The cleavage position in the reading frame for cloning sites is (where 3 = between triplets): EcoRI-1; SacI-2; KpnI-2; SmaI-1; BamHI-1; XbaI-1; SalI-1; PstI-2; SphI-2; HindIII-1. The EcoRI, SacI, and KpnI sites are not unique. Restriction digests of the clone give the following sizes (kb): EcoRI--4.1, 2.9; SalI--7.0; PstI--7.0; HindIII--7.0. One of 3 promoter-cloning, YI type shuttle vectors (ATCC 37752 - 37754) 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'GAA TTC GAG CTC GGT ACC CGG GGA TCC TCT AGA GTC GAC CTG CAG GCA TGC AAG CTT GCT CCC 3', from nucleotide 1 of the MCS through CCC for amino acid 8 of beta-galactosidase. |
