Figure 2.

RF-Cloning.org input page. (1) Optional information about the project. (2) ‘Orientation’ refers to complementarity of the insert sequence relative to the destination plasmid, and ‘arrow’ refers to how the insert will be drawn in the Savvy map. (3) Collapsible form fields where the default primer length and annealing temperature parameters can be modified. (4) Destination plasmid sequence can be entered manually (numbers and FASTA comments are ignored) or by selecting popular plasmids from the dropdown menu. (5) Insert sites within the plasmid sequence should be designated numerically in the fields provided (the very front of the plasmid is position 0), or by placing exclamation points (!) directly into the sequence. (6) Insert sequences must be entered manually (numbers and FASTA comments are ignored). (7) Plasmid maps are dynamically generated based on destination plasmid sequence. (8) When ready, the project can be submitted to the server.