Difference between revisions of "Part:BBa K2332052:Design"
Paola handal (Talk | contribs) (→Design Notes) |
Paola handal (Talk | contribs) (→Source) |
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===Source=== | ===Source=== | ||
− | + | Intimin protein sequence obtained from UniProtKB: [http://www.uniprot.org/uniprot/P43261#sequences P43261], reverse translated and codon optimised for E. coli. | |
− | Intimin protein sequence obtained from: http://www.uniprot.org/uniprot/P43261#sequences | + | Intimin for the cell surface display of our tag was truncated according to Wentzel et al., 2001. |
+ | SpyTag was obtained from: [https://parts.igem.org/Part:BBa_K1159201 BBa_K1159201]. | ||
+ | GFPmut3b was obtained from: [https://parts.igem.org/Part:BBa_E0040 BBa_E0040]. | ||
+ | The DNA sequence was synthesised by Integrated DNA Technologies (IDT) | ||
===References=== | ===References=== |
Revision as of 23:38, 22 October 2017
Intimin'-GFP-SpyTag (constitutive)
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 2826
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal SpeI site found at 2826 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 2826
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 2826
Illegal NgoMIV site found at 862
Illegal NgoMIV site found at 1603 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2697
Design Notes
We decided to use Intimin' as this has been previously proven to function effectively for cell surface display of up to 30kDa passenger proteins (Wentzel et al., 2001). We included a linker between Intimin' and GFP and between GFP and SpyTag to facilitate mobility of GFP and SpyTag on the cell surface. The stop codon in GFP was removed for the generation of this fusion protein.
Source
Intimin protein sequence obtained from UniProtKB: [http://www.uniprot.org/uniprot/P43261#sequences P43261], reverse translated and codon optimised for E. coli. Intimin for the cell surface display of our tag was truncated according to Wentzel et al., 2001. SpyTag was obtained from: BBa_K1159201. GFPmut3b was obtained from: BBa_E0040. The DNA sequence was synthesised by Integrated DNA Technologies (IDT)