Restriction digests of the clone give the following sizes (kb): PstI--4.4; BglII--4.4; HindIII--3.1, 1.3; EcoRI--4.4; XbaI--4.4. Contains a HIS4C/URA3 fusion cassette that can be used to make fusions to other proteins of interest. Also includes a M13 sequencing primer approximately 300 bp from the HIS4C/URA3 fusion junction. Encodes 93 amino acids of HIS4C and the complete 267 amino acids of URA3. Can be used for promoter/enhancer trap screens. Constructed by inserting a 1.4 kb HindIII fragment from pNKY2029 containing the HIS4/URA3 fusion into pNKY47, which is a derivative of pGC-1 with the original HindIII site destroyed and a new one created at the BamHI site of the MCS. The order of the major features in this plasmid is: M13 primer - ClaI - XhoI - SalI - XbaI (stop) - BamHI - HindIII - 90 aa HIS4C bridge/URA3 - BamHI - HindIII - BamHI - BglII - NcoI - EcoRI - M13 ori - pMB1 ori - ampR. |
