The help file for this program is available here

Enter the name of gene to delete

Enter desired length of primer target sequence

i.e. the length of primer excluding plasmid specific sequence.

Which plasmid will you use as a PCR template?

Five pairs of primers will be suggested to you. The primers are directly upstream of the ORF (not including the ORF sequence). The remaining pairs are positioned away from the ORF in user-defined increments. How far away (base pairs from ORF) would you like each subsequent primer set to be? Minus values acceptable, which will result in primers encroaching into the ORF.

Primer increment

Long strings of identical bases are highlighted in colour.