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| 产品名称： | pUN65 pNN345 |
|---|---|
| 商品货号： | TS196844 |
| Designations： | pUN65 pNN345 |
| Depositors： | SJ Elledge |
| Biosafety Level： | 1 |
| Vector Information： | Size (kb): 5.6999998092651370 Vector: pUN65 (plasmid) Promoters: Promoter lac Construction: pUC18, pBluescript KS+ Marker(s):URA3,ampR,SUP11 Construct size (kb): 5.699999809265137 Features: insert detection: lacZ marker(s): SUP11 marker(s): URA3 marker(s): ampR promoter: lac promoter for in vitro transcription: T3 promoter for in vitro transcription: T7 replicon: ARS1 replicon: pMB1 MCS: KpnI...SacI centromere: CEN4 |
| Applications： | YC-type (centromeric) shuttle vector mutation detection shuttle vector vector permitting RNA synthesis in vitro vector permitting visual detection of recombinants beta-galactosidase beta-D-galactosidase |
| Comments： | Restriction digests of the clone give the following sizes (kb): XhoI--5.8; EcoRI--5.8; NotI--5.8. SUP11 in the vector suppresses the red colony phenotype of ade2-101 ochre allele host strains, grown on limiting adenine media. In the absence of plasmid selection, plasmids are lost at a frequency of 1 to 3%, giving rise to red/white sectoring. One strategy for mutation scanning is to clone the normal allele of a gene into a SUP11 vector and transform into an ade2-101 host with a null mutation in the same gene, thus selecting for plasmid maintenance and producing white colonies. Vectors pUN20 (ATCC 77263), pUN25 (ATCC 77264), pUN60 (ATCC 77266) and pUN65 (TS196844) contain SUP11. A second, potentially mutated copy of the gene can then be cloned into a second centromeric shuttle vector (pUN30, ATCC 77265; pUN70, ATCC 77268; pUN90, ATCC 77269; pUN100, ATCC 77270) for transformation into the SUP11 plasmid containing host. A normal gene in plasmid #2 can relieve the selective pressure on the SUP11 plasmid, restoring the red/white sectoring; a null allele in plasmid #2 will result in SUP11 plasmid maintenance and white colonies. YC-type shuttle vector useful for the sectoring-shuffle mutagenesis assay. Contains promoters for in vitro RNA synthesis and permits visual detection of recombinants by lacZalpha complementation. |
| Media： | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| Growth Conditions： | Temperature: 37.0°C |
| References： | Elledge SJ, Davis RW. A family of versatile centromeric vectors designed for use in the sectoring-shuffle mutagenesis assay in Saccharomyces cerevisiae. Gene 70: 303-312, 1988. PubMed: 3063604 |
| Shipped： | freeze-dried |