Difference between revisions of "Part:BBa K2611003"
 
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We added a spacer sequence and a NGG site closely before the promoter (J23101). This spacer sequence can be recognized by the sgRNA we designed to direct dCas9. Once dCas9 binds with the spacer sequence, the expression of RFP will be repressed.To be used in pSB1C3.
 
We added a spacer sequence and a NGG site closely before the promoter (J23101). This spacer sequence can be recognized by the sgRNA we designed to direct dCas9. Once dCas9 binds with the spacer sequence, the expression of RFP will be repressed.To be used in pSB1C3.
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We selected part J23101-GFP ([https://parts.igem.org/Part:BBa_K364000 BBa_K364000]) as a reporter to test the function of the modified promoter and measure the repression level. The modified promoter does not influence the function of RFP gene as the RFP fluorescence intensity of the modified part is even stronger than the original one (Figure 3).
+
 
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We selected part J23101-RFP ([https://parts.igem.org/Part:BBa_K516132 BBa_K516132]) as a reporter to test the function of the modified promoter and measure the repression level. The modified promoter does not influence the function of RFP gene as the RFP fluorescence intensity of the modified part is even '''stronger''' than the original one (Figure 3).
  
 
[[File:SCU_China-2018_spacer_J23101_RFP(1).png]]
 
[[File:SCU_China-2018_spacer_J23101_RFP(1).png]]

Latest revision as of 02:16, 18 October 2018


Spacer J23101-RFP

We added a spacer sequence and a NGG site closely before the promoter (J23101). This spacer sequence can be recognized by the sgRNA we designed to direct dCas9. Once dCas9 binds with the spacer sequence, the expression of RFP will be repressed.To be used in pSB1C3.

We selected part J23101-RFP (BBa_K516132) as a reporter to test the function of the modified promoter and measure the repression level. The modified promoter does not influence the function of RFP gene as the RFP fluorescence intensity of the modified part is even stronger than the original one (Figure 3).

SCU China-2018 spacer J23101 RFP(1).png

SCU China-2018 spacer J23101 RFP(2).png

SCU China-2018 spacer J23101 RFP(3).png


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 30
    Illegal NheI site found at 53
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 640
    Illegal AgeI site found at 752
  • 1000
    COMPATIBLE WITH RFC[1000]