Team:Hong Kong HKUST/data

How our system works:

Data For Our Favorite New Parts:

• 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 parts with qualified sample in the Part Registry:

• Lock 1, BBa_J01008 (Golden Bear, 2005): BioBricked version of Isaacs' riboregulator trans-activating key, taR12
• Lock3c, BBa_J23031 (Team Berkeley iGEM, 2006): lock3c.
• Encodes a short RNA sequence that "unlocks" the biobrick part J01122 (lock3) which is locked RFP., BBa_J23008 (Berkeley iGEM, 2006): Key 3c

Data and submitted physical DNA for parts without qualified sample in the Part Registry:

• 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.
• Key for lock of medium (K175031) RBS (B0032) from the lock/key library TUD09, BBa_K175032: This biobrick generates a mRNA which forms a secondary structure that opens LMR (K175031).
• 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.

  
  
  Data and submitted physical DNA for constructs used in characterization:

  • RBS(Lock 3c), BBa_K1379015: B0032 derivate for Lock 3C.
  • Lock 1-GFP-B0015, BBa_K1379020: Cis-repressed GFP Generator Lock 1
  • Lock 3-GFP-B0015, BBa_K1379024: Cis-repressed GFP Generator Lock 3C
  • K175031-GFP-B0015, BBa_K1379046: GFP generator using a Lock 1 modified RBS.
  • RBS(Lock 3c)-GFP-B0015, BBa_K1379048: Repression measurement device 1 with trans activator.
  
  
  

We also constructed the following new parts:

• HKUST Lock 1, BBa_K1379018: Cis-Repressor HKUST Lock 1.
• HKUST Key 1, BBa_K1379019: Trans-Activator for HKUST Key 1.
• HKUST Lock 1-GFP-B0015, BBa_K1379028: Cis-repressed GFP Generator HKUST Lock 1

We constructed the following intermediates/measurement devices during characterization:

• Key 3-B0015, BBa_K1379023
• K175032-B0015, BBa_K1379027
• HKUST Key 1-B0015, BBa_K1379029
• J23102-Lock 1-GFP-B0015, BBa_K1379030
• J23102-Lock 3-GFP-B0015, BBa_K1379032
• pBad/araC-Key 3, BBa_K1379033
• J23102-Lock 3c-GFP-B0015, BBa_K1379034
• pBad/araC-Key 3c, BBa_K1379035
• J23102-K175031-GFP-B0015, BBa_K1379036
• pBad/araC-K175032, BBa_K1379037
• J23102-HKUST Lock 1-GFP-B0015, BBa_K1379038
• pBad/araC-HKUST Key 1, BBa_K1379039