微信小陈
微信小章
产品简介
购买须知
| 产品名称： | pSLF173 |
|---|---|
| 商品货号： | TS152522 |
| Designations： | pSLF173 |
| Depositors： | SL Forsburg |
| Biosafety Level： | 1 |
| Vector Information： | Size (kb): 8.5000000000000000 Vector: pSLF173 (phagemid) Promoters: Promoter for expression nmt1 (full strength) Construction: REP4X Marker(s):ampR,ura4+ Construct size (kb): 8.5 Features: marker(s): ampR marker(s): ura4+ promoter for expression: nmt1 (full strength) replicon: ars1 replicon: f1 replicon: pMB1 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): BamHI--8.5; EcoRI--7.3, 1.2; PstI--8.5. The fission yeast tagging vectors, pSLF173 (TS152522), pSLF273 (ATCC 87613) and pSLF373 (ATCC 87614), 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 1) amplification by PCR with primers designed to flank the triple HA tag in Bluescript-HA and to modify the polylinker, 2) gel purification of the PCR product and digest with XhoI and 3) ligation into REP4X cleaved with XhoI and SmaI. |
| 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 |
| Shipped： | frozen |