This is one of 3 promoter-cloning, YE-type shuttle vectors (ATCC® No. 37737-37739™) with URA3 selection in Saccharomyces cerevisiae, a beta-galactosidase reporter gene and multiple cloning sites differing in reading frame. The sequence and reading frame of the multiple cloning sequence is: 5’ AAGCT TGC ATG CCT GCA GGT CGA CTC TAG AGG ATC CCC GGG TAC CGA GCT CGA ATT CCC AGC TTC GAT CCC 3’ from nucleotide 1 of the MCS through CCC for amino acid 8 of beta-galactosidase. Cloning into the HindIII, SphI, PstI SalI or XbaI sites leads to a TAG stop codon within the downstream XbaI site of the multiple cloning region.
The cleavage position in the reading frame for cloning sites is (where 3= between triplets): HindIII-2; SphI-3; PstI-3; SalI-2; XbaI-2; BamHI-2; SmaI-3; KpnI-3; SacI-3; EcoRI-2. The SacI site is not unique.
Restriction digests of the clone give the following sizes (kb): HindIII--8.0; EcoRI--8.0; BamHI--8.0; PstI--8.0; SalI--8.0.
Media
ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37°C
References
Myers AM, et al. Yeast shuttle and integrative vectors with multiple cloning sites suitable for construction of lacZ fusions. Gene 45: 299-310, 1986. PubMed: 3026915
