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Module 2 : Gene Sequences and Locations for PCR

G4 : RcsB

G5 : B3023

G6 : WaaL Ligase

G7 + G8 : OtsA and OtsB

Promoter 1

Promoter 1 (P1): BBa_I0500

Specs: Inducible

Notes: Induced by L-arabinose

Present in the 2009 kit

Use modelling to determine how much arabinose should be used


Promoter 2

Promoter 2 (P2): BBa_I714889

Specs: Constitutive & Repressible

Notes: Works but not present in the 2009 kit


Promoter 2 (P2): BBa_R0040

Specs: Constitutive & Repressible

Notes: Repressed by TetR. Present in the 2009 kit


Promoter 2 (P2): BBa_R0053

Specs: Repressible with p22 cII repressor protein(BBa_C0053)

Notes: Not compatible with RBS B0030. This combination yielded very low gfp expression(BBa_I7108)
Present in 2009 kit


Promoter 2 (P2): BBa_K113001

pT7->cI434 operator ->GFP Specs: Repressible operator

Notes: cI434 operator can repress the gene expression once the concentration of cI434 reaches certain level (threshold detector?) But experience is none and not present in kit


Promoter 3

Promoter 3 (P3): lacI regulated BBa_R0010

Specs: Repressible

Notes: Present in the 2009 kit


Promoter 3 (P3): lambda cI regulated BBa_R0051

Specs: Repressible

Notes: lambda cI regulated, cI binding results in repression of transcription
Present in the 2009 kit


Promoter 3 (P3): lacI regulated BBa_R0011

Specs: Repressible

Notes: lacI regulated, lambda pL hybrid. repressed by LacI,nd induced by IPTG. (Same as P3)


Promoter 3 (P3): pLacIQ1 promoter BBa_K091112

Specs: Repressible

Notes: Reduces leaky transcription.
Present in the 2009 kit 2


Module 3 : Killing strategy


Promoter 4 (nonleaky)

T7 promoter system


RBS 1: BBa_B0034 (Strength = 1) or BBa_B0035 (Strength = 1.124)

Specs: As strong as possible

RBS 2: BBa_B0033 (Strength = 0.01-0.002)

Specs: Weak

RBS 3: BBa_B0035 (Strength = 1.124)

Specs: As strong as possible

RBS 4: BBa_B0034 (Strength = 1)

Specs: Strong


Gene 1 (G1)

Specs: Protein of interest (e.g. Cellulase or PAH)

Gene 2 (G2) BBa_C0056

Specs: P2 repressor

Notes: Present in the 2009 kit


Gene 3 (G3) BBa_C0012

Specs: P3 repressor

Notes: Present in the 2009 kit, LacI repressor


Gene 3 (G3) BBa_C0051

Specs: P3 repressor

Notes: Present in the 2009 kit, cI repressor


Do you want an RBS?

If you do, follow this link http://partsregistry.org/Ribosome_Binding_Sites/Prokaryotic/Constitutive/Community_Collection