Reporter

Part:BBa_K542005

Designed by: Anthony Vuong   Group: iGEM11_Lethbridge   (2011-09-23)
Revision as of 00:38, 29 September 2011 by Dustin (Talk | contribs)

pTet Regulated Arg-tagged ECFP and EYFP (FRET Reporter)

Made by assembling BBa_K331033 with BBa_K331035.

The pTet promoter is constitutively "on"; thus, Arg-tagged ECFP and Arg-tagged EYFP will be produced constitutively. By placing this part downstream of BBa_K542003, expression of the fluorescence proteins maybe "turned off" in the presence of arabinose.

Fluorescence/Förster Resonance Energy Transfer(FRET) is a distance-dependent phenomenon in which the excitation of a donor fluorophore leads to emission by an acceptor fluorophore; this occurs if the two fluorophores are within a certain distance to each other. The distance is dependent on the FRET pair used. The emission spectrum of the donor fluorophore must overlap with the excitation spectrum of the acceptor fluorophore for FRET to occur. FRET is explained in further detail on the [http://2009.igem.org/Team:Lethbridge/Project#FRET Lethbridge 2009 Wiki]. This phenomenon will allow for characterizing co-localization within the Lumazine Synthase microcompartment.

This part was characterized in BBa_K542008 in E. coli strain DH5alpha demonstrating this is a functional intermediate in that construct. The data sheet for BBa_K542008 is shown below.

UoflECdatasheet.png

Sequence and Features


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]


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