BACKBONE：

Vector backbone： pET

Backbone size (bp)：6465

Vector type ：Bacterial Expression

GENE/INSERT：

Gene/Insert name： None

Tag / Fusion Protein
               His6-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 Enzyme
• 5′ 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 reverse

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. 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/