Restriction digests of the clone give the following sizes (kb): EcoRI--6.8; PstI--4.6, 1.7, 0.6; HindIII--3.7, 1.6, 1.5; SmaI--3.7, 3.2; BglII--5.8, 1.0. Shuttle vector to facilitate construction of gene disruptions in E. coli. The gene of interest is cleaved at an appropriate site, made flush, and ligated to the SmaI fragment from this plasmid that contains kanR and the yeast selectable marker. The appropriate product is selected for by kanR in E. coli. The order of the major features in this plasmid is: TRP1 - kanR - EcoRI/MCS/HindIII - f1 ori - ampR - pMB1 ori. |
