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URL: http://www.addgene.org/22287
Proper Citation: RRID:Addgene_22287
Insert Name: REV1
Organism: Saccharomyces cerevisiae
Bacterial Resistance: Ampicillin
Defining Citation: PMID:19528320
Vector Backbone Description: Backbone Size:6112; Vector Backbone:YCplac111; Vector Types:Yeast Expression; Bacterial Resistance:Ampicillin
Comments: The REV1 ORF plus 1050 nucleotides (nt) of its upstream region that include the REV1 promoter and 500nt of the downstream region that include the REV1 transcription terminator was cloned in the vector YCplac111, which carries the LEU2 gene, the autonomous replicating sequence ARS1, and the centromeric CEN4 region. A BglII restriction site was added by site-directed mutagenesis right before the stop codon of REV1. A 33-mer oligonucleotide duplex containing the HA-coding sequence and a BamHI and a BglII restriction site was inserted into the BglII restriction site, adding an HA tag in fusion with REV1, and regenerating a unique BglII restriction site. This step was repeated three times in order to get four HA tags in fusion with REV1.
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Source: Addgene
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