MacVectorTip: Designing Primers for Gibson Assembly

You can use MacVector to design primers for multi-fragment Gibson Assembly, and also generate the predicted recombinant DNA molecule resulting from the assembly. All you have to do to get started is choose the File->New->Gibson/Ligase-independent Assembly… menu item. From there, you can choose the type of assembly (it doesn’t have to be the usual 5’ exonuclease Gibson approach) and follow the instructions. If you are familiar with the New England Biolabs NEBuilder interface then you will love this. The algorithm is very similar, but you can add your annotated MacVector nucleic acid files and the final molecule will retain all of their features and annotations. In addition, the interface lets you view the translations around the junctions so you can be absolutely sure that your primer will create that perfect fusion protein.


(Note the gray CC residues that were inserted to ensure that the adjacent ATG start codon is maintained in-frame)

We do not currently have a dedicated tutorial for Gibson/Ligase-independent Assembly, but there is a useful section with examples you can follow in the Whats New in MacVector Workshop manual.

..and don’t forget that if you prefer Ligase Independent Cloning or other similar techniques the tool will do those too.

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