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

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===Source===
 
===Source===
  
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Obtained synthetically.
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Obtained synthetically. The RNA thermosensor sequence was obtained and adapted from [4] (u6 synthetic RNA thermometer). The mCherry coding sequence [1] was optimized for ''E. coli'' codon usage.
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The RNA thermosensor sequence was obtained and adapted from [1] (u6 synthetic RNA thermometer).  
+
 
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The RFP variant employed is the same that is found in part BBa_E1010, which is a monomeric variant of the red fluorescent protein found in the coral ''Discosoma striata''.
+
This composite part use the TetR sequence designed by June Rhee, Connie Tao, Ty Thomson and Louis Waldman (BBa_R0040) in 2003.
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The pTet promoter is identical to the sequence found in part BBa_R0040.
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===References===
 
===References===
  
−
Elowitz, MB and Leibler, S, (2000). A synthetic oscillatory network of transcriptional regulators, ''Nature'', 20;403(6767):335-8.
+
1. Shaner NC, Steinbach PA, and Tsien RY. (2005). A guide to choosing fluorescent proteins. ''Nat Methods''. 2(12):905-9.
 +
 
 +
2. Elowitz MB, and Leibler S. (2000). A synthetic oscillatory network of transcriptional regulators. ''Nature'', 20;403(6767):335-8.
 +
 
 +
3. 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.
  
−
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.
+
4. 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.

Latest revision as of 20:51, 28 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 [4] (u6 synthetic RNA thermometer). The mCherry coding sequence [1] 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.

References

1. Shaner NC, Steinbach PA, and Tsien RY. (2005). A guide to choosing fluorescent proteins. Nat Methods. 2(12):905-9.

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

3. 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.

4. 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.