Difference between revisions of "Part:BBa K2243009"
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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. | 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=== | ||
+ | |||
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Latest revision as of 13:11, 31 October 2017
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 82
Illegal NheI site found at 105 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]