Difference between revisions of "Part:BBa K1442023:Design"

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===Design Notes===
 
===Design Notes===
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The EMCV IRES had to be made.  
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<p>An AscI and BsrGI restriction site were added at the beginning and end of the sequence respectively in order to clone this part into our testing modules. </p>
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<p>'''AscI''' is compatible with MluI </p>
−
 
+
<p>'''BsrGI'''is compatible with BsiWI and TatI </p>
  
 
===Source===
 
===Source===
  
−
The IRES came from the EMCV virus.
+
<p> This Internal Ribosome Entry Site (IRES) was derived from the Encephalomyocarditis Virus genome. It is commonly used in replicon experiments in HeLa cells. Specifically this sequence was taken from the detailed genome used in the Lohmann (1999) HCV replicon experiment. </p>
 +
<p></p>
 +
<p> Lohmann V. et al. (1999). Replication of Subgenomic Hepatitis C Virus RNAs in a Hepatoma Cell Line. Science. 285 (5424), 110-113. </p>
 +
<p> The exact sequence of the plasmid used in these experiments can be found on NCBI: http://www.ncbi.nlm.nih.gov/nuccore/5441831 </p>
  
 
===References===
 
===References===
  
−
[1] Bochkov and Palmenberg, 2006, Translational efficiency of EMCV IRES in bicistronic vectors is dependent upon IRES sequence and gene location, BioTechniques, 41:283-292 (September 2006)
+
<p>'''[1]'''Bochkov and Palmenberg, 2006, Translational efficiency of EMCV IRES in bicistronic vectors is dependent upon IRES sequence and gene location, BioTechniques, 41:283-292 (September 2006) </p>
−
http://www.biotechniques.com/multimedia/archive/00002/BTN_A_000112243_O_2391a.pdf
+
<p> http://www.biotechniques.com/multimedia/archive/00002/BTN_A_000112243_O_2391a.pdf </p>
 +
<p></p>
 +
<p>'''[2]'''Lohmann V. et al. (1999). Replication of Subgenomic Hepatitis C Virus RNAs in a Hepatoma Cell Line. Science. 285 (5424), 110-113.</p>
 +
<p>http://www.sciencemag.org/content/285/5424/110.long</p>

Latest revision as of 11:10, 6 October 2014

EMCV IRES


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
    COMPATIBLE WITH RFC[1000]


Design Notes

An AscI and BsrGI restriction site were added at the beginning and end of the sequence respectively in order to clone this part into our testing modules.

AscI is compatible with MluI

BsrGIis compatible with BsiWI and TatI

Source

This Internal Ribosome Entry Site (IRES) was derived from the Encephalomyocarditis Virus genome. It is commonly used in replicon experiments in HeLa cells. Specifically this sequence was taken from the detailed genome used in the Lohmann (1999) HCV replicon experiment.

Lohmann V. et al. (1999). Replication of Subgenomic Hepatitis C Virus RNAs in a Hepatoma Cell Line. Science. 285 (5424), 110-113.

The exact sequence of the plasmid used in these experiments can be found on NCBI: http://www.ncbi.nlm.nih.gov/nuccore/5441831

References

[1]Bochkov and Palmenberg, 2006, Translational efficiency of EMCV IRES in bicistronic vectors is dependent upon IRES sequence and gene location, BioTechniques, 41:283-292 (September 2006)

http://www.biotechniques.com/multimedia/archive/00002/BTN_A_000112243_O_2391a.pdf

[2]Lohmann V. et al. (1999). Replication of Subgenomic Hepatitis C Virus RNAs in a Hepatoma Cell Line. Science. 285 (5424), 110-113.

http://www.sciencemag.org/content/285/5424/110.long