Composite

Part:BBa_K4421029

Designed by: Shuyang Zhu   Group: iGEM22_NMU_China   (2022-07-15)
Revision as of 01:42, 7 October 2022 by Juejue (Talk | contribs)

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NFAT_RE-Gal4-KRAB

Gal4-KRAB(BBa_K2446037) transcription inhibitor was constructed under the control of the NFAT-RE promoter(BBa_K4421021).

Special Note

To simplify the transfection process and improve the transfection efficiency, we decided to transfect pSV40-UAS-iCASP9-2A-GFP and NFAT_RE-Gal4-KRAB on the same plasmid. But the KRAB protein has been demonstrated to be capable of inhibiting all promoters within at least 3 kB and the forward construct would therefore inhibit the NFAT-RE promoter, we generated the opposite construct, in which the two expression cassettes were cloned in such a manner that both promoters were at opposite ends and at a long distance.

  • The structure of NFAT_RE-Gal4-KRAB

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 478
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 478
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 478
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 478
    Illegal NgoMIV site found at 263
    Illegal NgoMIV site found at 398
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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