Basic product information:
BACKBONE:
Vector backbone: pET
Backbone size (bp):6465
Vector type :Bacterial Expression
GENE/INSERT:
Gene/Insert name: None
Tag / Fusion ProteinHis6-MBP-TEV (N terminal on backbone)
GROWTH IN BACTERIA:
acterial Resistance(s):Kanamycin
Growth Temperature:37°C
Growth Strain(s):DH5alpha
Copy number:Low Copy
CLONING INFORMATION:
Cloning method: Restriction Enzyme5′ cloning site: LIC tag (destroyed during cloning)3′ cloning site: LIC tag (destroyed during cloning)5′ sequencing primer: MBP forward (5'ggtcgtcagactgtcgatgaagcc)3′ sequencing primer: T7 reverseThis 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. MBP can enhance your protein's solubility and expression. It can also be used as an affinity tag
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/