Restriction digests of the clone give the following sizes (kb): EcoRV--8.8, 2.15; PstI--5.6, 3.1, 2.3; HindIII--9.4, 1.6; PvuI--8.8, 2.15; BamHI--uncut. This construct is available in two forms: in Escherichia coli as the plasmid (ATCC 39344) and integrated into the Bacillus chromosomal DNA (ATCC 39345). The depositor suggests streaking for single colonies on L broth + 1.5% agar + 100 ug/ml ampicillin + 5 ug/ml chloramphenicol and using a single colony. The vector contains the replicon and beta-lactamase gene from pBR322, a galactokinase gene, a CAT gene from pC194, and a transcription terminator from bacteriophage lambda. It also contains chromosomal sequences from Bacillus subtilis for integration. |