Part:BBa_K2243008
phiC31 attB_J23119_phiC31 attP
The constitutive promoter J23119 is between phiC31 specific sites and will be inverted by the phiC31 recombinase, thus leading to the direction of promoter and change of gene expression. This unit can be used as standard reporter of recombinase phiC31 characterization.
Usage
We constructed this part to characterize the recombination efficiency of the phiC31 integrase. It consists of a constitutive promoter (BBa_J23119) flanked by attB and attP sites of the phiC31 integrase. Upon recombination, the orientation of the constitutive promoter change. As a result, expression of downstream sequence is shut down, and upstream sequence is transcribed.
Biology
J23119 is the consensus sequence of a combinatorial constitutive promoter library family. Integrase phiC31 from phage phiC31 can recognize and bind the attP sequence on phage DNA and the attB sequence on the host chromosomal DNA, and promotes unidirectional recombination to integrate the phage DNA into the host. We obtained the promoter, attB and attP sites by oligo synthesis.
Figure 2. The standard testing system used to characterize the recombinases.
Design Notes
We construct this structure by Gibson Assembly.
Source
The Streptomyces phage
Characterization
We used this part to characterize the recombination efficiency of the integrase phiC31.
References
J. Bonnet, P. Subsoontorn, D. Endy, Rewritable digital data storage in live cells via engineered control of recombination directionality. Proc. Natl. Acad. Sci. U.S.A. 109, 8884–8889 (2012).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 86
Illegal NheI site found at 109 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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