Difference between revisions of "Part:BBa K3510005"

Revision as of 12:15, 23 October 2020

Anderson-Promoter-Mn-Riboswitch-FAST-Phytochelatine-Terminator

Important: This construct does not include a RBS upstream of the FAST2 sequence. When building on our design please make sure you add it in order to allow for translation

This composite part is a manganese inducible expression system of a phytochelatin (BBa_K1321005) tagged with FAST2 (BBa_K3510000). Expression is controlled by two individual elements: A strong Anderson promoter (BBa_J23102) and a manganese riboswitch (BBa_K902074). With this promoter choice transcription will constitutively happen, but translation initiation is dependent on manganese binding to the riboswitch. Transcriptional termination occurs through the activity of the reliable double terminator (BBa_B0015). This construct allows to assess the efficiency of the riboswitch by itself, without the support of the manganese promoter (contained inBBa_K3510003).

Usage: In our project we coupled this manganese sensing device with a phytochelatin in order to create a bifunctional biosensor, that is not only able to detect manganese, but also directly binds it. With our suggested implementation this would contribute to the increase of water quality. The FAST2 tag allows a quantitative measurement of the fluorescence, which should provide information about the manganese concentration in the sample.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 147

@@ Line 1: / Line 1: @@
 <partinfo>BBa_K3510005 short</partinfo>
-This composite part is a manganese inducible expression system of a phytochelatin (BBa_K1321005) tagged with FAST2 (BBa_K3510000). Expression is controlled by two individual elements: A strong Anderson promoter (BBa_J23102) and a manganese riboswitch (BBa_K902074). With this promoter choice transcription will constitutively happen, but translation initiation is dependent on manganese binding to the riboswitch. Transcriptional termination occurs through the activity of the reliable double terminator (BBa_B0015). This construct allows to determine the efficiency of the riboswitch by itself, without the support of the manganese promoter.
+<b><font color= red>Important: This construct does not include a RBS upstream of the FAST2 sequence. When building on our design please make sure you add it in order to allow for translation</font color= red></b>
-<b>Usage:</b> In our project we coupled this manganese sensing device with a phytochelatin in order to create a bifunctional biosensor, that is not only able to detect manganese, but also directly binds it. With our suggested implementation this would contribute to the increase of water quality. Unlike the biosensor with the chromoprotein, the FAST2 tag allows a quantitative measurement of the fluorescence, which should provide information about the manganese concentration in the sample.
+This composite part is a manganese inducible expression system of a phytochelatin (BBa_K1321005) tagged with FAST2 (BBa_K3510000). Expression is controlled by two individual elements: A strong Anderson promoter (BBa_J23102) and a manganese riboswitch (BBa_K902074). With this promoter choice transcription will constitutively happen, but translation initiation is dependent on manganese binding to the riboswitch. Transcriptional termination occurs through the activity of the reliable double terminator (BBa_B0015). This construct allows to assess the efficiency of the riboswitch by itself, without the support of the manganese promoter (contained inBBa_K3510003).
+<b>Usage:</b> In our project we coupled this manganese sensing device with a phytochelatin in order to create a bifunctional biosensor, that is not only able to detect manganese, but also directly binds it. With our suggested implementation this would contribute to the increase of water quality. The FAST2 tag allows a quantitative measurement of the fluorescence, which should provide information about the manganese concentration in the sample.