Difference between revisions of "Part:BBa K4488009"

Revision as of 03:50, 1 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_K1321014) 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_K4488013 for improved construct with higher fluorescence.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 163
1000
COMPATIBLE WITH RFC[1000]

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 <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 (BBa_K1321014) 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_K4488011 for improved construct with higher fluorescence.
+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_K1321014) 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_K4488013 for improved construct with higher fluorescence.
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