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golden-gate-assembly

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Design and validate Type IIS scarless DNA assembly (Golden Gate, MoClo) using Biopython Bio.Restriction. Screens parts for internal BsaI/BsmBI/BbsI/SapI sites (domestication), previews the fusion overhangs a digest exposes, and validates a fusion-overhang set for distinctness and fidelity. Use when designing a Golden Gate or MoClo assembly, domesticating a part by removing internal Type IIS sites, or choosing and checking fusion overhangs for one-pot assembly.

When to use

• Designing Golden Gate or MoClo assembly
• Domesticating parts by removing internal Type IIS sites
• Choosing and checking fusion overhangs
• Validating fusion-overhang set for distinctness and fidelity
• Screening parts for internal BsaI/BsmBI/BbsI/SapI sites
• Previewing fusion overhangs exposed by digest

When not to use

• Classic restriction-ligation (Type IIP)
• Gibson assembly
• One-off two-piece clone with convenient sites
• Assemblies where parts cannot be domesticated

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