Difference between revisions of "Part:BBa K4497030"
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[[File:MUC cycle.png|800px]]
 
[[File:MUC cycle.png|800px]]
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Figure 4. Flow Cytometry Histograms comparing the usage of EYFP and miRFP in our loop system. (A): Gating for FITC positive cells expressing either the miRFP or EYFP reporter. Both cell samples were also transfected with the heterodimeric MESA system G1TA & M1TEV. 500 ng/mL GFP-mCherry were added to the samples. (B) Gated FITC Histogram of the EYFP reporter sample. It is not possible to differentiate the GFP-mCherry receptor binding from the induced reporter activation. (C) Gated FITC Histogram of the miRFP reporter sample. GFP binding is visible by the signal shift of the right wing of the curve. (D) Gated APC-A Histogram of the miRFP reporter sample. The reporter signal is visible in the APC-A channel separated from the FITC signal.
 
Figure 4. Flow Cytometry Histograms comparing the usage of EYFP and miRFP in our loop system. (A): Gating for FITC positive cells expressing either the miRFP or EYFP reporter. Both cell samples were also transfected with the heterodimeric MESA system G1TA & M1TEV. 500 ng/mL GFP-mCherry were added to the samples. (B) Gated FITC Histogram of the EYFP reporter sample. It is not possible to differentiate the GFP-mCherry receptor binding from the induced reporter activation. (C) Gated FITC Histogram of the miRFP reporter sample. GFP binding is visible by the signal shift of the right wing of the curve. (D) Gated APC-A Histogram of the miRFP reporter sample. The reporter signal is visible in the APC-A channel separated from the FITC signal.
  

Revision as of 15:42, 12 October 2022


tTA Induceable miRFP Reporter Components

This part consists of the bidirectional Promoter Pbi-1 (BBa_K4497027), that is induced by the Transcription Factor tetracycline-inducable Transactivator (tTA). The promoter is made of Tetracycline repeat elements (TRE) with a minimal CMV promoter on both sides. The promoter induces expression of the downstream miRFP (BBa_K4497029), allowing for an miRFP signal (Ex661/Em680) readout on tTA induction of the promoter.

MUC cycle.png

Figure 4. Flow Cytometry Histograms comparing the usage of EYFP and miRFP in our loop system. (A): Gating for FITC positive cells expressing either the miRFP or EYFP reporter. Both cell samples were also transfected with the heterodimeric MESA system G1TA & M1TEV. 500 ng/mL GFP-mCherry were added to the samples. (B) Gated FITC Histogram of the EYFP reporter sample. It is not possible to differentiate the GFP-mCherry receptor binding from the induced reporter activation. (C) Gated FITC Histogram of the miRFP reporter sample. GFP binding is visible by the signal shift of the right wing of the curve. (D) Gated APC-A Histogram of the miRFP reporter sample. The reporter signal is visible in the APC-A channel separated from the FITC signal.



Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 465
    Illegal XbaI site found at 484
    Illegal SpeI site found at 490
    Illegal PstI site found at 471
    Illegal PstI site found at 919
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 465
    Illegal SpeI site found at 490
    Illegal PstI site found at 471
    Illegal PstI site found at 919
    Illegal NotI site found at 476
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 465
    Illegal XhoI site found at 841
    Illegal XhoI site found at 847
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 465
    Illegal XbaI site found at 484
    Illegal SpeI site found at 490
    Illegal PstI site found at 471
    Illegal PstI site found at 919
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 465
    Illegal XbaI site found at 484
    Illegal SpeI site found at 490
    Illegal PstI site found at 471
    Illegal PstI site found at 919
    Illegal NgoMIV site found at 622
    Illegal NgoMIV site found at 1104
  • 1000
    COMPATIBLE WITH RFC[1000]