..5465)
                     /gene="bla"
                     /label=AmpR promoter