The cleavage position in the reading frame for cloning sites is (where 3 = between triplets): EcoRI-3; SacI-1; KpnI-1; SmaI-3; BamHI-3; XbaI-3; SalI-3; PstI-1; SphI-1; HindIII-3. The EcoRI, SacI, and Kpn I sites are not unique. Restriction digests of the clone give the following sizes (kb): HindIII--8.4; BamHI--8.4; SalI--8.4; PstI--8.4; EcoRI--4.25, 4.15. One of 3 promoter-cloning, YE type shuttle vectors (ATCC 37734 - 37736) 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'G AAT TCG AGC TCG GTA CCC GGG GAT CCT CTA GAG TCG ACC TGC AGG CAT GCA AGC TTC GAT CCC 3', from nucleotide 1 of the MCS through CCC for amino acid 8 of beta-galactosidase. |
