Part:BBa_K1660010

RFP+RBS+attB+P+attP+RBS+GFP+Terminator

Bxb1 gp35 is a serine integrase in Mycobacterium phage Bxb1. This intergrase can exclusively catalyze site-specific recombination. The dismiss of light singal can drive the expression of integrase, and results in the reverse of the sequence （there it means the promoter Pcon (BBa_J23110)） between attB and attP, changing the two sites to attL and attR at the meantime. As a consequence, the promotor moves on to express the toxic protein instead of the attractant.

We insert the gp35 gene into the downstream region of the PompC promotor by means of 3A Assembly so that the expression of integrase can be regulated by light. In addition, we add a 6N promotor between PompC and gp35 to decrease basal integrase expression above flipping threshold level which could lower the translation efficiency of gp35. Meanwhile, we insert the recombined locus of attB and attP into the flanking sequences of our constitutive promotor (BBa_J23110) and so far we have accomplished our reverse promotor system (Fig.1, 2).

Fig.1 The biobrick designed for bidirectional transcriptional system.

Fig.2 The biobrick for trap and kill system.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 933
  Illegal NheI site found at 956
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 880
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 900
  Illegal BsaI.rc site found at 983
  Illegal BsaI.rc site found at 1684