If desired, a T-vector for TA cloning may be constructed manually and then added to the Cloning Vector Catalog prior to beginning the cloning workflow. To make the vector recognizable for TA cloning, a specific overhang must be created on both strands at the 3’ ends.

  1. Use File > Open to open the file or SeqBuilder project containing the vector sequence.
  1. Select Cloning > Change Overhangs.

    In this dialog, you can use the Trim Blunt or Fill In buttons to simulate modification of overhangs for incompatible ends, or manually manipulate the overhangs by dragging the arrows located above and below the two strands. The Revert button lets you return to the default overhangs.

    1. Use the arrow sliders to leave one overhanging thymine (T) residue on the bottom left and another on the top right.



    2. Click Change.
  1. Choose File > Save.
  1. Add the new vector to the Cloning Vector Catalog so it can be used with future projects.

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