Difference between revisions of "Part:BBa K864444"
(New page: __NOTOC__ <partinfo>BBa_K864444 short</partinfo> Use this part to insert any 5’ÚTR of your gene of interest to screen for downregulation of your gene with small RNA. PCR amplify your 5...) |
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Use this part to insert any 5’ÚTR of your gene of interest to screen for downregulation of your gene with small RNA. PCR amplify your 5’UTR with EcoRI and BamHI and clone into any backbone carrying this part. Digest whis part with EcoRI and BamHI and as RFP is replaced, this part functions as a red/white screening system. This allows you to create a reporter system with a fluorescent marker for screening for functional small RNAs. | Use this part to insert any 5’ÚTR of your gene of interest to screen for downregulation of your gene with small RNA. PCR amplify your 5’UTR with EcoRI and BamHI and clone into any backbone carrying this part. Digest whis part with EcoRI and BamHI and as RFP is replaced, this part functions as a red/white screening system. This allows you to create a reporter system with a fluorescent marker for screening for functional small RNAs. | ||
| − | SYFP2 is situated downstrean of the RFP and is connected in frame with a 12 aa glycine-serine repeated linker <partinfo>BBa_J18922</partinfo>, to prevent folding | + | SYFP2 <partinfo>BBa_K864100</partinfo> is situated downstrean of the RFP <partinfo>BBa_J04450</partinfo> and is connected in frame with a 12 aa glycine-serine repeated linker <partinfo>BBa_J18922</partinfo>, to prevent folding errors in the YFP or the 5’UTR. The first 6 nt of the glycine-serine linker is a BamHI restriction site (GGATCC), which translates into glycine-serine in E.Coli. |
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| + | When screening for working small RNAs its recommended to include the first 15 codons of your gene of interest. | ||
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Revision as of 01:11, 26 September 2012
RFP-Linker-SYFP2
Use this part to insert any 5’ÚTR of your gene of interest to screen for downregulation of your gene with small RNA. PCR amplify your 5’UTR with EcoRI and BamHI and clone into any backbone carrying this part. Digest whis part with EcoRI and BamHI and as RFP is replaced, this part functions as a red/white screening system. This allows you to create a reporter system with a fluorescent marker for screening for functional small RNAs.
SYFP2 BBa_K864100 is situated downstrean of the RFP BBa_J04450 and is connected in frame with a 12 aa glycine-serine repeated linker BBa_J18922, to prevent folding errors in the YFP or the 5’UTR. The first 6 nt of the glycine-serine linker is a BamHI restriction site (GGATCC), which translates into glycine-serine in E.Coli.
When screening for working small RNAs its recommended to include the first 15 codons of your gene of interest.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1070
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 781
Illegal AgeI site found at 893 - 1000COMPATIBLE WITH RFC[1000]