Difference between revisions of "Part:BBa K2285012"
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constitutive promoter J23119 followed by a stem-loop structure Target3, with chromoprotein eforRed as the reporter gene. | constitutive promoter J23119 followed by a stem-loop structure Target3, with chromoprotein eforRed as the reporter gene. | ||
The original part of eforRed chromoprotein is presented on <partinfo>BBa_K592012</partinfo>. What should be noticed is, the original part is a coding part and doesn't have its own RBS and Term. Therefore, to achieve greater convenience fo our Star-chromoprotein reporter system, we add the RBS which is on the upstream of sfGFP(<partinfo>BBa_K515005</partinfo>) as well as a short terminator <partinfo>BBa_B1006</partinfo> to all chromoproteins we used in this project via PCR. | The original part of eforRed chromoprotein is presented on <partinfo>BBa_K592012</partinfo>. What should be noticed is, the original part is a coding part and doesn't have its own RBS and Term. Therefore, to achieve greater convenience fo our Star-chromoprotein reporter system, we add the RBS which is on the upstream of sfGFP(<partinfo>BBa_K515005</partinfo>) as well as a short terminator <partinfo>BBa_B1006</partinfo> to all chromoproteins we used in this project via PCR. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
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To achieve further control of a this reporter, we simply add a stem-loop structure(Target3) upstream of it. To activate reporter's expression, Antisense3 <partinfo>BBa_K2285010</partinfo>, which is the complementary sequence to part of Target3, is needed neither in the same plasmid or even another plasmid. | To achieve further control of a this reporter, we simply add a stem-loop structure(Target3) upstream of it. To activate reporter's expression, Antisense3 <partinfo>BBa_K2285010</partinfo>, which is the complementary sequence to part of Target3, is needed neither in the same plasmid or even another plasmid. | ||
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| + | ===Characterization=== | ||
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[[Image:eforRed.JPG|420px]] | [[Image:eforRed.JPG|420px]] | ||
Revision as of 08:52, 1 November 2017
J23119+Target3+eforRed(with RBS and Term added)
constitutive promoter J23119 followed by a stem-loop structure Target3, with chromoprotein eforRed as the reporter gene. The original part of eforRed chromoprotein is presented on BBa_K592012. What should be noticed is, the original part is a coding part and doesn't have its own RBS and Term. Therefore, to achieve greater convenience fo our Star-chromoprotein reporter system, we add the RBS which is on the upstream of sfGFP(BBa_K515005) as well as a short terminator BBa_B1006 to all chromoproteins we used in this project via PCR.
Usage and Biology
The coding region of eforRed was originated from iGEM11_Uppsala-Sweden.
To achieve further control of a this reporter, we simply add a stem-loop structure(Target3) upstream of it. To activate reporter's expression, Antisense3 BBa_K2285010, which is the complementary sequence to part of Target3, is needed neither in the same plasmid or even another plasmid.
Characterization
iGEM17_SJTU_BioX_Shanghai: Left picture: Chromoprotein eforRed in EP
Right picture: E.coli with eforRed expression
During experiments, we found that expression of chromoproteins amilCP BBa_K1357009, amilGFP BBa_K592010, eforRed BBa_K592012 and cjBlue BBa_K592011 took nearly 24 hours to see the colors with naked eyes on the LB plate. But they are still great reporters when it comes to build visualized systems which do not need to depend on extra equippments.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]