Purpose(Empty Backbone) Destination vector for Gateway based on vector #395 from the Tol2Kit, containing a gRNA scaffold under the control of a zebrafish U6 promoter, and cmlc2:GFP as a transgenesis marker.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||63156||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backbonepDestTol2CG2 (Tol2Kit#395)
- Backbone size (bp) 8267
Modifications to backboneMutated the BseRI site present in GFP. Added a zebrafish U6 promoter driving a gRNA scaffold containing BseRI sites to clone the seed.
Vector typeCRISPR ; Tol2 destination vector for Gateway
Growth in Bacteria
Bacterial Resistance(s)Chloramphenicol and Ampicillin
Growth Strain(s)ccdB Survival
Growth instructionsMay need to grow for 2 days in liquid media to achieve turbid culture
- Cloning method Restriction Enzyme
- 5′ sequencing primer GAGGATCATAATCAGCCATA (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
Reference for the Tol2Kit:
Kwan, K.M., Fujimoto, E., Grabher, C., Mangum, B.D., Hardy, M.E., Campbell, D.S., Parant, J.M., Yost, H.J., Kanki, J.P., and Chien, C.B. (2007). The Tol2kit: a multisite gateway-based construction kit for Tol2 transposon transgenesis constructs. Dev Dyn 236, 3088-3099.
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:pDestTol2CG2-U6:gRNA was a gift from Leonard Zon (Addgene plasmid # 63156 ; http://n2t.net/addgene:63156 ; RRID:Addgene_63156)
For your References section:A CRISPR/Cas9 Vector System for Tissue-Specific Gene Disruption in Zebrafish. Ablain J, Durand EM, Yang S, Zhou Y, Zon LI. Dev Cell. 2015 Mar 4. pii: S1534-5807(15)00075-1. doi: 10.1016/j.devcel.2015.01.032. 10.1016/j.devcel.2015.01.032 PubMed 25752963