PlasmidLab / Guides / Gibson assembly

Design a Gibson assembly

Gibson assembly joins pieces that share their ends, so the junction carries no scar. You can ask for the construct in words, or build it from the menus.

Ask the agent

Open the vector and the file the insert comes from, then say what you want in the AI panel (View > Show AI Panel).

Get the GFP from the template into the vector, right behind the CMV promoter, without leaving a scar.

The PlasmidLab window with vector.gb and template.gb open and the request
Two files open, and the request typed in the panel.

The agent reads both files and designs the assembly on them. In this run it noticed the vector already carries a Kozak sequence right after the CMV promoter and put the ATG there, so neither junction gained a base. It wrote the four primers, made the 3,192 bp product as a new tab, and said what it had not checked.

The finished reply in the AI panel next to the new product tab pPL-CMV-GFPmut3b.dna, 3,192 bp, whose map shows GFPmut3b behind the CMV promoter and the four designed primers.
The reply, and the product it made as a new tab.
The agent works through PlasmidLab's own cloning tools, so what it makes is an ordinary file you can open, edit and save. Your sequences go to the AI provider you pick; the privacy page says what is sent.

Or do it yourself

Choose Clone > Gibson Assembly®…. Pick a mode for one insert, several inserts or fragments on their own, then give each piece its source file, the bases to use and the orientation. For a circular vector, choose the enzymes that open it, or the region to replace.

The Clone menu open over a plasmid map, with Gibson Assembly listed among the cloning methods.
Where the manual route starts.

The Primers tab holds the designed primers, the overlaps and any warnings; the Product tab holds the construct. Name the file and click Assemble. The window takes up to ten pieces, and each fragment has to be at least 20 bases.

Before you order

Read the product before you spend money on oligos: the sequence at each junction, the reading frame through a coding fusion, and the feature orientation. A design is not experimental validation, and the agent can be wrong.

Both routes are offline except the agent itself. Help > Try a Cloning Design… opens this exercise with its files.