OGS211
PSF-CMV-NH2-HIS-EKT-NCOI - N-TERMINAL HIS TAG PLASMID
plasmid vector for molecular cloning
Synonym(s):
cloning vector, expression vector, molecular cloning vector, plasmid, plasmid vector, snapfast vector, vector
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About This Item
UNSPSC Code:
12352200
NACRES:
NA.85
tag
6-His tagged
form
buffered aqueous solution
mol wt
size 6152 bp
bacteria selection
kanamycin
Origin of replication
pUC (500 copies)
Peptide cleavage
EKT
Peptide tag location
N-terminal
Promoter
Promoter name: CMV
Promoter activity: constitutive
Promoter type: mammalian
reporter gene
none
shipped in
ambient
storage temp.
−20°C
General description
This vector adds a His epitope tag to the N-terminus of a protein that is encoded within the multiple cloning site. This tag allows the purification of a tagged protein by binding to metal matrices such as nickel or cobalt. There is an enterokinase cleavage site (DDDDK) immediately downstream of the His tag that can be used to remove the His tag from a purified protein. It cleaves after the lysine residue.
Promoter Expression Level: This plasmid contains the mammalian CMV promoter to drive gene expression. We have tested all of our mammalian promoters in a range of cell types and CMV is consistently the strongest in those we have studied. However there are many reports of the CMV promoter demonstrating silencing by methylation in long-term culture.
Promoter Expression Level: This plasmid contains the mammalian CMV promoter to drive gene expression. We have tested all of our mammalian promoters in a range of cell types and CMV is consistently the strongest in those we have studied. However there are many reports of the CMV promoter demonstrating silencing by methylation in long-term culture.
Application
Multiple cloning site notes: The his tag is in frame with the ATG start codon that is within the NcoI restriction site allowing fusion with any genes that we sell in the MCS that contain an NcoI at the 5 prime. This tag is positioned immediately upstream of the NcoI site to minimise the amount of extra amino acids that are added to the N-terminus of your gene.
The start codon that is within the NcoI restriction site is in frame with and adjacent to the coding sequence for the His/EKT peptide tag. Shine-Dalgarno sequences and KOZAK sequences are positioned correctly at the start of the His signal peptide. The ClaI to NheI sites have other functions such as adding peptide tags or IRES elements. The BsgI and BseRI restriction sites cleave within the stop codon in the XbaI site and allow fusion with our C-terminal peptide tags provided that your genes stop codon is also positioned here.
The start codon that is within the NcoI restriction site is in frame with and adjacent to the coding sequence for the His/EKT peptide tag. Shine-Dalgarno sequences and KOZAK sequences are positioned correctly at the start of the His signal peptide. The ClaI to NheI sites have other functions such as adding peptide tags or IRES elements. The BsgI and BseRI restriction sites cleave within the stop codon in the XbaI site and allow fusion with our C-terminal peptide tags provided that your genes stop codon is also positioned here.
Sequence
To view sequence information for this product, please visit the product page
Analysis Note
To view the Certificate of Analysis for this product, please visit www.oxgene.com
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Description
Pricing
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Regulatory Information
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