Contains the following sites separated by (bp): EcoRI-764 -AhaIII-97 -ScaI-112 -PvuI-2101 -AccI-1848 -ScaI-55 -AccI-2039 -BstEII-372 -PvuII-93 -PvuII-94 -HpaI-320 -BstEII-182 -MluI-736 -EcoRI. Restriction digests of the clone give the following sizes (kb): AccI--3.9, 3.0, 1.9; BamHI--8.8; HindIII/PvuII--7.6, 1.3. In the pMMB66 (ATCC 37620, 37621) and pMMB67 (ATCC 37622, 37623) vector series, the EH and HE in the name designate the orientation of the multiple cloning site to the tac promoter. An autoregulated high-level expression vector utilizing the tac promoter, rrnB terminators and lacIq for lac repression. Concentration of induced gene product may be regulated by varying the lac inducer concentration. Can be mobilized by conjugative IncP helper plasmids [e.g. pRK2013 (ATCC 37159), pUB307] into Klebsiella aerogenes, Proteus mirabilis, Pseudomonas aeruginosa, Serratia marcescens, and other gram-negative bacteria. The sequence of RSF1010 is given in this reference. A 1246 bp lacIQ HindIII fragment was inserted at XmaIII/BamHI of pKK223-3. The 2995 bp NruI/AhaIII fragment of the resulting plasmid was cloned between PstI (bp 7768) and PvuII (bp 1951) of RSF1010. |
