pACYC-lacZ1

This vector allows simplified selection of recombinant clones using blue/white selection on X-Gal, IPTG - containing media.

This vector is a small, medium copy number, E.coli plasmid, 2172 bp in length.

The plasmid contains:
1) the p15A replicon (ori) responsible for plasmid replication (source - plasmid pACYC184);
2) the Cm(R) gene, coding for chloramphenicol acetyl transferase, which confers resistance to chloramphenicol (source - plasmid pACYC184);
3) the region of E.coli lac operon containing a CAP protein binding site, promoter Plac, lac repressor binding site, and the 5’-terminal part of the lacZ gene encoding the N-terminal fragment of beta-galactosidase. This fragment, whose synthesis can be induced by IPTG, is capable of intra-allelic (alpha) complementation with a defective form of beta-galactosidase encoded by the host (mutation delta(lacZ)M15). In the presence of IPTG, bacteria synthesize both fragments of the enzyme and form blue colonies on media with X-Gal. Insertion of DNA into the Multiple Cloning Sites (MCS) located within the lacZ gene inactivates the N-terminal fragment of beta-galactosidase and abolishes alpha-complementation. Bacteria carrying recombinant plasmids therefore give rise to white colonies.

The map shows enzymes that cut pACYC-lacZ'1 once.

The plasmid is fully compatible with those carrying the pMB1 and ColE1 replicons.