I have a pET41a+ vector in which the SacB-Cat gene is cloned. After digestion with SphI and XhoI, I have amplified this (SacB-Cat) cassette with the desired primer followed by PCR purification and can be further used for transforming into electrocompetent cells which already have recombinase gene in helper plasmid.
This cassette is generally used for Homologous Recombination or mutant preparation.
The amplified product is near 2500 base pair. But the problem is after getting an electric pulse for 5.5 ms I did not even get a single transformed colony.