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| 产品名称： | pRS401 plasmid in E. coli |
|---|---|
| 商品货号： | TS163569 |
| Designations： | pRS401 plasmid in E. coli |
| Depositors： | JD Boeke |
| Biosafety Level： | 1 |
| Host： | Distribution host: Escherichia coli HB101 (ATCC 33694) |
| Vector Information： | Size (kb): 4.893 DESCRIPTION OF VECTOR: Intact vector size: 4.893 Type of vector: phagemid Cloning sites: SacI BstXI SacII EagI NotI SpeI BamHI SmaI PstI ClaI SalI XhoI ApaI KpnI Polylinker sites: SacI BstXI SacII EagI NotI XbaI SpeI BamHI SmaI PstI EcoRI EcoRV HindIII ClaI SalI XhoI ApaI KpnI Construction: pJK142 Host range: Saccharomyces cerevisiae; Escherichia coli Features (with orientation and position when available): marker(s): MET15, ->, 421-1270 replicon: f1, promoter for in vitro transcription: T7, ->, 2486-2505 MCS: KpnI...SacI, ->, 2513-2619 promoter for in vitro transcription: T3, promoter: lac, coding sequence: lacZ, replicon: pMB1, 3075-3075 marker(s): ampR, Vector: pRS401 (phagemid) Promoters: Promoter for in vitro transcription T7 Construction: pJK142 Marker(s):ampR,MET15 Construct size (kb): 4.893 Features: marker(s): MET15 marker(s): ampR promoter: lac promoter for in vitro transcription: T3 promoter for in vitro transcription: T7 replicon: f1 replicon: pMB1 MCS: KpnI...SacI coding sequence: lacZ |
| Applications： | YI-type (integrating) shuttle vector vector permitting visual detection of recombinants O-acetylhomoserine sulfhydrylase |
| Comments： | Restriction digests of the clone give the following sizes (kb): BamHI--4.9; EcoRI--3.2, 1.7; XbaI--2.9, 2.0. met15 phenotype produces brown colonies when grown on Pb containing media. This requires two approx. 60nt PCR primers; the 20nts of sequence at the 3 ends of each primer is specific for amplifying the MET15 gene from pRS401, and the 40nts of sequence at the 5 ends matches the genomic sequences flanking the gene of interest. These same primers can be used to amplify an ADE2 marker gene disruption product from pRS402 (ATCC 87477). MET15, MET17 and MET25 are synonymous. pRS401 can be used to generate a gene specific MET15 marker gene disruption cassette for tranformation in gene knockout experiments. The 20nt PCR primer sequences for generating the MET15 marker from pRS401 are: 5-CTGTGCGGTATTTCACACCG-3 (left primer) and 5-AGATTGTACTGAGAGTGCAC-3 (right primer). |
| Media： | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| Growth Conditions： | Temperature: 37.0°C |
| References： | Cost GJ, Boeke JD. A useful colony colour phenotype associated with the yeast selectable/counter-selectable marker MET15. Yeast 12: 939-941, 1996. PubMed: 8873447 Brachmann CB, et al. Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications. Yeast 14: 115-132, 1998. PubMed: 9483801 |
| Related Products： | Component of:ATCC 87476 Component of: ATCC 87538 Component of:ATCC 87529 Component of:ATCC 87536 |
| Shipped： | frozen |
| Shipping Information： | Distributed: freeze-dried |