Purpose(Empty Backbone) This is a Gateway destination vector for secreted expression in Drosophila cell culture by induction via CuSO4. Inserts will be C-terminally tagged with Fc, V5 and hexahistidine tags.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||47032||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backbonepMT/BiP/V5/His A
- Backbone size (bp) 4349
Vector typeInsect Expression ; Secreted expression in Drosophila Culture
- Promoter Metallothionein (Copper-Inducible)
/ Fusion Proteins
- HRV 3C Protease Cleavage site (C terminal on backbone)
- Fc(human IgG1) (C terminal on backbone)
- V5 (C terminal on backbone)
- Hexahistidine (C terminal on backbone)
Growth in Bacteria
Bacterial Resistance(s)Chloramphenicol and Ampicillin
Growth Strain(s)ccdB Survival
Copy numberHigh Copy
- Cloning method Gateway Cloning
- 5′ sequencing primer MT-Upstream (Common Sequencing Primers)
Inducible with 0.8 uM Copper Sulfate. Targets proteins to the extracellular millieu using the Drosophila melanogaster BiP signal peptide. Expresses in Drosophila melanogaster cell lines, such as Schneider 2 (S2) cells. Published in Ozkan et al, Cell 154(1): 228 (2013); PMID: 23827685; doi: 10.1016/j.cell.2013.06.006
After recombination, transform and grow in DH5alpha. The plasmid is no longer chloramphenicol resistant after recombination.
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:pECIA2 was a gift from Chris Garcia (Addgene plasmid # 47032 ; http://n2t.net/addgene:47032 ; RRID:Addgene_47032)
For your References section:An Extracellular Interactome of Immunoglobulin and LRR Proteins Reveals Receptor-Ligand Networks. Ozkan E, Carrillo RA, Eastman CL, Weiszmann R, Waghray D, Johnson KG, Zinn K, Celniker SE, Garcia KC. Cell. 2013 Jul 3;154(1):228-39. doi: 10.1016/j.cell.2013.06.006. 10.1016/j.cell.2013.06.006 PubMed 23827685