LOCUS       Exported                8447 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCD-F-HLH-MK-bioC, complete sequence.
ACCESSION   LT726942
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8447)
  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  JOURNAL   Unpublished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 8447)
  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 8447)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8447
                     /organism="Mammalian expression vector pCD-F-HLH-MK-bioC"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 5681."
                     /db_xref="taxon:1944907"
     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"
     misc_feature    923..946
                     /label=FLAG epitope
                     /note="FLAG epitope"
     CDS             923..946
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      962..971
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             968..1447
                     /codon_start=1
                     /note="unnamed protein product; HLH dimerization domain"
                     /protein_id="SJL87022.1"
                     /translation="MALLGDPLQALPPSAAPTGPLLAPPAGATLNRLREPLLRRLSYAS
                     STPLHVPVPRALRMEEDSIRLPAHLRLQPIYWSRDDVAQWLKWAENEFSLRPIDSNTFE
                     MNGKALLLLTKEDFRYRSPHSGDVLYELLQHILKQRKPRILFSPFFHPGNSIHTQP"
     misc_feature    971..973
                     /label=Kozak
                     /note="Kozak"
     regulatory      1457..1466
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1463..3934
                     /codon_start=1
                     /note="unnamed protein product; hMLT"
                     /protein_id="SJL87023.1"
                     /translation="MALLGDPLQALPPSAAPTGPLLAPPAGATLNRLREPLLRRLSELL
                     DQAPEGRGWRRLAELAGSRGRLRLSCLDLEQCSLKVLEPEGSPSLCLLKLMGEKGCTVT
                     ELSDFLQAMEHTEVLQLLSPPGIKITVNPESKAVLAGQFVKLCCRATGHPFVQYQWFKM
                     NKEIPNGNTSELIFNAVHVKDAGFYVCRVNNNFTFEFSQWSQLDVCDIPESFQRSVDGV
                     SESKLQICVEPTSQKLMPGSTLVLQCVAVGSPIPHYQWFKNELPLTHETKKLYMVPYVD
                     LEHQGTYWCHVYNDRDSQDSKKVEIIIGRTDEAVECTEDELNNLGHPDNKEQTTDQPLA
                     KDKVALLIGNMNYREHPKLKAPLVDVYELTNLLRQLDFKVVSLLDLTEYEMRNAVDEFL
                     LLLDKGVYGLLYYAGHGYENFGNSFMVPVDAPNPYRSENCLCVQNILKLMQEKETGLNV
                     FLLDMCRKRNDYDDTIPILDALKVTANIVFGYATCQGAEAFEIQHSGLANGIFMKFLKD
                     RLLEDKKITVLLDEVAEDMGKCHLTKGKQALEIRSSLSEKRALTDPIQGTEYSAESLVR
                     NLQWAKAHELPESMCLKFDCGVQIQLGFAAEFSNVMIIYTSIVYKPPEIIMCDAYVTDF
                     PLDLDIDPKDANKGTPEETGSYLVSKDLPKHCLYTRLSSLQKLKEHLVFTVCLSYQYSG
                     LEDTVEDKQEVNVGKPLIAKLDMHRGLGRKTCFQTCLMSNGPYQSSAATSGGAGHYHSL
                     QDPFHGVYHSHPGNPSNVTPADSCHCSRTPDAFISSFAHHASCHFSRSNVPVETTDEIP
                     FSFSDRLRISEK"
     misc_feature    1466..1468
                     /label=Kozak
                     /note="Kozak"
     misc_feature    2948..2950
                     /label=stability creator
                     /note="stability creator"
     misc_feature    3410..3412
                     /label=PCR artefact
                     /note="PCR artefact"
     misc_feature    3941..3964
                     /label=SGSSGSSG linker
                     /note="SGSSGSSG linker"
     misc_feature    3965..4009
                     /label=bio-tag
                     /note="bio-tag"
     CDS             3965..4009
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     regulatory      4044..4268
                     /regulatory_class="terminator"
                     /note="BGH polyA"
     polyA_signal    4044..4268
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4314..4742
                     /label=f1 ori
                     /note="f1 ori"
     regulatory      4747..5028
                     /regulatory_class="promoter"
                     /note="SV40 early promoter"
     promoter        4756..5085
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4936..5071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4985..5060
                     /label=SV40 ori
                     /note="SV40 ori"
     CDS             5152..5946
                     /codon_start=1
                     /note="unnamed protein product; Tn5 neomycin resistance"
                     /protein_id="SJL87024.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    6120..6241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(6120..6194)
                     /regulatory_class="terminator"
                     /note="SV40 polyA late"
     regulatory      6120..6194
                     /regulatory_class="terminator"
                     /note="SV40 polyA early"
     primer_bind     complement(6290..6306)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6314..6330
                     /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(6333..6416)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(6338..6368)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6383..6404
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6508..6910)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(6692..7280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7451..8311)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL87025.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8312..8416)
                     /gene="bla"
                     /label=AmpR promoter