Part:BBa_K3380608
No scaffold TXO ArsR regulated construct under T7 promoter (BBa_K3380103) expressing DIR2s-Apt
The Edinburgh iGEM team 2020 designed a construct comprising a fluorescent RNA aptamer (iSpinach BBa_K3380150) under an ArsR transcription factor regulated promoter (BBa_K3380103). As opposed to BBa_K3380602, this does not have a scaffold. It was designed to test the ArsR regulated promoter and to assess the impact of the tRNA scaffold absence on the RNA aptamer fluorescence. The construct is capable of exhibiting fluorescence being a transcription only construct. Simultaneously we tested the efficiency of transcription in the absence of a terminator in a cell-free extract. The T7 RNA polymerase is capable of synthesizing transcripts via run-off transcription in the absence of a terminator (because the DNA template is linear). Figure 1 illustrates a schematic design of the construct.
Figure 1: Construct BBa_K3380608 design. The DIR2s-Apt fluorescent RNA aptamer (shown in green) was expressed under the BBa_K3380103 promoter (shown in black). The promoter has an ArsR binding site (shown in yellow) that represses the transcription in the absence of Arsenic.
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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