Part:BBa_K2243009

phiC31 attL_J23119_phiC31 attR

The constitutive promoter J23119 is between phiC31 RDF specific sites and will be inverted by the phiC31 recombinase and its RDF(or their fusion protein), thus leading to the direction of promoter and change of gene expression. This unit can be used as standard reporter of recombinase phiC31-RDF fusion protein characterization.

Usage

We construct this part to characterize the recombination efficiency of the excisionase phiC31. It consist of a constitutive promoter flanked with the attL and attR sites of excisionase phiC31. Once the attL and attR sites recombined by phiC31, the orientation of the constitutive promoter change, which influence the expression of downstream sequence.

Biology

J23119 is the consensus sequence of a combinatorial constitutive promoter library family. The attL and attR sites are generated from attB and attP site when integrase phiC31 from phage phiC31 promotes unidirectional recombination to integrate the phage DNA into the host. With phiC31 integrase and its RDF, attL and attR can be recognized and undergo recombination. We obtained the promoter, attB and attP sites by oligo synthesis.

Design Notes

We construct this structure by Gibson Assembly.

Source

The Streptomyces phage

Characterization

We plan to use this part to characterize the recombination efficiency of the phiC31 integrase with RDF. However, the characterization have not done yet.

References

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 82
  Illegal NheI site found at 105
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]