Difference between revisions of "Part:BBa K3510004"

Revision as of 17:06, 22 October 2020

Anderson-Promoter-Mn-Riboswitch-FAST-Chromoprotein-Terminator

This composite part is a manganese inducible expression system of a blue chromoprotein (BBa_K864401) additionally 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.

Usage: In our project we coupled this manganese sensing device with a chromoprotein to have an easy-to-use biosensor that gives a qualitative signal by color. This allows in field testing without the necessity of a laboratory providing a fluorescence plate reader. Therefore, this construct could be really beneficial to roughly examine water quality in areas with poor infrastructure. Additionally, this construct was designed as a control for the effect of phytochelatin on the sensing mechanism in the construct BBa_K351005. Unlike the phytochelatin the chromoprotein should not interfere with manganese.

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

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 This composite part is a manganese inducible expression system of a blue chromoprotein (BBa_K864401) additionally 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.
-In our project we coupled this manganese sensing device with a chromoprotein to have an easy-to-use biosensor that gives a qualitative signal by color. This allows in field testing without the necessity of a laboratory providing a fluorescence plate reader. Therefore, this construct could be really beneficial to roughly examine water quality in areas with poor infrastructure.
+<b>Usage:</b> In our project we coupled this manganese sensing device with a chromoprotein to have an easy-to-use biosensor that gives a qualitative signal by color. This allows in field testing without the necessity of a laboratory providing a fluorescence plate reader. Therefore, this construct could be really beneficial to roughly examine water quality in areas with poor infrastructure.
 Additionally, this construct was designed as a control for the effect of phytochelatin on the sensing mechanism in the construct BBa_K351005. Unlike the phytochelatin the  chromoprotein should not interfere with manganese.