Composite

Part:BBa_K1077006:Design

Designed by: Mike Ferguson   Group: iGEM13_Michigan   (2013-09-16)

nat fim switch b0034 GFP OFF orientation


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1048


Design Notes

This switch half is in the "ON" orientation.

Design and Construction of the new natural fim switch:

Given the importance of the LRP and IHF binding sites in the switch, and the lack of these sites in the engineered fim switch we created last year, we decided to copy the natural fim switch as close as possible. The sequence of the switch varies slightly from strain to strain in E. coli. We chose to use the natural switch sequence from E. coli CFT073 because that is the strain that has the most characterization data on hbiF.


Source

Synthesized based on E. coli CFT073 genomic sequence.

References

1. Schwan WR. Regulation of fim genes in uropathogenic Escherichia coli. World J Clin Infect Dis 2011; 1(1): 17­25.

2. I. C. Blomfield, D. H. Kulasekara and B. I. Eisenstein. Integration host factor stimulates both FimB- and FimE-mediated site-specific DNA inversion that controls phase variation of type 1 fimbriae expression in Escherichia coli. Molecular Microbiology (1997) 23(4), 705–717.

3. M. P. McCusker, E. C. Turner and C. J. Dorman. DNA sequence heterogeneity in Fim tyrosine-integrase recombinase-binding elements and functional motif asymmetries determine the directionality of the fim genetic switch in Escherichia coli K-12. Molecular Microbiology, 67, 171–187.

4. Rice PA, Yang S, Mizuuchi K, Nash HA. Crystal structure of an IHF-DNA complex: a protein-induced DNA U-turn. Cell. 1996 Dec 27;87(7):1295-306.

5. Wang Q, Calvo JM. Lrp, a major regulatory protein in Escherichia coli, bends DNA and can organize the assembly of a higher-order nucleoprotein structure. EMBO J. 1993 Jun;12(6):2495-501.

6. Jerome Bonnet, Pakpoom Subsoontorn, and Drew Endy. Rewritable digital data storage in live cells via engineered control of recombination directionality. PNAS. 2012 Apr 6.

7. D. L. Gally, J. Leathart and I. C. Blomfield. Interaction of FimB and FimE with the fim switch that controls the phase variation of type 1 fimbriae in Escherichia coli K-12. Molecular Microbiology (1996) 21(4), 725–738.

8. Ham et al. A Tightly Regulated Inducible Expression System Utilizing the fim Inversion Recombination Switch. Biotechnology and Bioengineering, Vol. 94, No. 1, May 5, 2006.

9. Jerome Bonnet et al. Amplifying Genetic Logic Gates. Science 3 May 2013, Vol. 340 no. 6132 pp. 599-603.

10. Ham TS, Lee SK, Keasling JD, Arkin AP. Design and Construction of a Double Inversion Recombination Switch for Heritable Sequential Genetic Memory. PLoS ONE, 2008, 3(7): e2815. doi:10.1371/journal.pone.0002815