IMPORTANT CHANGES: We have introduced two main modifications in our domestication tools to better accommodate them to the common standard agreed by several Plant laboratories (see Patron et al., New Phytologist 2015).

a. The domestication with the pUPD is no longer supported by gbcloning software. From now on all GBparts will be automatically domesticated in pUPD2 with chloramphenicol resistance. Please, visit this page for an overview of the differences between pUPD and pUPD2. In anticipation to this move, our lab distributed pUPD2 a few months ago to all our registered GB users. If you did not received pUPD2 vector, you can request a sample to Addgene or directly to our lab.

b. The new domestication tool incorporates an option to select additional domestication enzymes (except on the 'Phylogeny Search'). Domestication for BsaI and BsmBI is mandatory to work with GB. However, you can decide to additionally domesticate your parts for BpiI (BbsI) in order to make them fully compatible with MoClo. You can also decide to waive BtgZI domestication. In that case, remember that your new TUs need to be always assembled first in alpha level plasmids.


The first step in the GoldenBraid assembly strategy is to adapt your DNA building block to the GB standard. It implies the removal of the internal restriction sites for the enzymes used in GB (BsaI, BsmBI and BtgZI) and the addition of the appropriate 4-nt flanking overhangs .

This process is referred to as domestication and we offer three different domestication strategies:

1. The GB Domesticator uses PCR mutagenesis followed by typeIIS-based assembly of mutagenized fragments to domesticate the sequence.

2. The Phylogenetic Search, searches the databases for the closest orthologous genes containing minimum internal sites.

3. The Synthetic strategy provides the domesticated sequence in a single DNA fragment, ready to order it and clone it into the pUPD vector.