Difference between revisions of "Part:BBa K3814004:Design"
m
 
Line 8: Line 8:
 
===Design Notes===
 
===Design Notes===
  
−
We aim to introduce this part into the iGEM registry to increase its access to researchers around the world. For USYD 2021's team, we planned on using this to qualitatively verify the expression levels of our Psal variants via an assay.  
+
We aim to introduce this part into the iGEM registry to increase its visibility and access to researchers around the world. For our USYD 2021 iGEM project, we planned on using this fluoroprotein to quantify expression levels from novel variants of a salicylate-inducible promoter; these changes to the promoter were made to try to generate varying expression levels.  (see BBa_K3814002/BBa_K3814005/BBa_K3814067/BBa_K3814068). By using the expression of fuGFP as a metric for the expression levels driven by the promoters, we would be able to determine the viability of these promoter mutations for our other experiments on natural transformation.
  
−
We designed a project that uses an inducible promoter that varying expression levels depending on some base changes (BBa_K3814002/BBa_K3814005). One of the first steps in our lab work would be to order two structures (BBa_K3814067/BBa_K3814068) from TWIST, which would contain fuGFP gene regulated by these promoters. By using the expression of fuGFP as a metric for the expression levels driven by the promoters, we would be able to determine the viability of these mutations in the real experiments.
 
−
 
−
Restriction enzymes were removed to minimise off-target effects. Substitute bases were chosen to most closely match the natural codon frequency in bacteria.
 
  
 
===Source===
 
===Source===

Latest revision as of 04:48, 14 June 2022


free-use GFP (fuGFP)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 151
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We aim to introduce this part into the iGEM registry to increase its visibility and access to researchers around the world. For our USYD 2021 iGEM project, we planned on using this fluoroprotein to quantify expression levels from novel variants of a salicylate-inducible promoter; these changes to the promoter were made to try to generate varying expression levels. (see BBa_K3814002/BBa_K3814005/BBa_K3814067/BBa_K3814068). By using the expression of fuGFP as a metric for the expression levels driven by the promoters, we would be able to determine the viability of these promoter mutations for our other experiments on natural transformation.


Source

Coleman Lab


References