Plasmid pKQ-GFPuv was digested with BglII and Mung Bean Nuclease, purifed on a zymo column, and then digested with EcoRI. The material was run on a gel resulting in two bands. The smaller of the bands was gel purified to give fragment A. Meanwhile, plasmid pSC101 was digested with HindIII, purified, and then added to a reaction containing polymerase buffer, 0.2 mM dNTPs, and T4 DNA polymerase. After purifying the DNA on a zymo column, the material was digested with EcoRI and then subjected to gel electrophoresis. The larger of two bands was purified from the gel affording fragment B. Fragment A and fragment B were combined in a reaction with ligase buffer and T4 DNA ligase, introduced into E. coli by transformation, and spread on LB plates supplemented with Ampicillin.