The help file for this program is available here

Enter the name of gene to delete

Enter desired length of target sequence (i.e. length of primer excluding plasmid specific sequence)

The reverse primer is designed to allow controlled expression or N-terminal tagging of full length proteins. Five forward primer sequences will be suggested to you. The first primer is directly upstream of the stop codon. The remaining forward 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

Which N-Terminal tag would you like to use?

Long strings of identical bases are highlighted in colour.