Part:BBa_K4365021

SP-SUMO-turboRFP

SP-SUMO-tRFP is a reporter device which could be used to test protein secretion in yeast. Moreover, if tRFP is exchanged for the protein of interest, after protein purification with His-tag you can scarlessly remove the tag from the protein with a SUMO protease. This part is optimized for expression in Saccharomyces cerevisiae.

SP-SUMO-tRFP consists of:

• FIG1 promoter K4365000
• signal peptide (alpha mating factor signal peptide) K4365019
• glycine linker K4365005
• 6x His-Tag K3033006
• SUMO K4365004
• codon optimized turboRFP K4365020
• terminator tCYC1 K849009

Figure 1: illustration of SP-SUMO system for secretion and scar-less protein production in yeast. The SP-SUMO is attached to the cargo protein turboRFP as this was the construct we tested during our experiments. Created with Biorender.com.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal SpeI site found at 1001
  Illegal PstI site found at 1036
  Illegal PstI site found at 1335
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 1001
  Illegal PstI site found at 1036
  Illegal PstI site found at 1335
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2168
  Illegal BamHI site found at 1007
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal SpeI site found at 1001
  Illegal PstI site found at 1036
  Illegal PstI site found at 1335
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal SpeI site found at 1001
  Illegal PstI site found at 1036
  Illegal PstI site found at 1335
  Illegal NgoMIV site found at 172
  
• 1000
  COMPATIBLE WITH RFC[1000]