Restriction digests of the clone give the following sizes (kb): EcoRI--2.7, 2.1; HindIII--4.8; PstI--4.8. Used with mini-Tn10 constructions for insertional mutagenesis of yeast and bacterial genes. pBR322 was modified by deletion of bp 75-2352 and destruction (by filling in) of the HindIII site. 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. |
