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URL: http://www.addgene.org/17772
Proper Citation: RRID:Addgene_17772
Insert Name: miR-1003 seed mutant perfect site
Organism: Drosophila melanogaster
Bacterial Resistance: Ampicillin
Defining Citation: PMID:17589500
Vector Backbone Description: Backbone Marker:David Sabatini Lab; Backbone Size:0; Vector Backbone:pMT-puro; Vector Types:Insect Expression, Luciferase, Drosophila metallothionein gene promoter drives expression; Bacterial Resistance:Ampicillin
Comments: Luciferase-reporter inserts were made by annealing oligonucleotides with their reverse complements, leaving overhangs for the indicated restriction sites (lower case): miR-1003-psm (gagctcCTGTGAATATGTAAAAGAGTGAactagt). 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.
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Source: Addgene
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