pET His6 MBP Asn10 TEV LIC cloning vector (1C)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||29654||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size (bp) 6507
Vector typeBacterial Expression
Growth in Bacteria
Copy numberLow Copy
/ Fusion Protein
- His6-MBP-Asn10-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 MBP forward (5'ggtcgtcagactgtcgatgaagcc)
- 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. MBP can enhance the expression and solubility of your protein of interest. The long poly-Asn linker may help avoid steric clashes between your protein and MBP. If a shorter linker is desired, please use vector 1M.
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 MBP Asn10 TEV LIC cloning vector (1C) was a gift from Scott Gradia (Addgene plasmid # 29654 ; http://n2t.net/addgene:29654 ; RRID:Addgene_29654)