The cleavage position in the reading frame for cloning sites is (where 3 = between triplets): EcoRI-2; SmaI-3; BamHI-3; SalI-3; PstI-1; HindIII-2. The EcoRI site is not unique. Restriction digests of the clone give the following sizes (kb): EcoRI--4.2, 2.8; BamHI--7.0; SalI--7.0; HindIII--7.0; PstI--7.0. One of 3 promoter-cloning, YI type shuttle vectors (ATCC 37746 - 37748) with LEU2 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' GA ATT CCC GGG GAT CCG TCG ACC TGC AGC CAA GCT TGC GAT CCC 3'. The sequence begins with the first nucleotide of the MCS and ends with the CCC for amino acid 8 of beta-galactosidase. |
