Part:BBa_K1041002:Design

AntG Promoter + RFP Coding Device

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 683
  Illegal AgeI site found at 795
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

Team NRP-UEA_Norwich 2013 created this part using biobricks BBa_K1041000 and BBa_K1041001. These biobricks both contain an Nde1 site after their promoter sequence, enabling a restriction digest to be performed. The RFP coding gene was excised from BBa_K1041000 and ligated downstream of the AntG promoter of BBa_K1041001 to create this new biobrick.

Source

This part was designed by our team to aid our project in identifying new strains of Antimycin-Producing Actinomycetes. The 14 known antimycin biosynthetic gene clusters contain four operons: antAB, antCDE, antFG and antHIJKLMNO. The antA gene encodes a unique ECF RNA polymerase sigma factor, referred to as σAntA, which has the sole function of regulating antimycin synthesis by activating transcription of the antFG and antHIJKLMNO genes [1]. Homologues of the AntA sigma factor, the key regulatory protein in antimycin biosynthesis, are present in all known gene clusters [2]. Due to this property a biosensor was designed with the AntA-regulated promoter (antGp) controlling the expression of the reporter neomycin resistance gene. This part was produced by performing restriction digests of the parts BBa_K1041000 and BBa_K1041001.

References

1.Speike, R., Barke, J., Brearley, C., Hill, L., Yu, D., Goss, R & Hutchings, M (2011) A single Streptomyces Symbiont Makes Multiple Antifungals to Support the Fungus Farming Ant Acromyrmex Octospinosus, PlosOne, Volume: 6, Issue: 8.
2.Sandy, M., Rui, Z., Gallagher, J & Zhang, W (2012) Enzymatic Synthesis of the Dilactone Scaffold of Antimycins, American Chemical Society