|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||47945||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector backbonepBluescript SK+
- Backbone size w/o insert (bp) 2958
- Total vector size (bp) 8380
Vector typeWorm Expression, CRISPR
Growth in Bacteria
Growth instructionsPlasmid isolation from standard DH5alpha results in low yield. We isolate this plasmid from BL21 cells, which results in normal yields.
Copy numberHigh Copy
Insert Size (bp)4254
- Promoter eef-1A.1 (eft-3)
/ Fusion Proteins
- 3xFLAG (N terminal on insert)
- SV40 NLS (N terminal on insert)
- egl-13 NLS (C terminal on insert)
- Cloning method Unknown
- 5′ sequencing primer M13F
- 3′ sequencing primer M13R (Common Sequencing Primers)
To generate the unique SpeI site, a short, 37 bp, linker was inserted into an existing NdeI site. This linker seems to have been inserted 2 or more times, creating a short repeat region that we have been unable to sequence across. Sequencing with the M13 reverse primer will fail.
For more information on Boxem Lab CRISPR Plasmids please refer to: http://www.addgene.org/crispr/boxem/
Please note: eft-3 has officially been changed to eef-1A.1 Please see the eef-1A.1 WormBase entry: http://www.wormbase.org/species/c_elegans/gene/WBGene00001168#05-9g-3
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:pMB63 was a gift from Mike Boxem (Addgene plasmid # 47945 ; http://n2t.net/addgene:47945 ; RRID:Addgene_47945)
For your References section:CRISPR/Cas9-Targeted Mutagenesis in Caenorhabditis elegans. Waaijers S, Portegijs V, Kerver J, Lemmens BB, Tijsterman M, van den Heuvel S, Boxem M. Genetics. 2013 Aug 26. 10.1534/genetics.113.156299 PubMed 23979586