Team:Technion/Attributions

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* We have created[https://2012.igem.org/Team:Technion/Project/Reporter new reporter system] to test our YES gates and hope that it will become standard system for testing other parts as well. We used it to characterize [https://2012.igem.org/Team:Technion/Project/RNAPs#WT_polymerases T7 RNAP] activity, the results can be seen [https://2012.igem.org/Team:Technion/Project/Reporter#Characterization_of_the_Assays here].
* We have created[https://2012.igem.org/Team:Technion/Project/Reporter new reporter system] to test our YES gates and hope that it will become standard system for testing other parts as well. We used it to characterize [https://2012.igem.org/Team:Technion/Project/RNAPs#WT_polymerases T7 RNAP] activity, the results can be seen [https://2012.igem.org/Team:Technion/Project/Reporter#Characterization_of_the_Assays here].
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* We have managed to combine RS with T7, T7*(T3), T7*(K1F), T7*(N4) by using fusion PCR and put the resulting construct under control of pLac, more details can be seen at [https://2012.igem.org/Team:Technion/Project/RS RS section]. Due to lack of time we hadn't transferred RS+RNAP constructs under pLux control, as was planned at the beginning.
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* We have added MCS to pSB1C3 plasmid backbone. This backbone is very convenient to work with, however some of our parts had the same restriction sites as standard prefix and suffix found in the backbone, so we couldn't insert them into the pSB1C3 plasmid. Addition of MCS helps to circumvent this problem.
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* We had produced a working [https://2012.igem.org/Team:Technion/Project/YES_gates#YES_gate_.231 YES gate #1] with T7* RNAP

Revision as of 14:15, 26 September 2012



Attributions

During our project we have used parts that were given to us by different labs or were taken from the registry/distribution kit.

Donated parts

  • Ann K. Ganesan - WT T7 RNAP
  • Changwon Kang - WT SP6 RNAP
  • Christopher A. Voight - T7*, T7*(T3), T7*(N4), T7*(K1F) RNAPs with suitable induced promoters: pT7, pT3, pK1F, pN4
  • Gallivan Lab (Gallivan Justin P., Lynch Sean A.) - Theophylline riboswitch and pTAC
  • Roee's Lab (Roee Amit) - Cerulean FP under control of pLac/Ara, mCitrin FP and mCherry under control of pTetO.

Parts from Distribution Kit

Name Description
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 BBa_B0015] Double terminator
[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] TetR controllable LuxR and lux pR genes
[http://partsregistry.org/Part:BBa_I0462 BBa_I0462]** LuxR protein generator
[http://partsregistry.org/Part:BBa_I13522 BBa_I13522]** GFP under pTetO control
[http://partsregistry.org/Part:BBa_J06504 BBa_J06504]** mCherry FP
[http://partsregistry.org/Part:BBa_J06702 BBa_J06702] mCherry, bacterial with RBS and forward terminator
[http://partsregistry.org/Part:BBa_J23119 BBa_J23119] Constitutive promoter
[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] Promoter (luxR & HSL regulated -- lux pR)
[http://partsregistry.org/Part:pSB1C3 pSB1C3] High copy BioBrick assembly plasmid
[http://partsregistry.org/Part:pSB3C5 pSB3C5] Low to medium copy BioBrick standard vector

Notes: Parts followed by ** are parts that haven't worked because distribution kit contained some other DNA in the well where the part should be.

Contributions

  • We have submitted 25 BioBricks to the registry. Due to lack of time we haven't managed to put all the parts inside the shipping plasmid.
  • We have managed to successfully clone 4 out of 8 parts that phage's genome had been divided into.
  • We have creatednew reporter system to test our YES gates and hope that it will become standard system for testing other parts as well. We used it to characterize T7 RNAP activity, the results can be seen here.
  • We have managed to combine RS with T7, T7*(T3), T7*(K1F), T7*(N4) by using fusion PCR and put the resulting construct under control of pLac, more details can be seen at RS section. Due to lack of time we hadn't transferred RS+RNAP constructs under pLux control, as was planned at the beginning.
  • We have added MCS to pSB1C3 plasmid backbone. This backbone is very convenient to work with, however some of our parts had the same restriction sites as standard prefix and suffix found in the backbone, so we couldn't insert them into the pSB1C3 plasmid. Addition of MCS helps to circumvent this problem.
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