Difference between revisions of "Part:BBa K2235003"

(Usage and Biology)
 
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<partinfo>BBa_K2235003 short</partinfo>
 
<partinfo>BBa_K2235003 short</partinfo>
  
<partinfo>BBa_K2235003 SequenceAndFeatures</partinfo>
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This composite part encodes all the necessary regulatory elements for a functional chemically inducible system, the cumate system. In particular, there is an expression cassette encoding the reporter gene, mTagBFP, <partinfo>K592100</partinfo> and there are two additional multiple cloning sites (MCS) flanking the reporter (check the design page for more information). The repressor of the system, CymR <partinfo>K415202</partinfo> , is located upstream of the expression cassette and it is constitutively expressed. The RNA spinach, <partinfo>K1330000</partinfo>, is an additional reporter protein following CymR aiming to detect its expression upon addition of its cognate fluorophore.
  
  
 
===Usage and Biology===
 
===Usage and Biology===
  
 +
The cumate system have been previously described as a versatile and tightly regulated expression system induced by 4-isopropylbenzoic acid (cumate) in various strains of E.coli (1). It can be employed for the regulation and production of heterologous proteins in a dose-dependent manner. Moreover, the induction is achieved quickly within minutes therefore, a robust and homogenous expression of any protein can be achieved upon addition of cumate.
  
  
  
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2235003 SequenceAndFeatures</partinfo>
<partinfo>BBa_K2235004 SequenceAndFeatures</partinfo>
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===References===
 
===References===
  
 
1. Choi Y, Morel L, Le Francois T, Bourque D, Bourget L, Groleau D et al. Novel, Versatile, and Tightly Regulated Expression System for Escherichia coli Strains. Applied and Environmental Microbiology. 2010;76(15):5058-5066.
 
1. Choi Y, Morel L, Le Francois T, Bourque D, Bourget L, Groleau D et al. Novel, Versatile, and Tightly Regulated Expression System for Escherichia coli Strains. Applied and Environmental Microbiology. 2010;76(15):5058-5066.

Latest revision as of 12:53, 23 October 2017


Cumate gene switch expressing BFP

This composite part encodes all the necessary regulatory elements for a functional chemically inducible system, the cumate system. In particular, there is an expression cassette encoding the reporter gene, mTagBFP, BBa_K592100 and there are two additional multiple cloning sites (MCS) flanking the reporter (check the design page for more information). The repressor of the system, CymR BBa_K415202 , is located upstream of the expression cassette and it is constitutively expressed. The RNA spinach, BBa_K1330000, is an additional reporter protein following CymR aiming to detect its expression upon addition of its cognate fluorophore.


Usage and Biology

The cumate system have been previously described as a versatile and tightly regulated expression system induced by 4-isopropylbenzoic acid (cumate) in various strains of E.coli (1). It can be employed for the regulation and production of heterologous proteins in a dose-dependent manner. Moreover, the induction is achieved quickly within minutes therefore, a robust and homogenous expression of any protein can be achieved upon addition of cumate.



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
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 418
    Illegal XhoI site found at 199
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 101

References

1. Choi Y, Morel L, Le Francois T, Bourque D, Bourget L, Groleau D et al. Novel, Versatile, and Tightly Regulated Expression System for Escherichia coli Strains. Applied and Environmental Microbiology. 2010;76(15):5058-5066.