Part:BBa_K2797002
Composite part for streptomycin resistance
This composite part confers resistance to streptomycin. The part contains the constitutive BBa_J23119 promoter, the strongest promoter of the J23100 through to J23119 promoter family
Part Characterisation
Figure 1 E. coli strain DH5α transformed with streptomycin resistance part BBa_K2797002 growing on LB agar containing streptomycin at a concentration of 50 μg/ml.
Figure 2 Untransformed E. coli strain DH5α unable to grow on LB agar containing streptomycin at a concentration of 50 μg/ml.
Figure 3 E. coli strain DH5α transformed with streptomycin resistance part BBa_K2797002 growing on LB agar containing chloramphenicol at a concentration of 25 μg/ml due to resistance gene in pSB1C3 vector.
Figure 4 Optical density at 600nm of three replicate wells containing LB and incrementally increasing streptomycin concentrations inoculated with E. coli strain DH5α untransformed and transformed with part BBa_K2797002 (SmRp) during incubation at 37 C over 24 hours. Control wells contained untransformed E. coli DH5α and an antibiotic concentration of zero. Error bars show standard error mean.
Usage and Biology
A selection of resistance genes are required for selection of transformants when inserting foreign DNA into a chassis organism. Newcastle iGEM 2018 focused on developing a new chassis organism capable of colonising plant roots. The selected organism, Pseudomonas fluorescens DSM 25356 was found to be resistant to a range of commonly used antibiotics. Streptomycin was one of two antibiotics P. fluorescens was found not to be resistant to. The team therefore decided to design a streptomycin resistance part that could in future be used for selection of transformed P. fluorescens.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 861 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 709
- 1000COMPATIBLE WITH RFC[1000]
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