Regulatory

Part:BBa_K4044010:Design

Designed by: Kalinichenko Andrei   Group: iGEM21_LMSU   (2021-10-19)
Revision as of 21:47, 19 October 2021 by Registry (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)


p119_G4 (Pj23119-based Gal4 dependant bacterial promoter)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The promoter contains the sequence of CGG-N11-CCG between GACA-box and TATA-box, which can be recognized by Gal4 protein. Eleven nucleotides in the middle are presented of the consensus sequence of Gal4 binding locus of Saccharomyces cerevisiae. When binding to promoter Gal4 domain inhibits transcription activity completely. This promoter was tested to be active in E.coli and comparable with Anderson promoter pj23119 baseline transcriptional activity. Promoters were tested in construction containing YFP under promoter that is tested with strong RBS used (BBa_J34801). The results of QPAS1-Gal4 inhibition show a significant decrease of fluorescence and result in a close to zero level of transcription.

  • fluorescence graf*

We also have shown with a computer simulation that HTH domains of Gal4 binds exactly to CGG-N11-CCG site of the promoter.

  • modeling of QOAS-Gal4 with DNA*


Source

Promoter designed on the base of Anderson pj23119 promoter (BBa_J23119).

References