pET MBP His6 LIC cloning vector (2Cc-T)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||37237||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 5898
Vector typeBacterial Expression
- Promoter T7
/ Fusion Proteins
- MBP (C terminal on backbone)
- His6 (C terminal on backbone)
Growth in Bacteria
Growth Strain(s)XL1 Blue
Copy numberLow Copy
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer T7 promoter
- 3′ sequencing primer MBP 226R 5'ttgagcgtagccaccaaag (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. For more details, see the MacroLab vector cloning manual.
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).
2Cc-T has a TEV-cleavable C-terminal MBP tag to enhance solubility, as well as a His6 tag to ease purification. The TEV site is created after performing LIC cloning described below.
To clone into this vector, add LIC v3 tags to the 5' end of your PCR primers.
Forward - 5'TTTAAGAAGGAGATATAGTTC(ATG)3'
Reverse - 5'GGATTGGAAGTAGAGGTTCTC3'
Linearize the plasmid with HpaI 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/
For your Materials & Methods section:pET MBP His6 LIC cloning vector (2Cc-T) was a gift from Scott Gradia (Addgene plasmid # 37237 ; http://n2t.net/addgene:37237 ; RRID:Addgene_37237)