Part:BBa_K3380501
Broccoli fluorescent RNA aptamer construct under T7 RNA polymerase promoter (BBa_z0251)
The Edinburgh iGEM team 2020 designed a construct comprising a fluorescent RNA aptamer (Broccoli BBa_K3380153) flanked by a tRNA scaffold (F30 BBa_K3380101 and BBa_K3380102) under a strong class III T7 RNA polymerase promoter (BBa_z0251). It was designed to test the fluorescence intensity of the Broccoli aptamer flanked by F30 tRNA scaffold when binding to the DFHBI-1T fluorophore. The construct is capable of exhibiting fluorescence being a transcription only construct. The T7 RNA polymerase is capable of synthesizing transcripts via run-off transcription in the absence of a terminator. Figure 1 illustrates the detailed design of the construct.
Usage and Biology
The construct functioning solely on transcription and having a small length of only 154 nucleotides has multiple advantages over the conventional fluorescent proteins. The time required for the fluorescence formation is much shorter than compared to the time needed for the synthesis of the fluorescent proteins. Moreover, there is less burden exhibited on the cell when expressing the transcription only construct.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 25
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