Difference between revisions of "Part:BBa K2244009:Design"
 
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LEV1 is a light repressor with an E. coli lexA DNA-binding domain, which binds to its cognate operator sequence on promoter when dimerized upon light irradiation. LEV1 is a fusion protein of VVD and LexA: VVD contains a light-oxygen-voltage (LOV) domain & N cap was added to the N-terminal of DNA-binding domain of LexA. The C-terminal domain of lexA repressor was removed. The expression of Lev1 is under a constitutive promoter so that it is constantly expressed.  
 
LEV1 is a light repressor with an E. coli lexA DNA-binding domain, which binds to its cognate operator sequence on promoter when dimerized upon light irradiation. LEV1 is a fusion protein of VVD and LexA: VVD contains a light-oxygen-voltage (LOV) domain & N cap was added to the N-terminal of DNA-binding domain of LexA. The C-terminal domain of lexA repressor was removed. The expression of Lev1 is under a constitutive promoter so that it is constantly expressed.  
  
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ColE promoter, with an intrinsic RBS, is obtained from E. coli Col1 plasmid and contains a cognate operator sequence for LexA. Expression of target gene (in this case, mCherry) is under the control of ColE promoter, thus is also regulated by LEV1 binding and light irradiation. mCherry sequence is codon optimized for E. coli expression. T1 terminator is the most commonly used terminator in E. coli.
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ColE promoter, with an intrinsic RBS, is obtained from E. coli ColE1 plasmid and contains a cognate operator sequence for LexA. Expression of target gene (in this case, mCherry) is under the control of ColE promoter, thus is also regulated by LEV1 binding and light irradiation. mCherry sequence is codon optimized for E. coli expression. T1 terminator is the most commonly used terminator in E. coli.

Latest revision as of 03:53, 2 November 2017

ColE promoter+mCherry gene+T1 terminator+Constitutive promoter+Lev1 gene+T1terminator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1015
    Illegal NheI site found at 1038
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 845
    Illegal AgeI site found at 1753
  • 1000
    COMPATIBLE WITH RFC[1000]


Source

LEV1 is a light repressor with an E. coli lexA DNA-binding domain, which binds to its cognate operator sequence on promoter when dimerized upon light irradiation. LEV1 is a fusion protein of VVD and LexA: VVD contains a light-oxygen-voltage (LOV) domain & N cap was added to the N-terminal of DNA-binding domain of LexA. The C-terminal domain of lexA repressor was removed. The expression of Lev1 is under a constitutive promoter so that it is constantly expressed.

ColE promoter, with an intrinsic RBS, is obtained from E. coli ColE1 plasmid and contains a cognate operator sequence for LexA. Expression of target gene (in this case, mCherry) is under the control of ColE promoter, thus is also regulated by LEV1 binding and light irradiation. mCherry sequence is codon optimized for E. coli expression. T1 terminator is the most commonly used terminator in E. coli.