pAM-Red-Spe9

This plasmid can be used to modify genomes of host Escherichia coli cells as well as present in these cells episomes via mechanism of homologous recombination. When needed, this plasmid can be easy eliminated from host cells.

This vector is medium to high copy number E.coli plasmid, 8197 bp in length.

The plasmid contains:
1) the ColE1 replicon (ori) placed under control of lactose regulatory elements (promorer Plac and lactose repressor LacI);
2) the Spc(R) sequence confers resistance to spectinomycin;
3) the chimera operon encoding components of bacteriophage lambda DNA recombination machinery (Gam, Bet and Exo) under control of bacteriophage lambda promoter (Pr) and regulated by temperature-sensitive CI repressor.

Due to its structure, this vector directs synthesis of bacteriophage lambda recombination machinery only when temperature of host propagation is 37^oC or higher. Also, this vector is efficiently replicates when host cells are propagated in culture media supplemented with IPTG or lactose and is efficiently lost from such cells in the absence of these inducers.

The plasmid is fully compatible with those carrying the p15A replicon or other replicons compatible that of ColE1. Alternatively, this plasmid can co-maintained with other plasmids carrying ColE1 origin and encoding different antibiotic resistance in the same host cells by applying selective pressure by both corresponding antibiotics (Please see vector pAM-Red-Ap3 for the alternative choice of antibiotic resistance) .