CLC Main Workbench offers tools to perform in silico Gateway cloning (Thermo Fisher Scientific), including Multi-site Gateway cloning.
The three tools for doing Gateway cloning in the CLC Main Workbench mimic the procedure followed in the lab:
- First, attB sites are added to a sequence fragment
- Second, the attB-flanked fragment is recombined into a donor vector (the BP reaction) to construct an entry clone
- Finally, the target fragment from the entry clone is recombined into an expression vector (the LR reaction) to construct an expression clone. For Multi-site gateway cloning, multiple entry clones can be created that can recombine in the LR reaction.
For more information about the Gateway technology, please visit http://www.thermofisher.com/us/en/home/life-science/cloning/gateway-cloning/gateway-technology.html. To perform these analyses in CLC Main Workbench, you need to import donor and expression vectors. These can be found on the Thermo Fisher Scientific's website: find the relevant vector sequences, copy them, and paste them in the field that opens when you choose New | Sequence in the workbench. Fill in additional information appropriately (enter a "Name", check the "Circular" option) and save the sequences in the Navigation Area.