Chlorite Dismutase Composite Part [Cld(SP-)]
See the full characterization of CLD(SP-) on the BioBrick Registry
Here we have the Cld+sp and Cld-sp g-blocks from the previous slide digested with BsaI, resulting in the respective sticky ends. These sticky ends are complementary to the sticky ends of the CPB-38-441 expression vector which is shown below. This particular vector contains the Kanamycin Resistance gene which we used as a selectable marker when we plated our transformed cells in media treated with kanamycin. This plasmid also contains a LacI repressor which acts as the on/off switch for gene expression. Upon induction with IPTG, the repressor releases its bind on the lac operators, allowing RNA polymerase to begin transcription. The plasmid also includes the “Tinsel” gene from DNA 2.0’s “Paintbox” chromophore collection. The expression of this chromogenic protein will play an important part in the selection process of our transformation.
It is important to note that our BioBrick Submission was sent on an iGEM Registry standard backbone, different from the plasmid shown in the image above.