Difference between revisions of "Part:BBa K4046840:Design"

 
(→‎Design Notes)
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===Design Notes===
 
===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.
 
<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.
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===Source===
 
===Source===

Revision as of 21:51, 21 October 2021


CMV - BS #1 with spacer - BS #1 with spacer - BS #1 - Kozak - cLuc - bghA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 614
    Illegal BglII site found at 1311
    Illegal BglII site found at 1370
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

dhdO sequences were designed and optimized for binding to the DhdR protein. Furthermore, a promoter, Kozak sequence, and a terminator were added.

Source

This includes identified binding site for the DhdR gene, as described by Xiao et al, 2021. This gene was synthesized commercially from the published sequence (IDT). The DhdR gene is a transcriptional repression factor that is derived from the bacteria Achromobacter denitrificans. The dhdO 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 V. hilgendorfii , 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