Difference between revisions of "Part:BBa K3380618"

Revision as of 13:17, 25 October 2020

No scaffold TXO ArsR regulated iSpinach construct under T7 P(BBa_K3380103) and (BBa_K3137010) T

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_K3380600, 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. Simultaneously we tested the efficiency of transcription in a cell-free extract when adding a T7 terminator (BBa_K3137010) as compared to the "run off" transcription (BBa_K3380606) from a linear DNA with no terminator. The construct is capable of exhibiting fluorescence being a transcription only construct. The T7 RNA polymerase can synthesize 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_K3380618 design. The iSpinach fluorescent RNA aptamer (shown in green) was expressed under the BBa_K3380103 promoter (shown in black) and BBa_K3137010 T7 terminator (shown in black). The promoter has an ArsR binding site (shown in yellow) that represses the transcription in the absence of Arsenic.

Usage and Biology

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
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COMPATIBLE WITH RFC[12]
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COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 25

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 <partinfo>BBa_K3380618 short</partinfo>
-The Edinburgh iGEM team 2020 designed a construct comprising a fluorescent RNA aptamer (iSpinach BBa_K3380150) under an ArsR transcription factor regulated promoter (BBa_K190015). As opposed to BBa_K3380603, 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.
+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_K3380600, 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. Simultaneously we tested the efficiency of transcription in a cell-free extract when adding a T7 terminator (BBa_K3137010) as compared to the "run off" transcription (BBa_K3380606) from a linear DNA with no terminator. The construct is capable of exhibiting fluorescence being a transcription only construct. The T7 RNA polymerase can synthesize 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.
 [[File:Construct618v1.png|500px|]]
-<!-- Add more about the biology of this part here
+'''Figure 1: Construct BBa_K3380618 design.''' The iSpinach fluorescent RNA aptamer (shown in green) was expressed under the BBa_K3380103 promoter (shown in black) and BBa_K3137010 T7 terminator (shown in black). The promoter has an ArsR binding site (shown in yellow) that represses the transcription in the absence of Arsenic.
 ===Usage and Biology===