Part:BBa_K3132105
PIP-KRAB-Gal4-VP64
This part is transformed based on the part: PIP_KRAB(BBa_K2446038) of iGEM17_Fudan.The part is one of our synthetic transcription factors (SynTFs) based on 2 parts, PIP-KRAB and Gal4-VP64(BBa_K3132000).Its parental form PIP-KRAB, which was developed by iGEM_Fudan, serves as an inhibitory transcription factor because of the KRAB domain; while the added part Gal4-VP64, an improved part of our team this year, serves as an activating transcription factor because both of the Gal4 and VP64 domain.This is why we call it “bifunctional transcription factor”. PIP-KRAB-Gal4-VP64 contains six core domains from N-terminal to C-terminal: PIP DNA binding domain, nuclear location sequence, KRAB transcription regulating domain, GAL4 DNA binding domain, another nuclear location sequence and VP64 transcription regulating domain[1]. And 2 (G4S) linkers were added between DBD and NLS for providing region flexibility[2]. PIP-DBD and Gal4-DBD enable binding to specific DNA sequences, together with KRAB and VP64 constitute the inhibitory and activating part, respectively. So that we can use it as a bifunctional transcription factor to either activate or repress the expression of our downstream synthetic promoter elements and minimalCMV when binding to specific DNA sequences . Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1085
Illegal NgoMIV site found at 1220 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 475
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