Difference between revisions of "Part:BBa K4421029"

 
 
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<partinfo>BBa_K4421029 short</partinfo>
 
<partinfo>BBa_K4421029 short</partinfo>
  
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Gal4-KRAB transcription inhibitor was constructed under the control of the NFAT-RE promoter.
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Gal4-KRAB(<partinfo>BBa_K2446037</partinfo>) transcription inhibitor was constructed under the control of the NFAT-RE promoter(<partinfo>BBa_K4421021</partinfo>).
  
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===Special Note===
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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.
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<div><ul>
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<center>
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  <li style="display: inline-block;">  [[File:Cicuit section1.jpeg|thumb|none|400px|<b></b> The structure of NFAT_RE-Gal4-KRAB]]
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</li>
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</center>
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    </ul></div>
 
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<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
  
 
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<span class='h3bb'>Sequence and Features</span>
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===Sequence and Features===
 
<partinfo>BBa_K4421029 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4421029 SequenceAndFeatures</partinfo>
  

Latest 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.

  • 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]