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| 产品名称： | pPW264 |
|---|---|
| 商品货号： | TS174573 |
| Designations： | pPW264 |
| Depositors： | F Puta |
| Biosafety Level： | 1 |
| Vector Information： | Size (kb): 8.1070003509521480 Vector: pPW264 (plasmid) Promoters: Promoter lambda PR Construction: pML25, pPW263 Marker(s):URA3,ampR,cmlR Construct size (kb): 8.107000350952148 Features: marker(s): URA3 marker(s): ampR marker(s): cmlR operator: lac promoter: lambda PR promoter: tetR replicon: 2 micron replicon: pMB1 repressor gene: cI transcription terminator: tandem fd terminators |
| Applications： | vector permitting positive selection for inserts |
| Comments： | Restriction digests of the clone give the following sizes (kb): HindIII--8.2; EcoRI--7.2, 0.96; BglII--8.2. The cytoplasmic chloramphenicol resistance protein does not negatively affect cell growth when overproduced (in contrast to cloning vectors that overexpress membrane associated tetR protein). The level of resistance to chloramphenicol differs substantially depending on the stage at which they are tested. The chloramphenicol resistance of the plasmid should be tested before cloning inserts. YE-type shuttle vector allowing positive selection of inserts by conferring chloramphenicol resistance (recommended concentration 10 ug/ml) when a sequence has been inserted into the cI repressor gene. A pair of phage fd rho-independent strong terminators downstream of the cmlR gene prevents transcription through the origin of replication. This stabilizes recombinant plasmid copy number when the PR promoter for the cmlR gene is derepressed. |
| Media： | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| Growth Conditions： | Temperature: 37.0°C |
| References： | Puta F, Wambutt R. Construction of a new Escherichia coli-Saccharomyces cerevisiae shuttle plasmid cloning vector allowing positive selection for cloned fragments. Folia Microbiol. 37: 193-198, 1992. PubMed: 1505881 Puta F, et al. Direct selection shuttle plasmid vector, pPW264, used for cloning the alpha-amylase gene of Schwanniomyces occidentalis. Folia Microbiol. 39: 255-260, 1994. PubMed: 7729761 Frantisek Puta, personal communication |
| Shipped： | freeze-dried |