Team:UPO-Sevilla/Project/Improving Flip Flop/Data Page

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                          <p>The <strong>Improved Flip Flop</strong> (Part: <a href="http://partsregistry.org/Part:BBa_K510019" target="_blank">BBa_K510019</a> and Part: <a href="http://partsregistry.org/Part:BBa_K510036" target="_blank">BBa_K510036</a>) is based on the basic flip flop (Part: <a href="http://partsregistry.org/Part:BBa_K177038" target="_blank">BBa_K177038</a>) and was built with devices which are in the Registry of Standard Biological Parts. As for the different sequences used in our construction, here it is a brief description of each one:</p>
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<li><p> <strong>cI promoter </strong>(Part: <a href="http://partsregistry.org/Part:BBa_R0051" target="_blank">BBa_R0051</a>): The cI regulated promoter is based on the pR promoter from bacteriophage lambda. The promoter has two DNA binding sites for lambda cI repressor BBa_C0051. cI binding results in repression of transcription. The specific sequence used here is based on the cI repressible promoter used in the Elowitz repressilator (and references therein).</p></li>
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<li><p><strong>Lac promoter </strong>(Part: <a href="http://partsregistry.org/Part:BBa_R0010" target="_blank">BBa_R0010</a>): This part is an inverting regulator sensitive to LacI and CAP. It contains two protein binding sites. The first binds CAP protein, which is generally present in E.coli and is associated with cell health and availability of glucose. The second one binds LacI protein.</p> </li>
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                          <p>As for the different sequences used in our construction, here it is a brief description of each one:</p>
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<li><p><strong>RBS.3 medium </strong> (Part: <a href="http://partsregistry.org/Part:BBa_B0032" target="_blank">BBa_B0032</a>):This part is a weak RBS based on Ron Weiss thesis.</p> </li>
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<li><p><strong>LacI:</strong> Our LacI was taken from E. coli strain K-12 substrain MG1655 chromosome genome and it has a total of 1083 bp.</p></li>
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<p>- <strong>cI promoter (Part: BBa_R0051):</strong> The cI regulated promoter is based on the pR promoter from bacteriophage lambda. The promoter has two DNA binding sites for lambda cI repressor BBa_C0051. cI binding results in repression of transcription. The specific sequence used here is based on the cI repressible promoter used in the Elowitz repressilator (and references therein).</p>
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<li><p><strong>GFP </strong>(Part: <a href="http://partsregistry.org/Part:BBa_E0040" target="_blank">BBa_E0040</a>): Green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP.</p></li>
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<p>- <strong>Lac promoter (Part: BBa_R0010):</strong> This part is an inverting regulator sensitive to LacI and CAP. It contains two protein binding sites. The first binds CAP protein, which is generally present in E.coli and is associated with cell health and availability of glucose. The second one binds LacI protein.</p>
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<li><p><strong>DAS+4:</strong> Short tag at the end of the proteins. For more information see Figure 2 of Proteolysis section.</p></li>
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<li><p><strong>Temperature sensitive cI </strong>(Part: <a href="http://partsregistry.org/Part:BBa_K177050" target="_blank">BBa_K177050</a>): Thermosensitive version of the lambda phage repressor. Functions in 30°C and it's inactivated in 42°C</p></li>
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<p>    o In the absence of LacI and CAP proteins, this part promotes transcription.</p>
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<li><p><strong>SspB:</strong> Taken from <a href="http://www.ncbi.nlm.nih.gov/pubmed/16762842" target="_blank">McGinness et al. (2006)</a>. For more information, see Proteolysis section. The OmpN::SspB fusion is detailed in Figure 5 of the asRNA section.</p> </li>
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<p>    o In the presence of LacI and CAP proteins, this part inhibits transcription.</p>
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<li><p><strong>RFP </strong>(Part: <a href="http://partsregistry.org/Part:BBa_E1010" target="_blank">BBa_E1010</a>): Highly engineered mutant of red fluorescent protein from Discosoma striata (coral). Monomeric RFP with its excitation peak at  584 nm and its emission peak at 607 nm. The OmpN::RFP fusion is detailed in Figure 5 of the asRNA section.</p></li>
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<p>    o LacI can be inhibited by IPTG.</p>
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<li><p><strong>Double terminator </strong>(Part <a href="http://partsregistry.org/Part:BBa_B0015" target="_blank">BBa_B0015</a>): Double terminator consisting of <a href="http://partsregistry.org/Part:BBa_B0010" target="_blank">BBa_B0010</a> and <a href="http://partsregistry.org/Part:BBa_B0012" target="_blank">BBa_B0012</a>.This is the most commonly used terminator and it seems to be reliable.</p></li>
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<p>- <strong>RBS.3 (medium):</strong> This part is a weak RBS based on Ron Weiss thesis.</p>
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<li><p><strong>RybB asRNA:</strong> Taken from <a href="http://www.ncbi.nlm.nih.gov/pubmed/19111662"_blank">Bouvier et al.(2008)</a>. For further information see Figure 2 of the asRNA section.</p></li>
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<p>- <strong>LacI:</strong> Our LacI was taken from E. coli strain K-12 substrain MG1655 chromosome genome and it has a total of 1083 bp.</p>
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<p>- <strong>GFP (Part: BBa_E0040):</strong> Green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP.</p>
+
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<p>- <strong>DAS+4:</strong> Short tag at the end of the proteins. For more information see Figure 2 of Proteolysis section.</p>
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<p>- <strong>Temperature sensitive cI (Part: BBa_K177050):</strong> Thermosensitive version of the lambda phage repressor. Functions in 30°C and it's inactivated in 42°C</p>
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<p>- <strong>SspB:</strong> Taken from McGinness et al. (2006). For more information, see Proteolysis section. The OmpN::SspB fusion is detailed in Figure 5 of the asRNA section.</p>
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<p>- <strong>RFP (Part: BBa_E1010):</strong> Highly engineered mutant of red fluorescent protein from Discosoma striata (coral). Monomeric RFP with its excitation peak at  584 nm and its emission peak at 607 nm. The OmpN::RFP fusion is detailed in Figure 5 of the asRNA section.</p>
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<p>- <strong>Double terminator (Part BBa_B0015):</strong> Double terminator consisting of BBa_B0010 and BBa_B0012.This is the most commonly used terminator and it seems to be reliable.</p>
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<p>- <strong>RybB asRNA:</strong> Taken from Bouvier et al. (2008). For further information see Figure 2 of the asRNA section.</p>
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Latest revision as of 22:14, 27 October 2011

Grey iGEM Logo UPO icon

Data Page


The Improved Flip Flop (Part: BBa_K510019 and Part: BBa_K510036) is based on the basic flip flop (Part: BBa_K177038) and was built with devices which are in the Registry of Standard Biological Parts. As for the different sequences used in our construction, here it is a brief description of each one:


  1. cI promoter (Part: BBa_R0051): The cI regulated promoter is based on the pR promoter from bacteriophage lambda. The promoter has two DNA binding sites for lambda cI repressor BBa_C0051. cI binding results in repression of transcription. The specific sequence used here is based on the cI repressible promoter used in the Elowitz repressilator (and references therein).

  2. Lac promoter (Part: BBa_R0010): This part is an inverting regulator sensitive to LacI and CAP. It contains two protein binding sites. The first binds CAP protein, which is generally present in E.coli and is associated with cell health and availability of glucose. The second one binds LacI protein.

  3. RBS.3 medium (Part: BBa_B0032):This part is a weak RBS based on Ron Weiss thesis.

  4. LacI: Our LacI was taken from E. coli strain K-12 substrain MG1655 chromosome genome and it has a total of 1083 bp.

  5. GFP (Part: BBa_E0040): Green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP.

  6. DAS+4: Short tag at the end of the proteins. For more information see Figure 2 of Proteolysis section.

  7. Temperature sensitive cI (Part: BBa_K177050): Thermosensitive version of the lambda phage repressor. Functions in 30°C and it's inactivated in 42°C

  8. SspB: Taken from McGinness et al. (2006). For more information, see Proteolysis section. The OmpN::SspB fusion is detailed in Figure 5 of the asRNA section.

  9. RFP (Part: BBa_E1010): Highly engineered mutant of red fluorescent protein from Discosoma striata (coral). Monomeric RFP with its excitation peak at 584 nm and its emission peak at 607 nm. The OmpN::RFP fusion is detailed in Figure 5 of the asRNA section.

  10. Double terminator (Part BBa_B0015): Double terminator consisting of BBa_B0010 and BBa_B0012.This is the most commonly used terminator and it seems to be reliable.

  11. RybB asRNA: Taken from Bouvier et al.(2008). For further information see Figure 2 of the asRNA section.