|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||26117||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerSGC Oxford
- Backbone size (bp) 7218
Vector typeBacterial Expression
- Promoter T7-lacO
/ Fusion Protein
- His (C terminal on backbone)
Growth in Bacteria
Copy numberHigh Copy
- 5′ sequencing primer T7
- 3′ sequencing primer T7-term (Common Sequencing Primers)
pET expression vector with C-terminal His6 tag. Includes sites for LIC cloning, and a “stuffer” fragment that includes the SacB gene, allowing negative selection on 5% sucrose. GenBank accession number: EF199843
Primers for LIC cloning:
Add the following 5’ extensions to the PCR primers:
Upstream: TTAAGAAGGAGATATACTATG (ATG-initiation codon)
The purified PCR fragments are treated with T4 DNA polymerase and dGTP, then annealed to the treated vector. See supplemental document for more information.
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:pNIC-CH was a gift from Opher Gileadi (Addgene plasmid # 26117 ; http://n2t.net/addgene:26117 ; RRID:Addgene_26117)