From 2012.igem.org

Data page: Summary of what we have done this year (so far!)

New frog plasmids

Fig 1: Eucaryotic expression plasmid having both transcriptional and translational reagulational elements.

• K812000: PCS2+ plasmid backbone for eukaryotic expression system containing regulational elements crucial for the efficacy of such system

The auxin system

The auxin receiver system

Fig 2: schemetic representation of the Auxin-receiver system

• K812010:GFP fusion with the ubuquitinase E3 OsTirI recoginition domain which a main part of the degron system
• K812012:OsTirI Ubiquitinase E3 for AID tagged protein degradation in the presence of auxin
• K812013:
  The Auxin reciever system golden gate-assembled containing GFP-AID OsTirI polysistronic system for auxin detection in tadpole

The auxin emission system

Fig 3: Schematic representation of the Auxin two-step production cassette.

• K812020:IAAH enzyme that catalyzes the second and last step towards auxin production
• K812021:IAAM enzyme that catalyses the transformation of Tryptophan to Inodle-3-acetamide
• K812014:Auxin production cassette containg both enzymes above which is monocystronic

Reporters and promotors

Summery

In our Project we planned to screen a series of different reporters along with different tissue specific promotors. This was done to validate cross-talk between the emitter-receiver system and and screen various reporters to elucidate the efficiency of different reporters in Tadpole manipulation.

• K812030:Citrine reporter with a Kozak sequence for eukaryotic expression
• K812031:sfGFP with kozak sequence for eukaryotic expression
• K812032:mCFP with kozak sequence for eukaryotic expression
• K812200:Biobricked pSC2+ plasmid with Elastase promoter for frogs and chicken
• K812300:Biobricked pSC2+ plasmid with HSP70 promoter for frogs and chicken
• K812233:sfGFP in pCS2+ downstream to elastase promoter
• K812331:sfGFP in pCS2+ downstream to HSP70 promoter

Characterization

the information on characterized part can be found herein:

• FrenchFrog: ,/ul>

  Goldenbricks

  Summary

  

  

  Fig 4: Summary of the GoldenBrick procedure

  The GoldenBrick is a new assembly method for the partsregistry. If you want to know more, see this page.

  Our favourites parts

  • K812050: A GoldenBricked version of pSB1C3 with J04450 as negative cloning control
  • K812051: A GoldenBricked version of pSB1K3 with J04450 as negative cloning control

  The other parts we have created

  • K812050: A GoldenBricked version of pSB1C3 with J04450 as negative cloning control
  • K812051: A GoldenBricked version of pSB1K3 with J04450 as negative cloning control
  • K812053: A GoldenBricked version of the strong RBS B0034
  • K812054: A GoldenBricked version of the RFP E1010
  • K812055: A GoldenBricked version of the terminator B0015
  • K812056: A GoldenBricked version of the pLac R0010 promoter
  • K812057: A GoldenBricked of an sfGFP protein
  • K812058: A GoldenBricked of medium strenght RBS J61107
  • K812059: A GoldenBricked of week RBS J61117

  say we are testing them

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