Composite
Part:BBa_K3519011
Designed by: Omkar Mohapatra Group: iGEM20_IISER-Tirupati_India (2020-10-07)
araC-araBAD promoter-mRFP
This part contains the RFP downstream to the engineered arabinose inducible promoter (BBa_K3519010). This has been designed to characterise the part BBa_K3519010 for a tightly regulated 'kill switch'. Look at the part BBa_K3519012 for the 'kill switch'. The RFP gene here is expressed on the addition of arabinose to the media. Excess glucose in the media represses the expression of RFP. All data presented below have been adapted from the literature.
Usage and Biology
Figure 1: The ‘native’ genetic circuit used for characterisation of the designed arabinose inducible promoter system. The araC is constitutively transcribed in the opposite direction.
Figure 2: The ‘active’ genetic circuit. The system here is switched ‘on’ in addition of arabinose to the media. The araC is constitutively transcribed in the opposite direction. The promoter is activated on recruitment of the cAMP-CAP complex and the arabinose-AraC complex to the regulatory regions in the promoter, leading to increase in the transcription of the mRFP gene. The designed arabinose inducible promoter system is characterised by quantifying mRFP using fluorescence measurements.
Figure 3: The ‘inactive’/repressed genetic circuit. The system here is switched ‘off’ in absence of arabinose in the media. The araC gene is constitutively transcribed in the opposite direction. The AraC dimer binds to the regulatory regions of the promoter and represses the transcription of mRFP, leading to very low fluorescence.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1043
Illegal NheI site found at 1066 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1388
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1223
Illegal AgeI site found at 2030
Illegal AgeI site found at 2142 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 1205
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Categories
Parameters
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