pK19mobsacB plamid in E. coli SCS110
Construct size (kb): 5.66
Vector type: plasmid
Cloning sites: HindIII SphI PstI SalI XbaI BamHI SmaI EcoRI
Construction: pK19, pSUP102 (RP4 mob) sacB; the sacB gene was inserted into the pK19mob vector.
Genome: Bacillus subtilis
Gene name: levansucrase
Insert end: Ecl136II
Insert end: XbaI (modification: blunt ended)
Insert size (kb): 1.9
Complete coding sequence ?: Y
Vector size (kb): 3.76
Type of vector: plasmid
Vector ends: AsuII (modification: blunt ended)
Host range: Escherichia coli ; Salmonella sp. ; Serratia sp.
Features (with orientation and location, if known):
Marker: sacB (sucrose sensitivity)
Insert detection: lacZ’,
Produces protein levansucrase
Vector containing primer sites useful for sequencing
Vector permitting visual detection of recombinants
Restriction digests of the clone give the following sizes (kb): EcoRI--5.6; HindIII--5.6; PstI--5.6.
After mobilization, the plasmid can be maintained by integration into the host chromosome via homologous recombination.
Excision of the intervening plasmid sequence by a double cross-over event can be faciliated by selection on medium containing 10 percent sucrose.
The sacB gene has been modified to eliminate the HindIII and EcoRI sites in the coding region.
Differs from pK18mobsacB (ATCC 87097)
only in the orientation of the polylinker.
Cloning vector allowing mobilization into a wide range of Gram- and Gram+ bacteria.
ATCC Medium 1065 (see below) plus kanamycin (50 mcg/ml) ATCC Medium 1065: Tryptone (Difco 0123), 10.0 g Yeast Extract (Difco 0127), 5.0 g NaCl, 10.0 g Distilled water, 1.0 L
Simon R, et al. A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram negative bacteria. Bio-Technology 1: 784-791, 1983.
Schafer A, et al. Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene 145: 69-73, 1994. PubMed: 8045426