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. | ||
+ | We selected part J23101-GFP (BBa_J364000) 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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+ | [[File:SCU_China-2018_spacer_J23101_RFP(1).png]] | ||
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+ | [[File:SCU_China-2018_spacer_J23101_RFP(2).png]] | ||
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+ | [[File:SCU_China-2018_spacer_J23101_RFP(3).png]] | ||
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Revision as of 06:48, 17 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-GFP (BBa_J364000) 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).
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 30
Illegal NheI site found at 53 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 640
Illegal AgeI site found at 752 - 1000COMPATIBLE WITH RFC[1000]