Fungal GB Domesticator
The first step in the GoldenBraid assembly strategy is to adapt your DNA building block to the GB grammar and to remove the internal restriction sites for the enzymes used in GoldenBraid.
This process is referred to as domestication and is computer assisted by the GB Domesticator software tool. Here you can domesticate your DNA sequence to convert it into a GBpart/superpart. For this, you can either select one of the standard GBpart categories from the dropdown menu or you can introduce the prefix and suffix of your choice (4 nts barcodes).
For the assembly of gene disruption cassettes with GB, selection markers including a promoter, a coding sequence and a terminator are domesticated as a single part in the pUPD2. Please, make sure that you use lowercase letters for non coding regions and mark the 'The noncoding sequence is in lowercase' tickbox in order to domesticate marker parts.
Markers should be domesticated with the same orientation as they will be assembled in the final construct. Therefore positive marker sequences (forward) should be uploaded as (5'- 3') while negative marker sequences (reverse) should be uploaded as (3'-5').