S. cerevisiae ORF sequences

Genomic and coding DNA sequences of ORFs in S. cerevisiae strain S288C were retrieved from the Saccharomyces Genome Database at: http://downloads.yeastgenome.org/sequence/S288C_reference/orf_dna/

Plasmid sets:

• pFA6

Longtine MS, McKenzie A 3rd, Demarini DJ, Shah NG, Wach A, Brachat A, Philippsen P, Pringle JR. Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Yeast. 1998. [PubMed]

• pYM

Janke C, Magiera MM, Rathfelder N, Taxis C, Reber S, Maekawa H, Moreno-Borchart A, Doenges G, Schwob E, Schiebel E, Knop M. A versatile toolbox for PCR-based tagging of yeast genes: new fluorescent proteins, more markers and promoter substitution cassettes. Yeast. 2004. [PubMed]

• pCg / pKl

These plasmids contain selection markers from Candida glabrata and Kluyveromyces lactis.  

Kitada K, Yamaguchi E, Arisawa M. Cloning of the Candida glabrata TRP1 and HIS3 genes, and construction of their disruptant strains by sequential integrative transformation. Gene. 1995. [PubMed]

DAmP (Decreased Abundance by mRNA Perturbation)

The DAmP library was built using a pFA6-KAN construct, although the primers that were used were not the pFA6 generic amplification sequences (F1 & R1), rather new sequences found internaly (in respect to the F1&R1), that we refer to as DAmP F&R (see in information page). Therefore you may choose to perform DAmP in this way, or to use the generic pFA6 or pYM plasmid sequences to amplify the selection cassette.

Schuldiner M, Collins SR, Thompson NJ, Denic V, Bhamidipati A, Punna T, Ihmels J, Andrews B, Boone C, Greenblatt JF, Weissman JS, Krogan NJ. Exploration of the function and organization of the yeast early secretory pathway through an epistatic miniarray profile. Cell. 2005. [Pubmed]

Batch primers design

Frank M You, Naxin Huo, Yong Qiang Gu, Ming-cheng Luo, Yaqin Ma, Dave Hane, Gerard R Lazo, Jan Dvorak and Olin D Anderson. BatchPrimer3: a high throughput web application for PCR and sequencing primer designi. BMC Bioinformatics. 2008. [PubMed] [BatchPrimer3]