Understanding GoldenBraid Assembly

GoldenBraid is a modular cloning syntax generally used in plants. The first time you use it, it may take a second to understand how it works. This page aims to help with that, and how to use it in OpenCloning.

GoldenBraid parts

GoldenBraid parts are domesticated into vectors related to pUPD. Their main feature is that they have two type IIS restriction sites (for BsaI and BtgZI) flanking the part, so regardless of which enzyme you use to cut the vector, you get the same overhangs (1234 and 5678 in the example below).

Adapted from 10.1104/pp.113.217661

GoldenBraid Backbones and sequential assembly

• GoldenBraid is a "binary" assembly system, basically meaning that you assemble two things at a time.
• Backbones are classified into categories:
  • alpha / omega:
    • alpha has kanamycin resistance, and you use BsaI to clone into it.
    • omega has streptomycin resistance, and you use BsmBI (and BtgZI for TU, see below) to clone into it.
  • 1 / 2:
    • 1 will be on the left side of the assembly, and 2 will be on the right side.
  • R / not R (reversed, not reversed).
    • The R indicates that the Transcription Unit (TU) is reversed.

Not all kits support TU cloning into alpha and omega, for example.