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| 产品名称： | pNKY1009 |
|---|---|
| 商品货号： | TS214557 |
| Designations： | pNKY1009 |
| Depositors： | E Alani |
| Biosafety Level： | 1 |
| Host： | Distribution host: Escherichia coli FD 27747 (ATCC 35673) |
| Vector Information： | Size (kb): 9.60 DESCRIPTION OF VECTOR: Intact vector size: 9.600 Type of vector: plasmid Cloning sites: Polylinker sites: Other unique sites: PvuII Construction: YRp7, pNKY51 Host range: Saccharomyces cerevisiaeCandida robusta; Escherichia coli Features (with orientation and position when available): restriction site: EcoRI coding sequence: 3 TRP1, coding sequence: hisG, -> marker(s): URA3, -> coding sequence: hisG, -> coding sequence: 5 TRP1, restriction site: BglII coding sequence: ROP, -> replicon: pMB1 marker(s): ampR, replicon: ARS1, -> Vector: pNKY1009 (plasmid) Construction: YRp7, pNKY51 Marker(s):URA3,ampR Construct size (kb): 9.60 Features: marker(s): URA3 marker(s): ampR replicon: ARS1 replicon: pMB1 restriction site: BglII restriction site: EcoRI coding sequence: 3 TRP1 coding sequence: 5 TRP1 coding sequence: ROP coding sequence: hisG |
| Applications： | contains sequence ATP phosphoribosyltransferase contains sequence phosphoribosylanthranilate isomerase marker deletion vector phosphoribosylanthranilate isomerase produces protein uridine monophosphate synthetase UMP synthase, orotate phosphoribosyltransferase, orotidine 5-phosphate decarboxylase, orotate phosphoribosyltransferase 1 |
| Comments： | Restriction digests of the clone give the following sizes (kb): BglII--9.6; EcoRI--5.2, 4.4; BglII/EcoRI--4.6, 4.4, 0.6. The two step selection process requires a ura3 transformation host (this host can be created using pJL164 (ATCC 87471)). After transformation with the EcoRI/BglII digested plasmid, URA3 integrants are selected on ura- plates. The deletion strain is then recovered by selection on 5-FOA plates (loss of URA3 marker by a homologous recombination event between the two hisG repeats). E. coli containing this plasmid should be grown on media lacking pyrimidines to select for URA3-containing cells. This deleter vector is used to create yeast strains with a trp1 auxotrophic marker deletion. The 4.6 kb EcoRI/BglII insert contains two direct repeats of the Salmonella hisG gene flanking URA3 plus TRP1 sequences flanking the hisG-URA3-hisG sequence. The plasmid was constructed by inserting the 3.8 kb BamHI-BglII hisG-URA3-hisG fragment into the modified EcoRV site within the TRP1 gene of YEp7. |
| Media： | ATCC® Medium 2057: M9 salts with supplements |
| Growth Conditions： | Temperature: 37°C |
| References： | Alani E, et al. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Genetics 116: 541-545, 1987. PubMed: 3305158 Jef D Boeke, personal communication |
| Related Products： | component of:ATCC 87472 |
| Shipped： | frozen |
| Shipping Information： | Distributed: freeze-dried |