Restriction digests of the clone give the following sizes (kb): BamHI--12.0; EcoRI--4.4, 4.1, 2.6, 0.8; HindIII--4.6, 3.4, 2.7, 1.3; PstI--12.0. LEU2-d is a promoter-defective allele of the yeast LEU2 gene, which increases plasmid copy number to 200-300 copies in the presence of LEU2 selection (host grown in medium lacking leucine). High-copy YE-type shuttle vector. Consists of the entire 2 micron circle genome cloned, at the EcoRI site of the small unique region, into the EcoRI site of pBR322. The leu2-d sequence (approximately 1.4 kb) is inserted into the PstI site of the 2 micron circle. |
