pHPS9

Cloning into the NdeI, NheI, BamHI, SmaI or EcoRI sites inactivates lacZalpha. The NcoI site interrupts the chloramphenicol resistance sequence. [1] Permits alpha-complementation to identify recombinants when used with Bacillus subtilis 6GM15, and alpha-complementation with plasmid marker rescue when used with Bacillis subtilis 6GM15[pHPS9R] (ATCC 37818).[3] The cat86::lacZalpha fusion is in-frame. Expression is controlled by the P59 promoter from Lactococcus lactis subsp. cremoris Wg2. [1] This is the preferred strain for isolating the plasmid, because the copy number is higher in E.coli than in B. subtilis. (personal communication) Restriction digests of the clone give the following sizes (kb): BamHI–5.7, EcoRI–5.7, PstI–4.9, 0.7. (ATCC staff) Medium is -1 1065 plus erythromycin (200 ug/ml). Plasmid was not verified by restriction analysis. Strain was checked for the following phenotypes:showed growth on LB + chloramphenicol (5 ug/ml, but not 10 ug/ml); blue colonies on IPTG + Xgal (80 ug/ml); (ATCC staff) blue colonies on IPTG + Xgal (80 ug/ml) + erythromycin (150 ug/ml). (ATCC staff) Medium is -1 1065 plus chloramphenicol (5 ug/ml). Hosts: E.coli NM522, Bacillus subtilis, E.coli, Bacillus subtilis 6GM15 [ATCC 47051].