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MultiSite Gateway Kit
(Kit # 1000000050 )

Depositing Labs:   Anna Planas, Tomas Santalucia

This is a versatile cloning toolkit for constructing fully-customizable three-part fusion proteins, based on the MultiSite Gateway® cloning kit from Life Technologies.

This kit consists of 12 individually packaged plasmids, which are shipped as bacterial stabs at room temperature. Individual plasmids can be ordered at $65 per plasmid, and will be shipped as bacterial stabs.

Add to Cart
$350 USD + shipping

Available to academics and nonprofits only.

Original Publication

A plasmid toolkit for cloning chimeric cDNAs encoding customized fusion proteins into any Gateway destination expression vector. Buj R, Iglesias N, Planas AM, Santalucia T. BMC Molecular Biology 7, 2013, August 20th; 14(1):18. PubMed

Description

This is a versatile cloning toolkit for constructing fully-customizable three-part fusion proteins, based on the MultiSite Gateway® cloning kit from Life Technologies. The fusion protein components (i.e. an ORF of interest plus two flanking modules) are encoded in the three plasmids integral to the original MultiSite Gateway® kit. These can recombine with any purposely-engineered destination vector that uses a heterologous promoter external to the MultiSite Gateway® cassette, leading to the in-frame cloning of a cDNA encoding a fusion protein. In its current form, the toolkit contains a set of 12 promoter-less Gateway® entry clones (mostly encoding fluorescent protein cDNAs: mKate2, EGFP, ECFP, EYFP, but also an epitope tags cassette and a polyA signal) that provide N-terminal and C-terminal flanking modules when used in MultiSite Gateway reactions with other entry clones encoding the ORFs of interest.

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How to Cite this Kit

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.

For your Materials and Methods section:
"The plasmid kit used for constructing fusion proteins was a gift from Anna Planas and Tomas Santalucia (Addgene kit #1000000050)"

For your Reference section:
A plasmid toolkit for cloning chimeric cDNAs encoding customized fusion proteins into any Gateway destination expression vector. Buj R, Iglesias N, Planas AM, Santalucia T. BMC Molecular Biology 7, 2013, August 20th; 14(1):18. PubMed

Kit Contents

This kit consists of these plasmids:

ID Name
48344 pDONR P4-P1R-mKate2
48345 pDONR P2R-P3-mKate2
48346 pDONR P4-P1R-V5-6xHis
48347 pDONR P2R-P3-V5-6xHis
48348 pDONR P4-P1R-EGFP
48349 pDONR P2R-P3-EGFP
48350 pDONR P4-P1R-EYFP
48351 pDONR P2R-P3-EYFP
48352 pDONR P4-P1R-ECFP
48353 pDONR P2R-P3-ECFP
48354 pDONR P2R-P3-IRES_ECFP
48627 pDONR P2R-P3-SV40 polyA signal
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