pEF1/ V5- HisA Plasmid

$216.00

  • Model: PVT1035
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pEF1/ V5-HisA

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pEF1/V5-HisA,Plasmid pEF1/V5-HisA,pEF1/V5-HisA vector

 

pEF1/ V5-HisA Information

Promoter: EF-1 alpha

Replicator: F1 ori, pUC ori

Terminator: BGH poly (A) signal

Plasmid classification: mammalian cell carrier; protein overexpression plasmid.

Plasmid size: 6174bp

Plasmid label: 6X His, V5

Prokaryotic resistance: ampicillin Amp

Screening markers: neomycin Neo

Cloned strain: DH5 alpha

Culture conditions: 37 centigrade, aerobic, LB

Expression host: mammalian cells

Culture conditions: 37 centigrade, aerobic, LB

5'sequencing primers: T7 Fwd: 5

 

pEF1/ V5-HisA Description

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pEF1-V5-HisA-vector

 

 

pEF1/ V5-HisA Sequence

LOCUS       Exported                6174 bp ds-DNA     circular SYN 19-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pEF1 V5-HisA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6174)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 2016-9-19 from SnapGene Viewer 2.8.1
 
FEATURES             Location/Qualifiers
     source          1..6174
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        473..1651
                     /note="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     intron          704..1642
                     /note="EF-1-alpha intron A"
                     /note="intron upstream of the start codon of human
                     EF-1-alpha"
     promoter        1668..1686
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1805..1846
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /note="V5 tag"
                     /translation="GKPIPNPLLGLDST"
     CDS             1856..1873
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     polyA_signal    1902..2126
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2172..2600
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2614..2943
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      2794..2929
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             3010..3804
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3980..4101
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4150..4166)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    4174..4190
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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(4198..4228)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4243..4264
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4541..5129)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5300..6160)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
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