Restriction digests of the clone give the following sizes (kb): BamHI--7.2; EcoRI--5.4, 2.0; SacI--7.2. The GAL1 and GAL10 promoters are of similar strength, but the GAL10 promoter has a disrupted RNA start site. Unless a start site is provided by the insert, little or no message should be made from the GAL10 side of the promoter cassette. Do not use this side of the promoter unless you can insure that mRNA will be properly initiated by providing an RNA start site. Note that no transcription terminators are provided. Avoid palindromic restriction enzyme recognition sequences between the transcription start point and the start of translation by placing the AUG of the gene to be expressed as close to the promoter cassette as possible. Check for inappropriate initiation or termination codons 5' to the cloned insert. The polylinker sites listed may not be unique in this vector. YE-type, high copy number, galactose-inducible expression shuttle vector containing a divergent GAL1/GAL10 promoter cassette. The cassette is a 685 bp EcoRI/BamHI fragment with the GAL1 promoter adjacent to the EcoRI site. Can be used to produce ssDNA. Contains promoters for in vitro RNA synthesis, priming sites useful for sequencing, and yeast REP3 and FRT sequences. |
