Option D – Use part of an open SeqBuilder Pro sequence file as the insert

1. Use File > Open to open the file or SeqBuilder Project (.sbd) containing the sequence fragment you wish to clone.

2. Use any of the following methods to select the sequence fragment to use as the insert:

• • Manually select a portion of sequence by dragging the mouse across it.
    
    
  • Click on an existing feature to select the corresponding sequence.
    
    
  • Use the method described in Using a PCR product of the current primer pair as the insert
    
    
  • Use the method described in Using restriction enzyme cut sites to create the insert. Note that you do not actually need use a restriction enzyme cut site as the sequence fragment. The sequence fragment can be anything you want: an open reading frame (ORF), multiple cloning site (MCS; polylinker), feature, or a random selection.

3. Choose Cloning > Clone Selected Fragment, or Cloning > Clone Selected Fragment From Opposite Strand, as needed. The latter command selects the opposite orientation of the sequence.

The Project window opens with the Clone tab active. The selection from Steps 1-2 is placed in the Inserts folder in the Project tab, as well as in the Fragments section near the bottom right of the Clone tab.

Proceed to Step 2: Specify the method and options.