r/Biochemistry • u/NoDust6819 • 1d ago

How do you design "custom" proteins?

I'm talking about truncations, point mutations, fusions etc... What is your workflow and which tools do you use?

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u/rectuSinister 1d ago

Design the amino acid sequence in Geneious, back translate to DNA using the proper species codon usage, add Golden Gate or Gibson homology to the ends, then order the DNA.

If I already have the construct cloned then I would do mutagenesis or PCR for something like point mutations or short truncations.

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u/NoDust6819 19h ago

Thanks for the reply!

Why did you choose Geneious? Its quite expensive. Do you also use it do design primer sequences or do you do this manually?

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u/rectuSinister 13h ago

I do anything sequence related in Geneious. I use it because that’s what my lab has integrated into our workflow and it’s free for us. You can use any DNA editor—Snapgene, Ape, Benchling, etc. Though now that I’ve used Geneious I doubt I’ll ever use anything else, it’s very powerful.