Difference between revisions of "Part:BBa K389012:Design"

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===Design Notes===
 
===Design Notes===
 
* medium strong constitutive promoter
 
* medium strong constitutive promoter
−
* double terminator (forward) to keep basal expression of kanamycin resistance low  
+
* The ''virG'' gene is mutated so it works without an ''rpoA'' subunit from ''Agrobacterium tumefaciens'' (YC Jung ''et al.'', 2004)
 +
* double terminator (forward) to keep expression of luciferase gene by the constitutive promoter low  
 
** double terminator <partinfo>BBa_B0017</partinfo> instead of <partinfo>BBa_B0015</partinfo> because of problems mentioned with <partinfo>BBa_B0012</partinfo>
 
** double terminator <partinfo>BBa_B0017</partinfo> instead of <partinfo>BBa_B0015</partinfo> because of problems mentioned with <partinfo>BBa_B0012</partinfo>
 
* luciferase gene to show the activity of the ''vir'' promoter
 
* luciferase gene to show the activity of the ''vir'' promoter
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===Source===
 
===Source===
  
−
parts.igem, synthetic gene by Mr. Gene, pGL4.10luc2 by Promega, TI-plasmid from ''Agrobacterium tumefaciens'' C58
+
* ''virG'' gene synthesized by Mr. Gene (<partinfo>BBa_K389002</partinfo>)
 +
* ''vir'' promoter from ''A. tumefaciens'' C58 (<partinfo>BBa_K389003</partinfo>)
 +
* luciferase gene from Promega's pGL4.10[luc2] vector (<partinfo>K389004</partinfo>)
 +
* constitutive promoter and double terminator from parts.igem (<partinfo>BBa_J23110</partinfo>, <partinfo>BBa_B0017</partinfo>)
  
 
===References===
 
===References===
 +
YC Jung ''et al.'' (2004) Mutants of ''Agrobacterium tumefaciens virG'' Gene That Activate Transcription of ''vir'' Promoter in ''Escherichia coli'', ''Current Microbiol'' 49:334-340.

Latest revision as of 14:36, 17 October 2010

VirA reporter system luc


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1195
    Illegal NgoMIV site found at 2539
    Illegal NgoMIV site found at 2560
    Illegal AgeI site found at 2263
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2445


Design Notes

  • medium strong constitutive promoter
  • The virG gene is mutated so it works without an rpoA subunit from Agrobacterium tumefaciens (YC Jung et al., 2004)
  • double terminator (forward) to keep expression of luciferase gene by the constitutive promoter low
  • luciferase gene to show the activity of the vir promoter

Source

References

YC Jung et al. (2004) Mutants of Agrobacterium tumefaciens virG Gene That Activate Transcription of vir Promoter in Escherichia coli, Current Microbiol 49:334-340.