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Plasmid 30183: pET GFP domain 11 LIC cloning vector (GFP C-11)
Comments: 

This plasmid is an empty vector. Your gene can be inserted with a LIC cloning protocol.

This vector adds a short GFP domain 11 tag to the C-terminus of your protein. After expression and purification, your protein can be mixed with a "biosensor" molecule (GFP domains 1-10), and fluorescence will develop. This technique can be useful if directly fusing the entire GFP molecule interferes with the function of your protein.

To clone into this vector, add LIC tags to the 5' end of your PCR primers.

Forward - 5' TACTTCCAATCCAATGCA3'

Reverse - 5' CTCCCACTACCAATGCC 3'

Do NOT include a stop codon with your reverse primer.

Linearize the plasmid with SspI, then gel purify.

When digesting the DNA with T4 polymerase, use dCTP for the insert and dGTP for your linearized vector.

More information on this vector can be found through http://qb3.berkeley.edu/qb3/macrolab/

Addgene has sequenced a portion of this plasmid for verification. Full plasmid sequence is available only if provided by the depositing laboratory.

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 30183" in your Materials and Methods section.

Price: US $65

Available to academic and non-profits only