pND201


Vector IG Sequence Link :
General : plasmid ds-DNA 4013 BP
Functions : (cloning)
Selection : ()
Copy Number :
Hosts : (E.coli AN1459)(E.coli)
Suppliers : (ATCC)
Misc.Comments : Because cI857 is expressed by the plasmid itself, from its natural promoter PM, the vector may be used in virtually any E.coli strain. Constructed by inserting an oligonucleotide containing a HpaI site and a ribosome-binding site between the BamHI and SmaI sites of pCE30. 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. The sequence surrounding the cloning sites has been used to design primers for direct sequencing, including the M13 universal primer. Restriction digests of the clone give the following sizes (kb): BamHI--4.0; BamHI/EcoRI--4.0; BglI--2.9, 1.15. (ATCC staff) Expression vector containing a strong ribosome-binding site and primer sites useful for sequencing. Encodes cI857. (ATCC staff) Medium is 1227 LB plus ampicillin.
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