vector permitting positive selection for integration
vector permitting visual detection of recombinants
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--3.1, 2.8; HindIII--3.9, 2.1; BamHI--6.0.
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.
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 - EcoRI/MCS/HindIII - P15A - traJ - oriT - sacB.
Media
ATCC® Medium 1885: LB medium (ATCC medium 1065) with 15 mcg/ml gentamycin
Growth Conditions
Temperature: 37.0°C
References
Quandt J, Hynes MF. Versatile suicide vectors which allow direct selection for gene replacement in gram-negative bacteria. Gene 127: 15-21, 1993. PubMed: 8486283
