OGS1175
PSF-CMV-PURO-NH2-FLAG®-6HIS-EKT - N-TERMINAL FLAG® AND 6 HIS DUAL TAG MAMMALIAN 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
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tag
6-His tagged
FLAG® tagged
form
buffered aqueous solution
mol wt
size 6206 bp
bacteria selection
kanamycin
mammalian cells selection
puromycin
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 plasmid is designed to express tagged proteins in mammalian cells either by transient transfection or by creating stable cell lines. It contains a puromycin resistance expression cassette using the human Ubiquitin promoter to drive expression and allow for the selection of cells containing the plasmid.
About the Peptide Tag:This plasmid contains an n-terminal Hexa-Histidine (6His) affinity tag that can be fused to a gene of interest to allow protein detection and/or purification. The sequence of the tag is: HHHHHH. This plasmid also contains a secondary Flag protein tag. The sequence of this tag is: DYKDDDDKWe provide a range of dual peptide tag plasmids. This is because some peptide tags provide specific biological properties (e. g., small molecule affinity new epitopes solubility or protein secretion) that are not provided by others.
About the Cleavage Tag: This plasmid also encodes a protease cleavage site that is designed to be positioned between your gene of interest and the tag to allow the removal of the tag following protein purification or isolation. This plasmid contains a EKT cleavage tag. The protein sequence of the cleavage tag is: DDDDK. Enterokinase (EKT) protease cleaves after the Lysine residue. It can cleave at other basic residues but this is dependent on protein confirmation. If a proline follows the site it will not cut. None of our products contain a proline after the site.
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.
About the Peptide Tag:This plasmid contains an n-terminal Hexa-Histidine (6His) affinity tag that can be fused to a gene of interest to allow protein detection and/or purification. The sequence of the tag is: HHHHHH. This plasmid also contains a secondary Flag protein tag. The sequence of this tag is: DYKDDDDKWe provide a range of dual peptide tag plasmids. This is because some peptide tags provide specific biological properties (e. g., small molecule affinity new epitopes solubility or protein secretion) that are not provided by others.
About the Cleavage Tag: This plasmid also encodes a protease cleavage site that is designed to be positioned between your gene of interest and the tag to allow the removal of the tag following protein purification or isolation. This plasmid contains a EKT cleavage tag. The protein sequence of the cleavage tag is: DDDDK. Enterokinase (EKT) protease cleaves after the Lysine residue. It can cleave at other basic residues but this is dependent on protein confirmation. If a proline follows the site it will not cut. None of our products contain a proline after the site.
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.
Sequence
Please select the file type you require. For reference most cloning programs will import a .gb (Genbank) file and will show all of the plasmids features automatically when downloaded and imported.
Genebank Vector Sequence File
FASTA Vector Sequence File
Full Plasmid Map
Genebank Vector Sequence File
FASTA Vector Sequence File
Full Plasmid Map
Analysis Note
To view the Certificate of Analysis for this product, please visit www.oxfordgenetics.com.
Other Notes
Looking for more vector options to move your experiments forward faster? Consider a custom cloning vector designed and built by Oxford Genetics™. Find out more at Oxford Genetics - Sigma's partner for cloning and expression vectors for molecular biology and synthetic biology applications.
Legal Information
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
Oxford Genetics is a trademark of Oxford Genetics Ltd
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A range of forward and reverse sequencing primers that allow you to sequence any insert that you make into a particular position within any plasmid. Where possible, the binding sites for each of these primers is conserved.
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