Difference between revisions of "Part:BBa K4421029"
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Revision as of 01:42, 7 October 2022
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.
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 478
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 478
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 478
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 478
Illegal NgoMIV site found at 263
Illegal NgoMIV site found at 398 - 1000COMPATIBLE WITH RFC[1000]