Difference between revisions of "Part:BBa K3380603"
| Line 2: | Line 2: | ||
__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K3380603 short</partinfo> | <partinfo>BBa_K3380603 short</partinfo> | ||
| − | + | The Edinburgh iGEM team 2020 designed a construct comprising a fluorescent RNA aptamer (iSpinach BBa_K3380150) flanked by a tRNA scaffold (F30 BBa_K3380101 and BBa_K3380102) under an ArsR transcription factor regulated promoter (BBa_K190015). As opposed to BBa_K3380600, this construct has the binding ArsR binding site before the promoter. It was designed to test the ArsR regulated promoter. 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''' illustrates a schematic design of the construct. | '''Figure 1''' illustrates a schematic design of the construct. | ||
[[File:Construct 603.png|500px|]] | [[File:Construct 603.png|500px|]] | ||
| + | |||
| + | '''Figure 1: Construct BBa_K3380603 design.'''The iSpinach fluorescent RNA aptamer (shown in green) was flanked by the F30 upstream and downstream scaffolds (shown in blue) to protect it from the RNAse degradation (from the cell free extract), it was expressed under the BBa_K190015 promoter (shown in black). The promoter has an ArsR binding site (shown in yellow) that represses the transcription in the absence of Arsenic. | ||
| + | |||
Revision as of 10:25, 25 October 2020
TXO ArsR regulated construct under T7 promoter (BBa_K190015) expressing iSpinach
The Edinburgh iGEM team 2020 designed a construct comprising a fluorescent RNA aptamer (iSpinach BBa_K3380150) flanked by a tRNA scaffold (F30 BBa_K3380101 and BBa_K3380102) under an ArsR transcription factor regulated promoter (BBa_K190015). As opposed to BBa_K3380600, this construct has the binding ArsR binding site before the promoter. It was designed to test the ArsR regulated promoter. 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 illustrates a schematic design of the construct.
Figure 1: Construct BBa_K3380603 design.The iSpinach fluorescent RNA aptamer (shown in green) was flanked by the F30 upstream and downstream scaffolds (shown in blue) to protect it from the RNAse degradation (from the cell free extract), it was expressed under the BBa_K190015 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]
- 1000COMPATIBLE WITH RFC[1000]