Coding
TF-KRAB-1

Part:BBa_K2446037

Designed by: Yijie Pan   Group: iGEM17_Fudan   (2017-10-21)
Revision as of 15:40, 29 October 2017 by Pyj6060 (Talk | contribs)


ZF_GAl4_KRAB

This part is one of our mammalian synthetic transcription factors (SynTFs) based on Gal4 DNA binding domain [1]. Gal4-KRAB (or TF-KRAB-1 in our project) containing three core domains from N-terminal to C-terminal: GAL4 DNA binding domain, nuclear location sequence and KRAB transcription regulating domain [2].And a (G4S) linker was added between DBD and NLS for providing region flexibility [3]. GAL4DBD enable binding to specific DNA sequences, so that we can use Gal4-KRAB as a specific transcription factors to repress the expression of our mammalian synthetic promoter Sv40-UAS (BBa_K2446036).

The information of other SynTF-SynPro is showed in the table below.

SynTFs SynPros
Gal4-KRAB(TF-KRAB-1) BBa_K2446037 Sv40-UAS(Sv40-UAS) BBa_K2446036
ZF_PIP_KRAB(TF-KRAB-2) BBa_K2446045 SV40_2/4/8_PIP BBa_K2446033/BBa_K2446034/BBa_K2446035
ZF_21-16KRAB(TF-KRAB-3) BBa_K2446039 SV40_8_ZF_21-16 BBa_K2446030
ZF_42-10_KRAB(TF-KRAB-4) BBa_K2446040 SV40_8_ZF_42-10 BBa_K2446025
ZF_43-8_KRAB(TF-KRAB-5) BBa_K2446041 SV40_2/4/8_ZF_43-8 BBa_K2446026/BBa_K2446027/BBa_K2446028
ZF_54-8_KRAB(TF-KRAB-6) BBa_K2446042 SV40_8_ZF_54-8 BBa_K2446029
ZFHD1_KRAB(TF-KRAB-7) BBa_K2446043 SV40_4_ZFHD1 BBa_K2446032

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 256
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 256
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 256
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 256
    Illegal NgoMIV site found at 41
    Illegal NgoMIV site found at 176
  • 1000
    COMPATIBLE WITH RFC[1000]


References

[1] Morsut, L. et al. Engineering Customized Cell Sensing and Response Behaviors Using Synthetic Notch Receptors. Cell 164, 780--791 (2016).

[2] Witzgall, R., O'Leary, E., Leaf, A., Onaldi, D. & Bonventre, J. V. The Krüppel-associated box-A (KRAB-A) domain of zinc finger proteins mediates transcriptional repression. Proceedings of the National Academy of Sciences 91, 4514-4518 (1994).

[3] Chen, X., Zaro, J. L. & Shen, W.-C. Fusion protein linkers: property, design and functionality. Advanced drug delivery reviews 65, 1357-1369 (2013).

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