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| 产品名称： | pRS314 plasmid in E. coli |
|---|---|
| 商品货号： | TS134761 |
| Designations： | pRS314 plasmid in E. coli |
| Depositors： | P Hieter |
| Biosafety Level： | 1 |
| Host： | Distribution host: Escherichia coli HB101 (ATCC 33694) |
| Vector Information： | Size (kb): 4.785 DESCRIPTION OF VECTOR: Intact vector size: 4.785 Type of vector: phagemid Cloning sites: KpnI ApaI XhoI SalI ClaI EcoRI PstI SmaI BamHI SpeI EagI NotI SacII SacI NaeI Polylinker sites: KpnI ApaI XhoI SalI ClaI HindIII EcoRV EcoRI PstI SmaI BamHI SpeI XbaI EagI NotI BstXI SacII SacI Construction: pRSS56 pBluescript KS+, pBS(+) Host range: Saccharomyces cerevisiaeCandida robusta; Escherichia coli Features (with orientation and position when available): marker(s): TRP1, ->, 468-1142 replicon: f1, ->, 1463-1562 insert detection: lacZ, promoter for in vitro transcription: T7, ->, 1862-1881 MCS: SacI...KpnI, ->, 1889-1996 promoter for in vitro transcription: T3, promoter: lac, replicon: pMB1, 2451-2451 marker(s): ampR, centromere: CEN6, 4211-4327 replicon: ARSH4, 4328-4702 Vector: pRS314 (phagemid) Promoters: Promoter for in vitro transcription T7 Construction: pRSS56 pBluescript KS+, pBS(+) Marker(s):TRP1,ampR Construct size (kb): 4.785 Features: insert detection: lacZ marker(s): TRP1 marker(s): ampR promoter: lac promoter for in vitro transcription: T3 promoter for in vitro transcription: T7 replicon: ARSH4 replicon: f1 replicon: pMB1 MCS: SacI...KpnI centromere: CEN6 |
| Applications： | YC-type (centromeric) shuttle vector shuttle vector vector containing primer sites useful for sequencing vector permitting RNA synthesis in vitro vector permitting production of single-stranded DNA vector permitting visual detection of recombinants |
| Comments： | Restriction digests of the clone give the following sizes (kb): EcoRI--4.8; BamHI--4.8; PvuII--4.2, 0.5. One of a series of pBluescript-based centromere vectors (ATCC 77142-77145, 77157-77158) differing in the yeast selectable marker gene. YC-type centromere vector permitting visual detection of recombinants and production of ssDNA in E. coli. Contains promoters for in vitro RNA synthesis, priming sites useful for sequencing, and encodes the lacZ alpha (lacZ) peptide. pRSS56, constructed by ligating a PvuI fragment (bp 498-2412) of pBluescript KS+ to a PvuI fragment (bp 2850-730) of pBS(+), contains the KS MCS from pBluescript KS+ and the unique NdeI and AatII sites between bla and f1 origin of pBS(+). A genomic HincII/PstI fragment (1.002 kb) containing the TRP1 gene was inserted into the NdeI site and a cassette containing CEN6 and the ARS associated with histone 4 (ARSH4) was inserted into the AatII site of pRSS56. All ends were blunted. An EcoRI site in the TRP1-containing fragment (external to the coding sequence) was destroyed. The order of the major features in this plasmid is: TRP1 - f1 ori (NaeI) - T7 promoter - lacZ/MCS - T3 promoter - pMB1 ori - bla - CEN6 - ARSH4. |
| Media： | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| Growth Conditions： | Temperature: 37.0°C |
| References： | Sikorski RS, Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122: 19-27, 1989. PubMed: 2659436 |
| Related Products： | component of:ATCC 77190 |
| Shipped： | frozen |
| Shipping Information： | Distributed: freeze-dried |