pCGS990


Vector IG Sequence Link :
General : plasmid ds-DNA 14000 BP
Functions : (cloning)(shuttle)
Selection : ()
Copy Number :
Hosts : (E.coli HB101)(Saccharomyces cerevisiae)
Suppliers : (ATCC)
Misc.Comments : Conversion vector to insert copy number control elements into existing pYAC4-derived clones by targeted homologous recombination. pCGS990 should be linearized with SalI before transformation of the YAC-carrying strain and Lys+ clones selected. Homologous recombination between pCGS990 and the target YAC results in replacement of the centric pYAC4 arm with sequences from pCGS990. Recombinant clones will be Trp-. Approximately 0.5-2.5 percent of transformants will have the desired phenotype (Lys+, Ura+, Trp-). These clones may be grown in liquid MST medium for 2-5 days to increase the YAC copy number. The cis-acting elements necessary for amplification are the thymidine kinase gene and a conditional CEN4 regulated by GAL1. The order of the major features in the plasmid is: BamHI - Tetrahymena telomere - ampR - thymidine kinase - EcoRI - LYS2 - ARS1 - GAL1 promoter - CEN4 - ClaI - SalI. [1] Medium is LB plus ampicillin (ATCC medium number 1227) 37C Deposited by: D.T.Moir
Parents : ()
Siblings : ()
Descendents : ()


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