Comments
Restriction digests of the clone give the following sizes (kb): BamHI--3.2, 2.2, 1.4; EcoRI--3.1, 2.0, 1.8; KpnI--5.2, 1.8; PvuI--5.0, 2.0.
Vector contains the following restriction sites: EcoRV--1.01,3.20,5.64; BstXI--1.04,3.15; EcoRI--1.12,5.94; KpnI--1.51,3.23; ClaI--1.60,3.30; NacI--2.79; PvuI--2.97,4.88; PvuII--3.00,3.45; ScaI--3.12,4.99; AatII--5.44.
Two-step replacement in budding yeast. LEU2 gene provides positive selection for the integrative step; CYH2S provides negative selection for mitotic recombinants that have excised the plasmid sequences. Can be used for modification of YACs.
The fragments were ligated in the plasmid: LEU2 -> f1(+) -> lacZ -> T7/T3 -> ori -> ampR -> CYH2 ...
