Difference between revisions of "Part:BBa K3128008:Design"
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Two restriction sites were added on both sides of RFP gene as such : BglII_RFP_BamHI
 
Two restriction sites were added on both sides of RFP gene as such : BglII_RFP_BamHI
  
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From <cite>Campbell</cite>:  
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From Campbell:  
 
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).
 
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).
  
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[http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=Retrieve&db=nucleotide&dopt=GenBank&list_uids=21464837 Genbank accession AF506027]
 
[http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=Retrieve&db=nucleotide&dopt=GenBank&list_uids=21464837 Genbank accession AF506027]
  
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<biblio>
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#Campbell pmid=12060735
 
#Campbell pmid=12060735
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</biblio>
+
 
 
[http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=12060735 URL]
 
[http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=12060735 URL]
  

Revision as of 07:14, 16 September 2019

**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. Two restriction sites were added on both sides of RFP gene as such : BglII_RFP_BamHI

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

[http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=Retrieve&db=nucleotide&dopt=GenBank&list_uids=21464837 Genbank accession AF506027]


  1. Campbell pmid=12060735

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

References

<biblio>

  1. Zhang pmid=12461557

</biblio>