Difference between revisions of "Part:BBa K2319004:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | A stop codon (TAA) the nucleotide G are added immediately upstream of BBa_K731721 (T7 terminator). This extra stop codon (assuming the fusion protein sequence has its own) ensures that translation is halted and prevents any translational read-through. The extra nucleotide G is added for a more subtle purpose: when placed just before the T7 terminator sequence (5'-CTAGC...TTTTG-3'), it forms the NheI restriction site (G\CTAGC). This allows any fusion protein to be inserted into our expression backbone using the HindIII and NheI sites. | + | A stop codon (TAA) and the nucleotide G are added immediately upstream of BBa_K731721 (T7 terminator). This extra stop codon (assuming the fusion protein sequence has its own) ensures that translation is halted and prevents any translational read-through. The extra nucleotide G is added for a more subtle purpose: when placed just before the T7 terminator sequence (5'-CTAGC...TTTTG-3'), it forms the NheI restriction site (G\CTAGC). This allows any fusion protein to be inserted into our expression backbone using the HindIII and NheI sites. |
===Source=== | ===Source=== | ||
Latest revision as of 10:10, 1 November 2017
TAAG
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
A stop codon (TAA) and the nucleotide G are added immediately upstream of BBa_K731721 (T7 terminator). This extra stop codon (assuming the fusion protein sequence has its own) ensures that translation is halted and prevents any translational read-through. The extra nucleotide G is added for a more subtle purpose: when placed just before the T7 terminator sequence (5'-CTAGC...TTTTG-3'), it forms the NheI restriction site (G\CTAGC). This allows any fusion protein to be inserted into our expression backbone using the HindIII and NheI sites.
Source
This short sequence was added using PCR, with primers having 5'-overhangs corresponding to these nucleotides.