|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||33144||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerAddgene plasmid #33143
Modifications to backboneCEN6ARSH4 element from pRS415 (Christianson et al., 1992) was amplified using primers flanked with AatII sites, digested with AatII and cloned into the AatII site of parent vector carrying 256 copies of LacO (Straight et al., 1998).
Vector typeYeast Expression
Growth in Bacteria
Gene/Insert nameLacO (256 copies)
Insert Size (bp)10000
Gene/Insert nameGAL-GFP-kd200-GALpA reporter
MutationGFP is mutated such that amino acids 64-73 (FSYGVQCFSR) are replaced with IYKIHNQQLANDYYLYVL such that the fluorophore is non-functional, but the protein is still recognized by antibody
- Promoter bidirectional GAL1-GAL10 promoter
Cloning Information for Gene/Insert 2
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer GFP-kd200-R
- 3′ sequencing primer GFP forward (Common Sequencing Primers)
Terms and Licenses
Addgene cannot verify or guarantee the presence of all 256 copies of LacO. This element is unstable.
For use in two-plasmid-based gene-nuclear periphery tethering assay with either plasmids pAK67 (Addgene #33142).
pVS2 contains 256 LacO repeats and a GAL-GFP-GALpA-reporter gene (pVS1 [Addgene #33143] is identical but does not contain the reporter gene). The reporter was previously characterized and contains a GFP open reading frame flanked by the GAL1 promoter and GAL1 3′ UTR ([Abruzzi et al., 2006] and [Dower et al., 2004]).
pKA67 expresses both LacI-GFP and Nup49-GFP to mark the first plasmid as well as the nuclear periphery with GFP.
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:pVS2 was a gift from Michael Rosbash (Addgene plasmid # 33144 ; http://n2t.net/addgene:33144 ; RRID:Addgene_33144)
For your References section:The nuclear exosome and adenylation regulate posttranscriptional tethering of yeast GAL genes to the nuclear periphery. Vodala S, Abruzzi KC, Rosbash M. Mol Cell. 2008 Jul 11;31(1):104-13. 10.1016/j.molcel.2008.05.015 PubMed 18614049
Map uploaded by the depositor.