Part:BBa_K1140003

pCons + 37 oC RNA thermometer + LacI (LVA)

A construction that has a gene coding for the LacI transcription factor, which also has a LVA degradation tag. The LacI gene is transcribed through a constitutive promoter and a thermolabile RBS with an optimal translation temperature of 37°C. Two termination sites were added at the 3’ end of the gene.

Usage and Biology

RNA thermometers (RNAT) are temperature sensing RNA sequences that regulate translation by preventing the ribosome from binding the transcript until higher temperatures shift it to an open structure. Several naturally occurring RNA thermometers have been described and synthetic sequences that emulate them have been designed and proved to regulate genetic expression at different temperature ranges. The RNAT herein implemented, designed and dubbed u6 by Neupert et al. [2], is a synthetic sequence that allows translation only at temperatures over 37 °C. This part can be incorporated into a genetic circuit to integrate transcriptional and post-transcriptional regulation, through Tet Repressor and u6-RNAT, respectively.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 44
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]