pET GFP domain 11 LIC cloning vector (GFP C-11)
- Backbone size (bp) 5422
Vector typeBacterial Expression
Growth in Bacteria
Copy numberLow Copy
/ Fusion Proteins
- GFP domain 11 (C terminal on backbone)
- Biotin-H6-TEV recognition (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site LIC site vGFP (destroyed during cloning)
- 3′ cloning site LIC site vGFP (destroyed during cloning)
- 5′ sequencing primer T7 forward
- 3′ sequencing primer T7 reverse (Common Sequencing Primers)
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/
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pET GFP domain 11 LIC cloning vector (GFP C-11) was a gift from Scott Gradia (Addgene plasmid # 30183)
Map generated by Addgene from full sequence supplied by depositor.