My group has an issue using pTi for a particular cloning because of the BbsI sites in it, so I’m going to remove them. I logged this issue on the Reclone github:
While we’re at it are there any other Type IIS sites that could be taken out to remove any that are used in MoClo/SEVA @FernanFederici@Isaac_N?
SEVA removed the following from the WT backbone (then added some back so origin of replication is flanked by FseI and AscI; antibiotic marker by SwaI and PshA; and the cargo by PacI and SpeI): HindIII, PstI, XbaI, BamHI, SmaI, KpnI, SacI, SalI, EcoRI, SfiI, SphI, AvrII, PshAI, SwaI, AscI, FseI, PacI, SpeI, SanDI and NotI.
I assume that the following are safe to remove (or at least to try and check for deleterious effects):
BbsI
BsmBI
BtgZI
HindIII
KpnI
PshAI
PstI
SacI
SapI (except for the ones to make it uLoop compatible)
SpeI
Are the following used for anything? I’m guessing not but they are directly in between parts, so just checking…
XbaI
AvrII