biosketchharvard igem 2005 1 biobrick: aggregate parts quad part inverter (qpi) rbs codingregion...
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BioSketchBioSketch Harvard iGEM 2005Harvard iGEM 2005
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BioBrick: Aggregate PartsBioBrick: Aggregate Parts
Quad Part Inverter (QPI)Quad Part Inverter (QPI)
RBSRBSCodingCodingRegionRegion
TerminatorTerminator
PromoterPromoter RBSRBS TerminatorTerminator
ReporterReporterCodingCodingRegionRegion
ReporterReporter
BioSketchBioSketch Harvard iGEM 2005Harvard iGEM 2005
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QPIs and ReportersQPIs and Reporters
LacI QPILacI QPI
GFP (mut3b)GFP (mut3b)
Lambda cI QPILambda cI QPI
434 cI QPI434 cI QPI
HaveHave
LacI(ts) QPILacI(ts) QPI
VenusVenus
Lambda cI857 QPILambda cI857 QPI
NeedNeed
mCherrymCherry
BioSketchBioSketch Harvard iGEM 2005Harvard iGEM 2005
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Test ConstructsTest ConstructsGFP (mut3b)GFP (mut3b)
LacI QPILacI QPI
GFP (mut3b)GFP (mut3b)
Lambda cI QPILambda cI QPI
GFP (mut3b)GFP (mut3b)
434 QPI434 QPI
PPlambdalambda
PPlambdalambda
PPLacLac
GFP (mut3b)GFP (mut3b)
LacI(ts) QPILacI(ts) QPI
GFP (mut3b)GFP (mut3b)
857 QPI857 QPI
PP434434
PPlambdalambda
LacI QPILacI QPI
PPlambdalambda
GFP (mut3b)GFP (mut3b)
Lambda cI QPILambda cI QPI
PPlambdalambda
GFP (mut3b)GFP (mut3b)
Lambda cI QPILambda cI QPI
LacI(ts) QPILacI(ts) QPI
PP434434
GFP (mut3b)GFP (mut3b)
434 QPI434 QPI
857 QPI857 QPI
BioSketchBioSketch Harvard iGEM 2005Harvard iGEM 2005
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BioBricks TestingBioBricks TestingConditioConditionn
Temp (Temp (°° C) C) 3030 3030 3030 3030 4040 4040 4040 4040
IPTGIPTG (-)(-) (-)(-) (+)(+) (+)(+) (-)(-) (-)(-) (+)(+) (+)(+)
UVUV (-)(-) (+)(+) (-)(-) (+)(+) (-)(-) (+)(+) (-)(-) (+)(+)
QPIsQPIs LacILacI (-)(-) (-)(-) (+)(+) (+)(+) (-)(-) (-)(-) (+)(+) (+)(+)
lambdalambda (-)(-) (+)(+) (-)(-) (+)(+) (-)(-) (+)(+) (-)(-) (+)(+)
434434 (-)(-) (+)(+) (-)(-) (+)(+) (-)(-) (+)(+) (-)(-) (+)(+)
857857 (-)(-) (-)(-) (-)(-) (-)(-) (+)(+) (+)(+) (+)(+) (+)(+)
LacI(ts) LacI(ts) (-)(-) (-)(-) (+)(+) (+)(+) (+)(+) (+)(+) (+)(+) (+)(+)
ConditioConditionn
StimulusStimulus UVUV heatheat IPTGIPTG nonenone
Initial StateInitial State (-)(-) (+)(+) (-)(-) (+)(+) (-)(-) (+)(+) (-)(-) (+)(+)
Toggle Toggle SwitcheSwitchess
LacI (Collins)LacI (Collins) (+)(+) (+)(+) (-)(-) (+)(+) (-)(-) (-)(-) (-)(-) (+)(+)
857857 (+)(+) (+)(+) (-)(-) (-)(-) (-)(-) (+)(+) (-)(-) (+)(+)
LacI(ts) LacI(ts) (+)(+) (+)(+) (-)(-) (-)(-) (-)(-) (-)(-) (-)(-) (+)(+)
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Things We Want to KnowThings We Want to KnowMinimal exposure time to UV to switch stateMinimal concentration of IPTG to switch stateMinimal exposure time to heat to switch stateAmount of time a given state is maintainedTime delay between input and reportingEffect of varying cell densitiesEffect of competing inputsDoes SOS work in stationary phaseEffectiveness of UV laser pointer
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Schedule - Week 1Schedule - Week 1Week 1
Mutation
Begin introducing ts mutation into LacI coding region and LacI QPI
BioBrick Assembly
RBS + 857 T + Plambda
mCherry + T Venus + TPlambda + QPILacI QPIlambda + GFP
QPI434 + GFP
Collins
BioSketchBioSketch Harvard iGEM 2005Harvard iGEM 2005
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Schedule - Week 2Schedule - Week 2Week 2
Mutation
Continue mutations
BioBrick Assembly
RBS-857 + T-Plambda (QPI857)
Plambda-QPILac + QPIlambda-GFP
Plambda-QPILac + GFP
Plac + QPIlambda-GFP
Plambda + QPI434-GFP
RBS + mCherry-T RBS + Venus-T
Collins
BioSketchBioSketch Harvard iGEM 2005Harvard iGEM 2005
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Schedule - Week 3Schedule - Week 3Week 3 Mutation Sequence mutations
BioBrick Testing
Plambda-QPILac-GFP
Plac-QPIlambda-GFP
Plambda-QPI434-GFP
Plambda-QPILac-QPIlambda-GFP (Collins)
BioBrick Assembly
P434 + QPI857
Collins
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Schedule - Week 4Schedule - Week 4Week 4 BioBrick
Testing IIPlambda-QPILac-GFP
Plac-QPIlambda-GFP
Plambda-QPI434-GFP
Plambda-QPILac-QPIlambda-GFP (Collins)
BioBrick Assembly
P434-QPI857 + GFP
P434-QPI857 + QPI434-GFP
Plambda + QPILac(ts)
Collins
BioSketchBioSketch Harvard iGEM 2005Harvard iGEM 2005
1010
Schedule - Week 5Schedule - Week 5Week 5 BioBrick
TestingP434-QPI857-GFP
P434-QPI857-QPI434-GFP (857 system)
BioBrick Assembly
Plambda-QPILac(ts) + GFP
Plambda-QPILac(ts) + QPIlambda-GFP
Collins
BioSketchBioSketch Harvard iGEM 2005Harvard iGEM 2005
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Schedule - Week 6Schedule - Week 6Week 6 BioBrick
Testing IIP434-QPI857-GFP
P434-QPI857-QPI434-GFP (857 system)
BioBrick Testing
Plambda-QPILac(ts)-GFP
Plambda-QPILac(ts)-QPIlambda-GFP (Lac(ts) system)
Collins