Difference between revisions of "Part:BBa K4174002"
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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).
 
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).
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===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.
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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).
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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.
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===Source===
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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
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===References===
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Pédelacq, J. D., Cabantous, S., Tran, T., Terwilliger, T. C., & Waldo, G. S. (2006). Engineering and characterization of a superfolder green fluorescent protein. <i>Nature biotechnology</i>, 24(1), 79-88.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K4174002 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4174002 SequenceAndFeatures</partinfo>
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===Functional Parameters===
 
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<partinfo>BBa_K4174002 parameters</partinfo>
 

Revision as of 20:04, 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]