How do I insert a back-translated protein into a DNA sequence?
Open a DNA Sequence File and Define the Insertion Point
Open a DNA sequence and switch to Sequence view.
Click at the position you wish to insert the sequence, or select 1 or more nucleotides if you wish to replace a region with the feature. In this example, the eGFP CDS stop codon will be replaced with the incoming sequence.
Specify the Protein Sequence and Back Translation Parameters
Click menu Edit → Insert → Reverse Translation....
- Type or paste the desired protein sequence into the field provided.
- Use the drop-down menu to specify the genetic code to use for back-translation.
- Choose whether alternate synonymous codons are chosen a) Randomly, b) with degeneracies, or c) using a specific codon usage table.
- Choose whether to add the new sequence as a translated feature.
Click Replace (or Insert) to add the new back translated sequence to your sequence.
If the option to "Add a translated feature" was checked, then give the feature a name and click OK.
View the Newly Added Sequence
Select the "Show Colors" button in the side toolbar to highlight (in red) the newly added sequence.
Click File → Save the modified sequence.
If you wish to add multiple sequence elements to an existing sequence then consider using Tools → Design Synthetic Construct...