LOCUS       Exported                7087 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian vector pdcDNA-FLAGMyc, complete sequence.
ACCESSION   LT727165
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7087)
  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  JOURNAL   Unpublished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 7087)
  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 7087)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7087
                     /organism="Mammalian vector pdcDNA-FLAGMyc"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 4705."
                     /db_xref="taxon:1945171"
     regulatory      27..610
                     /regulatory_class="promoter"
                     /note="hCMV-IE promoter and enhancer"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        655..673
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      655..671
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    696..719
                     /label=FLAG epitope
                     /note="FLAG epitope"
     CDS             705..719
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     regulatory      complement(729..853)
                     /regulatory_class="attenuator"
                     /note="attR1"
     protein_bind    729..853
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      861..913
                     /regulatory_class="promoter"
                     /note="lacUV5 promoter"
     promoter        878..908
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             962..1621
                     /codon_start=1
                     /note="unnamed protein product; chloramphenicol resistance"
                     /protein_id="SJL87637.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1963..2268
                     /codon_start=1
                     /note="unnamed protein product; ccdB"
                     /protein_id="SJL87638.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     regulatory      2309..2433
                     /regulatory_class="attenuator"
                     /note="attR2"
     protein_bind    complement(2309..2433)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2438..2467
                     /label=Myc epitope
                     /note="Myc epitope"
     CDS             2438..2467
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(2504..2522)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      complement(2506..2522)
                     /regulatory_class="promoter"
                     /note="SP6 promoter"
     regulatory      2548..2772
                     /regulatory_class="terminator"
                     /note="BGH polyA"
     polyA_signal    2548..2772
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2818..3246
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      3251..3460
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        3299..3509
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3321..3517
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3368..3503
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      3417..3492
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             3584..4378
                     /codon_start=1
                     /note="unnamed protein product; Tn5 neomycin resistance"
                     /protein_id="SJL87639.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4552..4673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(4552..4626)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      4552..4626
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     primer_bind     complement(4722..4738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4746..4762
                     /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(4765..4848)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(4770..4800)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4815..4836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4940..5342)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(5124..5712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5883..6743)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL87640.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6744..6848)
                     /gene="bla"
                     /label=AmpR promoter