Difference between revisions of "Part:BBa K2243007"

 
 
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The constitutive promoter J23119 is between Bxb1 gp35 specific sites and will be inverted by the Bxb1 gp35 recombinase and its RDF bxb1 gp47(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 bxb1 gp35-gp47 fusion protein characterization
 
The constitutive promoter J23119 is between Bxb1 gp35 specific sites and will be inverted by the Bxb1 gp35 recombinase and its RDF bxb1 gp47(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 bxb1 gp35-gp47 fusion protein characterization
  
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===Usage and Biology===
 
===Usage and Biology===
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We constructed this part to characterize the recombination efficiency of the excisionase Bxb1 gp35-gp47 fusion protein (BBa_K2243013). It consists of a constitutive promoter (BBa_J23119) flanked by attL and attR sites of recombinase Bxb1 gp35. Upon recombination, the orientation of the constitutive promoter change. As a result, expression of downstream sequence is shut down, and upstream sequence is transcribed.
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===Biology===
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J23119 is the consensus sequence of a combinatorial constitutive promoter library family. In nature, attL and attR is generated when Mycobacteriophage Bxb1 integrates its DNA at the attB site of the Mycobacterium smegmatis genome using the viral attP site. With RDF Bxb1 gp47, attL and attR can be recognized and undergo recombination. We obtained the promoter, attL and attR sites by oligo synthesis.
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===Design note===
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This part is constructed by Gibson Assembly.
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===Characterize===
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We plan to use this part to characterize the recombination efficiency of the integrase Bxb1 gp35 (BBa_K2243013). However, the characterization have not done yet.
  
 
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Latest revision as of 12:56, 31 October 2017


Bxb1 attL_J23119_Bxb1 attR

The constitutive promoter J23119 is between Bxb1 gp35 specific sites and will be inverted by the Bxb1 gp35 recombinase and its RDF bxb1 gp47(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 bxb1 gp35-gp47 fusion protein characterization


Usage and Biology

We constructed this part to characterize the recombination efficiency of the excisionase Bxb1 gp35-gp47 fusion protein (BBa_K2243013). It consists of a constitutive promoter (BBa_J23119) flanked by attL and attR sites of recombinase Bxb1 gp35. 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. In nature, attL and attR is generated when Mycobacteriophage Bxb1 integrates its DNA at the attB site of the Mycobacterium smegmatis genome using the viral attP site. With RDF Bxb1 gp47, attL and attR can be recognized and undergo recombination. We obtained the promoter, attL and attR sites by oligo synthesis.

Design note

This part is constructed by Gibson Assembly.

Characterize

We plan to use this part to characterize the recombination efficiency of the integrase Bxb1 gp35 (BBa_K2243013). However, the characterization have not done yet.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 96
    Illegal NheI site found at 119
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 41
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 61
    Illegal BsaI.rc site found at 160