pCN61

Shuttle vector permitting maintaining recombinants at low copy number and, if necessary, curing the host of recombinants. (ATCC staff) Constructed by deleting a 0.6 kb HindIII fragment from pCN60 (ATCC 77101) which included the multiple cloning site & 0.2 kb (nt 1615-1823) from the pPS10 replicon. 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 & 1500), with the site at nt 536 between the repA promoter and coding sequences. (personal communication) Retains the mobilization site of pBR322-derived vectors and can be mobilized if suitable functions are provided in trans. 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. (ATCC staff) The order of the major features in this plasmid is:KpnI – repA – strR – PstI – SmaI – BamHI – HindIII – bla – pMB1 ori. Medium is 1227 LB plus ampicillin. Hosts: E.coli HB101, Pseudomonas aeruginosa, Pseudomonas putida, E.coli, Pseudomonas aeruginosa PAO1024, Pseudomonas aeruginosa PAO1162, P.putida KT2440.