Part:BBa_K1526009:Experience

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Creating the regulator LacRS

In order to characterize the parts we have created a new BioBrick compatible with our chassis. As E.coli DH5α does not have a lac repressor system we have assembled together basic parts J32100, B0033, C0012, B0010, B0012 and R0010 to create the composite regulator Lac Repressor System (LacRS)

We have successfully assembled the parts together by step-wise 3A Assemblies, transformations and gel checking for expected sizes. <p><p>We have successfully cloned and tested the functionality of LacRS in pSB1A3 (<a href="https://parts.igem.org/Part:BBa_K1526009">BBa_K1526009</a>) into E.coli DH5α

Our first functionalization test coupling LacRS upstream the Green Fluorescent Protein (GFP) Biobrick (E0040) showed expression of GFP under UV light when the system is previously induced by IPTG.

Left plate: E.coli DH5α transformed with BBa_K1526009 in PSB1A3 and plated on LB-Agar+AMP+0.5mM IPTG; Right plate: same treatment without IPTG

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