Part:BBa_K2549029

8*ZF21.16-CMV

This part is one of the response elements of our amplifier, also executing the combiner function. 8*ZF21.16 binding sites is assembled using two 4*ZF21.16 binding sites (Part:BBa_K2446008) with a biobrick scar between them. CMV (Part:BBa_K2549050) is a promotor which has a high-level constitutive expression. This part can switch off the expression of gene downstream after induced by our zinc finger-based transcription repressor.

This part is also served as an improvement version of Part:BBa_K2446030 of iGEM-team [http://2017.igem.org/Team:Fudan/Composite_Part Fudan 2017]. We placed responsive elements of zinc finger-based transcription factors on the 5' end of the promotor to lower the impact they may have on the basal expression of the promotor. Besides, CMV promotor performs a higher-level expression than SV40 promotor. 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]

Biology

It works as we designed

interaction between ZF21.16-KRAB and 8*ZF21.16-CMV promotor

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

Khalil AS et al have reported several synthetic promotor operators which can interact with artificial zinc finger-based transcription factors with high specificity and high orthogonality.

Khalil AS 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.