Part:BBa_K1041001:Design

Neomycin Resistance Coding Device + AntG promoter

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 NgoMIV site found at 757
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 606
  Illegal SapI.rc site found at 816
  

Design Notes

Team NRP-UEA_Norwich 2013 designed this part to contain the promoter sequence AntG, where the unique sigma factor AntA binds to activate transcription of the neomycin resistance gene. There is a Nde1 resticition site between the promoter and gene to allow either to be removed and exchanged for a different promoter or gene. This facilitated cloning of the biobrick Bba_K1041002

Source

This part was designed by our team to aid our project in identifying new strains of Antimycin-Producing Actinomycetes.The 14 known 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.

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