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| 产品名称： | pCGS990 CGE1684 |
|---|---|
| 商品货号： | TS185003 |
| Designations： | pCGS990 CGE1684 |
| Depositors： | DT Moir |
| Biosafety Level： | 1 |
| Vector Information： | Size (kb): 14.0000000000000000 Vector: pCGS990 (plasmid) Promoters: Promoter GAL1 Construction: LYS2, CEN4, pBR322, pCGS986, pYTCa-1 Marker(s):ampR,LYS2 Construct size (kb): 14.0 Features: marker(s): ampR, LYS2 promoter: GAL1, HSV TK replicon: pMB1, ARS1 centromere: CEN4 |
| Applications： | YR-type (replicating) shuttle vector shuttle vector vector permitting positive selection for integration |
| Comments： | Restriction digests of the clone give the following sizes (kb): EcoRI--14.0; BamHI--10.0, 4.4; XbaI--14.0. 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-. The cis-acting elements necessary for amplification are the thymidine kinase gene and a conditional CEN4 regulated by GAL1. 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. Conversion vector to insert copy number control elements into existing pYAC4-derived clones by targeted homologous recombination. The order of the major features in the plasmid is: BamHI - Tetrahymena telomere - ampR - thymidine kinase - EcoRI - LYS2 - ARS1 - GAL1 promoter - CEN4 - ClaI - SalI. |
| Media： | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| Growth Conditions： | Temperature: 37.0°C |
| References： | Smith DR, et al. Incorporation of copy-number control elements into yeast artificial chromosomes by targeted homologous recombination. Mamm. Genome 4: 141-147, 1993. PubMed: 8439726 |
| Shipped： | freeze-dried |