Difference between revisions of "Part:BBa K1140006:Design"

Revision as of 21:52, 5 October 2013

pTet + 37 oC RNA thermometer + mCherry (LVA)

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 63
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 676
Illegal AgeI site found at 788
1000
COMPATIBLE WITH RFC[1000]

Design Notes

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

Source

Obtained synthetically. The RNA thermosensor sequence was obtained and adapted from [3] (u6 synthetic RNA thermometer). The mCherry coding sequence was optimized for E. coli codon usage.

This composite part use the TetR sequence designed by June Rhee, Connie Tao, Ty Thomson and Louis Waldman (BBa_R0040) in 2003.

This Promoter is constitutively ON and repressed by TetR transcription factor. The TetR repression is inhibited by the addition of tetracycline or its analog, aTc to the media.

References

1. Elowitz MB, and Leibler S, (2000). A synthetic oscillatory network of transcriptional regulators. Nature, 20;403(6767):335-8.

2. Duval-Valentin G, and Ehrlich R, (1986). Interaction between E. coli RNA polymerase and the tetR promoter from pSC101: homologies and differences with other E. coli promoter systems from close contact point studies. Nucleic Acids Res., 14(5):1967-83.

3. Neupert J, Karcher D, Bock R, (2008). Design of simple synthetic RNA thermometers for temperature- controlled gene expression in Escherichia coli. Nucleic Acids Res, 36(19):e124.

@@ Line 10: / Line 10: @@
 ===Source===
-Obtained synthetically.
+Obtained synthetically. The RNA thermosensor sequence was obtained and adapted from [3] (u6 synthetic RNA thermometer). The mCherry coding sequence was optimized for E. coli codon usage.
-The RNA thermosensor sequence was obtained and adapted from [2] (u6 synthetic RNA thermometer).
-The mCherry coding sequence was optimized for ''E. coli'' codon usage.
+This composite part use the TetR sequence designed by June Rhee, Connie Tao, Ty Thomson and Louis Waldman (BBa_R0040) in 2003.
-The pTet promoter is identical to the sequence found in part BBa_R0040.
+This Promoter is constitutively ON and repressed by TetR transcription factor. The TetR repression is inhibited by the addition of tetracycline or its analog, aTc to the media.
 ===References===