PP2393

Sigma-Aldrich

Mammalian FLAG® Tag Vector Set

plasmid vectors for molecular cloning

Synonym(s):
expression vector, molecular cloning vector, vector, snapfast vector, plasmid vector, cloning vector, plasmid
NACRES:
NA.85
Pricing and availability is not currently available.

form

buffered aqueous solution

conjugate

FLAG® tagged

Origin of replication

pUC (500 copies)

Peptide cleavage

TEV
no cleavage

Peptide tag location

C-terminal
N-terminal

Promoter

Promoter name: CMV
Promoter activity: constitutive
Promoter type: mammalian

bacteria selection

kanamycin

mammalian cells selection

puromycin

shipped in

ambient

storage temp.

−20°C

General description

Molecular cloning often benefits from optimizing the vector used for expression.

This pack enables you to compare placing FLAG epitope tags at either the N or C terminus of your gene of interest (inserted into the MCS, under transcriptional control of the CMV promoter) with, and also without a TEV (Tobacco Etch Virus) protease cleavage site. The TEV site enables removal of the FLAG tag from the protein after production. Comparing these four configurations should enable you to evaluate how best to express and detect your gene of interest from mammalian cells. We also provide many other functional tags and cleavage sites, if required.This plasmid set has been designed to be compatible with a range of cloning techniques. The multiple cloning site contains a range of standard commonly used restriction sites for cloning. Using these sites genes can be inserted using standard cloning methods with DNA ligase. Other methods such as ligase independent cloning (LIC) Gibson Assembly InFusionHD or Seamless GeneArt can also be used and because all of our plasmids are based on the same backbone the same method can be used for cloning into all of our catalogue vectors.

Multiple cloning site notes: There are a few important sites within the MCS. These include the NcoI site the XbaI site and the BsgI and BseRI sites. The NcoI site contains a start codon that is immediately downstream of both a Kozak and Shine-Dalgarno ribosomal binding site. These allow for optimal positioning of genes when the start codon is placed in this location. If this is not required and you wish to use a downstream site for gene cloning you can remove the NcoI site by cleaving the plasmid with KpnI. The XbaI site contains a stop codon. This stop codon is positioned in a specific position in relation to the BsgI and BseRI sites that are immediately downstream. When either BseRI or BsgI cleave the plasmid they produce a TA overhang from the stop codon in the XbaI site that is compatible with all of our peptide tag plasmids cut with the same sites. BseRI and BsgI sites are non-palindromic and cleave a defined number of bases away from their binding site. Whenever we clone a gene into our multiple cloning site we always position the start and stop codon in the same positions in the MCS. If the start and ends of the genes are not compatible with NcoI and XbaI we extend the sequence to the nearest external sites but keep the start and stop codons locations consistent.

Transcription Termination: These plasmids contains three alternative transcription terminators for mammalian bacterial and bacteriophage (T7) expression. This means that only the promoter needs to be changed to alter the expression system you are using. We sell multiple promoters that can be used in each of these systems. The presence of each terminator does not reduce expression in the alternative systems.

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

Legal Information

These plasmids are sold free of reach-through rights and can be used to make commercial products. However the plasmids themselves (or derivatives) cannot be sold.
FLAG is a registered trademark of Sigma-Aldrich Co. LLC

Kit Components Also Available Separately

Product No.
Description
SDS

  • OGS3213PSF-CMV-PURO-NH2-FLAG® - N-TERMINAL FLAG® TAG MAMMALIAN PLASMID, plasmid vector for molecular cloning

  • OGS1124PSF-CMV-PURO-COOH-TEV-FLAG® - C-TERMINAL FLAG® TAG MAMMALIAN PLASMID, plasmid vector for molecular cloning

  • OGS3423PSF-CMV-PURO-COOH-FLAG® - C-TERMINAL FLAG® TAG MAMMALIAN PLASMID, plasmid vector for molecular cloning

  • OGS1123PSF-CMV-PURO-NH2-FLAG®-TEV - N-TERMINAL FLAG® TAG MAMMALIAN PLASMID, plasmid vector for molecular cloning

RIDADR

NONH for all modes of transport

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis
Certificate of Origin
Jin-Gyoung Jung et al.
PLoS genetics, 10(10), e1004751-e1004751 (2014-10-31)
The Notch3 signaling pathway is thought to play a critical role in cancer development, as evidenced by the Notch3 amplification and rearrangement observed in human cancers. However, the molecular mechanism by which Notch3 signaling contributes to tumorigenesis is largely unknown....
Diana Romero et al.
Carcinogenesis, 37(1), 18-29 (2015-10-28)
Dickkopf-3 (Dkk-3) is a secreted protein whose expression is downregulated in many types of cancer. Endogenous Dkk-3 is required for formation of acini in 3D cultures of prostate epithelial cells, where it inhibits transforming growth factor (TGF)-β/Smad signaling. Here, we...
Geoffrey M Lynn et al.
Nature biotechnology, 33(11), 1201-1210 (2015-10-27)
The efficacy of vaccine adjuvants such as Toll-like receptor agonists (TLRa) can be improved through formulation and delivery approaches. Here, we attached small molecule TLR-7/8a to polymer scaffolds (polymer-TLR-7/8a) and evaluated how different physicochemical properties of the TLR-7/8a and polymer...
Alexander C Cerny et al.
PLoS genetics, 11(10), e1005578-e1005578 (2015-10-29)
Recycling of signaling proteins is a common phenomenon in diverse signaling pathways. In photoreceptors of Drosophila, light absorption by rhodopsin triggers a phospholipase Cβ-mediated opening of the ion channels transient receptor potential (TRP) and TRP-like (TRPL) and generates the visual...

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