Part:BBa_K3245013
BBa_K3245013:
BBa_K3245013 is a composite part that expresses sfGFP (K3245008) with regulative promoter R0040.
Usage and biology:
BBa_K3245013 is one of a collection of parts designed for characterizing ptetR (R0040). This part expresses EsfGFP (BBa_K3245008) at high level strength when not inhibited by tetR. BBa_K3245013 is RFC10 compatible so using it should comply with biobrick assembly standard process.
Design:
BBa_K3245013 is designed to serve as the positive control of BBa_K3245003, BBa_K3245012 and BBa_K3245011. Since our process of characterization involves usage of tetracycline, it’s essential for us to measure and control its antibiotic nature. Therefore, we came up with the idea of measuring comparative strength of BBa_K3245003, BBa_K3245012 and BBa_K3245011 against BBa_K3245013 instead of their absolute strength when characterizing R0040. Through this method, we believe we can eliminate the effect of tet to the utmost extent.
Characterization:
As the positive control of BBa_K3245003, BBa_K3245012 and BBa_K3245011, BBa_K3245013 was cloned to a medium copy vector (p15A ori), transferred into E.coli DH10B, and incubated overnight. The culture was diluted with LB to 1/500 before adding tet. Altogether 11 different tet concentrations were tested. Each group were then incubated for 7hours. Data are collected and analyzed according to iGEM standard data analysis form after 7 hours of induction.
Fig.1-Fig.3 Reaction curves of BBa_K3245013 against different tet concentration.
TetR regulated reporter that constitutively expresses GFP when not repressed by tetR.
a
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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