Part:BBa_K734002
Spinach Aptamer with Stabilizing tRNA Scaffold
The fluorescent Spinach Aptamer (SpA) created by the Jaffrey Lab at Cornell University, designed to bind to the GFP-like synthetic fluorophore, 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI). SpA generates a green fluorescence upon binding DFHBI after transcription into mRNA. As reported by Paige et al.; excitation wavelength: 469 nm, emission wavelength: 501 nm, and exhibits 53% EGFP fluorescent strength.
Usage and Biology
UT Austin 2012 iGEM team used this part to evaluate a constitutive T7 promoter. Sequence: TAATACGACTCACTATAGGGT, differing from Bba_J64997 by two bases.
This image shows a fluorescent kinetic trial taken in a Tecan F200 plate reader. The RFU of spinach aptamer is measured at 501nm over time, and in a dilution series of DFHBI concentrations. (Note: 3.125uM DFHBI works, but was removed from the graph because there was an issue with the specific well plate used.) We found that 200uM DHFBI in cell culture is the ideal concentration to maximize RFU and OD600 without slowing cell growth significantly.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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