Part:BBa_K303001:Design

Ptet/lac with J23106

Assembly Compatibility:

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

  Illegal NheI site found at 32
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

The part was synthesized as linear DNA and was placed inside its vector using biobrick assembly.

Two sets of promoters were designed to respond to tetracycline and lactose, tetracycline and arabinose, and lactose and arabinose, using J23106 and J23110 as bases. According to literature, a promoter with an activation site that is not activated is not entirely inactive. RNA polymerase can still bind, only the interaction is weaker. Promoters J23106 and J23110 were chose because they are of relatively moderate strength. It is predicted that until arabinose is present in the system, the relative strength of these promoters will be fairly weak. When coupled with weak ribosomal binding sites, the rate at which proteins are translated should be negligible, preventing false positives.

Source

The constitutive promoter J23106 was used to construct this part. The promoter was modified by adding two new operator sites.

References

Tet:

1. Lutz R and Bujard H. Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res 1997 Mar 15; 25(6) 1203-10. pmid:9092630. PubMed HubMed [Lutz-1997]

Lac:

1. Gilbert, Walter; Maxam, Allen. (1973). The Neculeotide Sequence of the lac Operator. Proc. Natl. Acad. Sci. USA. 70(12):3581-84