View source for Part:BBa K4046840:Design


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<partinfo>BBa_K4046840 short</partinfo>

<partinfo>BBa_K4046840 SequenceAndFeatures</partinfo>


===Design Notes===
<i>dhdO</i> sequences were designed and optimized for binding to the  <i>DhdR</i> protein. Furthermore, a promoter, Kozak sequence, and a terminator were added.

===Source===

This includes identified binding site for the <i>DhdR</i> gene, as described by Xiao et al, 2021. This gene was synthesized commercially from the published sequence (IDT). The <i>DhdR</i> gene is a transcriptional repression factor that is derived from the bacteria <i>Achromobacter denitrificans</i>. The <i>dhdO</i> binding sites were designed and modified to improve binding behavior of the DhdR protein to the sequence. 

cLuc is an enzyme isolated from the genomic sequence of <i> V. hilgendorfii </i>, a marine crustacean. This gene was obtained through PCR of a commercially available plasmid pIND-CLucZ (Addgene, 53224). In normal function, cLuc is a luciferase enzyme that produces luminescence upon contact with its substrate, vargulin.

CMV is a constitutive reporter associated with the cytomegalovirus. This gene was obtained through the pcDNA5 backbone that we were using (Thermo Fischer, V103320). In normal function, the CMV promoter allows for high levels of expression of associated gene products.

===References===
Xiao, D., Zhang, W., Guo, X., Liu, Y., Hu, C., Guo, S., Kang, Z., Xu, X., Ma, C., Gao, C., & Xu, P. (2021). A D-2-hydroxyglutarate biosensor based on specific transcriptional regulator DHDR. https://doi.org/10.1101/2021.02.18.430539 
 
Cambridge, S. (2014). Photoswitching proteins: Methods and protocols (Vol. 1148). Humana Press.