Part:BBa_K1041004:Design
AntG Promoter + Gus gene
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 498
Illegal NgoMIV site found at 630
Illegal NgoMIV site found at 1227 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 896
Design Notes
Team NRP-UEA_Norwich 2013 desgined this part using biobrick BBa_K1041001. This biobrick contains an Nde1 site after its promoter sequence, enabling a restriction digest to be performed. The Neomycin resistance coding gene would be excised from BBa_K1041001 and a gene encoding Gus ligated downstream of the promoter of BBa_K1041001 to create a new biobrick. The protein product of the GUS gene provides a reporter that can indicate expression of the gene.
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 Gus. This part was produced by performing restriction digests of the part 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