Part:BBa_K1727000:Design

Signiferin with signal peptide

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 7
  Illegal XbaI site found at 174
  Illegal PstI site found at 168
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 7
  Illegal PstI site found at 168
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 7
  Illegal BamHI site found at 162
  Illegal XhoI site found at 1
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 7
  Illegal XbaI site found at 174
  Illegal PstI site found at 168
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 7
  Illegal XbaI site found at 174
  Illegal PstI site found at 168
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

We had change the originally sequence into E.coli codon usage.

We treated signal peptide upstream of the N-terminal of Signiferin to facilitate the peptide production.To make sure the secretion system is work we had modified the amino acid sequence by attached an Ala at the N-terminal of Signiferin.

We use PQE 60 as vector,so we design Nco I enzyme site at 5' end and BamH I enzyme at 3' end.

Source

Artificial synthesis.

References

TU_Delf, i.t. 2013; Available from: http://2013.igem.org/Team:TU-Delft.

Tokuyasu, K., et al., Production of a recombinant chitin deacetylase in the culture medium of Escherichia coli cells. FEBS Lett, 1999. 458(1): p. 23-6. Fujii, T. and K. Miyashita, Multiple domain structure in a chitinase gene (chiC) of Streptomyces lividans. J Gen Microbiol, 1993. 139(4): p. 677-86.

Fujii, T. and K. Miyashita, Multiple domain structure in a chitinase gene (chiC) of Streptomyces lividans. J Gen Microbiol, 1993. 139(4): p. 677-86.