2. Cloning Vectors Enable DNA molecules to be replicated inside host (e.g., bacteria) cells. Features: 1. Origin of replication (ORI) 2. Cloning sites = Restriction enzyme recognition sites 3. Selection = Identify bacteria that contain recombinant DNA (i.e., vector + insert)

3. Types of cloning vectors 1. Plasmids: Autonomously replicating circular DNA molecules. 2.Bacteriophage: Small viruses that infect bacteria cells. 3. Vectors for large DNA inserts: Examples: Cosmids PACs BACs

4. Plasmid Vectors Origin of Replication  -lactamase: breaks down ampicillin Multiple cloning sites  -galactosidase Ligate DNA

5. Transformation Bacteria take up DNA from surrounding liquid colony

6. pUC18 Ampicillin: Selects for bacteria cells that contain plasmid X-Gal: Allows you to select bacteria that have plasmids with an insert.  -galactosidase X-Gal Blue Pigment “intact” lacZ gene disrupted lacZ gene