The cleavage position in the reading frame for cloning sites is (where 3 = between triplets): HindIII-3; SphI-1; PstI-1; SalI-3; XbaI-3; BamHI-3; SmaI-1; KpnI-1; SacI-1; EcoRI-3. The EcoRI, KpnI, and SacI sites are not unique. Restriction digests of the clone give the following sizes (kb): HindIII-8.4; BamHI-8.4; PstI-8.4. One of 3 promoter-cloning, YE type shuttle vectors (ATCC 37740 - 37742) 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'A AGC TTG CAT GCC TGC AGG TCG ACT CTA GAG GAT CCC CGG GTA CCG AGC TCG AAT TCC CAG CTT GCT CCC3', from nucleotide 1 of the MCS through CCC for amino acid 8 of beta-galactosidase. |
