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NACRES:
NA.85
UNSPSC Code:
12352200
Promoter:
Promoter name: CMV
Promoter activity: constitutive
Promoter type: mammalian
Promoter activity: constitutive
Promoter type: mammalian
Origin of replication:
pUC (500 copies)
Bacteria selection:
kanamycin
Reporter gene:
none
Peptide cleavage:
no cleavage
tag
Hemagglutinin A (HA) tagged
form
buffered aqueous solution
mol wt
size 4265 bp
bacteria selection
kanamycin
origin of replication
pUC (500 copies)
peptide cleavage
no cleavage
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 plasmid adds a HA epitope tag to the N-terminus of a protein that is encoded within the multiple cloning site. This tag allows the detection and purification of a tagged protein using antibodies raised against the Influenza HA epitope. The HA tag coding sequence is YPYDVPDYA. There is an enterokinase cleavage site (DDDDK) immediately downstream of the HA tag that can be used to remove the HA 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
Cloning in a gene: This vector has been designed to allow the addition of a peptide tag to the end of a protein of interest using standard cloning techniques.Multiple Cloning Site Notes:
There is a start codon in the NcoI site can be removed by digestion with KpnI if required. The MCS for gene insertions extends from NotI to XbaI however the tag resides between the NotI and HindIII sites. There are Shine-Dalgarno sequences and KOZAK sequences aligned with the start codon of the peptide tag.
The ClaI to NheI sites have other functions such as adding C-terminal peptide tags second promoters or IRES expression components. The BsgI and BseRI restriction sites cleave within the stop codon in the XbaI site and allow the retrospective fusion of C-terminal peptide tags sequences if the stop codon is placed in this position.
There is a start codon in the NcoI site can be removed by digestion with KpnI if required. The MCS for gene insertions extends from NotI to XbaI however the tag resides between the NotI and HindIII sites. There are Shine-Dalgarno sequences and KOZAK sequences aligned with the start codon of the peptide tag.
The ClaI to NheI sites have other functions such as adding C-terminal peptide tags second promoters or IRES expression components. The BsgI and BseRI restriction sites cleave within the stop codon in the XbaI site and allow the retrospective fusion of C-terminal peptide tags sequences if the stop codon is placed in this position.
Analysis Note
To view the Certificate of Analysis for this product, please visit www.oxgene.com
Other Notes
To view sequence information for this product, please visit the product page
Clase de almacenamiento
12 - Non Combustible Liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
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