|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||35538||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerWitold Filipowicz
- Backbone size w/o insert (bp) 6380
- Total vector size (bp) 8144
Modifications to backboneRL was inserted into pcI-neo(Promega) to form the plasmid pcI-neo-Rl by Witold Filipowicz. Pillai R, Bhattacharyya S, Artus C, Zoller T, Cougot N, Basyuk E, Bertrand E, Filipowicz W (2005) Inhibition of translational initiation by let-7 microRNA in human cells. Science 309:1573-1576
Vector typeMammalian Expression, Luciferase
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Gene/Insert name3'UTR of ZEB2 gene
Alt nameHRIHFB2411, HSPC082, SIP-1, SIP1, SMADIP1, ZFHX1B
SpeciesH. sapiens (human)
Insert Size (bp)1264
Mutation6 Mutations in the miR-200b seed regions of the ZEB2 3'UTR
Entrez GeneZEB2 (a.k.a. HSPC082, SIP-1, SIP1, SMADIP1, ZFHX1B)
- Cloning method Restriction Enzyme
- 5′ cloning site XbaI (destroyed during cloning)
- 3′ cloning site ZbaI (destroyed during cloning)
- 5′ sequencing primer unknown
- 3′ sequencing primer unknown (Common Sequencing Primers)
Mutations in the miR-200b seed regions of the ZEB2 3'UTR were generated using the QuikChange Multi site-directed mutagenesis kit.
The ZEB2 3'UTR was cloned into the plasmid pCIneo-RL which was obtained from Witold Filiowicz.
Pillai R, Bhattacharyya S, Artus C, Zoller T, Cougot N, Basyuk E, Bertrand E, Filipowicz W (2005) Inhibition of translational initiation by let-7 microRNA in human cells.
Please note that Addgene's sequencing results identified a single nucleotide deletion at bp# 2440 when compared to the full plasmid sequence provided by the depositing laboratory. Addgene's sequencing results are correct as the extra base in the sequence from the depositing laboratory was an error due to a sequencing artifact.
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:pCI-neo-RL-ZEB2 200bmut was a gift from Greg Goodall (Addgene plasmid # 35538 ; http://n2t.net/addgene:35538 ; RRID:Addgene_35538)
For your References section:The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G, Vadas MA, Khew-Goodall Y, Goodall GJ. Nat Cell Biol. 2008 May;10(5):593-601. Epub 2008 Mar 30. 10.1038/ncb1722 PubMed 18376396