Difference between revisions of "Part:BBa K4488008"
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<partinfo>BBa_K4488008 short</partinfo> | <partinfo>BBa_K4488008 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 CBDcex. The fuGFP sequence is towards the N terminus of the protein with CBDcex ( | + | The construct can be cloned into an expression vector such as pET28c in E.coli to produce a fusion protein of fuGFP with CBDcex. The fuGFP sequence is towards the N terminus of the protein with CBDcex (BBa_K4488023) 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_K4488012 for improved construct with higher fluorescence. |
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Latest revision as of 04:59, 9 October 2022
Fusion of free-use GFP with CBDcex (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 CBDcex. The fuGFP sequence is towards the N terminus of the protein with CBDcex (BBa_K4488023) 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_K4488012 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]