Purpose(Empty Backbone) Backbone for lentiviral gene silencing with monomeric Kusabira-Orange2 expression
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||85208||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backbonepLKO.1 - TRC cloning vector
Backbone manufacturerDavid Root (Addgene #10878)
- Backbone size (bp) 7032
Modifications to backbonePuromycin N-acetyltransferase resistance was removed with BamHI and KpnI and inserted monomeric Kusabira-Orange2 for fluorescence expression.
Vector typeMammalian Expression, Lentiviral, RNAi
- Promoter RNA polymerase III promoter for human U6 snRNA for shRNA and hPGK promoter for mKO2
Growth in Bacteria
Growth Strain(s)NEB Stable
Copy numberHigh Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer GACTATCATATGCTTACCGT (Common Sequencing Primers)
The 1.9kb stuffer can be released with AgeI and EcoRI and replaced with your shRNA sequence of choice.
Also, see Addgene's pLKO.1 protocol http://www.addgene.org/plko on how to use the pLKO.1 vector.
Plasmid grows more slowly than standard plasmids.
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:pLKO.1-TRC.mKO2 was a gift from Sandra Martha Gomes Dias (Addgene plasmid # 85208 ; http://n2t.net/addgene:85208 ; RRID:Addgene_85208)