Extended Set of GoldenBraid Compatible Vectors for Fast Assembly of Multigenic Constructs and Their Use to Create Geminiviral Expression Vectors

• DOI: 10.3389/fpls.2020.522059
• Publication Date: 2020-10-22T04:21:13Z
• Authors: Jakub Dusek; Helena Plchova; Noemi Cerovska; Zuzana Poborilova; Oldrich Navratil; Katerina Kratochv...; Cornelius Gunter; Raygaana Jacobs; Inga I. Hitzeroth; Edward P. Rybicki; Tomas Moravec
• Keywords: Synthetic Biology; Cloning (programming)

Methods for simple and fast assembly of exchangeable standard DNA parts using Type II S restriction enzymes are becoming more popular in plant synthetic molecular biology. These methods enable routine construction large complex multigene structures. Two available frameworks emphasize either high cloning capacity (Modular Cloning, MoClo) or simplicity (GoldenBraid, GB). Here we present a set novel α-level plasmids compatible with the GB convention that extend ability to rapidly assemble genetic constructs, while maintaining compatibility all existing as well most MoClo modules. With use our new plasmids, can be assembled into assemblies containing 1, 5, 10 up theoretically 50 units each successive level infinite loop assembly. Assembled constructs also combined conventional binary GB-assemblies (1, 2, 4, 8… units). We demonstrate usefulness framework on single tube replicating expression based geminivirus Bean yellow dwarf virus (BeYDV).