Restriction digests of the clone give the following sizes (kb): BamHI--4.0; BamHI/EcoRI--4.0; BglI--2.9, 1.15. Potential inserts should be trimmed with Bal31 exonuclease to remove 5' sequences to within 10 bp of the start codon, and blunt-end ligated into the HpaI site downstream of a strong ribosome-binding site. Because cI857 is expressed by the plasmid itself, from its natural promoter PM, the vector may be used in virtually any E. coli strain. The sequence surrounding the cloning sites has been used to design primers for direct sequencing, including the M13 universal primer. Expression vector containing a strong ribosome-binding site and primer sites useful for sequencing. Encodes cI857. Constructed by inserting an oligonucleotide containing a HpaI site and a ribosome-binding site between the BamHI and SmaI sites of pCE30. |
