Team:Hong Kong HKUST/data

How our system works:

• σ^x, BBa_K1379004: gene sequence of competence protein ComX which produces σ^x factor. σ^x factor induces promoter containing Com-Box sequence (P_celA and P_comFA).
• P_celA promoter, BBa_K1379000: a σ^x-inducible promoter (containing Com-Box sequence) which initiates the transcription of competence CelA protein.
• P_comFA promoter, BBa_K1379001: a σ^x-inducible promoter (containing Com-Box sequence) which initiates the transcription of ComFA protein.

Data for Pre-existing Parts:

• Strong promoter, strong RBS combination, BBa_K880005 (Team Michigan iGEM, 2012): combination of strong constitutive promoter (BBa_J23100) and strong RBS (BBa_B0034) was used as a backbone for a very high level of expression of ComX protein (σ^x factor).
• GFP generator, BBa_E0240 (Antiquity Boston University iGEM, 2004): green fluorescent protein reporter (BBa_E0040) with RBS (BBa_B0032) and double terminator (BBa_B0015) was used as a reporter for characterization of P_celA and P_comFA promoters.

We Also Built and Characterized the Following Parts:

• σ^x Generator, BBa_K1379006: comX gene sequence was assembled with constitutive promoter, RBS and double terminator so ComX protein is produced constitutively.
• P_celA-GFP Generator, BBa_K1379002: σ^x-inducible P_celA promoter was assembled with GFP generator.
• P_comFA-GFP Generator, BBa_K1379003: σ^x-inducible P_comFA promoter was assembled with GFP generator.
• σ^x Generator, P_celA-GFP Generator, BBa_K1379005: σ^x-generator was assembled with P_celA-GFP generator. Following the assemblage of both constructs, GFP was successfully expressed.
• σ^x Generator, P_comFA-GFP Generator, BBa_K1379007: σ^x-generator was assembled with P_comFA-GFP generator. Following the assemblage of both constructs, GFP was successfully expressed.

We Built but did not Characterize the Following Parts:

• comE, BBa_K1379051: gene sequence of response regulator ComE which induces the P_comCDE promoter.

Data For Our Favorite New Parts:

• Lock 3C RBS, BBa_K1379015: B0032 derivate for Lock 3C.
• HKUST Lock 1, BBa_K1379018: Cis-Repressor HKUST Lock 1.
• HKUST Key 1, BBa_K1379019: Trans-Activator for HKUST Key 1.
• Cis-repressed GFP Lock 1, BBa_K1379020: Cis-repressed GFP Generator Lock 1
• Cis-repressed GFP Generator Lock 3, BBa_K1379024: Cis-repressed GFP Generator Lock 3C
• Cis-repressed GFP Lock MCR, BBa_K1379028: Cis-repressed GFP Generator HKUST Lock 1
• Lock 1 RBS GFP generator, BBa_K1379047: GFP generator using a Lock 3C modified RBS.

Parts that we constructed and submitted:

• Inducible pBad/araC promoter, BBa_I0500: Inducible pBad/araC promoter.
• Riboregulator Lock 1, BBa_J01010:Biobrick version of Isaacs' riboregulator cis repressed lock, crR12.
• Lock3, BBa_J01080: Biobrick version of Isaacs' riboregulator cis repressed lock, crR12, and modified version of Riboregulator Lock 1.
• Key3, BBa_J01086: Biobricked version of Isaacs' riboregulator trans activating key, taR12, and modified version of Riboregulator Key 1.
• Lock for medium RBS (B0032) from the lock/key library TUD09, BBa_K175031: This biobrick generates a mRNA which forms a secondary structure blocking the (medium) RBS.
• B0034 derived RBS to match spacing of J01122, BBa_J23080: RBS spacing modification to [RBS] such that the spacing between the shine-delgarno and the ORF is the same as it is in J01122.
• Key for lock of medium (K175031) RBS (B0032) from the lock/key library TUD09, BBa_K175031: This biobrick generates a mRNA which forms a secondary structure that opens LMR (K175031).

Parts that already exist in Parts Registry and are currently under characterization:

• Riboregulator Lock 1, BBa_J01008 (Golden Bear, 2005): BioBricked version of Isaacs' riboregulator trans-activating key, taR12
• Lock 3c, BBa_J23031 (Team Berkeley iGEM, 2006): lock3c.
• Key 3c, BBa_J23008 (Berkeley iGEM, 2006): Encodes a short RNA sequence that "unlocks" the biobrick part J01122 (lock3) which is locked RFP.

Parts that we designed but did not submit:

• Key 1 Double Terminator, BBa_K1379021: Trans-Activator with terminator for lock 1.
• Key 3 double terminator, BBa_K1379023: Trans-Activator with terminator for lock 3.
• Key 3c double terminator, BBa_K1379025: Trans-Activator with terminator for lock 3C
• MTA double terminator, BBa_K1379027: Trans-Activator with terminator for MCR.
• HKUST Key 1 double terminator, BBa_K1379029: Trans-Activator with terminator for HKUST Lock 1.
• Cis-Repressing device 1, BBa_K1379030: Constitutive promoted, Cis-repressed GFP Coding device Lock 1.
• AraC Key 1, BBa_K1379031: AraC inducible Trans-Activator device, Key 1
• Cis-Repressing device 3, BBa_K1379032: Constitutive promoted, Cis-repressed GFP Coding device Lock 3.
• AraC Key 3, BBa_K1379033: AraC inducible Trans-Activator device, Key 3
• Cis-Repressing device 3C, BBa_K1379051: Constitutive promoted, Cis-repressed GFP Coding device Lock 3C.
• AraC Key 3C, BBa_K1379035: AraC inducible Trans-Activator device for, Key 3C.
• Cis-Repressing device MCR, BBa_K1379036: Constitutive promoted, Cis-represed GFP Coding device Lock MCR
• AraC MTA, BBa_K1379037: AraC inducible Trans-Activator device, MTA
• Cis-Repressing device HKUST 1, BBa_K1379038: Constitutive promoted, Cis-repressed GFP Coding device HKUST Lock 1.
• AraC HKUST Key 1, BBa_K1379039: AraC inducible Trans-Activator device, HKUST Key 1.
• Repression device 1, BBa_K1379040: Repression measurement device 1.
• Repression device 3, BBa_K1379041: Repression measurement device 3.
• Repression device 3C, BBa_K1379042: Repression measurement device 3C.
• Repression device Medium, BBa_K1379043: Repression measurement device Medium.
• Cis-Repression device HKUST 1, BBa_K1379044: Repression measurement device HKUST 1.
• Cis repressed GFP lock 1, with Trans-activator key 1, BBa_K1379049: Cis repressed GFP lock 1, with Trans-activator key 1.