CJ40 (let-7 seed mutant perfect site)
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||17773||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerDavid Sabatini Lab
Vector typeInsect Expression, Luciferase ; Drosophila metallothionein gene promoter drives expression
Growth in Bacteria
Gene/Insert namelet-7 seed mutant perfect site
SpeciesD. melanogaster (fly)
Entrez Genelet-7 (a.k.a. Dmel_CR32968, CR32968, Dmel\CR32968, Let-7, dme-let-7, let7)
/ Fusion Protein
- renilla luciferase (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ sequencing primer MT forward (Common Sequencing Primers)
Luciferase-reporter inserts were made by annealing oligonucleotides with their reverse complements, leaving overhangs for the indicated restriction sites (lower case): let-7-psm (gagctcACTATACAACCTACAAGCACAactagt). Annealed oligos were ligated into SacI/SpeI-cleaved pIS2. These plasmids were linearized with HindIII, polished with Klenow enzyme to create blunt ends, and digested with NotI to excise the Renilla luciferase gene with the modified UTR from the remainder of pIS2. The gel-purified Renilla gene fragment was then ligated into pMT-puro between EcoRV and NotI sites for copper-induced expression in S2 cells.
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:CJ40 (let-7 seed mutant perfect site) was a gift from David Bartel (Addgene plasmid # 17773 ; http://n2t.net/addgene:17773 ; RRID:Addgene_17773)
For your References section:Intronic microRNA precursors that bypass Drosha processing. Ruby JG, Jan CH, Bartel DP. Nature. 2007 Jul 5. 448(7149):83-6. 10.1038/nature05983 PubMed 17589500
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