pET MBP mCitrine LIC cloning vector (MBP-mCitrine)
- Backbone size (bp) 7276
Vector typeBacterial Expression
Growth in Bacteria
Copy numberLow Copy
/ Fusion Proteins
- Biotin - His6 - MBP -TEV (N terminal on backbone)
- mCitrine (C 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 GFP reverse (5'cagctcgaccaggatgggc3') (Common Sequencing Primers)
This plasmid is an empty vector. Your gene can be inserted with a LIC cloning protocol.
mCitrine has a excitation max of 515 nm and an emission max of 529 nm. A TEV-cleavable MBP will be added to the N-terminal side of your protein to enhance 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 mCitrine LIC cloning vector (MBP-mCitrine) was a gift from Scott Gradia (Addgene plasmid # 29749)
Map generated by Addgene from full sequence supplied by depositor.
Map uploaded by the depositor.