pET His6 MBP prescission LIC cloning vector (HMPKS)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||29721||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
- Backbone size (bp) 8106
Vector typeBacterial Expression
Growth in Bacteria
Bacterial Resistance(s)Kanamycin, 50 μg/mL
Copy numberLow Copy
/ Fusion Protein
- His6-MBP-prescission (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site SLIC site w/ stuffer (destroyed during cloning)
- 3′ cloning site SLIC site w/ stuffer (destroyed during cloning)
- 5′ sequencing primer MBP forward (5'ggtcgtcagactgtcgatgaagcc)
- 3′ sequencing primer T7 reverse (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
This plasmid is an empty vector. Your gene can be inserted with a SLIC cloning protocol.
Prescission is a highly specific protease that cleaves LEVLFQ/GP. It is generally more active than TEV protease, and many users have found that when their protein inhibits TEV cleavage, switching to a prescission vector has proved worthwhile.
To clone into this vector, add SLIC tags to the 5' end of your PCR primers.
Forward - 5'GTGCTGTTCCAGGGTCCGAAT3'
Reverse - 5'TGGTGGTGGTGGTGCTCGA(TTA)3'
Linearize the plasmid with SspI and XhoI, then gel purify. There is a 1650 bp stuffer sequence that will be visible on a gel.
When digesting the DNA with T4 polymerase, use no nucleotides for either your insert or the 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 His6 MBP prescission LIC cloning vector (HMPKS) was a gift from Scott Gradia (Addgene plasmid # 29721 ; http://n2t.net/addgene:29721 ; RRID:Addgene_29721)