Yeast artificial chromosomes
Yeast artificial chromosomes (YACs) are synthetic double stranded linear constructs containing the elements necessary for replication in yeast. These elements are:
- an autonomous replication sequence (ARS): ARS1, chromosome III ARS, ARSH4
- a centromere: CEN4
- a telomeric sequence at each end
Typically the chromosome also contains a selection marker such as Lys2 or Ura3.
A common tool for constructing YACs is a shuttle plasmid such as pYAC4 which replicates in E. coli, has a multiple cloning site, and a pair of telomeres which can be cleaved to form a linear fragment. Available as an E.coli plasmid ATCC 67379, sequence at U01086. Yeast host AB1380 is available as ATCC 204682 (MATa ura3-52 trp1 lys2-1 ade2-1 can1-100 his5) (Burke87).
The pRS313- pRS316 plasmids use the CEN6 + ARSH4 cassette (Sikorski89).
pJS97 / pJS98 plasmids are available at ATCC 77191 as a two-plasmid kit. pJS98 has a functional TRP1 gene and promoter.
The pCGS966 plasmid (Smith90, Smith92, Moir93) has ARS1 on both arms, Gal inducible extra copy production, NeoR for mammalian expression. Functional promoter for the Trp gene, unlike pYAC4.
Minimal size for a YAC is between 50kb and 100kb, while maximum sizes are 1Mb to 3Mb.
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