Part:BBa_K257001:Design

Gene 3 protein of filamentous phages without the 18 first amino acids

Assembly Compatibility:

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

  Illegal BamHI site found at 592
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

• In our project we use g3p as a fusion to OmpA-Linker (BBa_K103996) which need SacI restriction site for inframe fusion.
• So we design g3p with SacI site at the N-terminal. SacI (GAGCT^C) site is shared with XbaI (T^CTAGA) in order to have SacI site for fusion and standard sites.
• Moreover we decide to suppres the signal peptide (18 first amino acids) which is cleaved in order to conserve the N-ter fusion.

PFec is inserted into PSB1A3.

Source

• g3p could be found in filamentous bacteriophages like M13, fd, f1, etc... or in phage helper like M13KO7, etc...

References

• The Mechanism of Bacterial Infection by Filamentous Phages Involves Molecular Interactions between TolA and Phage Protein 3 Domains. Fredrik Karlsson, Carl A. K. Borrebaeck, Nina Nilsson, and Ann-Christin Malmborg-Hager
• Interdomain interactions within the gene 3 protein of philamentous phage. Jean Chatellier, Oliver Hartley, Andrew D. Grifths, Alan R. Fershta, Greg Wintera, Lutz Riechmannb
• A prokaryotic membrane anchor sequence: Carboxyl terminus of bacteriophage fl gene III protein retains it in the membrane. Jef D. Boeke AND Peter Model