Part:BBa_K653001:Design

GFP reporter plus Double Terminator

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 644
  

Design Notes

This GFP part was derived from the reporter device used by Elowitz's repressilator : GFPmut3* has a couple of mutations from wtGFP - S2R and S65G and S72A. Emerald and EGFP have S65T and multiple other mutations. The S2R mutation should have no effect.

The Double terminator consisting of BBa_B0010 and BBa_B0012. This is the most commonly used terminator. It seems to be reliable.

The assembling of this two parts was performed using the 3A Assembly Method.

Source

This BBa_E0040part was derived from the reporter device used by Elowitz's repressilator [1]. Elowitz obtained the GFP tagged sequence (gfpmut3*) published by Andersen and coworkers [2] to build the reporter device. Andersen constructed those GFP variants based on a gfpmut3b containing plasmid that was a gift from R. H. Valdivia [3]. The PCR method used in construction by Andersen changed aa residue 2 from serine to arginine with no detectable phenotypic effect in their experiments.

The BBa_B0015 Double Terminator part is the most commonly used terminator. It seems to be reliable.

Note, however, that Part:BBa_B0014 is a better part for forward and reverse termination. The part was designed by Reshma Shetty

References

<biblio>

1. Elowitz pmid=10659856
2. Andersen pmid=9603842
3. Valdivia pmid=8707053
4. Double Terminator by Reshma Shetty BBa_B0015

</biblio>