Comments
Restriction digests of the clone give the following sizes (kb): SstI--7.9; KpnI--6.0, 1.9; BamHI--7.9.
Replicates only in enterobacteria, and so functions as a suicide vector in other Gram negative hosts.
A single BglII site within the gentamicin resistance gene (gtmR) allows insertion of other resistance genes (e.g. as a BamHI cassette).
When induced by media containing 5% sucrose, sacB is lethal in a wide range of Gram negative bacteria, and thus permits selection for loss of the vector.
IMPORTANT: To prevent amplification of a rearranged and/or deleted cosmid, we recommend streaking on LB + amp plates at 30C and picking small colonies for liquid culture.
Suicide vector permitting gene replacement and mobilization into a wide range of Gram negative bacteria. Allows positive selection for integration.
The order of the major features of the plasmid is: gtmR - SstI/MCS/KpnI - P15A - cos - traJ - oriT - sacB.
