Difference between revisions of "Part:BBa K1031804"

Latest revision as of 13:27, 27 September 2013

Pu-B0032-sfGFP-Terminator (XylR)

Structure

Pu promoter which is activated by XylR, is σ-54 dependent. It is composed of three elements. The UBS (Upstream Binding Site) site which is responsible for interacting with XylR transcriptional factor. The IHF binding site which allows IHF to participate, enhancing transcription efficiency. -24 and -12 region interact with σ-54 factor of RNA polymerase, enabling the formation of open complex. (Fig 1)

Fig 1 Structure of Pu promoter. The UAS of this promoter shown as blue sequence in the blue frame interacts with DmpR. IHF binding site is shown in green in the green frame. The orange sequence indicates σ54 binding site as -24 region and -12 region. The G in red represents +1 site.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 167
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 218

Construction

Pc promoter J23114 is selected to initiate the transcription of XylR. Based on this circuit, we constructed a library of RBS (Ribosome Binding Site) including B0031[1], B0032[2], B0033[3] and B0034[4] for tunning for expression level of reporter gene sfGFP. K1031804 consists of Pu promoter, RBS B0032 and reporter gene sfGFP (Fig 2).

Fig 2 Construction of reporter circuit Pu-B0032-sfGFP. The orange arrow represents Pu promoter for XylR. The green oval stands for RBS B0032. sfGFP coding sequence is shown with dark blue, while terminator B0015[5] is in dark red.

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K1031804 short</partinfo>
-Pu-32-sfGFP-TT (XylR)
+<!-- -->
+== '''Structure''' ==
+''Pu'' promoter which is activated by XylR, is σ-54 dependent. It is composed of three elements. The UBS (Upstream Binding Site) site which is responsible for interacting with XylR transcriptional factor. The IHF binding site which allows IHF to participate, enhancing transcription efficiency. -24 and -12 region interact with σ-54 factor of RNA polymerase, enabling the formation of open complex. ('''Fig 1''')
+<html>
+<img src="https://static.igem.org/mediawiki/igem.org/thumb/1/16/Peking2013_part_XylR_promoter.png/800px-Peking2013_part_XylR_promoter.png", width=700px; />
+</html>
+'''Fig 1'''  Structure of ''Pu'' promoter. The UAS of this promoter shown as blue sequence in the blue frame interacts with DmpR. IHF binding site is shown in green in the green frame. The orange sequence indicates σ54 binding site as -24 region and -12 region. The G in red represents +1 site.
+== '''Sequence and Features''' ==
-<!-- Add more about the biology of this part here
-===Usage and Biology===
 <!-- -->
 <span class='h3bb'>Sequence and Features</span>
 <partinfo>BBa_K1031804 SequenceAndFeatures</partinfo>
+== '''Construction''' ==
+''Pc'' promoter J23114 is selected to initiate the transcription of XylR. Based on this circuit, we constructed a library of RBS (Ribosome Binding Site) including B0031[https://parts.igem.org/Part:BBa_B0031], B0032[https://parts.igem.org/Part:BBa_B0032], B0033[https://parts.igem.org/Part:BBa_B0033] and B0034[https://parts.igem.org/Part:BBa_B0034] for tunning for expression level of reporter gene sfGFP. K1031804 consists of Pu promoter, RBS B0032 and reporter gene sfGFP ('''Fig 2''').
+<html>
+<img src="https://static.igem.org/mediawiki/igem.org/5/5b/Peking2013_part_K1031804.png", width=400px; />
+</html>
+'''Fig 2''' Construction of reporter circuit ''Pu''-B0032-sfGFP. The orange arrow represents ''Pu'' promoter for XylR. The green oval stands for RBS B0032. sfGFP coding sequence is shown with dark blue, while terminator B0015[https://parts.igem.org/Part:BBa_B0015] is in dark red.
 <!-- Uncomment this to enable Functional Parameter display
 ===Functional Parameters===
-<partinfo>BBa_K1031804 parameters</partinfo>
+<partinfo>BBa_K1035201521 parameters</partinfo>
 <!-- -->