Difference between revisions of "Part:BBa K3132001"

Latest revision as of 15:48, 21 October 2019

ZF43-8-VP64

This part is transformed from the part: ZF_43_8_KRAB(BBa_K2446041) of iGEM17_Fudan. It is a synthetic transcription factor (SynTF) that use Cys2-His2 zinc finger 43-8 (ZF_43-8) as its protein chassis. Its parental form ZF_43_8_KRAB(BBa_K2446041), which developed by iGEM_Fudan, serves as an inhibitory transcription factor because of the KRAB domain. To turn this TF into an activating one, we replaced the KRAB domain with a VP64. And this approach successfully reversed its effect as we had expected. ZF_43-8_VP64 contains three core domains from N-terminal to C-terminal: a ZF_43-8 DNA binding domain, a nuclear location sequence and a VP64 transcription regulating domain. And a (G4S) linker was added between DBD and NLS for providing region flexibility. ZF_43-8-DBD enable binding to specific DNA sequences, together with VP64 constitutes the activating-form SynTF. So that we can use ZF_43-8-VP64 as a specific transcription factor to activate the expression of our downstream synthetic promoter 8×ZF_43-8 elements and minimalCMV.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 269
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 152
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K3132001 short</partinfo>
-This part is transformed from the part: ZF_43_8_KRAB(BBa_K2446041) of iGEM17_Fudan. It is a synthetic transcription factor (SynTF) that use Cys2-His2 zinc finger 43-8 (ZF_43-8) as its protein chassis. Its parental form ZF_43_8_KRAB(BBa_K2446041), which developed by iGEM_Fudan,  serves as an inhibitory transcription factor because of the KRAB domain. To turn this TF into an activating one, we replaced the KRAB domain with a VP64. And this approach successfully reversed its effect as we had expected. ZF_43-8_VP64 contains three core domains from N-terminal to C-terminal: a ZF_43-8 DNA binding domain, a nuclear location sequence and a VP64 transcription regulating domain. And a (G4S) linker was added between DBD and NLS for providing region flexibility. ZF_43-8-DBD enable binding to specific DNA sequences, together with VP64 constitutes the activating-form SynTF. So that we can use ZF_43-8-VP64 as a specific transcription factor to activate the expression of our downstream synthetic promoter 8×ZF_43-8 elements and minimalCMV.
+This part is transformed from the part: ZF_43_8_KRAB(<partinfo>BBa_K2446041</partinfo>) of iGEM17_Fudan. It is a synthetic transcription factor (SynTF) that use Cys2-His2 zinc finger 43-8 (ZF_43-8) as its protein chassis. Its parental form ZF_43_8_KRAB(<partinfo>BBa_K2446041</partinfo>), which developed by iGEM_Fudan,  serves as an inhibitory transcription factor because of the KRAB domain. To turn this TF into an activating one, we replaced the KRAB domain with a VP64. And this approach successfully reversed its effect as we had expected. ZF_43-8_VP64 contains three core domains from N-terminal to C-terminal: a ZF_43-8 DNA binding domain, a nuclear location sequence and a VP64 transcription regulating domain. And a (G4S) linker was added between DBD and NLS for providing region flexibility. ZF_43-8-DBD enable binding to specific DNA sequences, together with VP64 constitutes the activating-form SynTF. So that we can use ZF_43-8-VP64 as a specific transcription factor to activate the expression of our downstream synthetic promoter 8×ZF_43-8 elements and minimalCMV.
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