Part:BBa_K3766112

sfGFP-LVAtag expression cassette under the control of pTacI promoter

This part is an sfGFP-LVAtag (BBa_K2675006) expression cassette under the control of the pTacI promoter BBa_K864400.

Usage and Biology

In this composite part sfGFP-LVAtag (BBa_K2675006) was equipped by a custom made RBS (BBa_K2675017) and placed under the control of the pTacI promoter BBa_K864400 and of the strong L3S2P56 Terminator (BBa_K2675030.

The LVA degradation tag (AANDENYALVA), that equips the superfolder GFP (sfGFP) in this expression cassette, is a ssrA tag that accelerates protein degradation in Escherichia coli at 37°C [1]. It is used in synthetic biology to rapidly degrade reporter genes and thus finely observe the synthetic gene networks dynamics.

pTacI is a strong IPTG inducible promoter, regulated by the LacI transcription factor [2].

References

[1] Purcell O, Grierson CS, Bernardo Md, Savery NJ. Temperature dependence of ssrA-tag mediated protein degradation. J Biol Eng (2012) 6, 10.

[2] de Boer HA, Comstock LJ, Vasser M. The tac promoter: a functional hybrid derived from the trp and lac promoters. Proceedings of the National Academy of Sciences of the United States of America (1983) 80: 21–25.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 116
  
• 1000
  COMPATIBLE WITH RFC[1000]