Protein_Domain
Intein

Part:BBa_K300004:Design

Designed by: Sara Bisi, Paolo Magni   Group: iGEM10_UNIPV-Pavia   (2010-10-03)

Engineered pH-inducible intein (codon optimized for E. coli) - internal domain


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 SapI.rc site found at 460


Design Notes

Codon optimized for E. coli. The amino acid sequence comes from [1].

This part has been designed as an internal domain: it lacks start/stop codons and it has Prefix and Suffix sequences compatible with RFC 23 in order to enable protein fusions upstream and downstream.

Source

The original sequence of this intein comes from M. tuberculosis genomic DNA (RecA gene) that can be found in InBase (http://www.neb.com/neb/inteins.html , NEB), but it has been highly engineered (according to [1]) to create a pH-inducible protein domain.

References

[1] Wood DW, Wu W, Belfort G, Derbyshire V, Belfort M (1999), A genetic system yields self-cleaving inteins for bioseparations. Nature Biotechnology, Vol. 17, September, 889-892.

[2] Banki MR, Gerngross TU, Wood DW (2005), Novel and economical purification of recombinant proteins: Intein-mediated protein purification using in vivo polyhydroxybutyrate (PHB) matrix association. Protein Sci. June; 14(6): 1387–1395. doi: 10.1110/ps.041296305.