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 from each plasmid page and will be shipped as bacterial stabs.
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.
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 |