Difference between revisions of "Part:BBa K1463000"

Revision as of 12:52, 15 October 2014

Recombinase Switch With GFP and RFP

A recombinase switch with GFP one side of the switch and RFP on the other.
PhiC31 integrase flips the orientation of the DNA between attP and attB, changing the direction of the promoter, switching rfp off and gfp on.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 813
Illegal NheI site found at 836
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 793
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 34
Illegal AgeI site found at 146
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 711
Illegal BsaI.rc site found at 1668

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1463000 short</partinfo>
-A recombinase switch with GFP one side of the switch and RFP on the other.
+A recombinase switch with GFP one side of the switch and RFP on the other.<br>
+PhiC31 integrase flips the orientation of the DNA between attP and attB, changing the direction of the promoter, switching rfp off and gfp on.
 https://static.igem.org/mediawiki/2014/f/f6/Switch_diagram_sites_small2.jpg
-[[Image:GU_jake_gel_1.jpg|thumb|500px|right|'''Fig. 1:''' In vivo recombination of BBa_K1463000 switch using a plasmid (pBAD-Int) that expresses PhiC31 integrase under the control of the arabinose inducible Pbad promoter. Cells were grown for 16 hours with arabinose or glucose. Plasmid DNA was purified, cut with BamHI and run on a 1.2% agarose gel.The gel shows three replicates of the same experiment. 1.pBAD-Int on its own, 2.Switch #2 on its own, 3.Switch #2 + pBAD-Int glucose, 4.Switch #2 + pBAD-Int arabinose, 5.Switch #3 on its own, 6.Switch #3 + pBAD-Int glucose, 7.Switch #3 + pBAD-Int arabinose, 8.Switch #4 on its own, 9.Switch #4 + pBAD-Int glucose, 10.Switch #4 + pBAD-Int arabinose, 11.pBAD33 gvpAC (ignore)12.1kb+ marker.]]
+<!-- Add more about the biology of this part here
+===Usage and Biology===
+Recombination was tested in vivo by placing BBa_K1463000 on a low copy number pSC101 plasmid and introducing a second plasmid (pBAD-int) that only expresses integrase when arabinose is added.
+[[Image:GU_jake_gel_1.jpg|thumb|center|500px|'''Fig. 2:''' In vivo recombination of BBa_K1463000 switch using a plasmid (pBAD-Int) that expresses PhiC31 integrase under the control of the arabinose inducible Pbad promoter. Cells were grown for 16 hours with arabinose or glucose. Plasmid DNA was purified, cut with BamHI and run on a 1.2% agarose gel.The gel shows three replicates of the same experiment. 1.pBAD-Int on its own, 2.Switch #2 on its own, 3.Switch #2 + pBAD-Int glucose, 4.Switch #2 + pBAD-Int arabinose, 5.Switch #3 on its own, 6.Switch #3 + pBAD-Int glucose, 7.Switch #3 + pBAD-Int arabinose, 8.Switch #4 on its own, 9.Switch #4 + pBAD-Int glucose, 10.Switch #4 + pBAD-Int arabinose, 11.pBAD33 gvpAC (ignore)12.1kb+ marker.]]
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-<!-- Add more about the biology of this part here
-===Usage and Biology===
 <!-- -->