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Merck

E9783

Sigma-Aldrich

p3XFLAG-CMV-9 Expression Vector

Shuttle vector for transient or stable expression of secreted N-terminal 3xFLAG fusion proteins

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About This Item

UNSPSC Code:
12352200

tag

3X FLAG tagged

grade

for molecular biology

form

buffered aqueous solution

shipped in

dry ice

storage temp.

−20°C

General description

The p3XFLAG-CMV-9 Expression Vector is a 6.4 kb derivative of pCMV5 used to establish transient or stable expression of secreted N-terminal 3XFLAG fusion proteins in mammalian cells. The vector encodes three adjacent FLAG?epitopes (Asp-Tyr-Lys-Xaa-Xaa- Asp) upstream of the multiple cloning region. This results in increased detection sensitivity using ANTIFLAG M2 antibody. The third epitope includes the enterokinase recognition sequence, allowing cleavage of the 3XFLAG peptide from the purified fusion protein.

p3XFLAG-CMV-9 expression vector is a shuttle vector for E. coli and mammalian cells. Efficiency of replication and genomic integration is optimal when using an SV40 T antigen expressing host, such as COS cells. A related vector, p3XFLAG-CMV-3, has been used for stable transfection of HEK 293 cells. Efficiency of replication and genomic integration is optimal when using an SV40 T antigen-expressing host.

The p3XFLAG-CMV-7-BAP Control Plasmid is a 6.2 kb derivative of pCMV5 used for transient intracellular expression of N-terminal 3XFLAG bacterial alkaline phosphatase fusion protein in mammalian cells. The vector encodes three adjacent FLAG epitopes (Asp-Tyr-Lys-Xaa-Xaa-Asp) upstream of the multiple cloning region. This results in increased detection sensitivity using ANTI-FLAG M2 antibody. The third FLAG epitope includes the enterokinase recognition sequence, allowing cleavage of the 3XFLAG peptide from the purified fusion protein.

Vector Maps and Sequences

Components

  • p3XFLAG-CMV-9 Expression Vector 20 μg (E4276) is supplied as 0.5 mg/ml in 10 mM Tris-HCl (pH 8.0) with 1 mM EDTA.
  • p3XFLAG-CMV-7-BAP Control Plasmid 20 μg (C7472) is supplied as 0.5 mg/ml in 10 mM Tris-HCl (pH 8.0) with 1 mM EDTA.

Principle

The promoter-regulatory region of the human cytomegalovirus drives transcription of FLAG®-fusion constructs. The preprotrypsin leader sequence precedes the FLAG® sequence. The aminoglycoside phosphotransferase II gene (Neo-r) confers resistance to aminoglycosides such as G418 allowing for selection of stable transfectants.

Legal Information

This product is covered by the following patents owned by Sigma-Aldrich Co. LLC: US6,379,903, US7,094,548, JP4405125,EP1220933, CA2386471 and AU774216.
3xFLAG is a trademark of Sigma-Aldrich Co. LLC
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
p3xFLAG-CMV is a trademark of Sigma-Aldrich Co. LLC

Related product

Storage Class

10 - Combustible liquids


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A procedure for precise assembly of linear DNA constructs as long as 20 kb is proposed. The method, which we call long multiple fusion, has been used to assemble up to four fragments simultaneously (for a 10.8 kb final product)
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