Reporter

Part:BBa_K260017

Designed by: Kaj Bernhardt   Group: iGEM09_BIOTEC_Dresden   (2009-10-17)

P_F3_ZeoR_F3_RFP. [FLP]->RFP

This is a FLP recombinase reporter. The genes for RFP (mRFP1) and ZeoR have no Start-codon and RFP is silenced by the upstream terminator. If FLP recombinase levels are high enough, recombination between both F3 sites will delete the intervening ZeoR gene plus terminator. Now RFP replaces ZeoR and comes under the control of the constitutive promoter that was driving expression of an F3-ZeoR fusion protein before. The result is an F3-RFP fusion protein that turns the cell red.

Prom-F3-zeoR-F3-RFP.jpg

Since this part is available in plasmid backbones pTetFlp (BBa_K260002) and pRhaFlp (BBa_K260003), FLP expression and thus RFP fluorescence can be controlled by small molecules (anhydrotetracycline and L-rhamnose respectively). It is thus a PoPS measurement device, where tet- and rhamnose- promoters can be compared and FLP activity be titrated. PTetFlp-F3-ZeoR-F3-TT-RFP.jpg


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 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 420


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Categories
Parameters
None