Part:BBa_K2379009
Adaptor with constitutive RBS
This construct consists of the adaptor BBa_K2379008(BBa_K2379008) and is under the influence of the pLac promoter(BBa_R0011). In addition to the adaptor, a constitutive RBS (BBa_B0034) is used to regulate the transcription of the genes downstream. Following the adaptor is the reporter protein GFP (BBa_E0040) and the double terminator(BBa_B0014). Since it is regulated by a constitutive RBS, the circuit will be active at all times.
Usage and Biology
ABOUT THIS PART :
This part(BBa_K2379009)is an variant of adaptor which is regulated by constitutive RBS(BBa_B0034) placed downstream of pLac promoter(BBa_R0011). Since this part contains constitutive RBS, it shows similar levels of fluorescence expression irrespective of temperatures. The part works as expected and when GFP is placed downstream of this gene, similar levels of fluorescence was observed at 32ºC, 37ºC and 42ºC.
CHARACTERISATION STUDIES FOR THIS PART:
1.The above graph indicates the fluorescence expression at three different time periods (each recorded at an interval of 60 minutes
2.The cell cultures were grown initially at temperature 37ºC until it reached an OD600 of 0.3. Then the cultures were transferred and incubated at three different temperatures: 32ºC, 37ºC and 42ºC.
3.Normalised fluorescence was calculated by dividing observed fluorescence by cell density at OD600, background fluoresence was minimized using a negative control.
4.Triplets were used for characterisation and the error bars indicate the Standard deviation of their mean values.
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CONCLUSION:
The part works as expected. Since there is no temperature regulatory element, similar fluorescence was observed at all the three different temperatures irrespective of the time points.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1028
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