Part:BBa_K3187028

Sortase A7M (Ca²⁺-independent variant)

Profile

Name	Sortase A7M
Base pairs	470
Molecular weigth	17.85&nbspkDa
Origin	Staphylococcus aureus, synthetic
Parts	Basic part
Properties	calcium-independent, transpeptidase, linking sorting motif LPXTG to poly-glycine Tag

Usage and Biology

Sortase&nbspA7M (SrtA7M) is a calcium-independent transpeptidase derived from the enzyme Sortase&nbspA from Staphylococcus aureus. Similar to Sortase&nbspA, SrtA7M connects two peptides covalently, one with a poly-glycine (poly-G) at the N-terminus and one with LPXTG motif at the C-terminus. It acts by cleaving between Gly and Thr of the LPXTG motif and catalyzing the formation of an amide bond to the poly-G sequence.

Methods

The basic part Sortase&nbspA7M is produced and purified as the composite part BBa_K3187005(Sortase&nbspA7M). For the expression and purification of SrtA7M, a part containing T7 promoter, a lac operator and a ribosomal binding site BBa_K3187029 N-terminal and a 6xHis-Tag with a TGA stop codon (BBa_K3187024) C-terminal of the coding sequence were added. The production was performed in the E. Coli strain BL21 (DE3). The purification was done with the GE Healthcare ÄTKA Pure machine which is a machine for FPLC.

Results

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 445
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]