pET His6 GFP TEV LIC cloning vector (1GFP)
- Backbone size (bp) 6075
Modifications to backboneThe GFP is the version described in Pedelacq et. al. (Jan 2006, Nature Biotechnology). The following mutations are included: F100S, M154T, V164A, S30R, Y39N, N105T, Y145F, I171V and A206K. These mutations have been shown to enhance brightness and solubility, and to inhibit dimerization.
Vector typeBacterial Expression
Growth in Bacteria
Copy numberLow Copy
/ Fusion Protein
- His6-GFP-TEV (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site LIC tag (destroyed during cloning)
- 3′ cloning site LIC tag (destroyed during cloning)
- 5′ sequencing primer GFP forward (5'cagctcgccgaccacta)
- 3′ sequencing primer T7 reverse (Common Sequencing Primers)
This plasmid is an empty vector to be used with a LIC cloning protocol.
It has a TEV-cleavable His6 fusion tag on its N-terminus. GFP can enhance your protein's expression and solubility. It can also be used as a reporter gene.
To clone into this vector, add LIC fusion tags to the 5' end of your PCR primers.
Forward - 5'TACTTCCAATCCAATGCA3'
Reverse - 5'TTATCCACTTCCAATGTTATTA3'
Linearize the plasmid with SspI and gel purify.
When digesting the DNA with T4 polymerase, use dCTP for insert and dGTP for 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 His6 GFP TEV LIC cloning vector (1GFP) was a gift from Scott Gradia (Addgene plasmid # 29663)
Map generated by Addgene from full sequence supplied by depositor.
Map uploaded by the depositor.