Difference between revisions of "Part:BBa K2319004:Design"

 
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
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The extra nucleotide (G) is used to form the NheI restriction site when the T7 terminator (BBa_K731721) is added after it, which was then used in our subsequent assembly.
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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.
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===Source===
 
===Source===

Revision as of 10:09, 1 November 2017


TAAG


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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.

Source

This scar was added to our other part BBa_K2319000 (sfGFP-SpyCatcher under T7 expression system) using primers with overhangs.

References