pET MBP mRuby2 LIC cloning vector (MBP-mRuby2)
Purpose(Empty Backbone) pET LIC cloning vector for Biotin-His6-MBP-N10-tev-yORF-mRuby2
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||86933||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 7267
Modifications to backbonemClover is from AddGene plasmid 49089.
Vector typeBacterial Expression
- Promoter T7
/ Fusion Proteins
- Biotin (N terminal on backbone)
- His6 (N terminal on backbone)
- MBP (N terminal on backbone)
- Tev Recognition Sequence (N terminal on backbone)
- mRuby2 (C terminal on backbone)
Growth in Bacteria
Copy numberLow Copy
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer MBP F (5' ggtcgtcagactgtcgatgaagcc 3')
- 3′ sequencing primer T7 Reverse (Common Sequencing Primers)
This plasmid is an empty vector. Your gene can be inserted with a LIC cloning protocol. This plasmid can be used as a single-expression vector, and it is also compatible with our 2-series polycistronic destination vectors (2D, 2E, and 2Z), if co-expression with other genes is desired.
To clone into this vector, add LIC tags to the 5' end of your PCR primers.
Forward - 5' TACTTCCAATCCAATGCA 3'
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 dGTP for the insert and dCTP 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 mRuby2 LIC cloning vector (MBP-mRuby2) was a gift from Chris Jeans (Addgene plasmid # 86933 ; http://n2t.net/addgene:86933 ; RRID:Addgene_86933)