Difference between revisions of "Part:BBa E1010:Design"
(→‎Design Notes)
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Four silent mutations made to remove three EcoRI sites and one PstI site: A28G, A76G, A349G, G337A.
 
Four silent mutations made to remove three EcoRI sites and one PstI site: A28G, A76G, A349G, G337A.
  
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+
From <cite>Campbell</cite>:
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The final clone, designated mRFP1, contains a total of 33 mutations relative to DsRed of which 13 are internal to the &beta;-barrel (N42Q, V44A, V71A, K83L, F124L, L150M, K163M, V175A, F177V, S179T, V195T, S197I, and T217A). Of the 20 remaining external mutations, three are the aggregation-reducing mutations from T1 (R2A, K5E, and N6D), three are AB interface mutations (I125R, V127T, and I180T), ten are AC interface mutations (R153E, H162K, A164R, L174D, Y192A, Y194K, H222S, L223T, F224G, and L225A), and four are additional beneficial mutations (T21S, H41T, C117E, and V156A).
  
 
===Source===
 
===Source===

Revision as of 21:03, 16 July 2009

**highly** engineered mutant of red fluorescent protein from Discosoma striata (coral)


Barcodes are discontinued, but one was appended to the sequence of this part. Composite parts using this part will include the barcode. More ...

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 555
    Illegal AgeI site found at 667
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

TAATAA double stop codon added (DE).

Four silent mutations made to remove three EcoRI sites and one PstI site: A28G, A76G, A349G, G337A.

From Campbell: The final clone, designated mRFP1, contains a total of 33 mutations relative to DsRed of which 13 are internal to the β-barrel (N42Q, V44A, V71A, K83L, F124L, L150M, K163M, V175A, F177V, S179T, V195T, S197I, and T217A). Of the 20 remaining external mutations, three are the aggregation-reducing mutations from T1 (R2A, K5E, and N6D), three are AB interface mutations (I125R, V127T, and I180T), ten are AC interface mutations (R153E, H162K, A164R, L174D, Y192A, Y194K, H222S, L223T, F224G, and L225A), and four are additional beneficial mutations (T21S, H41T, C117E, and V156A).

Source

<biblio>

  1. Campbell pmid=12060735

</biblio> [http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=12060735 URL]

References

<biblio>

  1. Zhang pmid=12461557

</biblio>