Difference between revisions of "Part:BBa K2549033"

 
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__NOTOC__
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<partinfo>BBa_K2549033 short</partinfo>
  
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This part is one of the response elements of our amplifier, also executing the combiner function. 6*ZF21.16 binding sites and 2*ZF43.8 binding sites ([[Part:BBa_K2549051]]) can bind to different zinc finger-based transcription activator ZF21.16-VP64 ([[Part:BBa_K2549023]]) and zinc finger-based repressor ZF43.8-KRAB ([[Part:BBa_K2446041]]), respectively, with high orthogonality. Minimal CMV ([[Part:BBa_K2549050]]) is a promotor providing very low basal expression and high maximal expression after induction. This part is designed to construct our NIMPLY logic gate and test our multiple binding sites amplifier model<ref>http://2018.igem.org/Team:Fudan/Model#Transcriptional_Amplifer</ref>.
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<!-- Add more about the biology of this part here -->
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===Biology===
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====synthetic promotor operators regulated by artificial zinc finger-based transcription factors====
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JJ Collins et al have reported several synthetic promotor operators which can interact with artificial zinc finger-based transcription factors with high specificity and high orthogonality<ref>A synthetic biology framework for programming eukaryotic transcription functions. Khalil AS, Lu TK, Bashor CJ, ..., Joung JK, Collins JJ. Cell, 2012 Aug;150(3):647-58  PMID: 22863014; DOI: 10.1016/j.cell.2012.05.045</ref>.
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[[File:ZF2.jpg|none|540px|thumb|JJ Collins et al stated:''sTFs constructed from OPEN-engineered ZFs are orthogonal to one another. sTF43-8 activated noncognate Promoter21-16 due to the fortuitous creation of a sequence that is significantly similar to the binding sequence of 43-8, when the downstream BamHI restriction site is considered.'']]
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====Promotor operator numbers can have an impact on tuning output====
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[[File:ZF3.jpg|none|540px|thumb|JJ Collins et al stated:''Tuning up output strength by increasing ZF operator number in synthetic promoter (sTF43-8).'']]
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====different transcription factors interaction with multiple binding sites====
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2549033 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K2549033 parameters</partinfo>
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===References===

Revision as of 01:21, 11 October 2018

6*ZF21.16-minCMV-2*ZF43.8

This part is one of the response elements of our amplifier, also executing the combiner function. 6*ZF21.16 binding sites and 2*ZF43.8 binding sites (Part:BBa_K2549051) can bind to different zinc finger-based transcription activator ZF21.16-VP64 (Part:BBa_K2549023) and zinc finger-based repressor ZF43.8-KRAB (Part:BBa_K2446041), respectively, with high orthogonality. Minimal CMV (Part:BBa_K2549050) is a promotor providing very low basal expression and high maximal expression after induction. This part is designed to construct our NIMPLY logic gate and test our multiple binding sites amplifier model[1].

Biology

synthetic promotor operators regulated by artificial zinc finger-based transcription factors

JJ Collins et al have reported several synthetic promotor operators which can interact with artificial zinc finger-based transcription factors with high specificity and high orthogonality[2].

JJ Collins et al stated:sTFs constructed from OPEN-engineered ZFs are orthogonal to one another. sTF43-8 activated noncognate Promoter21-16 due to the fortuitous creation of a sequence that is significantly similar to the binding sequence of 43-8, when the downstream BamHI restriction site is considered.

Promotor operator numbers can have an impact on tuning output

JJ Collins et al stated:Tuning up output strength by increasing ZF operator number in synthetic promoter (sTF43-8).

different transcription factors interaction with multiple binding sites

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

  1. ↑ http://2018.igem.org/Team:Fudan/Model#Transcriptional_Amplifer
  2. ↑ A synthetic biology framework for programming eukaryotic transcription functions. Khalil AS, Lu TK, Bashor CJ, ..., Joung JK, Collins JJ. Cell, 2012 Aug;150(3):647-58 PMID: 22863014; DOI: 10.1016/j.cell.2012.05.045