Part:BBa_K779315

5' Hammerhead with loss-of-function mutation MammoBlock

This sequence, along with Part:BBa_K779317, is designed to test the functionality of a hammerhead ribozyme in vitro. Hammerhead ribozymes are self-cutting strands of catalytic RNA. After a hammerhead ribozyme is transcribed in a cell, it folds into a secondary structure, which causes self-cleavage. Because of their RNA-cleaving function, hammerheads may be used to regulate protein expression and to make other RNA parts. For more information on hammerheads in general, see http://en.wikipedia.org/wiki/Hammerhead_ribozyme .

This sequence is designed for in vitro transcription - a promoter and terminator corresponding to the in vitro transcription kit should be attached via PCR. After IVT, the RNA transcript should be analyzed on a gel, to determine whether cleavage occurred.

This sequence is called a 5' hammerhead because its cut site is near its 5' end. The corresponding loss-of-site mutation, which should not cleave, serves as a control.

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 4
  Illegal BsaI.rc site found at 182