Restriction digests of the clone give the following sizes (kb): EcoRI--9.2; HindIII--6.6, 2.5; PstI--5.2, 2.95, 0.98; XbaI--6.8, 2.1. Construct permitting translational fusions of kan to target genes. Kanamycin resistance is expressed after transpositions in the correct orientation and reading frame into an expressed target gene. Contains a 2.2 kb mini-Tn10 cassette conferring erythromycin resistance and a promoterless kan gene ('kan) gene, bounded by inverted repeats of the outermost 70 bp of IS10R and embedded (as an EcoRI/HindIII fragment) in hisG and hisD. Also contains the wild type transposase gene regulated by the Ptac promoter, inducible by IPTG, oriented opposite to 'kan, and separated from 'kan by 4 tandem repeats of the rrnB terminator. The order of the major features in the insert is: XbaI - HindIII - transposase - Ptac - rrnB terminators - HindIII - 'kan - erm - EcoRI - XbaI. |
