The help file for this program is available here

Enter the name of gene to delete

Enter desired length of primer target sequence

Which plasmid will you use as a PCR template?

The forward primer is designed to allow C-terminal tagging of full length proteins. Five reverse primers sequences will be suggested to you. The first primer is directly downstream of the stop codon. The remaining reverse primers are positioned away from the ORF in user-defined increments.

How far away (base pairs from ORF) would you like each subsequent primer to be?

Primer increment

Long strings of identical bases are highlighted in colour.