Part:BBa_K4720101

DspB RFC [1000] compatible

This is a theoretical part for DspB with the BsaI restriction site removed by site directed mutagenesis. The original part is BBa_K1659200 by iGEM Oxford 2015 [1].

DspB was chosen for our project as a treatment against biofilm formation. The original idea was to use bacteriophage M13 as the carrier of the gene. A method that had been described before with bacteriophage T7 [2]. This plan was later abandoned in favor of displaying DspB on the PIII protein (BBa_K4720102). The idea being that this would make the treatment more broad and not just E. coli specific.

The following primers were designed for the mutagenesis (5'to 3'):

DspB_Mut_Fwd: AAAATACTTACAAGGACTCAAGAGTCGCCAGGTGGACGATG

DspB_Mut_Rev: CTGGCGACTCTTGAGTCCTTGTAAGTATTTTACGCCACGATC

The resulting linear fragment can be circularized by Gibson Assembly (31 bp overlapping)

Due to time restraint we were unable to perform this mutagenesis, but we leave the information here for future iGEM teams.

[1]: https://parts.igem.org/Part:BBa_K1659200

[2]: Lu, T. K., & Collins, J. J. (2007). Dispersing biofilms with engineered enzymatic bacteriophage. Proceedings of the National Academy of Sciences, 104(27), 11197-11202.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 262
  
• 1000
  COMPATIBLE WITH RFC[1000]