Restriction digests of the clone give the following sizes (kb): BamHI--12.5; HindIII--12.5. The EcoRI site is available for cloning but the lacPO N-terminal region will be deleted. The polylinker is inserted in frame within the lacZ gene facilitating selection for blue and white colonies. Ligating lambda cosR adapters onto the termini of potential inserts and cosL adapters onto the linearized vector enhances the step of circularization of large molecules. This is a low copy number mini-F based plasmid vector with rare restriction sites in its polylinker for cloning inserts of up to 100 kb. The depositor recommends adding spectinomycin to the media at a concentration of 20-30 ug/ml at the first establishment of spcR transformants and 15 ug/ml thereafter. |
