Difference between revisions of "Part:BBa K1659088:Design"
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− | ===Design Notes=== | + | ===Design Notes and Sources=== |
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+ | We obtained the SMAP-29 peptide sequence from [1] and converted it into a nucleotide sequence using [https://www.idtdna.com/CodonOpt IDT's Codon Optimization Tool], setting the product type for gBlocks Gene Fragments and using the codon table for ''E. coli K12''. The sequence for T4 Endolysin was obtained from [https://parts.igem.org/Part:BBa_K112806 BBa_K112806], where nucleotide 20 onwards (the sequence coming after the start codon) was fused downstream of the SMAP-29 nucleotide sequence. | ||
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+ | A Hisx6 tag is added at the C-terminus for ease of protein purification using metal-affinity chromatography. | ||
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===References=== | ===References=== | ||
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+ | [1] Tack, B.F. et al., 2002. SMAP-29 has two LPS-binding sites and a central hinge. European Journal of Biochemistry, 269(4), pp.1181–1189. |
Latest revision as of 14:16, 10 September 2015
Artilysin Art-E, an antibacterial fusion protein
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 342
Illegal AgeI site found at 412 - 1000COMPATIBLE WITH RFC[1000]
Design Notes and Sources
We obtained the SMAP-29 peptide sequence from [1] and converted it into a nucleotide sequence using IDT's Codon Optimization Tool, setting the product type for gBlocks Gene Fragments and using the codon table for E. coli K12. The sequence for T4 Endolysin was obtained from BBa_K112806, where nucleotide 20 onwards (the sequence coming after the start codon) was fused downstream of the SMAP-29 nucleotide sequence.
A Hisx6 tag is added at the C-terminus for ease of protein purification using metal-affinity chromatography.
References
[1] Tack, B.F. et al., 2002. SMAP-29 has two LPS-binding sites and a central hinge. European Journal of Biochemistry, 269(4), pp.1181–1189.