Part:BBa_K5047040

This sequence enables constitutive Dicer expression in yeast.

The sequence consists of three main parts:

Constitutive Promoter (pTEF1): Sourced from the Saccharomyces cerevisiae gene TEF1, this promoter drives continuous expression in yeast cells across various growth conditions. (Mumberg, D et al. Yeast vectors for the controlled expression of heterologous proteins in different genetic backgrounds. Gene vol. 156,1 (1995): 119-22. doi:10.1016/0378-1119(95)00037-7).

DICR Protein Coding Sequence: Encodes the Dicer protein, a crucial component of the RNA-induced silencing complex (RISC), which is essential for the processing of double-stranded RNA into small interfering RNAs (siRNAs). (Tang, Kai-Fu, and Hong Ren. "The role of dicer in DNA damage repair." International journal of molecular sciences vol. 13,12 16769-78. 7 Dec. 2012, doi:10.3390/ijms131216769).

Terminator (tSTE2): Derived from the yeast gene STE2, the terminator ensures proper transcription termination and stabilises the resulting mRNA transcript, facilitating efficient protein expression. The Dicer protein is a key part of the RISC complex, which is absent in S. cerevisiae.

This construct enables the introduction and functional expression of RNA interference machinery in this yeast species.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1845
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 796
  Illegal BglII site found at 992
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1950
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 160