Difference between revisions of "Part:BBa K2332050:Design"

Revision as of 23:27, 22 October 2017

Intimin'-SpyTag (constitutive)

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 862
Illegal NgoMIV site found at 1603
1000
COMPATIBLE WITH RFC[1000]

Design Notes

We decided to fuse SpyTag to the N-terminal of a truncated version of 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 also included a linker between Intimin and SpyTag to facilitate mobility of SpyTag on the cell surface.

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. The DNA sequence was synthesised by Integrated DNA Technologies (IDT)

@@ Line 11: / Line 11: @@
 ===Source===
-All parts were obtained as synthetic gBlocks Intimin protein sequence obtained from: http://www.uniprot.org/uniprot/P43261#sequences And reverse translated using: http://www.bioinformatics.org/sms2/rev_trans.html Intimin for the cell surface display of our tag was truncated according to Wentzel et al., 2001 SpyTag: BBa_K1159201
+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: [https://parts.igem.org/Part:BBa_K1159201 BBa_K1159201]. The DNA sequence was synthesised by Integrated DNA Technologies (IDT)
 ===References===