Part:BBa_K568004

Optogenetical AND-Gate blue light - without first promoter - choose your first input

This is part of an AND-gate where one can choose the first promoter which controls the supDtRNA. The logical gate is based on amber stop-codon suppression via the non-canonical tRNA supD (BBa_K228001). The blue light sensor (BBa_K238013) induces the expression of a mRNA coding for a T7-polymerase with an amber mutation (BBa_K228000), that can only be translated by ribosomes if the correct supD tRNA is present.

Usage and Biology

This part can be used, when one wants an AND-gate where the T7ptag (T7 polymerase with amber stop codon mutation) is under the control of a bluelight sensor. The part which controls the supDtRNA can be chosen individually and designed for specific experimental purposes. Blue light leads to dimerisation of YcgF and binding of the repressor YcgE. The formation of the YcgE-YcgF complex leads to the unbinding of YcgE from the YcgZ promoter which activates the transcription of T7ptag (T7 polymerase with the amber stop codon mutation) if enough supD tRNA is available. SupDtRNA is only produced once a chosen promoter is cloned at the 5'-site of the part.

The so constructed AND-gate should ensure that expression of T7ptag is only induced when two signals (one of them blue light) hit the part.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 33
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 36
  

Functional Parameters

n/a	Optogenetical AND-Gate blue light - without first promoter - choose your first input

Induction with blue light at 465 nm

Functional Parameters: Austin_UTexas

BBa_K568004 parameters

Burden Imposed by this Part:

Burden Value: 7.0 ± 5.8%

Burden is the percent reduction in the growth rate of E. coli cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This BioBrick did not exhibit a burden that was significantly greater than zero (i.e., it appears to have little to no impact on growth). Therefore, users can depend on this part to remain stable for many bacterial cell divisions and in large culture volumes. Refer to any one of the BBa_K3174002 - BBa_K3174007 pages for more information on the methods, an explanation of the sources of burden, and other conclusions from a large-scale measurement project conducted by the 2019 Austin_UTexas team.

This functional parameter was added by the 2020 Austin_UTexas team.