Difference between revisions of "Part:BBa K2319006:Design"

 
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===Design Notes===
 
===Design Notes===
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HindIII was used because the site is useful in our assembly. The 6xHis tag was codon-optimized for our protein.
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<p>A HindIII restriction site, a start codon and a 6xHis tag are added immediately downstream of the T7 promoter+RBS. A number of design considerations motivated the choice of the HindIII site. The HindIII site (A\AGCTT) — sandwiched between the RBS and the start codon — has a sequence very similar to the optimal sequence predicted by the sequence logo of <i>E. coli</i> ribosome binding sites, which can be found <a href="https://parts.igem.org/Help:Ribosome_Binding_Site">here</a>. In fact, the HindIII sequence is closer to the optimal sequence than the typical 5'-TACTAG-3' mixed SpeI-XbaI restriction site formed by BioBrick assembly, improving the initial ribosome-mRNA binding and improves translation.</p>
  
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<p>The 6xHis tag was codon-optimized for our protein.</p>
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===Source===
 
===Source===

Latest revision as of 01:18, 2 November 2017


HindIII+ATG+6xHis tag


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 HindIII restriction site, a start codon and a 6xHis tag are added immediately downstream of the T7 promoter+RBS. A number of design considerations motivated the choice of the HindIII site. The HindIII site (A\AGCTT) — sandwiched between the RBS and the start codon — has a sequence very similar to the optimal sequence predicted by the sequence logo of E. coli ribosome binding sites, which can be found here. In fact, the HindIII sequence is closer to the optimal sequence than the typical 5'-TACTAG-3' mixed SpeI-XbaI restriction site formed by BioBrick assembly, improving the initial ribosome-mRNA binding and improves translation.

The 6xHis tag was codon-optimized for our protein.

Source

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

References