Part:BBa_K2926006

SUP4 yeast terminator

Usage and Biology

SUP4 is a natural terminator of Saccharomyces cerevisiae, belonging to the SUP4 tRNA^Tyr locus.

https://2019.igem.org/wiki/images/e/e8/T--Bielefeld-CeBiTec--IconAssembly.png As soon as the Cas13a:crRNA surveillance complex is formed, it can be activated by the detection of its target RNA. Aktivation leads to colleteral cleavage of all nearby RNA strands, resulting in cell death.

 As soon as the Cas13a:crRNA surveillance complex is formed, it can be activated by the detection of its target RNA.
   Aktivation leads to colleteral cleavage of all nearby RNA strands, resulting in cell death.
 

File:T--Bielefeld-CeBiTec--IconAssembly.png

As soon as the Cas13a:crRNA surveillance complex is formed, it can be activated by the detection of its target RNA. Aktivation leads to colleteral cleavage of all nearby RNA strands, resulting in cell death.

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
  COMPATIBLE WITH RFC[1000]

Plasmid Design

For the analysis and characterization of the SNR52 Promotor XXXX in combiantion with the SUP4 Terminaror XXXX, mCherry XXXX was cloned beteen them using Gibson Assembly. The SNR52 Promotor and SUP4 Terminator where optained as a gBlock from IDT.

Sequencing Results

The biobrick was analysed with Sanger Sequenceing to confirm its correct base sequence.

Characterisation

We created a new protocol for the calibration of mCherry measurements with TexasRed, based on the standard iGEM protocoll for GFP fluorescence calibration, We performed the analysis with E. coli DH5α as well as S. cerevisiae cultures.
See BBa_K2926072 and BBa_K2926073 for the characterization results.