Gateway cloning
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.
During this process, both the attB-flanked fragment and the entry clone can be saved.
Please see Gateway technology for more information. 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.
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