Part:BBa_M36056:Design

Thiosulfate Sensor (ThsS)

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1208
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 607
  Illegal BamHI site found at 655
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 1140
  

Design Notes

pThsS utilizes a pD441-CH backbone acquired from DNA 2.0 which contains a high copy origin of replication and Kanamycin resistance genes. Expression of ThsS is driven by a T5 IPTG inducible promoter, sourced from DNA2.0 (Parts registry number: BBa_M50075). Immediately upstream of the coding sequence is a Strong RBS (Parts registry number: BBa_M50080) which is also provided by DNA2.0. The sequence of codon-optimized ThsS was located in Addgene on pKD182 and pasted into our plasmid; this is the exact protein sequence used by Tabor’s lab. We also added a C-terminal 6X His tag (Parts registry number: BBa_K133133) from DNA2.0 to allow for checking for protein production with a western. Transcription of this DNA is terminated by Term_PhageT7 (Parts registry number: BBa_M50060).

Source

• T5 (IPTG Inducible): BBa_M50075
• Strong RBS: BBa_B0032
• 6xHis: BBa_K133133
• Thiosulfate sensor: BBa_M50094
• Term_PhageT7: BBa_M50060

References

• https://parts.igem.org/Part:BBa_M50075
• https://parts.igem.org/Part:BBa_B0032
• https://parts.igem.org/Part:BBa_K133133
• https://parts.igem.org/Part:BBa_M50094
• https://www.ncbi.nlm.nih.gov/pubmed/28373240
• https://parts.igem.org/Part:BBa_M50060