Part:BBa_K215001

Secretion Signal and Streptavidin Binding Tags

Any favorite protein (afp) can be inserted into the NheI (compatable with XbaI and SpeI, but make sure the resulting product stays in frame) which places the afp in between a series of tags which can be used with the [http://2009.igem.org/Team:Washington/Project/Display University of Washington 2009 Ideal Protein Purification system (IPP)]. The series of tags are, respectively: a Nano-Tag (binds to streptavidin), a 6x His tag (for IMAC purification), a TEV cute site (for removal of N-terminal tags), the NheI site (insertion of afp), TEV, 6x His tag, and the 181 C-terminal amino acids of prtB (secretion tag recognized by the secretion system: BBa_K215107). To use in E. coli we recommend using the composite part BBa_K215002, which already has a strong promoter and RBS attached. In order for the secretion tag to function, this part needs to be used in conjunction with BBa_K2150107.

Usage and Biology

This part works as designed. This is documented in the BioBrick page BBa_K215002, which adds a promoter and RBS to this part so that it can be tested.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 115
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]