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| 产品名称： | pSLF373 |
|---|---|
| 商品货号： | TS186847 |
| Designations： | pSLF373 |
| Depositors： | SL Forsburg |
| Biosafety Level： | 1 |
| Vector Information： | Size (kb): 8.5000000000000000 Vector: pSLF373 (phagemid) Promoters: Promoter for expression nmt1** (low strength; nmt1) Construction: REP82X Marker(s):ampR,ura4+ Construct size (kb): 8.5 Features: marker(s): ampR marker(s): ura4+ promoter for expression: nmt1** (low strength; nmt1) replicon: ars1 replicon: pMB1, f1 MCS: XhoI...SmaI epitope tag (N-terminal): hemagglutinin (HA) triple tag |
| Applications： | encodes an epitope tag for protein isolation or monitoring expression vector |
| Comments： | Restriction digests of the clone give the following sizes (kb): HindIII--5.6, 1.7, 1.0; EcoRI--7.4, 1.2; BamHI--8.5. The fission yeast tagging vectors, pSLF173 (ATCC 87612), pSLF273 (ATCC 87613) and pSLF373 (TS186847), contain three versions of the nmt1 promoter: full strength (nmt1), medium strength (nmt1*) and low strength (nmt1**), respectively. The weaker promoters (nmt1* and nmt1**) contain mutations that attenuate both repressed and induced levels of expression. Each version of the nmt1 promoter can be expressed at low or high levels in thiamine-free media. The vector was designed to tag expressed protein at N-terminus with triple HA tag, which contains an internal BamHI site. The vector lack a stop codon. The vector was constructed by cutting pSLF173 with SacI and XhoI, isolating the fragment containing the HA polylinker, and ligating into XhoI-SacI digested REP82X. |
| Media： | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| Growth Conditions： | Temperature: 37.0°C |
| References： | Forsburg SL, Sherman DA. General purpose tagging vectors for fission yeast. Gene 191: 191-195, 1997. PubMed: 9218719 Basi G, et al. TATA box mutations in the Schizosaccharomyces pombe nmt1 promoter affect transcription efficiency but not the transcription start point or thiamine repressibility. Gene 123: 131-131, 1993. PubMed: 8422997 |
| Related Products： | component of:ATCC 87685 component of:ATCC DummyKit |
| Shipped： | freeze-dried |