Composite

Part:BBa_K1140002:Design

Designed by: Janssel Reyes del Castillo   Group: iGEM13_UANL_Mty-Mexico   (2013-09-08)

pLac + 32 oC RNA thermometer + GFP(LVA)


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 797


Design Notes

The prefix sites are EcoRI and XbaI and the sufix sites are SpeI and PstI.


Source

Obtained synthetically. RNA thermometer is based on BBa__K115017 of team TUdelft [3]. The GFP sequence is similar to the one found in part BBa_K145015 and is a mutant derived from the green fluorescent protein found in jellyfish (Andersen et al, 1998). LVA makes proteins susceptible to degradation by the ClpX and ClpA proteases (McGiness, Baker and Sauer, 2007). The transcription termination sequences employed are two: part BBa_B0010 (derived from T1 of E. coli rrnB) and part BBa_B0012 (derived from TE of bacteriophage T7).

References

1. Andersen, JB, et al., (1998). New Unstable Variants of Green Fluorescent Protein for Studies of Transient Gene Expression in Bacteria, Applied and Environmental Microbiology, vol. 64 no. 6 2240-2246.

2. McGiness, KE; Baker, TA and Sauer, RT, (2007), Engineering Controllable Protein Degradation, Mol. Cell., 22, 701-707.

3. http://2008.igem.org/Team:TUDelft