Restriction digests of the clone give the following sizes (kb): BamHI--6.2, 3.1; HindIII/PstI--4.5, 3.4, 0.8; PvuII--4.4 (doublet); PvuII/KpnI--4.4, 2.5, 1.6. Contains the basic replicon of pPS10 which has been sequenced (EMBL accession number X58896). The repA portion is flanked by EcoRI sites (nt 536 and 1500), with the site at nt 536 between the repA promoter and coding sequences. Multiple selectable markers permit insertional activation to screen for recombinants (PstI site in ampR). Retains the mobilization site of pBR322-derived vectors and can be mobilized if suitable functions are provided in trans. Constructed by ligating pCN38 (ATCC 77095) and pBR322 which had been linearized with PvuII. The order of the major features in this plasmid is: PvuII - pBR322 (pMB1 ori - bla - tetR) - PvuII - BamHI - kanR - HindIII - repA. |
