Part:BBa_K3771098

PsoxS-Riboj-RCP-J23101-SoxR​

Description

This composite part consists of promoter P_soxS Mutant 3 (BBa_K2862010), genetic insulator RiboJ (BBa_K1679038), RCP (BBa_K592012), constitutive promoter J23101 (BBa_J23101), and the soxR gene (BBa_K223000).

The soxR and the soxS gene are components of the soxRS regulon, which is important for E. coli to sense and respond to the oxidants. When SoxR protein is fully oxidized, it becomes a powerful transcription activator of the soxS promoter (P_soxS) (up to 100-fold), leading to the expression of the downstream gene^[1]. RiboJ is a genetic insulator that can increase the expression of insulated genes. RCP serves as a reporter to help us pick up the correct colony.

Usage and Biology

This composite part was ligated with the pSAA vector and transformed into E. coli. We conducted colony PCR to verify whether E. coli uptake the correct plasmid. The size of the PCR product was as expected.

Fig. 1. The electrophoresis result of colony PCR. M: Marker; Lane 1: pSAA-P_soxS-sfgfp (990 bp); Lane 2, 3: pSAA-P_soxS-RiboJ-rcp-J23101-soxR (1103 bp).​

References

1. Pomposiello PJ, Demple B. Redox-operated genetic switches: the SoxR and OxyR transcription factors. Trends Biotechnol. 2001;19(3):109-114. doi:10.1016/s0167-7799(00)01542-0

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 890
  Illegal NheI site found at 913
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]