Difference between revisions of "Part:BBa K3380600"
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<partinfo>BBa_K3380600 short</partinfo>
 
<partinfo>BBa_K3380600 short</partinfo>
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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_K3380103). 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).
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'''Figure 1''' illustrates a schematic design of the construct.
  
  
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[[File:Part BBa K3380600.jpeg|700px|Construct tested in Metallidurans cell-free extract]]
 
  
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===Usage and Biology===
 
===Usage and Biology===
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The efficiency of the ArsR transcription factor repression was tested using the ''Cuprividus Metallidurans CH34'' cell extract under a class III strong promoter.
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'''Figure 2''' shows the  a schematic design of the construct.
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[[File:Part BBa K3380600.jpeg|700px|Construct tested in Metallidurans cell-free extract]]
  
 
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Revision as of 17:23, 10 October 2020


TXO ArsR regulated construct under T7 promoter (BBa_K3380103) 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_K3380103). 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.


iSpinach under ArsR repression


Usage and Biology

The efficiency of the ArsR transcription factor repression was tested using the Cuprividus Metallidurans CH34 cell extract under a class III strong promoter. Figure 2 shows the a schematic design of the construct. Construct tested in Metallidurans cell-free extract

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    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