Expression plasmid. The expression cassette is under transcriptional control of the tetracycline promoter/operator. The expressed recombinant protein will be localized in the cytoplasm.
Strep-Tactin® affinity tag (Strep-tag II®) for the purification of recombinant protein. The affinity tag is fused to the N-terminus of the recombinant protein and can be removed by cleavage with factor Xa.
Expression is induced upon addition of 200 ug anhydrotetracycline (order no.: 2 0401 001; 2 0401 002) per 1 liter E. coli shaking culture (A550= 0.5).
Any E. coli strain. The tet promoter works independently from the genetic background of E. coli.
Chloramphenicol. Note: The CamR resistance gene codes for homotetrameric chloramphenicol acetyltransferase (MW of the monomer = 26.6 kDa) which is predominantly expressed in the cytosol of E.colitransformed with this plasmid
5 ug, dissolved in 10 mM Tris/HCl pH 8.0, 1 mM EDTA; 20 ul.
4 °C for frequent usage, 20 °C for long term storage.