|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||11085||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerNew England Biolabs
- Backbone size (bp) 6000
Vector typeBacterial Expression
- Promoter tac promoter
/ Fusion Proteins
- 6xHis (N terminal on backbone)
- MBP (N terminal on backbone)
Growth in Bacteria
Bacterial Resistance(s)Chloramphenicol and Ampicillin
Growth instructionsplasmid must be grown in gyrA mutant resistant to the toxin CcdB
Copy numberLow Copy
- Cloning method Gateway Cloning
- 5′ sequencing primer MBP-F
- 3′ sequencing primer M13_pUC_fwd; M13-F20 (Common Sequencing Primers)
The MBP ORF can be sequenced using 5'-GAG CGG ATA ACA ATT TCA CAC AGG-3' as a tac promoter sequencing primer (in the forward direction). To sequence inserts fused to the C-terminus of MBP, the primers are 5'-GAT GAA GCC CTG AAA GAC GCG CAG-3' (forward) and 5'-GCA AGG CGA TTA AGT TGG GTA ACG C-3' (reverse).
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:pDEST-HisMBP was a gift from David Waugh (Addgene plasmid # 11085 ; http://n2t.net/addgene:11085 ; RRID:Addgene_11085)
For your References section:Gateway vectors for the production of combinatorially-tagged His6-MBP fusion proteins in the cytoplasm and periplasm of Escherichia coli. Nallamsetty S, Austin BP, Penrose KJ, Waugh DS. Protein Sci. 2005 Dec . 14(12):2964-71. 10.1110/ps.051718605 PubMed 16322578