pET MBP-FlAsH LIC cloning vector (MBP-FlAsH)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||30185||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 6568
Vector typeBacterial Expression
Growth in Bacteria
Copy numberLow Copy
/ Fusion Proteins
- FlAsH (C terminal on backbone)
- Biotin-H6-MBP-TEV recognition (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site LIC site vGFP (destroyed during cloning)
- 3′ cloning site LIC site vGFP (destroyed during cloning)
- 5′ sequencing primer MBP forward (5'ggtcgtcagactgtcgatgaagcc)
- 3′ sequencing primer T7 reverse (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
This plasmid is an empty vector. Your gene can be inserted with a LIC cloning protocol.
This vector adds a short FlAsH tag to the C-terminus of your protein. When your protein is mixed with a biarsenical reagent (see Invitrogen's website), fluorescence will develop. This technique may be useful when a larger fusion interferes with the function of your protein.
Additionally, the vector adds an MBP fusion to the N-terminus of your protein, which may help expression and solubility.
To clone into this vector, add LIC tags to the 5' end of your PCR primers.
Forward - 5' TACTTCCAATCCAATGCA3'
Reverse - 5' CTCCCACTACCAATGCC 3'
Do NOT include a stop codon with your reverse primer.
Linearize the plasmid with SspI, then gel purify.
When digesting the DNA with T4 polymerase, use dCTP for the insert and dGTP for your linearized 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 MBP-FlAsH LIC cloning vector (MBP-FlAsH) was a gift from Scott Gradia (Addgene plasmid # 30185 ; http://n2t.net/addgene:30185 ; RRID:Addgene_30185)