Difference between revisions of "Part:BBa K2443031"
 
Line 3: Line 3:
 
<partinfo>BBa_K2443031 short</partinfo>
 
<partinfo>BBa_K2443031 short</partinfo>
  
−
Release factor 3 (RF3) allows for the termination of translation through the recognition of the stop codon in an mRNA sequence. RF3 has been Codon optimized for use in <i>Escherichia coli</i> and contains a N-Terminal hexahistidine tag. Under the regulation of T7 Promoter, RBS and double terminator.  
+
Release factor 3 (RF3) allows for the termination of translation through the recognition of the stop codon in an mRNA sequence. RF3 has been Codon optimized for use in <i>Escherichia coli</i> and contains a N-Terminal hexahistidine tag. Under the regulation of T7 Promoter, RBS and double terminator. The expected size of RF3 is 61 kDa, which corresponds with the band size in lane 7 of the overexpression gel (below). Protein was overexpressed from BL21DE3 gold containing the construct in a pUC57-kan vector according to the protocol, and notebook on the iGEM 2017 Lethbridge wiki (https://static.igem.org/mediawiki/2017/2/2c/Protein_Overexpression_.pdf and https://static.igem.org/mediawiki/2017/0/0d/October_.pdf).
  
 +
https://static.igem.org/mediawiki/parts/b/bf/Lethbridge_Oct_22_gel.png
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 18:52, 1 November 2017


RF3 optimized for expression in E. coli

Release factor 3 (RF3) allows for the termination of translation through the recognition of the stop codon in an mRNA sequence. RF3 has been Codon optimized for use in Escherichia coli and contains a N-Terminal hexahistidine tag. Under the regulation of T7 Promoter, RBS and double terminator. The expected size of RF3 is 61 kDa, which corresponds with the band size in lane 7 of the overexpression gel (below). Protein was overexpressed from BL21DE3 gold containing the construct in a pUC57-kan vector according to the protocol, and notebook on the iGEM 2017 Lethbridge wiki (https://static.igem.org/mediawiki/2017/2/2c/Protein_Overexpression_.pdf and https://static.igem.org/mediawiki/2017/0/0d/October_.pdf).

Lethbridge_Oct_22_gel.png

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 NgoMIV site found at 919
    Illegal AgeI site found at 980
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1143