pET LIC cloning vector (2A-T)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||29665||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 4731
Vector typeBacterial Expression
Growth in Bacteria
Copy numberLow Copy
- Cloning method Ligation Independent Cloning
- 5′ cloning site LIC site v2 (destroyed during cloning)
- 3′ cloning site LIC site v2 (destroyed during cloning)
- 5′ sequencing primer T7 forward
- 3′ sequencing primer T7 reverse (Common Sequencing Primers)
This plasmid is an empty vector. Your gene can be inserted with a LIC cloning protocol. All 2-series vectors work as single-expression vectors, as well as transfer vectors for our polycistronic system.
The LIC cloning site is flanked by 5 pairs of restriction sites, so that your gene can easily be subcloned into our polycistronic destination vectors (2D, 2E, or 2Z).
2A-T has no fusion tags, for those who want to express native protein.
To clone into this vector, add LIC v2 tags to the 5' end of your PCR primers.
Forward - 5'TTTAAGAAGGAGATATAGATC3'
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.
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 LIC cloning vector (2A-T) was a gift from Scott Gradia (Addgene plasmid # 29665 ; http://n2t.net/addgene:29665 ; RRID:Addgene_29665)