Part:BBa_K2633000
proB + RBS + Pseudomonas acdS+ linker + GFPmut3b + term
This part includes promoter proB the acdS open reading frame from Pseudomonas sp. 1C2P-04, a 10 amino acid flexible linker (BBa_K105012), GFPmut3b (BBa_E0040) open reading frame, a LEHHHHHH tag, and T7 terminator (BBa_B0012). The part is carried on the pSB1C3 backbone. The promoter proB was modified with a single nucleotide change, position 82 T to A, from the proB used in (Bienick et al. 2014). This removed the XbaI restriction site and made the part legal for the iGEM Registry. Colonies observed were not fluorescent compared to the original promoter. It was observed that the restriction-illegal version of the part produced fluorescent colonies in Enterobacter ludwigii FCP2-01, a grass endophyte isolated by MichiganState 2018. This part is stable in bacterial inoculants in and on Brachypodium distachyon roots for at least 5 weeks post-inoculation. The gene acdS is new to the repository, and can be used in systems where it is desired to affect ethylene signalling.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 824
Illegal XhoI site found at 1882 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 601
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 39
Illegal BsaI.rc site found at 1811
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