Restriction digests of the clone give the following sizes (kb): PstI--7.1; HindIII--4.6, 1.4; EcoRI--4.3, 2.8. Is compatible with plasmids derived from pBR322. Used with mini-Tn10 constructions for insertional mutagenesis of yeast and bacterial genes. Contains the ats1 ats2 transposase gene that permits relaxed insertion specificity (altered target specificity, ATS). Expression is regulated by the Ptac promoter, inducible by IPTG. The transposase segment extends from IS10R to the EcoRI site at nt 3140 of Tn10, with a deletion of nucleotides 1329-1942 to remove the transposase binding site. Transcription from the Ptac promoter is in a direction opposite to that of amp. |
