Difference between revisions of "Part:BBa K4488009"
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<partinfo>BBa_K4488009 short</partinfo> | <partinfo>BBa_K4488009 short</partinfo> | ||
| − | The construct can be cloned into an expression vector such as pET28c in E.coli to produce a fusion protein of fuGFP with CBDcipA. The fuGFP sequence is towards the N terminus of the protein with CBDcipA ( | + | The construct can be cloned into an expression vector such as pET28c in E.coli to produce a fusion protein of fuGFP with CBDcipA. The fuGFP sequence is towards the N terminus of the protein with CBDcipA (BBa_K4488024) downstream followed by a stop codon. Recognition sites for BamHI and BsaI are present before the RBS allowing golden gate cloning. XhoI and BsaI are also present downstream of the stop codon. Notably, the assembled protein is insoluble. See BBa_K3188013 for improved construct with higher fluorescence. |
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Revision as of 05:02, 9 October 2022
Fusion of free-use GFP with CBDcipA (cellulose-binding domain) at the C-terminal end
The construct can be cloned into an expression vector such as pET28c in E.coli to produce a fusion protein of fuGFP with CBDcipA. The fuGFP sequence is towards the N terminus of the protein with CBDcipA (BBa_K4488024) downstream followed by a stop codon. Recognition sites for BamHI and BsaI are present before the RBS allowing golden gate cloning. XhoI and BsaI are also present downstream of the stop codon. Notably, the assembled protein is insoluble. See BBa_K3188013 for improved construct with higher fluorescence.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 163
- 1000COMPATIBLE WITH RFC[1000]