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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 33
- 1000INCOMPATIBLE 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
| n/a | Optogenetical AND-Gate blue light - without first promoter - choose your first input |