HA-MCS-pcDNA3.1 vector (V009926)

Basic Vector Information

      • Vector Name:
      • HA-MCS-pcDNA3.1
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 5448 bp
      • Type:
      • Mammalian expression vector
      • Source/Author:
      • Nakashima K, Song SY.

HA-MCS-pcDNA3.1 vector Vector Map

HA-MCS-pcDNA3.15448 bp60012001800240030003600420048005400hCMV-IE promoter / enhancerT7 promoterHA-tagMulti Cloning SiteSP6 promoterBGH (Bovine Growth Hormone) polyA signalf1 originSV40 early promoter and originaphSV40 early polyA signalM13 revlac operatorlac promoterCAP binding sitepUC originbla

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

HA-MCS-pcDNA3.1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

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  JOURNAL   Unpublished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)"

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