Restriction digests of the clone give the following sizes (kb): BamHI--5.4; PstI--5.4; PvuII--3.7, 1.7; SacI--4.5, 0.9. M13K07 helper phage improves the yield of ssDNA in E. coli with this vector. M13mp18 may also be used. The neo promoter was replaced by a synthetic promoter recognized by both hosts. IMPORTANT: To prevent amplification of a rearranged and/or deleted cosmid, we recommend streaking on LB + amp plates at 30C and picking small colonies for liquid culture. Cosmid shuttle vector permitting production of ssDNA and containing promoters for in vitro RNA synthesis. Terminators flanking the polylinker protect the vector from transcripts originating in cloned inserts. The ori of pJV1 from Streptomyces phaeochromogenes is compatible with plasmids derived from pIJ101. This vector can be used for complementation studies using multiple cloned genes in one host. The order of the major features in this plasmid is: rrnC terminator - SP6 - SphI/MCS/HindIII - T7 - trpA terminator - neo - synthetic promoter - pMB1 ori - cos - M13 ori - pJV1 ori. |
