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URL: http://www.addgene.org/42922

Proper Citation: RRID:Addgene_42922

Insert Name: Ce-age-1 promoter (873 bp)::Ce-age-1 cDNA (3549 bp)::Ce-unc-54 terminator (765 bp)

Organism: Caenorhabditis elegans

Bacterial Resistance: Ampicillin and Kanamycin

Defining Citation: PMID:22701676

Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:3956; Vector Backbone:pCR4-TOPO; Vector Types:Worm Expression; Bacterial Resistance:Ampicillin and Kanamycin

Comments: The construct Ce-age-1p::Ce-age-1(3,549bp)::Ce-unc-54t, contained in plasmid pPV454, was assembled by overlap extension PCR. An 873 bp segment of Ce-age-1 5′ region was amplified from genomic DNA and fused to 2,376 bp of the 5′ region of Ce-age-1, amplified from cDNA, using the overlapping primers Ceage1p-cDNA-F and -R (below). This was subsequently fused to the remainder of the Ce-age-1 cDNA with the plasmid Pdpy-30-age-1, a gift from Yuichi Iino (University of Tokyo, Tokyo, Japan), as template and using the overlapping primers Ceage1-Cat-F and -R (below). Finally, the promoter and full-length 3,549 bp Ce-age-1 cDNA were fused to 765 bp of the Ce-unc-54 3′ region (terminator), amplified from Pdpy-30-age-1 template, using the overlapping primers Ceage1-unc54-F and -R (below). The PCR product was T/A cloned into the pCR4-TOPO vector (Life Technologies) and the insert fully sequenced. Ceage1p-cDNA-F caagctttcattttaagattttaagATGTCTATGGGACGAAGCCCCTC Ceage1p-cDNA-R GAGGGGCTTCGTCCCATAGACATcttaaaatcttaaaatgaaagcttg Ceage1-Cat-F CCTCCGTGGAAATGAAGAGCACatcaagatcatcacccgacaag Ceage1-Cat-R cttgtcgggtgatgatcttgatGTGCTCTTCATTTCCACGGAGG Ceage1-unc54-F CCACGCAGTCAAACACTACTGAgataagagctccgcatcggccg Ceage1-unc54-R cggccgatgcggagctcttatcTCAGTAGTGTTTGACTGCGTGG

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