Difference between revisions of "Part:BBa K2120311"

Latest revision as of 18:11, 22 October 2016

B0015+tetR+B0034+pBAD-AraC+B0015+ptet+B0034+RFP+B0015

<Description>
This part is made of three elements:
(1) Arabinose induced promoter pBAD(BBa_K2120305) which we reserve the parts BBa_K808000.This part contains the promoter as well as the coding sequence for the repressor gene araC which is transcribed in the opposite direction(“upstream”). By binding to L(+)-arabinose, AraC changes its conformation. This causes the protein to diffuses from the DNA thereby inducing transcription.
(2)The pR promoter's inhibitor gene cI from bacteriophage lambda.
(3)RFP, as the report gene, can verify the pLac promoter's activity. We opposite the two promoters’ direction for reducing the influence of two promoters.We add B0015 between araC and pR promoter to make sure two promoters won't influence each other .

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 911
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 1076
Illegal AgeI site found at 2839
Illegal AgeI site found at 2951
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 1093

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2120311 short</partinfo>
-pBAD is a promoter induced by arabinose and arac is the repressor. We use it to test the expression of reporter gene in our validation circuit .And tetR is an inhibitor which is built to repress the expression of ptet promoter. We use it to control the “on” or ”off” state of toxin gene. RFP is the reporter gene. In order to test these do not interfere with each other , We reserve [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2120303 BBa_K2120303] .
+'''<Description>'''
+<br>This part is made of three elements:
+<br>(1) Arabinose induced promoter pBAD([https://parts.igem.org/Part:BBa_K2120305 BBa_K2120305]) which we reserve the parts [https://parts.igem.org/Part:BBa_K808000 BBa_K808000].This part contains the promoter as well as the coding sequence for the repressor gene ''araC'' which is transcribed in the opposite direction(“upstream”). By binding to L(+)-arabinose, AraC changes its conformation. This causes the protein to diffuses from the DNA thereby inducing transcription.
+<br>(2)The pR promoter's inhibitor gene ''cI'' from ''bacteriophage lambda''.
+<br>(3)''RFP'', as the report gene, can verify the pLac promoter's activity. We opposite the two promoters’ direction for reducing the influence of two promoters.We add B0015 between ''araC'' and pR promoter to make sure two promoters won't influence each other .
@@ Line 18: / Line 22: @@
 <partinfo>BBa_K2120311 parameters</partinfo>
 <!-- -->
-===Functional Parameters===
-P-Slackiller is the iGEM 2016 project of BIT-China. In industrial produce, plasmid plays an important role in produce because it carries function gene which codes something we wanted. But bacteria will lose the functional plasmid during the division then become a “Slacker” decreasing the efficiency. In order to kill the “Slacker”, we aim to design logical gene switch to sense the copy number variety.
-However, it is difficult to control the plasmid copy number.  So we choose [https://parts.igem.org/wiki/index.php?title=Part:BBa_K808000 BBa_K808000] whose expression level of downstream gene can vary continuously for imitating the change of plasmid copy numbers. And connect the logical switch downstream.
-The logical switch we select the relationship of inhibitor-promoter. In our building work, we finally decide two kinds of inhibitor-promoter, cI-pR and tetR-pTet. Then we add RFP in the downstream of promoter as a reporter for indicating “on” or “off”.
-First, we built K2120309. But during our test, we found the expression of araC would influence the expression of RFP. So we added another B0015 between araC and pR.
-Beside, we also have built a similar device, [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2120311 BBa_K2120311], using tetR-pTet. But we had no time to test it.