The cleavage position in the reading frame for cloning sites is (where 3 = between triplets): HindIII-1; SphI-2; PstI-2; SalI-1; XbaI-1; BamHI-1; SmaI-2; KpnI-2; SacI-2; EcoRI-1. The EcoRI, KpnI, and SacI sites are not unique. Restriction digests of the clone give the following sizes (kb): HindIII-7.8; BamHI-7.8; PstI-7.8. One of 3 promoter-cloning, YI type shuttle vectors (ATCC 37758 - 37760) 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'AAG CTT GCA TGC CTG CAG GTC GAC TCT AGA GGA TCC CCG GGT ACC GAG CTC GAA TTC CCA GCT TGC GAT CCC3', from nucleotide 1 of the MCS through CCC for amino acid 8 of beta-galactosidase. |
