pBad His6 MBP TEV LIC cloning vector (8C)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||37503||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 6848
Vector typeBacterial Expression
- Promoter araBAD arabinose
/ Fusion Proteins
- His6 (N terminal on backbone)
- MBP (N terminal on backbone)
- TEV recognition (N terminal on backbone)
Growth in Bacteria
Growth Strain(s)XL1 Blue
Copy numberLow Copy
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer MBP F (5'ggtcgtcagactgtcgatgaagcc)
- 3′ sequencing primer ARA Rv (5'ctgttttatcagaccgcttc) (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
This plasmid is an empty vector. Your gene can be inserted via LIC cloning.
8-series vectors are induced with L-arabinose for tighter control of expression. Glucose can be added to the medium to further inhibit leaky expression. The plasmid can be expressed in any E. coli line that lacks proteases.
8C adds a TEV-cleavable His6-MBP tag to the N-terminus of your protein. MBP can enhance the solubility of your protein of interest.
To clone into this vector, add LIC v1 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:pBad His6 MBP TEV LIC cloning vector (8C) was a gift from Scott Gradia (Addgene plasmid # 37503 ; http://n2t.net/addgene:37503 ; RRID:Addgene_37503)