Intermediate
rPlac+TT

Part:BBa_K093004:Design

Designed by: John Heil, Danielle Nash, Shira Davis   Group: iGEM08_Waterloo   (2008-10-27)


Convergent Promoter System: Reverse Module: rPlac+TT


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

The forward and reverse efficiencies of TTs in the registry had to be considered, especially for this part's convergent partner, BBa_K093003. R0011 was used instead of R0010 to avoid catabolite repression.

The idea is to place BBa_K093003 upstream of the gene of interest, that requires tight repression, and to place BBa_K093004 downstream. If the cells are grown in the presence of IPTG POlacI is induced and anti-sense mRNA is produced. Anti-sense mRNA will repress expression of the gene of interest through RNAi. This system also requires cI repressor (BBa_C0051) under control of POlacI (BBa_R0011). This system would be functional in a laqIq strain such as E. coli. TG1.

O'Connor,C.D., & Timmins, K.N. (1987). Highly repressible expression system for cloning genes that specify potentialy toxic proteins. Journal of Bacteriology. 169, 4457-4462.


Source

Synthetic

References