BEVA2.0: modular assembly of golden gate-compatible vectors with expanded utility for genetic engineering

Area of Science:

• Synthetic Biology
• Molecular Biology
• Genomics

Background:

• The Bacterial Expression Vector Archive (BEVA) is a modular platform for constructing bacterial expression vectors.
• Expanding modular systems is crucial for increasing the diversity and functionality of engineered plasmids.

Purpose of the Study:

• To expand the BEVA platform with new parts and modules.
• To enhance the capacity for producing diverse replicating plasmids.
• To enable genome manipulation in bacterial strains using the expanded BEVA system.

Main Methods:

• Introduction of 11 new BEVA parts, including cloning site variants, antibiotic resistance modules, origins of replication, and accessory modules.
• Development of BEVA-derived Golden Gate cloning plasmids for part validation.
• Integration of parts compatible with the CIDAR MoClo parts library.

Main Results:

• The BEVA system has doubled in size, significantly increasing its capacity for diverse replicating plasmid production.
• The expanded platform is now suitable for genetic engineering methods involving bacterial genome manipulation (deletions, integrations).
• New parts ensure compatibility with existing modular cloning systems like CIDAR MoClo.

Conclusions:

• The expanded BEVA platform offers greater versatility for constructing replicating plasmids and performing bacterial genome engineering.
• The new modules and compatibility features facilitate broader applications in synthetic biology and microbial engineering.
• BEVA-derived plasmids provide valuable tools for genetic engineering in proteobacteria and other bacterial species.