Difference between revisions of "Part:BBa K1341000"

Latest revision as of 13:33, 15 October 2014

switch device(LTT gfp)for our bio-compass

We use this device as a part of a switch device designed for our bio-compass.It can help us screen out the possible path way which include the start point,terminal point, and the particular intermediate nodes(such as the museum(No.5 node)in our story).It has a LTT Promoter,with elements of the tet, lac, they will compose as a device and is responsive to the commonly used inducers IPTG and aTc, producing GFP as an output signal.If this device and tetR(BBa_K1341003)&lacI(BBa_K1341002)successful inserted in the path way by using the typically enzyme site. It will drive expression of green fluorescent protein (GFP). This characteristic will help us to screen out the possible way we want to find.PS: We build a measurement system to test the screen functional about this device in our project(BBa_K1341001).

model

The x and y axes form a grid of 36 pairs of inducer concentrations. The concentrations tested were 0, 1, 10, 50,100, and 200 ng/mL for aTC and 0, 0.001, 0.002, 0.005,0.010, 0.100, and 1.000mM for IPTG behavior is depicted 6 h after induction. The plotted model values are the means of 1000 independent stochastic kinetic simulations, the source and the reference are in the design page.

characterization

Conditions are ±100 ng/mL aTc and ±1mM IPTG, This panel depicts an LTT promoter design, where the single lac operator site is upstream of the −35 sequence. In this case, LacI is no longer able to entirely suppress transcription and an intermediate level of induction (leakiness) occurs at conditions of high aTc but no IPTG. The fluorescence level still has an apparent change when we add aTc firstly, so the figure can explain why we first use IPTG for testing firstly.(All data is obtained 6 h after induction.)

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 75
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 732

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1341000 short</partinfo>
-We use this device as a part of a switch device designed for our bio-compass. It can help us screen out the possible path way which include the start point, terminal point, and the particular intermediate nodes(such as the museum(No.5 node)in our story).It has a LTT Promoter,with elements of the tet, lac, they will compose as a device and is responsive to the commonly used inducers IPTG and aTc, producing GFP as an output signal.
+[[File:Ahutk0006.png|right|300px]]
-===Usage and Biology===
-Model for the device(BBa_K1341000) with LTT promoter
-<!-- -->
-<span class='h3bb'>Sequence and Features</span>
-<partinfo>BBa_K1341000 SequenceAndFeatures</partinfo>
-===Functional Parameters===
-<partinfo>BBa_K1341000 parameters</partinfo>
-<!-- -->
-==Assembly standards==
+We use this device as a part of a switch device designed for our bio-compass.It can help us screen out the possible path way which include the start point,terminal point, and the particular intermediate nodes(such as the museum(No.5 node)in our story).It has a LTT Promoter,with elements of the tet, lac, they will compose as a device and is responsive to the commonly used inducers IPTG and aTc, producing GFP as an output signal.If this device and tetR(BBa_K1341003)&lacI(BBa_K1341002)successful  inserted in the path way by using the typically enzyme site. It will drive expression of green fluorescent protein (GFP). This characteristic will help us to screen out the possible way we want to find.PS: We build a measurement system to test the screen functional about this device in our project(BBa_K1341001).
-This part is based on Part [https://parts.igem.org/Part:BBa_K325219 BBa_K325219].  We resynthesised this part to remove three PstI restriction sites, making it compatible with BioBrick assembly standards 10 and 23.
-Unfortunately, in the process of mutagenesis to create the colour change a SpeI site was introduced at site 3044.  We have ordered primers to remove this site by a substitution from T->C at base 3046.  [[BBa K325259]] (planning) will be the successor to this part compatible with Assembly Standards 10 and 23.  Unfortunately we did not have time to complete this before the submission deadline, therefore the part will be submitted during November 2010.  In the meantime it is possible to use this part with <html>RFC 57</html>.
-==Sequences==
-<partinfo>BBa_K325259 SequenceAndFeatures</partinfo>
+===model===
+[[File:Ahutk0002.png|right|300px]]
+The x and y axes form a grid of 36 pairs of inducer concentrations. The concentrations tested were 0, 1, 10, 50,100, and 200 ng/mL for aTC and 0, 0.001, 0.002, 0.005,0.010, 0.100, and 1.000mM for IPTG behavior is depicted 6 h after induction. The plotted model values are the means of 1000 independent stochastic kinetic simulations, the source and the reference are in the design page.
+===characterization===
+[[File:Ahut_k10.jpg|right|300px]]
+Conditions are ±100 ng/mL aTc and ±1mM IPTG,  This panel depicts an LTT promoter design, where the single lac operator site is  upstream of the −35 sequence. In this case, LacI is no longer able to entirely suppress transcription and an intermediate level of induction (leakiness) occurs at conditions of high aTc but no IPTG. The fluorescence level still has an apparent change when we add aTc firstly, so the figure can explain why we first use IPTG for testing firstly.(All data is obtained 6 h after induction.)
+<!-- -->
+<span class='h3bb'>Sequence and Features</span>
+<partinfo>BBa_K1341000 SequenceAndFeatures</partinfo>