How can I insert a restriction fragment generated by partial digestion into a circular vector?
A restriction fragment can be inserted at a restriction site in the vector, or it can replace a restriction fragment in the vector.
Start the Restriction Cloning Tool
Click Actions → Restriction and Insertion Cloning → Insert Fragment.
Set the Vector
In the Vector tab, use the dropdown to browse and choose the vector for cloning.
Select a site or sites for insertion of the insert.
In the above example, a blunt insert will ligated into the vector-based SmaI site.
Define the Source Sequence
Switch to the Fragment tab, click the dropdown to set the "Source of Fragment" that contains the insert.
If required, click on the "Show Enzymes" button on the side toolbar to display the restriction enzyme site of interest. To learn more about displaying specific enzymes on a sequence, see the guide chapter on Enzymes.
Define the Partial Restriction Fragment
Enter the name of the enzyme of interest, or select the first site on the sequence.
Initially only fragments generated by complete digestion will be shown in the list of possible fragments.
Click the dropdown and choose "Cut partially" and a list of all possible partial fragments will be displayed.
Select the partial fragment of interest.
Alternatively, in Map or Sequence view, click on the first site, then shift-click on the second site to select the partial digestion fragment of interest.
Create the Product
Switch to the Product tab to preview the ligation of the partial insert into the vector.
Enter an appropriate name for the new product sequence.
Click Clone to create the new product sequence.