Restriction digests of the clone give the following sizes (kb): BamHI--6.0; XbaI--6.0; PstI--6.0; HindIII--6.0. This is a Escherichia coli/yeast episomal shuttle vector with multiple restriction sites for cloning. pUC18 was digested with NdeI and blunt-ended by filling in. A 2.95 BamHI/XbaI fragment of pC4B was blunt-ended (filled in) and ligated to the pUC18. The 2.95 kb fragment of pC4B includes the LEU2 gene (nt 286-2256 in the final construct, where the A of the ATG of lacZ' is number 1) and the 2 micron replication system (nt 2257-3241). |
