Part:BBa_K2319000

sfGFP-SpyCatcher under T7 expression system

This part generates a fusion protein of superfolder GFP (sfGFP) and SpyCatcher (a 15.2 kDa protein) when transformed into a T7 expression system like E. coli strain BL21 (DE3).

[[1]]

This part can be used as a T7 expression backbone.

Using the T7 expression backbone

Expressing any protein of interest

This T7 expression backbone can be used to express any protein if its coding sequence (with a start codon) is inserted using the HindIII and NheI sites. These sites can be added to the coding sequence using PCR with primers having 5'-overhangs.

Fusing a protein domain at the N-terminus of an existing protein

By inserting the coding sequence of a protein domain (including the start codon) using the HindIII and AgeI sites into the T7 expression backbone (which already contains a protein coding sequence), an N-terminal fusion can be performed.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1200
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 768
  Illegal XhoI site found at 1001
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 42
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 57