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
• UNSPSC Code: 12352200
• NACRES: NA.85

Properties

• 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

Description

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.

Sequence

Quick-reference Plasmid Map

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

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

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable