Restriction digests of the clone give the following sizes (kb): BamHI/KpnI--4.1, 3.7; HindIII--8.2; SmaI--8.2; BamHI/SmaI--6.7, 1.2. Copy number is 2-3 per genome in Pseudomonas spp. Stability is reduced compared to pCN40 (ATCC 77097). 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. Retains the mobilization site of pBR322-derived vectors and can be mobilized if suitable functions are provided in trans. Shuttle vector permitting maintaining recombinants at low copy number and, if necessary, curing the host of recombinants. Constructed by deleting a 0.6 kb HindIII fragment from pCN60 (ATCC 77101) which included the multiple cloning site and 0.2 kb (nt 1615-1823) from the pPS10 replicon. The order of the major features in this plasmid is: KpnI - repA - strR - PstI - SmaI - BamHI - HindIII - bla - pMB1 ori. |
