Difference between revisions of "Part:BBa K4174002:Design"
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===Design Notes===
 
===Design Notes===
 
Based on the 2006 MIT iGEM team's composite part BBa_J45995, with GFP replaced with sfGFP from Ceroni <i>et al.</i> 2015, RBS BBa_B0030 replaced with an RBS containing region from Ceroni <i>et al.</i> 2015, scar sequences removed, and UNS1 and UNS10 added to the ends.
 
Based on the 2006 MIT iGEM team's composite part BBa_J45995, with GFP replaced with sfGFP from Ceroni <i>et al.</i> 2015, RBS BBa_B0030 replaced with an RBS containing region from Ceroni <i>et al.</i> 2015, scar sequences removed, and UNS1 and UNS10 added to the ends.
 +
<ul>
 +
<li>We elected to use super-folder green fluorescent protein (sfGFP) as opposed to the original GFP, as it folds more readily in <i>Escherichia coli</i>, thus serving as a more effective assay (Pédelacq 2006).</li>
  
−
===Source===
+
<li>We switched the original RBS with an RBS containing region used with the sfGFP sequence in Ceroni (2015)'s paper. Our team had previously used those parts together successfully, so we elected to use them together again.</li>
  
 +
<li>We added UNS1 and UNS10 sequences to make this part compatible with Gibson Assembly with our backbone, as we also added UNS1 and UNS10 to our pSB1C3 backbone.</li>
 +
</ul>
 +
 +
===Source===
 
Ceroni, F., Algar, R., Stan, G., & Ellis, T. (2015). Quantifying cellular capacity identifies gene expression designs with reduced burden. <i>Nature Methods,</i> 12(5):415-418. Doi: 10.1038/nmeth.3339
 
Ceroni, F., Algar, R., Stan, G., & Ellis, T. (2015). Quantifying cellular capacity identifies gene expression designs with reduced burden. <i>Nature Methods,</i> 12(5):415-418. Doi: 10.1038/nmeth.3339
 +
 +
===References===

Revision as of 20:19, 7 October 2022

osmY-sfGFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 419
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Based on the 2006 MIT iGEM team's composite part BBa_J45995, with GFP replaced with sfGFP from Ceroni et al. 2015, RBS BBa_B0030 replaced with an RBS containing region from Ceroni et al. 2015, scar sequences removed, and UNS1 and UNS10 added to the ends.

  • We elected to use super-folder green fluorescent protein (sfGFP) as opposed to the original GFP, as it folds more readily in Escherichia coli, thus serving as a more effective assay (Pédelacq 2006).
  • We switched the original RBS with an RBS containing region used with the sfGFP sequence in Ceroni (2015)'s paper. Our team had previously used those parts together successfully, so we elected to use them together again.
  • We added UNS1 and UNS10 sequences to make this part compatible with Gibson Assembly with our backbone, as we also added UNS1 and UNS10 to our pSB1C3 backbone.

Source

Ceroni, F., Algar, R., Stan, G., & Ellis, T. (2015). Quantifying cellular capacity identifies gene expression designs with reduced burden. Nature Methods, 12(5):415-418. Doi: 10.1038/nmeth.3339

References