PurposemiRNA sensor used in Drosophila cell lines
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||40834||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerMurhpy Lab, Carnegie Institution for Science
- Backbone size w/o insert (bp) 7233
Vector typeInsect Expression ; miRNA reporter
Growth in Bacteria
Gene/Insert nameFour target sites perfectly complementary to let-7
SpeciesD. melanogaster (fly)
- Promoter Actin5C
/ Fusion Protein
- EGFP (N terminal on insert)
- Cloning method Gateway Cloning
- 5′ sequencing primer EGFP-C (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
The let-7 complimentary sequence was inserted in the XbaI site in the 3'UTR of EGFP using the following oligos:
s: CTA GAA CTA TAC AAC CTA CTA CCT CAT CTA GAA CTA TAC AAC CTA CTA CCT CAT
as: CTA GAT GAG GTA GTA GGT TGT ATA GTT CTA GAT GAG GTA GTA GGT TGT ATA GTT
This expression cassette was then sub-cloned into the Gateway vector pAW using GATEWAY cloning.
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:pAW-EGFP-4xlet-7 (pAW-AS27) was a gift from Phillip Zamore (Addgene plasmid # 40834 ; http://n2t.net/addgene:40834 ; RRID:Addgene_40834)
For your References section:Target RNA-directed trimming and tailing of small silencing RNAs. Ameres SL, Horwich MD, Hung JH, Xu J, Ghildiyal M, Weng Z, Zamore PD. Science. 2010 Jun 18;328(5985):1534-9. 10.1126/science.1187058 PubMed 20558712
Map uploaded by the depositor.