pBAD LIC cloning vector (8A)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||37501||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 5629
Vector typeBacterial Expression
- Promoter araBAD arabinose
Growth in Bacteria
Growth Strain(s)XL1 Blue
Copy numberLow Copy
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer ARA F (5'cattctgtaacaaagcggga)
- 3′ sequencing primer ARA Rv (5'ctgttttatcagaccgcttc) (Common Sequencing Primers)
Terms and Licenses
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.
8A has no fusion tags, appropriate for expressing untagged protein. If your protein doesn't express well in this vector, note that expression can usually be enhanced by adding a short N-terminal fusion sequence. If this is the case for your protein, try one of our other arabinose-inducible LIC vectors (e.g. 8B).
To clone into this vector, add LIC v2 tags to the 5' end of your PCR primers.
Forward - 5'TTTAAGAAGGAGATATAGATC(atg)3'
Reverse - 5'TTATGGAGTTGGGATCTTATTA3'
Linearize the plasmid with EcoRV and gel purify.
When digesting the DNA with T4 polymerase, use dGTP for insert and dCTP for vector.
Visit http://qb3.berkeley.edu/macrolab/ for more information on this vector
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 LIC cloning vector (8A) was a gift from Scott Gradia (Addgene plasmid # 37501 ; http://n2t.net/addgene:37501 ; RRID:Addgene_37501)