LOCUS       Exported                5448 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector HA-MCS-pcDNA3.1 DNA, complete sequence.
ACCESSION   LC311022
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5448)
  AUTHORS   Nakashima K, Song SY.
  TITLE     Expression Vector for Mammalian Cell
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5448)
  AUTHORS   Nakashima K, Song SY.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUL-2017) Contact:Kentaro Nakashima Tokushima Bunri 
            University, Institute of Neuroscience; 1314-1 Shido, Sanuki, Kagawa 
            769-2193, Japan
REFERENCE   3  (bases 1 to 5448)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5448
                     /organism="Mammalian expression vector HA-MCS-pcDNA3.1"
                     /mol_type="other DNA"
                     /db_xref="taxon:2021192"
     regulatory      232..819
                     /regulatory_class="promoter"
                     /note="hCMV-IE promoter / 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"
     regulatory      863..882
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    904..933
                     /label=HA-tag
                     /note="HA-tag"
     CDS             907..933
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    934..1000
                     /label=Multi Cloning Site
                     /note="Multi Cloning Site"
     promoter        complement(1004..1022)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      complement(1005..1022)
                     /regulatory_class="promoter"
                     /note="SP6 promoter"
     regulatory      1048..1272
                     /regulatory_class="polyA_signal_sequence"
                     /note="BGH (Bovine Growth Hormone) polyA signal"
     polyA_signal    1048..1272
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1318..1746
                     /label=f1 origin
                     /note="f1 origin"
     regulatory      1751..2094
                     /regulatory_class="promoter"
                     /note="SV40 early promoter and origin"
     promoter        1760..2089
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1940..2075
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2156..2950
                     /codon_start=1
                     /gene="aph"
                     /product="Tn5 aminoglycoside phosphotransferase"
                     /label=aph
                     /note="Neomycin resistance gene"
                     /protein_id="BBA12681.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            2156..2950
                     /gene="aph"
                     /label=aph
     regulatory      3124..3254
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 early polyA signal"
     polyA_signal    3124..3245
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3294..3310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3318..3334
                     /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(3342..3372)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3387..3408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3637..4307)
                     /direction=LEFT
                     /label=pUC origin
                     /note="pUC origin"
     rep_origin      complement(3696..4281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     gene            complement(4452..5353)
                     /gene="bla"
                     /label=bla
     CDS             complement(4452..5312)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="Ampicillin resistance gene"
                     /protein_id="BBA12682.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5313..5417)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(5320..5324)
                     /regulatory_class="ribosome_binding_site"
                     /gene="bla"
     regulatory      complement(5347..5353)
                     /regulatory_class="promoter"
                     /gene="bla"
                     /note="bla promoter (P3)"