Size (kb): 8.5000000000000000
Vector: pSLF373 (phagemid)
Promoters: Promoter for expression nmt1** (low strength; nmt1)
Construct size (kb): 8.5
Features: marker(s): ampR
promoter for expression: nmt1** (low strength; nmt1)
replicon: pMB1, f1
epitope tag (N-terminal): hemagglutinin (HA) triple tag
encodes an epitope tag for protein isolation or monitoring
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 (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 cutting pSLF173 with SacI and XhoI, isolating the fragment containing the HA polylinker, and ligating into XhoI-SacI digested REP82X.
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
component of:ATCC DummyKit