Difference between revisions of "Part:BBa K4174002"
(→‎Design Notes)
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This composite part is an improvement of the 2006 MIT iGEM team's composite part BBa_J45995. We have replaced GFP with sfGFP from Ceroni <i>et al.</i> 2015, replaced RBS BBa_B0030 with an RBS containing region from Ceroni <i>et al.</i> 2015, removed the scar sequences, and added UNS1 and UNS10 sequences to the ends.
 
This composite part is an improvement of the 2006 MIT iGEM team's composite part BBa_J45995. We have replaced GFP with sfGFP from Ceroni <i>et al.</i> 2015, replaced RBS BBa_B0030 with an RBS containing region from Ceroni <i>et al.</i> 2015, removed the scar sequences, and added UNS1 and UNS10 sequences 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 <i>Escherichia coli</i>, thus serving as a more effective assay (Pédelacq 2006).  
+
<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>
  
−
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>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>
  
−
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>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>
  
 
===Source===
 
===Source===

Revision as of 20:06, 7 October 2022

osmY-sfGFP

This composite part is composed of BBa_K2680553 (UNS1), BBa_J45992 (osmY promoter), BBa_K3773008 (RBS containing region), BBa_K3773003 (sfGFP), BBa_B0015 (terminator), and BBa_K2680554 (UNS10).

Design Notes

This composite part is an improvement of the 2006 MIT iGEM team's composite part BBa_J45995. We have replaced GFP with sfGFP from Ceroni et al. 2015, replaced RBS BBa_B0030 with an RBS containing region from Ceroni et al. 2015, removed the scar sequences, and added UNS1 and UNS10 sequences 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

    Pédelacq, J. D., Cabantous, S., Tran, T., Terwilliger, T. C., & Waldo, G. S. (2006). Engineering and characterization of a superfolder green fluorescent protein. Nature biotechnology, 24(1), 79-88.






    Sequence and Features


    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]