Composite

Part:BBa_K2683016

Designed by: Kristi Turton   Group: iGEM18_Lethbridge   (2018-10-10)


Release Factor 1 in pSB1C3

Release factor 1 (RF1) is a protein that terminates translation through the recognition of the stop codon in an mRNA sequence. RF1 was originally designed by Lethbridge iGEM team 2017 (http://2017.igem.org/Team:Lethbridge). The original page can be found here: https://parts.igem.org/Part:BBa_K2443029 . RF1 had been Codon optimized for use in Escherichia coli and contains a N-Terminal hexahistidine tag and is under the regulation of T7 Promoter, RBS and double terminator.

This year we have been able to place this composite part in the pSB1C3 standard plasmid and have sent it to the registry.


T--Lethbridge--RF1RF2.PNG

Figure 1- Here is a overexpression gel for Release Figure 1. The samples taken was time of induction and three hour increments thereafter.

T--Lethbridge--RF1pur.PNG

Figure 2- A 12% SDS-PAGE gel for the purification of Release Factor 1 using nickel affinity chromatography. The purified protein can be seen in elutions 1-5 at the 55kDa range.


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 675
  • 1000
    COMPATIBLE WITH RFC[1000]


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