Regulatory
Ptet*

Part:BBa_K4579000:Design

Designed by: Alexa Morton   Group: iGEM23_Austin-UTexas   (2023-09-17)
Revision as of 03:20, 22 September 2023 by Alexakmorton (Talk | contribs) (→‎Source)


PTet* promoter + RBS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

When designing our inducible promoter parts derived from the 'Marionette' plasmids, we decided to create universal primers that would allow us to amplify the promoter from any of the Marionette plasmids. We chose the binding sites for these primers by creating sequence alignments between multiple Marionette plasmids on Benchling, which allowed us to see the regions of plasmid homology that flanked both the promoter and regulatory gene sequences, which differed between Marionette plasmids. One big design consideration during this process was the fact that the Marionette YFP expressing plasmids contain a BsaI site in the region just upstream of the promoter sequence. We designed our primers to create a single point mutation in order to mutate out this illegal BsaI site.

Source

This part was sourced from pAJM.011 on AddGene (https://www.addgene.org/108529/), which originates from the paper "Escherichia coli 'Marionette' strains with 12 highly optimized small-molecule sensors" by Meyer et al., cited in the long description. pAJM.011 contains tetR and a YFP gene under inducible control by pTet.

References