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...>Testing the performance of phiC31 recombinase using the register assembly construct with the attachment sites PxB.</p>
...tutive expression of phiC31 in the plant. c) Graphic representation of the construct comprised by a promoter and a terminator in opposite directions flanked by
50 KB (6,682 words) - 03:33, 2 November 2017
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<br> (2) Endolysin construct finish
<br> (5) Backbone construct-3NSII, 5NSII,AMP, B0015, and ORI (PCR: dream taq)<br>
26 KB (3,130 words) - 16:14, 28 November 2017
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...rror, so instead we performed 2-step NEB Phusion HF PCR of pSB1C3 with RFP construct</p>
<li>Performed Phusion PCR of pSB1C3 with RFP construct using psB1C3 #2 and the pKDL071 FWD/RVS primers
76 KB (12,349 words) - 20:03, 14 December 2017
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...ycin. The enzyme of this step is BsmBI again. This level is useful, if the construct you are designing requires multiple genes to be transformed into one yeast
...Cross <i>et al.</i>, published in 2017, in which they present a synthetic construct of the Pex5 protein, partly <i>Arabidopsis thaliana</i> and partly <i>Physc
199 KB (29,417 words) - 03:20, 2 November 2017
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...ldenBraid multipartite assembly reaction was performed to create a genetic construct composed by the transcriptional factor, PIF6, fused to a DNA-binding domain
...ression along time for different treatments, including the light-inducible construct (Etr8) under white-light conditions, and a weak constitutive promoter and a
51 KB (6,067 words) - 19:12, 14 December 2017
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.... The construct used for cutting <i>STE2</i> is shown in Figure 2, and the construct used for cutting <i>STE3</i> in Figure 3.
<figcaption><b>Figure 2.</b> gRNA construct for deletion of <i>STE2</i>.
253 KB (35,104 words) - 15:59, 30 October 2017
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<li style="font-family: Rubik">Designed, and began to construct, a variety of framework compatible systems, including a synthetic promoter
...moter was added to enable expression to be under the control of IPTG. This construct was then submitted to IDT for synthesis as a gBlock.
184 KB (26,744 words) - 19:53, 1 November 2017
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...moter was added to enable expression to be under the control of IPTG. This
construct was then submitted to IDT for synthesis as a gBlock.
...replacing the IPTG sensing system in the original detector module with the
construct detailed above, creating part <a href="http://parts.igem.org/Part:BBa_K2205
185 KB (26,866 words) - 22:39, 1 November 2017
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...subject, and also they get information about yeast extraction and protein construct.",,,,Consulted experts to gain more information on cyanobacteria and mathem
...ext two exchanges mainly focused on cooperation</li><li> Help BNU-China to construct a part of the gene circuit, and meanwhile, BNU-China helps with the charact
1.09 MB (163,642 words) - 17:36, 30 October 2017
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<p>Assess robustness of a GFP reporter construct in a range of environments.</p>
...ly. The Golden Gate assembly method successfully produced the GFP reporter construct, however it involved more steps and was more time consuming than Gibson ass
20 KB (2,994 words) - 21:41, 1 November 2017
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...ed split-luciferase system characterized by the 2015 Peking iGEM team, our construct consists of a promoter reporter system that looks at two downstream product
...ia in their intestines are influencing on protein nutrition. We thought to construct a pathway to synthesize amino acids from nitrogen in E.coli, introduce it i
400 KB (63,299 words) - 15:37, 15 August 2018
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...(Kittleson, 2012), which was essential both to obtain P1 particles and to construct each complete CRISPR system.</p>
engineered phagemid would need to become a P1 particle and construct whichever CRISPR system we have inserted into it within an <em>E. coli</em>
25 KB (3,973 words) - 20:43, 25 November 2017
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...lls (control) (15min) or not mixed in the presence or absence of the EL222 construct. Cell mixtures would have then be passed through a particle sizer with a ra
...fluorescence microscopy. We also included a His-Tag within all SpyCatcher construct variants to enable in-vitro analysis of complexes formed with SpyTag varian
122 KB (15,402 words) - 02:31, 2 November 2017
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<td class="col0"><strong>Construct</strong></td><td class="col1">6.43 µg</td>
<td class="col0"><strong>Construct</strong></td><td class="col1">6.43 µg</td>
167 KB (25,036 words) - 21:56, 31 October 2017
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Construct standard structure of integrase int5 and int8.<br>
Construct the expression system of integrase TP901, int2, int3, int4, int5, int7, int
16 KB (1,672 words) - 13:45, 1 November 2017
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...ycin. The enzyme of this step is BsmBI again. This level is useful, if the construct you are designing requires multiple genes to be transformed into one yeast
...Baker <i>et al.</i>, published in 2017, in which they present a synthetic construct of the Pex5 protein, partly <i>Arabidopsis thaliana</i> and partly <i>Physc
105 KB (15,409 words) - 02:56, 2 November 2017
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...o take our chances by building a D.I.V.E.R.T. cassette as well as a helper construct (carrying the RT, FLP and the primer cassettes) according to the best of ou
...synthetic regulatory elements<sup>1–3</sup> to keep the synthesized construct short. The synthetic galactose-inducible minimal promoter we used (sequence
68 KB (10,146 words) - 17:37, 15 December 2017
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...ctroporated DNA (BRED) (Figure 1). In this technique a double stranded DNA construct containing the
...to be replaced. The host cell for recombineering of the phage and the DNA construct contains a plasmid conferring genes promoting recombination. The double-str
10 KB (1,666 words) - 23:47, 1 November 2017
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...ycin. The enzyme of this step is BsmBI again. This level is useful, if the construct you are designing requires multiple genes to be transformed into one yeast
...Baker <i>et al.</i>, published in 2017, in which they present a synthetic construct of the Pex5 protein, partly <i>Arabidopsis thaliana</i> and partly <i>Physc
98 KB (14,470 words) - 00:02, 2 November 2017
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<a href="#ConDPR" class="pageNavSm">
Construct Design</a>
<a href="#ConDBio" class="pageNavSm">
Construct Design</a>
55 KB (6,458 words) - 14:08, 1 November 2017
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<td>Simple, easiest to construct</td>
...espective promoter (PphlF), and a GFP reporter (BBa_K2477008). We grew our construct in different molarities of fusaric acid (0µM, 100µM, and 500 
232 KB (33,473 words) - 02:45, 2 November 2017
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<div class="hull-content">Our dCas13a - GFP construct was synthesized by Integrated DNA Technologies. The dCas13a gene sequence w
To improve imaging, the dCas13a - GFP construct was sequesteredTo reduce background noise, unbound dCas13a - GFP should be
24 KB (3,114 words) - 00:16, 2 November 2017
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construct.<br><br>
<li>Construct half of the flip-flop structure and express it in <i>E. coli.</i></li>
170 KB (16,709 words) - 06:32, 15 December 2017
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<div class="hull-content">Our dCas13a - GFP construct was synthesized by Integrated DNA Technologies. The dCas13a gene sequence w
To improve imaging, the dCas13a - GFP construct was sequesteredTo reduce background noise, unbound dCas13a - GFP should be
24 KB (3,037 words) - 23:41, 15 December 2017
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Waiting on sequencing primers for my construct
Ordered Cph8 construct partly gene, partly gblock
159 KB (17,948 words) - 20:12, 1 November 2017
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<h2 class="textheading1" style="float:center"><u> Full Construct Testing</u> </h2>
...of GFP, the data seemed to be a bit problematic. We took a few of our full construct colonies as well as a few wild type Kanamycin resistant colonies and we ino
12 KB (1,480 words) - 02:11, 2 November 2017
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...This figure shows restriction enzyme check electrophoresis result of NrtA construct.
<h4>Endolysin Construct</h4>
29 KB (4,011 words) - 19:07, 1 November 2017
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...licated many times and matured, this allows us to have more cells with the
construct that can produce more pigment faster.
...wasted on transcribing an overly-long mRNA transcript. The melA expression
construct is the improved part that we were able to characterize the most (see <a hre
12 KB (1,924 words) - 17:29, 1 November 2017
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...luorescence was measured 20 minutes after this pH shift. The asr-mCardinal construct was inducible by change from pH 7.0 to pH 5.0 and lower but no fluorescence
...s in LPM with pH 7 overnight. For expression control of our alx-mNeonGreen construct, we inoculated 20 ml LPM with pH 7.0, 8.0 and 8.5 to an OD<sub>600</sub> of
7 KB (1,003 words) - 03:29, 2 November 2017
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Waiting on sequencing primers for my construct
Ordered Cph8 construct partly gene, partly gblock
161 KB (18,127 words) - 22:23, 1 November 2017
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Waiting on sequencing primers for my construct
Ordered Cph8 construct partly gene, partly gblock
161 KB (18,092 words) - 09:44, 23 November 2017
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...se). Here we performed saturation mutagenesis on the <i> araC </i> gene to construct AraC mutant library with four randomized ligand-binding pocket amino acid r
...sors and may not show sufficient specificity to xylulose. Hence, we aim to construct an alternative regulatory system which exploits the L-arabinose operon.
39 KB (6,105 words) - 16:49, 17 November 2017
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...2 regulated systems. Herein we will refer to each promoter + split protein construct as a “module”, i.e. a full AND-gate will require the presence and activ
...er for split T7 RNAP function. We utilised a preexisting T7 promoter (pT7) construct from the BioBrick registry, [http://parts.igem.org/Part:BBa_K1321338 K13213
33 KB (5,136 words) - 02:41, 2 November 2017
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<h3>New plan for assembling the cas13 construct</h3>
<h4>New plan for assembling the cas13a construct: Plan A and Plan B</h4>
656 KB (66,631 words) - 09:22, 15 December 2017
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Waiting on sequencing primers for my construct
Ordered Cph8 construct partly gene, partly gblock
160 KB (18,135 words) - 23:14, 1 November 2017
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...ed AcrIIA4 from G-block and amplified pSB1C3 backbone from LacILOV-mCherry Construct in pSB1C3</td>
...ILOV-mCherry construct in pKDL071 and pSB1C3 backbone from LacILOV-mCherrt construct in pSCB1C3</td>
7 KB (797 words) - 19:53, 14 December 2017
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...5 variant R19 with the PTS1* and PTS*, co-transformed with the Pex13-mRuby construct − the localization of mTurquoise tagged with PTS1* is clear and due t
...ed a Zeiss Elyra PS microscope. For Pex15 we observed localization using a construct with mVenus fused to the C-terminus of the Pex15 version we used. The fluor
59 KB (9,038 words) - 03:50, 2 November 2017
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...ngs were obtained after 24 hours of incubation. This temperature sensitive
construct was working as expected in 10β cells, elucidating higher GFP expression at
<p id="imgdescription">Figure 2: BBa_K2447014
construct which was characterised in 10β cells (incubated overnight). At higher temp
20 KB (2,876 words) - 13:25, 1 November 2017
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...work in a better understood organism and led to higher copy numbers of our
construct. </p>
<p>Our final
construct, pGUIDE, consisted of Chloramphenicol and Kanamycin resistance genes, the C
29 KB (4,511 words) - 03:55, 2 November 2017
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...">Figure 1. Graphic representation of Plant-Human genetic circuit with the construct comprised by a promoter and a terminator in opposite directions flanked by
...protein. It represents the positive control of our experiment. d) Genetic construct that allows constitutive expression of phiC31 in the plant under the contro
19 KB (2,276 words) - 03:46, 2 November 2017
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...to xylose, xylulose, ribose and mannitol. </ref> combined with a reporter construct to detect the presence of <i>C. jejuni</i>.
mtlR coding sequence was ordered as a G-block construct from IDT and was ligated into pSB1C3. As this was to be under the control o
19 KB (3,023 words) - 12:18, 12 December 2017
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...n) is added, the activity of the enzyme can be checked.<br>So far the test construct have only been implemented in silico. Also all needed primers were designed
...ed and used for a test digest with NotI. The growth of the riboswitch test construct was not properly grown. So the Golden Gate reaction and the transformation
191 KB (30,171 words) - 16:09, 15 December 2017
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...5 variant R19 with the PTS1* and PTS*, co-transformed with the Pex13-mRuby construct − the localization of mTurquoise tagged with PTS1* is clear and due t
...ed a Zeiss Elyra PS microscope. For Pex15 we observed localization using a construct with mVenus fused to the C-terminus of the Pex15 version we used. The fluor
56 KB (8,614 words) - 02:20, 2 November 2017
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...s indicate where a band with a molecular mass corresponding to that of the construct should be.
...re or during the purification. It wasn’t clear from our results why this construct was unstable, but we reasoned that it was too unstable for our use, so no f
20 KB (3,047 words) - 03:51, 2 November 2017
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...hecked our constructs for secondary structures and we scrambled the 14-3-3 construct to make sure that we get 4 completely different monomers, to make mutation
...of the 14-3-3 construct did not work, but the Gibson Assembly of the CT33 construct showed promising results.</h6>
34 KB (5,513 words) - 17:18, 1 November 2017
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...nhibit pλR, obstructing the promotion of tsPurple. Therefore, the working construct would supply us with a concentration of translated tsPurple dependant upon
...ration verified the success of ligation. Despite lacking the full, genetic construct, we managed to obtain viable data from the Chrome-Q system using the proof
21 KB (3,001 words) - 16:46, 16 December 2017
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...es each containing cells included to express recombinase and a measurement construct. These cells contain
a measurement construct with orthogonal target sites and give little fluorescence</figcaption>
12 KB (1,529 words) - 23:00, 1 November 2017
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...se the yields of anthocyanin when paired with the genes in our anthocyanin
construct. We have added the sequence to the registry for use in our composite part <
<p class="center">The Promoter we used for our Melanin
Construct is T7 promoter, This allows us to control the production of Melanin. This
14 KB (2,401 words) - 17:43, 1 November 2017
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<section><h3>1. Cloning of module 1:</h3><h4>Construct 1 --> Ptet - RBS1 - T7 RNA polymerase - terminator 1 - terminator 2.
...id not get positive results for cloning, resulting in failure to clone the construct 1.
12 KB (1,800 words) - 03:21, 2 November 2017
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<section><h3>1. Cloning of module 1:</h3><h4>Construct 1 --> Ptet - RBS1 - T7 RNA polymerase - terminator 1 - terminator 2.
...id not get positive results for cloning, resulting in failure to clone the construct 1.
12 KB (1,743 words) - 04:15, 14 December 2017
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...d to turn the protein and its required guide sequence into a BioBrick. The
construct was delivered in three fragments that had to be assembled with the backbone
...extracted from <a href="#references">Gootenberg et al. 2017</a>. This DNA
construct (7.7 kb), was ordered in three separate parts, denoted part 1 (2.5 kb)
102 KB (13,845 words) - 16:15, 15 December 2017
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...eful vector pPIGBACK for microalgae transformation. pPIGBACK is a powerful construct with NSII, AmpR, double terminator, ORI, and PrbcL, and it is able to embar
<!--construct-->
13 KB (1,734 words) - 19:39, 1 November 2017
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<h2 class="major">Construct Model</h2>
...d COX-2 (Peking iGEM Team 2015, 2015). These products are assembled into a construct composed of fusion proteins as well as split luciferase fragments (COX-2 -
78 KB (7,559 words) - 15:53, 1 November 2017
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.... Suddenly I came up with an interesting idea that what if the scaffold is construct by protein. Then I realized that scaffold is such a regular network that we
... So now the problem is how do we construct the scaffold. We knew that there was many protein self-assembly system. Usu
22 KB (3,229 words) - 09:18, 29 August 2017
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<input class="tealButton" type="button" value="Construct Design" onclick="window.location.href='#anchor3'" />
<input class="tealButton" type="button" value="Validation Construct" onclick="window.location.href='#anchor7'" />
25 KB (3,716 words) - 02:10, 2 November 2017
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...s (Center Point and Binding Partner) are connected and form the <i>Binding Construct</i>. There is a <i>Center Point</i> and at this Center Point, four Binding
<li><span>Of the Scaffold Construct</span></li>
5 KB (769 words) - 21:13, 1 November 2017
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Through simulations of our intended construct with parameters defined a priori, we were able to draw some conclusions as
<h3 class="section-heading">Control construct design</h3>
12 KB (1,643 words) - 01:46, 2 November 2017
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<a class="item passlink" data-passage="consexp">Construct Expression Vector for NzFE</a>
<td>easy to construct</td>
21 KB (2,679 words) - 13:26, 15 December 2017
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● Build up deinking enzyme construct<br>
● Transform construct into yeast<br>
12 KB (1,855 words) - 03:13, 16 December 2017
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...RNA from the HTT115 strain witouth the siRNA
construct and with the siRNA
construct (BSLA-AWD) with different hybridization oligos and treatments in a 12% no-d
<p>Figure 2. RNA from the HTT115 strain with the siRNA
construct (BSLA-SOD, BSLA-WNT and BSLA-RACI) with different hybridization oligos and
42 KB (4,985 words) - 03:59, 2 November 2017
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<td>Construct for Curvacin A expression</td>
<td>Construct for Enterocin A expression</td>
19 KB (2,225 words) - 19:40, 1 November 2017
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...e to subclone the immunity protein 2 into the tryptophan regulatory system
construct and perform proliferation tests. Unfortunately, due to time constraints, we
...uccess rate of a study utilizing machinery for genome integration. The RFP
construct was cloned into the UPO-Sevilla 2011 plasmids (<a target="_blank" href="htt
48 KB (6,966 words) - 15:28, 15 December 2017
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...contain EutSMNLK, however we were unable to obtain pure plasmid from this construct containing all microcompartment subunits as the transformed colonies became
...Tris-glycine SDS page gels of soluble and insoluble Eut S, MN, SMN and LK construct proteins. (-) indicates
12 KB (1,634 words) - 03:48, 2 November 2017
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...olor: #808080; font-size: 14px;'>Figure 1: Schematic of a generic reporter construct used to test degradation rates. Analogous constructs were built with sfGFP
...tructs (Figure 3a). Further, if you plot the gene expression speed of each construct against its degradation rate then the shape of the curve resembles that of
19 KB (2,797 words) - 03:07, 15 December 2017
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<li><b>Fer/Hyd</b>- Fer and Hyd as described above in a new biobrick construct.</li>
...i> and <i>hyd G</i>, coding all three maturation enzymes in a new biobrick construct.</li>
55 KB (9,025 words) - 13:09, 31 October 2017
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...atase that converts the enoyl-CoA into (R)-hydroxybutyrate (Lu, 2003). Our construct also includes the <i>phaC1</i> gene from <i>Pseudomonas aeruginosa</i>, enc
<h2>Genetic construct</h2>
7 KB (1,176 words) - 01:03, 2 November 2017
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...last monomer, leading to a functional valency of 3. The structure of this
construct is shown schematically in <a href="#Figure_1">Figure 1.</a>
..., leading to our second
construct, which has thus a valency of 4. The CT33
construct is schematically shown in <a href="#Figure_2">Figure 2</a> below.
7 KB (1,092 words) - 22:03, 11 December 2017
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...te-temperature cascaded system with GFP reporter. Characterisation of this construct was conducted with varying phosphate concentrations under two different tem
...led plan: Culture the transformed cells ( the temperature sensitive system construct, is transformed into MG 1655) from glycerol and incubate for 36 h in 30 deg
17 KB (2,425 words) - 16:34, 1 November 2017
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...g a VCP1-GFP-VCP2 construct and attempting to get the algae to express the construct. <br> <br>
.../2017/b/be/UConn_vcp1gfp_gel.JPG" class="vcp1-gfp"> <br> First, a VCP1-GFP construct was assembled through digesting the GFP out of its original backbone and th
15 KB (1,886 words) - 03:25, 2 November 2017
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...d series of anderson promoters next to the RNA I gene and then placed this construct next to RNA II (RNA II-Anderson-RNA I).</p>
...le copy number system. We first had to make sure that at least part of our construct is well characterized and to so we chose the rhamnose promoter from the bio
58 KB (8,538 words) - 14:52, 26 November 2017
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recombinase expression construct in pET28b, a medium copy plasmid (10~15), another one being Promoter-LoxP-T
...mediate the recombination between the two target sites on the measurement construct, excising the transcriptional terminator. This leads to the expression of R
79 KB (9,298 words) - 20:57, 1 November 2017
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The construct is long (1,029 bp), so we got only part of it sequenced due to using only o
...k sample no. 13 from the check-up digestion (Figure 8) for sequencing. The construct is long (1029 bp), so we got only
53 KB (6,877 words) - 20:55, 11 December 2017
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<cite>Figure I: Scheme of the BBa_K2425000 construct. It is important to know that the number of bases pairs is just the coding
...f METE gen (Helliwell et al.,Plant physiology, 165(1), 388-397, 2014). The construct has 574 bp and contains the 5’ UTR and the sequence of the promoter. All
17 KB (2,489 words) - 01:44, 16 December 2017
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...of the highly sensitive parameter in the design. As the original phosphate
construct <a href="http://parts.igem.org/Part:BBa_K116404">BBa_K116404</a> (
construct
5 KB (689 words) - 03:09, 2 November 2017
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.... The second step was the preparation of a pTet-GFP/RFP and a pLac-GFP/RFP construct to test these promoters in L. lactis. From next week progress for this part
...ad><tr class="tableizer-firstrow"><th>Backbone</th><th>construct A</th><th>construct B</th><th>Product name</th></tr></thead><tbody>
50 KB (7,945 words) - 03:35, 2 November 2017
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Each construct was sent for sequencing with primers VR and VF
...<li>RPA of TEV could not produce the whole (~1300 bp) construct but would produce amplicons of shorter size (100-500 bp) which was anticipa
380 KB (41,086 words) - 11:45, 1 November 2017
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Each construct was sent for sequencing with primers VR and VF
...<li>RPA of TEV could not produce the whole (~1300 bp) construct but would produce amplicons of shorter size (100-500 bp) which was anticipa
380 KB (41,105 words) - 02:27, 2 November 2017
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.../static.igem.org/mediawiki/2017/2/2b/XylRproofofconcept.jpg" alt="xylR-GFP Construct" style="width:600px;height:175px;"></center>
...nts, we eventually arrived at construct BBa_K2451013, depicted above. This construct displays promising results for m-xylene induction. The xylR coding sequence
46 KB (5,986 words) - 02:38, 2 November 2017
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..."></div><div>w that iron had no effect on the production of amilCP in this construct and we compared that to the co-transformed bacteria (plasmids 1+2). Again,
...rmed cells (i.e. cells with the FUR repressed construct and LacI repressed construct) within 24 hours, although to a lesser extent than cells which lacked the f
12 KB (1,980 words) - 18:53, 1 November 2017
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<h4>Construct 1 --> Ptet - RBS1 - T7 RNA polymerase - terminator 1 - terminator 2.</h4>
<h4>Construct 2 --> P<sub>T7</sub> - RBS2 - Chromoprotein II - RBS3 - tetR - terminator3.
11 KB (1,384 words) - 04:14, 14 December 2017
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<p>When designing this construct, we found a paper by Chen et al. that determined the optimal spacing betwee
...the production of varied gas vesicle phenotypes. Expression of the gvp3.0 construct is under control of the lac promoter (pLac), allowing inducible expression
9 KB (1,385 words) - 03:58, 2 November 2017
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...find out the best combination of vector and E.coli strain for this sensing construct.
3 KB (501 words) - 04:56, 16 December 2017
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<b>Figure 3:</b> construct design for 2 Dronpa domains
...y altering its conformation. To test this hypothesis we made the following construct:
21 KB (2,411 words) - 03:11, 16 December 2017
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...to time constraints of the parts submission deadline, we were not able to construct a new biobrick compatible hCas9 operon. We encourage next iGEM teams to pic
As mentioned earlier we were not able to construct the biobrick compatible hCas9 operon. So, the experiment described next was
25 KB (3,964 words) - 03:48, 2 November 2017
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var reservedEventNames = /^(?:change|reset|teardown|update|construct|config|init|render|complete|unrender|detach|insert|destruct|attachchild|det
....error( 'Cannot use reserved event names (change, reset, teardown, update, construct, config, init, render, unrender, complete, detach, insert, destruct, attach
1.96 MB (302,682 words) - 17:51, 24 October 2017
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...<i>Clostridium thermocellum</i>. This part could be used with any other construct to bind to cellulose.
...l peptide in form of a fusion protein accompanied with His6x-Smt3 tag. The construct can be purified with commercially available and widely used affinity chroma
18 KB (2,363 words) - 20:51, 15 December 2017
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...n later constructs and successfully isolated those. Digested all parts for construct 1 so far and gel purified the adequate bands.
Transformed cells with construct 1 and created glycerol stocks from those. Prepared a protocol for the gener
7 KB (1,157 words) - 13:03, 31 October 2017
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.... The second step was the preparation of a pTet-GFP/RFP and a pLac-GFP/RFP construct to test these promoters in L. lactis. From next week progress for this part
...ad><tr class="tableizer-firstrow"><th>Backbone</th><th>construct A</th><th>construct B</th><th>Product name</th></tr></thead><tbody>
39 KB (6,407 words) - 01:03, 2 November 2017
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...icin. Successful cloning was confirmed by gel electrophoresis, and the DNA construct was sent for sequencing. Whilst waiting for sequencing results, we began ch
...o do this, we used the already existing BioBrick BBa_I6211, containing the construct osmosensitive promoter-RBS-YFP-double terminator in the pSB1C3 backbone. As
16 KB (2,484 words) - 19:09, 15 December 2017
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...RNA from the HTT115 strain witouth the siRNA
construct and with the siRNA
construct (BSLA-AWD) with different hybridization oligos and treatments in a 12% no-d
<p>Figure 2. RNA from the HTT115 strain with the siRNA
construct (BSLA-SOD, BSLA-WNT and BSLA-RACI) with different hybridization oligos and
14 KB (1,654 words) - 01:36, 2 November 2017
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...uorescence detection assay utilizing green fluorescent protein (GFP)-fused construct to BBa_I14034 promoter part.</p>
This construct is produced by combining BBa_114034 promoter part and a GFP coding sequence
3 KB (469 words) - 03:58, 2 November 2017
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...roceeded to insert the T2A-P2A based display vector into our final ToleGen construct.
Our final experiment was to test the ToleGen construct we created <strong>(figure 5)</strong>.
20 KB (2,612 words) - 02:51, 2 November 2017
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...ow exchanging of the separate parts. As a result, we decided to divide our construct into 3 gBlocks, which could be connected to each other and placed in a pET2
...e large gBlocks and large amount of inserts, so we decided to redesign our construct, by dividing it into two gBlocks: one containing the first two monomers and
8 KB (1,272 words) - 13:50, 10 December 2017
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...xpected size band of ~3.5 kb. <strong>C:</strong> Results of <em>VanA</em> construct amplification via PCR using amplifying primers at four temperatures, 66°
...hite MoClo colonies were checked for correct insertion of the <em>KPC</em> construct via digestion with AvrII (A). All bands were of the expected size and uncut
9 KB (1,446 words) - 23:48, 1 November 2017
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<input class="tealButton" type="button" value="Validation
Construct" onclick="window.location.href='#anchor4'" />
<p class="text12j">Both the tRNA<sup>phe</sup>-MS2 aptamer
construct and MS2BP were overexpressed individually in <i>E. coli</i> BL21-Gold (DE3)
20 KB (2,975 words) - 03:38, 2 November 2017
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<p>To characterize LacILOV, cells containing LacILOV-mCherry construct were grown under a consistent light intensity and the fluorescent output wa
...tivity of the LacILOV mCherry construct, the assays for assessing reporter construct use the same hardware devices for delivery of light and dark conditions. Th
16 KB (2,418 words) - 19:50, 14 December 2017
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...ry tag has on enzyme activity. Results from this will aid how we build the construct to target the enzyme into the microcompartment at a later stage.</p>
<th>Construct</th>
26 KB (3,896 words) - 01:44, 2 November 2017
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...on developing a proof of concept for the Chrome-Q and assembling a genetic construct to use the Chrome-Q to quantify data.
Several weeks were devoted to construct pλR LacI R011 RFP in the NEB 10-beta E. coli strain, with individual parts
26 KB (3,923 words) - 17:16, 12 December 2017
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...d sequencing results from Tri-I Biotech confirmed that our final <i>pR</i> construct is correct.
...<b>Figure 2-2 Construct expressing PR. </b> Our construct includes a strong promoter, strong RBS, the <i>pR</i> ORF, and a double ter
19 KB (1,932 words) - 13:20, 31 October 2017
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<input class="tealButton" type="button" value="Validation
Construct" onclick="window.location.href='#anchor3'" />
<p class="text12j">Both the tRNA<sup>Phe</sup>-MS2
construct and MS2BP were expressed individually in <i>E. coli</i> BL21-Gold (DE3) cel
9 KB (1,447 words) - 02:13, 2 November 2017
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...se the yields of anthocyanin when paired with the genes in our anthocyanin construct. We have added the sequence to the registry for use in our composite part <
<h2><b> Melanin Construct</b></h2>
10 KB (1,730 words) - 17:37, 1 November 2017
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...engineer <i>E. coli</i> to synthesize PHB. We designed a pET29b(+)-phaC1J4 construct to allow our engineered <i>E. coli </i> to employ the beta-oxidation pathwa
...nfirm the ability of <i>E. coli</i> transformed with our pET29B(+)-phaC1J4 construct to produce PHB, we grew O/N cultures for 24 hours and induced them with 0.1
34 KB (5,337 words) - 04:28, 16 December 2017
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...sequence is identified as integral in determining whether the single chain construct has activity.</h4><br>
...ney_Australia/Patent_Law">patents</a>. With this in mind - we designed our construct to also have 12 amino acids.<br>
66 KB (8,543 words) - 03:14, 2 November 2017
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...ttps://static.igem.org/mediawiki/2017/d/d1/T--Bielefeld-CeBiTec--composite-construct.png">
...fot the mRFP transcript is downstream the CDS of the gene of interest. In construct 2 the CDS of the mRFP is downstream a T3 promoter and the CDS coding for th
27 KB (4,095 words) - 01:18, 2 November 2017
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...titutive promoter psaD (and its respective terminator) The detail of these
construct are shown as follows in the next image:</p>
<p id="FigureText">Fig. 1: Scheme that shows the fusion protein
construct to make the first transformation of <i>C. reinhardtii</i>.</p>
8 KB (1,220 words) - 00:35, 30 October 2017
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In order for our construct to work, it had to be transfected into HEK2935, a mammalian cell line. RNA
We transfected our construct (BBa_K2340000) with a guide RNA plasmid (BBa_K2340001 - BBa_K2340011) (1:2
21 KB (2,470 words) - 04:21, 16 December 2017
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...e our chances of successful assembly. We did succeed in inserting the melA construct into pJET (Figure 1).
...l of melA_pJET digested with EcoRI and PstI. Plasmids 1 and 3 had the melA construct successfully insert into pJET without disruption of the cut sites.</i></p>
8 KB (1,262 words) - 13:54, 1 November 2017
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In addition, the tested construct was approximately 1500 bp long, which is also quite far from RPA's optimum
construct of 200 bp, one larger in size and less intense and one smaller in size and
11 KB (1,840 words) - 21:57, 31 October 2017
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...titutive promoter psaD (and its respective terminator) The detail of these construct are shown as follows in the next image:</p>
<cite>Fig. 1: Scheme that shows the fusion protein construct to make the first transformation of C. reinhardtii.</cite>
9 KB (1,363 words) - 03:56, 2 November 2017
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...l peptide in form of a fusion protein accompanied with His6x-Smt3 tag. The construct can be purified with commercially available and widely used affinity chroma
The construct can be purified with commercially available and widely used affinity chroma
10 KB (1,538 words) - 20:53, 15 December 2017
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<p class="PP">This year, we aim to construct a diseases-monitoring system for automatic agriculture (AA) in which biolog
...ce<sup>[2]</sup>. What’s more, DAPG can also serve as signal molecule to construct a transcriptional switch that is similar to Tet on/off system<sup>[3]</sup>
31 KB (3,735 words) - 15:38, 1 November 2017
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...;\n }\n if (arguments.length < 1) {\n throw new TypeError(\"Failed to construct 'SockJS: 1 argument required, but only 0 present\");\n }\n EventTarget.ca
...the same order.\n // 5. Return the result of calling the [[Construct]] internal\n // method of target providing args as the arg
932 KB (107,677 words) - 07:59, 24 October 2017
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...;\n }\n if (arguments.length < 1) {\n throw new TypeError(\"Failed to construct 'SockJS: 1 argument required, but only 0 present\");\n }\n EventTarget.ca
...the same order.\n // 5. Return the result of calling the [[Construct]] internal\n // method of target providing args as the arg
932 KB (107,677 words) - 08:00, 24 October 2017
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...and plated. Preliminary test of RFP expression with BBa_K1966004 (2016 Pu construct)
- Colony PCR on xylB construct – success (right)
66 KB (8,136 words) - 00:10, 2 November 2017
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...aled for both species. Only the <i>gambiense</i> gene was used for further construct building. It was gel purified and digested with both KpnI and SacI. The bac
The construct building was repeated, since it failed in week 3. First, the ligation was r
8 KB (1,182 words) - 12:59, 31 October 2017
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<p><b>Superior experiment:</b> Gaining more of the construct pSB1C3-AraC-chiA</p>
<li style="font-size: 15px;"> The mutation still remains in the construct</li>
96 KB (13,160 words) - 21:18, 1 November 2017
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* IPTG Induction of BL21 promoter construct
* Transformation of BL21 with pick 4 of promoter B1C3 construct
10 KB (1,332 words) - 17:46, 31 October 2017
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...P fluorescence obtained from BL21(DE3) with the pETDuet-GST-f-p+pET28a-GFP construct cultured in LB medium (10X40).</span>
...fluorescence obtained from BL21(DE3) with the pETDuet-GST-f-p+pET28a-EGFP construct cultured in MOPS medium (10X40).</span>
26 KB (3,823 words) - 18:49, 1 November 2017
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...ts were run on a gel and gel purified in order to isolate the promoter-RFP construct. These constructs were then ligated using T4 ligase into pCAMBIA-MCS which
..., however there were colonies on the LB+Kan plates of <i>virB1</i>-pCAMBIA construct for both plasmid backbones, BBa_K1357010 and BBa_J04650.
19 KB (2,909 words) - 03:17, 2 November 2017
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...ing. The construct was tested in HEK293FT cells, targeting five genes. The construct was transfected concurrently with a GFP donor.</p>
...ge of the knock-in event for GAPDH gene. The data were normalized to empty construct. Error bar is +- s.d.
20 KB (3,198 words) - 03:53, 2 November 2017
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...he vector and insert, assembling the construct, transforming the assembled construct into DH5α cells, and verifying the assembly success via colony PCR and seq
...the DNA polymerase and DNA ligase lysate to assemble their PelB-saCas9-His construct using optimal ratios that we established. The success of the assembly would
8 KB (984 words) - 03:44, 2 November 2017
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...LuxR will result in a significant amount of GFP production. The goal is to construct a variation on this part that no longer has spontaneous activation, but can
...sults support the strategy of using aiiA to tune the sensitivity of the QS construct and indicate that, in principle, the functionality we want to engineer can
20 KB (2,280 words) - 21:40, 1 November 2017
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<li>Cells engineered with our Display construct present epitopes on their membrane MHC complexes.</li>
We came up with two possible ways to adequately test our construct:
18 KB (2,317 words) - 01:45, 2 November 2017
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Standard Genetic Construct for Pigment Production
...i> ribosome binding site (RBS) B0034 (with the exception of the zeaxanthin construct, which uses <i>Pantoea ananatis</i> native RBSs), the coding sequences for
12 KB (1,702 words) - 15:49, 1 November 2017
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...eriment, overnight cultures of MG1655 ΔLacI cells containing the reporter construct were diluted and further cultivated in 5 mL of culture in 15 mL falcon tube
...s with the same CRISPRi system and the anti-CRISPR protein AcrIIA4. If our construct functions as expected, the anti-CRISPR protein should bind to pdCas9 and pr
22 KB (2,497 words) - 18:54, 14 December 2017
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...n in lanes 2-6 show clean single bands that correspond to their respective construct sizes (Cytoplasmic Winsulin: 306bp, Ecotin Winsulin: 867bp, YncM Winsulin:
...n of Ecotin insulin due to time constraints, we did test our YncM Winsulin construct. YncM is an N-terminal tag which, when fused to a protein of interest and e
20 KB (2,972 words) - 23:39, 1 November 2017
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...parts listed below have all been functionally verified, and can be used to construct the systems
the validity of our measurement construct for testing if two recombinase sites are orthogonal. Indeed, it
26 KB (3,167 words) - 10:44, 1 November 2017
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...se the yields of anthocyanin when paired with the genes in our anthocyanin
construct. We have added the sequence to the registry for use in our composite part <
<p class="center">The Promoter we used for our Melanin
Construct is T7 promoter, This allows us to control the production of Melanin. This
8 KB (1,257 words) - 15:53, 1 November 2017
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<li>Make construct mix:<br>
<li>Pipette 2000 µl of the construct mix dropwise to the HEK293T cells</li>
30 KB (3,843 words) - 02:17, 2 November 2017
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<p>To begin with, we gave them
construct suggestions, communicated with their team adviser. In 24/3/2017, we were in
<p>1. NKU asked us to help them
construct a plasmid. Using pEX18Gm as the supporter, we ought to added Lyase gene seq
18 KB (2,589 words) - 16:09, 16 October 2017
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<h2><b> Melanin Construct</b></h2>
This construct allows us to make Melanin out of L-Tyrosine.
8 KB (1,277 words) - 17:41, 1 November 2017
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and were pleased to see that it worked well. This construct we called the
Figure A as a ssDNA molecule and tested it in our lysates. This construct
12 KB (1,870 words) - 02:18, 2 November 2017
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<li><span>Scaffold Construct (14-3-3 with GFP) + Binding Partner (CT33 with Strep-tag®II and mCherry) +
<li><span>Scaffold Construct (14-3-3 with GFP) + Binding Partner (CT33 with Strep-tag®II and mCherry) +
7 KB (1,058 words) - 13:47, 10 December 2017
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...2436001) was cloned successfully and submitted to the registry. The lipase construct was not cloned successfully. Our constructs were designed as follows.
...onstruct contains the chlorogenate esterase coding sequence, and the other construct has the lipase coding sequence.
6 KB (849 words) - 19:44, 19 November 2017
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...<sup>(5)</sup> or VP64<sup>(6)</sup> as transcription regulating domain to construct silencing- or activating-form of SynTFs, named SynTF(S)s or SynTF(A)s. Thei
...ensive usage in genome editing<sup>(10)</sup>. We applied this strategy to construct tunable SynTF-SynPro pairs. With more RE repeats, our SynTF has stronger in
28 KB (4,314 words) - 14:20, 14 December 2017
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<p class="PP">This year, we aim to construct a diseases-monitoring system for automatic agriculture in which hardware an
...Recently, it is demonstrated that 2,4-DAPG can serve as signal molecule to construct a transcriptional switch that is similar to Tet on/off system.</p>
41 KB (5,261 words) - 15:13, 1 November 2017
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...e estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of int
...e estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of int
8 KB (1,193 words) - 00:30, 2 November 2017
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...s which make AI-2, such as DS941 <i>E. coli</i>, when compared to the same construct in cells that don’t make AI-2, such as DH5α <i>E. coli</i>. The results
...et up from single colonies of strains containing P<sub>lsrA</sub> promoter construct.
15 KB (2,539 words) - 21:46, 1 November 2017
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Jurkat and HEK293T cells were transduced with
construct containing the suicide gene <i>herpes simplex virus 1</i> thymidine ki
<p><strong>Figure 11: Cloning strategy of knockdown
construct.</strong><br>
64 KB (8,680 words) - 03:27, 2 November 2017
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– Positive colonies carrying construct in tetracycline backbone from last week were re-streaked to check for viabi
...screening by digestion was done with modifications as the plasmids and the construct were of similar size therefore needed better resolution. <br>
28 KB (4,144 words) - 21:46, 1 November 2017
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<p>After achieving getting the signal for a particular VOC, we aim to construct logical gates and circuits by using CRISPRi system on plasmids to create we
...ration in their urine samples compared to healthy people. [4] The designed construct aims to get a correlated GFP signal with formaldehyde concentration.
27 KB (4,457 words) - 01:26, 2 November 2017
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Fragments obtained from IDT to construct our plasmid designs were not suitable for Gibson Assembly. Therefore, we or
...box shows amplification of a single fragment containing all the genes for construct K2268006 (MazF and cI repressor). The second yellow box shows the various f
7 KB (527 words) - 01:28, 2 November 2017
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...n any modifications (except for a substituted glycine on the A chain), our construct is outside the patent<o:p></o:p></span></p>
...ang="EN-AU" style="font-size:14.0pt;font-family:Quicksand">In our sequence construct </span><span lang="EN-AU" style="font-size:14.0pt;font-family:Quicksand;mso
294 KB (38,580 words) - 22:35, 22 November 2017
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...the fact that we were a lacking some reagents was key for us to decide to construct a constitutive circuit. At last, what we were trying to prove at this point
...uences at the NCBI database better characterized. Therefore, we decided to construct two new Biobricks with better experimental backgrounded proteins that will
7 KB (1,195 words) - 00:13, 28 October 2017
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...the parts registry page from last year was updated for clarity on the full construct design.
11 KB (1,383 words) - 03:52, 2 November 2017
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<p id="
CONSTRUCT-1-Part-BBa_K1163103-0"><img alt="File:Judd UK Updated Constructs.jpeg" src=
<h2 id="
CONSTRUCT-1-Part-BBa_K1163103-0">
CONSTRUCT 1: Part BBa_K1163103</h2>
4 KB (558 words) - 17:30, 1 November 2017
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...he fact that we were a lacking some reagents was vital for us to decide to construct a constitutive circuit. At last, what we were trying to prove at this point
...uences at the NCBI database better characterized. Therefore, we decided to construct two new Biobricks with better experimental backgrounded proteins that would
15 KB (2,493 words) - 03:40, 2 November 2017
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...(<a href="http://parts.igem.org/Part:BBa_K2447014">BBa_K2447014</a>). The
construct was functioning properly and elucidating higher GFP expression only when th
...(<a href="http://parts.igem.org/Part:BBa_K2447000">BBa_K2447000</a>). The
construct was functioning properly and elucidating higher GFP expression only when th
7 KB (963 words) - 11:59, 1 November 2017
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<p>Succeed
construct BBa_K2507004, BBa_K2507006, BBa_K2507008& BBa_K2507013.</p>
<p>
Construct BBa_K2507009, BBa_K2507010, BBa_K2507011, BBa_K2507012& BBa_K25070014, BBa_
10 KB (1,213 words) - 19:22, 1 November 2017
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...ffect. We want to find out if it is really the case in our warriors, so we
construct our warriors and carry out the killing test. <br><br></div>
<div class="myPara" > 1.
Construct our warriors and beggars by co-transformation. (the gene circuits are as fo
15 KB (2,229 words) - 15:55, 1 November 2017
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In order to test the mechanism, a strain containing a construct expressing GFP, LuxI and LuxR under the pLux promoter and the aiiA molecule
...<b>Figure B:</b> Gene sequence depicting the Inducible quorum sensing construct whose decrease in fluorescence is induced under the addition of anhydrotetr
64 KB (7,345 words) - 22:30, 1 November 2017
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...in (HBG) intron 2. We used this split mKate 2-exon
construct as a reporter
construct that we used to determine if our dCas13 or Ms2 systems were successful in s
<p>The figure above represents our
construct. The sequence in between the exons represents HBG intron 2. The 3' splice s
4 KB (655 words) - 01:31, 2 November 2017
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...Master Mix protocol. 7 colonies from each plate were run along with a RFP Construct postitive control and milliQ water negative control.
...Colonies from each 9/30/17 golden gate reaction were run along with a RFP Construct postitive control and milliQ water negative control.
64 KB (11,762 words) - 23:39, 1 November 2017
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<h2>Construct</h2>
...that the parts could be analyzed relative to other systems. Generally, the construct consisted of a constitutive promoter, the riboswitch - which includes a rib
9 KB (1,391 words) - 02:48, 16 December 2017
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...e estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of int
...e estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of int
7 KB (951 words) - 23:51, 1 November 2017
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...e estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of int
...e estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of int
7 KB (951 words) - 02:34, 2 November 2017
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...berg et al. 2017</a>. We did this in parallel with the assembly of our own construct. The activity of Cas13a was tested with fluorescence activity assays, which
...following unfoldables. See Figure 2 for a schematic overview of the entire construct.</p>
82 KB (10,812 words) - 22:11, 14 December 2017
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...ure 1. Scheme of the 3D printed device for the delivery of the HDT genetic construct with the use of bacteriophages. (A) Soft-impermeable membrane that allows p
...ualization of the 3D-printed prototype for the delivery of the HDT genetic construct with the use of synthetic bacteriophages (A) Closed tube with the Piercing
11 KB (1,735 words) - 02:40, 2 November 2017
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...ve types of culture at 37C. This is promising because it tells us that our construct has no negative impact on cell growth at working conditions. In theory, pro
...future applications of our design. In the future we would incorporate our construct into both pathogenic strains such as MRSA and non-pathogenic strains used i
19 KB (2,518 words) - 23:43, 1 November 2017
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...e estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of int
...e estimation of expression of their protein of interest. In addition, this construct will give an added advantage to tune the expression of their protein of int
31 KB (4,088 words) - 00:32, 2 November 2017
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1 KB (169 words) - 20:30, 1 March 2018
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The first construct containing the genes required for the light-induced dormancy was designed a
...ite between the LacI-regulated, lambda hybrid promoter and this particular construct under the tested conditions had formed a hotspot for transposons. In the li
384 KB (54,129 words) - 02:00, 2 November 2017
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...is. The graph below shows evidence for mf-Lon induced toxicity. The J23100 construct does not possess a pdt and fluorescence decreases despite that no GFP is ge
...se.) We started thinking about how exactly we'd want to design an mScarlet construct. Planning to clone mScarlet versions of J23100 + pTet parts (3K3 + 1C3).</d
30 KB (4,512 words) - 03:20, 2 November 2017
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...attached at the end of the tsPurple construct. BBa_K2353000, the tsPurple construct without any degradation tag, shows the baseline expression of tsPurple as a
11 KB (1,436 words) - 03:40, 2 November 2017
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...ssembled because it is the same promoter that is on the second part of our construct. Due to this, we moved back to pλR LacI and inoculated liquid cultures fro
In order to troubleshoot our pλR LacI construct, we decided to attempt to recreate the part from the iGEM Distribution Kits
17 KB (2,322 words) - 23:54, 1 November 2017
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The first construct containing the genes required for the light-induced dormancy was designed a
...ite between the LacI-regulated, lambda hybrid promoter and this particular construct under the tested conditions had formed a hotspot for transposons. In the li
424 KB (60,406 words) - 11:01, 20 November 2017
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...nstruct plasmids were all tested. The OD reflects the fluorescence of each construct tested in the presence of sugar. Again, ribose and sorbitol show the greate
...analysis compared three BL21 (DE3) colonies before (Pre) and after (Post) construct expression was induced with IPTG. Marker used was 1kb+ ladder (Invitrogen)
9 KB (1,325 words) - 01:43, 2 November 2017
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...he combination of up to 7 different transcription units yields a multigene
construct. </p>
<br><i><div style="font-size:80%;"> Figure 2: Different levels of
construct assembly (P: promotor, CDS: coding sequence, T: terminator)</div></i><br><b
57 KB (7,992 words) - 20:08, 1 November 2017
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<p>To quantify the amount of PHB made by our phaCBA
construct, we made 9 overnight cultures of <i>E. coli</i> BL21(DE3) that contained th
<p>The results showed that PhaJ was being expressed for our pET29b(+)-phaCJ
construct in the insoluble phase. After trouble-shooting our experiment, we decided t
189 KB (30,380 words) - 06:23, 16 December 2017
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...936003): Part of a red light-controlled synthetic genetic switch. Chimeric construct comprising the N-terminal fragment of A. thaliana PhyB (amino acids 1-650)
...etic switch. Designed module for the constitutive expression of a chimeric construct comprising a DNA binding protein fused to A. thaliana PIF6 (amino acids 1-1
26 KB (3,213 words) - 02:23, 2 November 2017
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...n of PhiC31 actuation in plant-human genetic circuit. A) Register assembly construct composed by 35s promoter inverted sequence and Mtb terminator flanked by tw
...">Figure 3. Graphic representation of Plant-Human genetic circuit with the construct comprised by a promoter and a terminator in opposite directions flanked by
41 KB (5,363 words) - 03:07, 2 November 2017
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...ation strategy, we plan to synthesize the remaining tRNA in our expression construct for submission into this collection.</p>
...><b>Validation:</b> Finally, we have also included our measurement control construct for the complete TX-TL system. This part expresses a RNA Spinach tagged EYF
24 KB (3,354 words) - 02:38, 2 November 2017
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...e <i>vox-ura3-terminator-vox</i> structure above. Therefore, it is easy to construct because we only need to change the <i>ura3</i> gene to the <i>yEmRFP</i> ge
...conveniently. To show the function of Mating Switcher more intuitively, we construct this<i> RFP</i> system to be a characterization.
52 KB (7,750 words) - 03:12, 2 November 2017
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All plasmids except pSB1C3-PtNTT2 were tagged with the same cMyc-GFP construct for imaging and western blots. cMyc was used as a linker to allow the targe
...-cMyc-GFP with a TAT signal peptide. No bands can be observed for the full construct, but a very weak band can be seen between 55 and 70 kDa for the fusion prot
56 KB (8,532 words) - 20:58, 29 October 2017
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At first, we designed our construct carrying our synthesized single guide RNA (sgRNA), which is essential for t
The PCR-construct responsible for the homologous recombination, and therefore the <i>cod</i>A
30 KB (4,719 words) - 03:58, 2 November 2017
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<h3> construct a biobrick </h3>
<h3> construct a biobrick </h3>
131 KB (18,470 words) - 03:56, 2 November 2017
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...choice of a basic strain of microorganism producers for expression of the
construct created.</p>
<p>When it was time to clone our
construct in the pSB1C3 vector that we had received from iGEM our team faced some dif
11 KB (1,746 words) - 12:18, 30 October 2017
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...the three functional 14-3-3 monomers. We also described that the Scaffold construct would be a tetramer, but because we mutated the last pocket, the pocket has
...ude a Strep-tag®II. The CT33 site can bind in the pockets of the Scaffold construct filled with inducer and the Strep-tag®II site can bind to Strep-Tactin®XT
4 KB (563 words) - 13:48, 10 December 2017
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<p>LB,
construct, Tet</p>
<p>-
Construct primers for G-blocks</p>
23 KB (3,938 words) - 20:58, 31 October 2017
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...he effect of different concentrations of luciferin on <i>Agrobacterium</i>-
construct infiltrated leaf disc samples. This can be seen on the <a href="https://201
2 KB (314 words) - 11:45, 30 October 2017
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...s why we could not synthesize all of the genes as a single
construct. Each
construct also contained promoter, ribosome-binding sites and the necessary homologie
<div style="padding-bottom:3%"> A
construct with beta-carotene hydroxylase gene <i>crtZ</i> and lycopene cyclase gene <
14 KB (1,977 words) - 21:23, 1 November 2017
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...l part of the dCas9QC with a gBlock containing the VRER mutations. The new construct was analyzed by sequencing the last part containing the gblock (figure 4).
...). These gRNA's were inserted into the plasmid pSB3C5 containing a pLacGFP construct as well. For dCas9 and dCas9VRER expression those parts were put into a pBa
21 KB (3,475 words) - 03:16, 2 November 2017
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Our CRISPR/dCas13a
construct functions as an “mRNA localization device” to investigate the fate of
<img src="https://static.igem.org/mediawiki/2017/d/d0/T--Kent--
construct.png">
15 KB (1,712 words) - 00:26, 16 December 2017
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...hich HheC-W249P achieve the best results, so We finally used the mutant to construct the plasmid.
...yme ratio of 1: 2: 4, so although EchA is expressed in the plant, we still construct the gene, hoping to let the plant overexpress the enzyme to a certain exten
20 KB (2,797 words) - 21:09, 30 October 2017
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<li> Make a label (clone ID # and date) for the construct.</li>
...<li> Determine the location (-80°C tower #, box #, row #) of the construct.</li>
28 KB (3,495 words) - 22:23, 1 November 2017
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We tested our
construct using <a target="_blank" href="https://2017.igem.org/Team:TECHNION-ISRAEL/c
...rding to a different strategy. Additionally, the protein order within each
construct was randomized to ensure the issue was not caused by specific proteins. In
14 KB (1,769 words) - 02:10, 2 November 2017
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...een populations in real ecosystems, we use methods of synthetic biology to
construct a more simple system using <i>E.coli</i>. The system involves two groups of
...<div class="myPara"> Therefore, our main task is to
construct three characters of the two groups. <br>
18 KB (2,437 words) - 03:33, 2 November 2017
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...e protein ACA3 (see figure 18). We were able to successfully assemble this construct (see gel electrophoresis), however we did not manage to successfully transf
...f our biobrick with 18bp overlaps for Gibson assembly. However, this large construct required 7 fragments (see figure 20) and we were unsuccessful in our attemp
64 KB (8,040 words) - 10:51, 31 October 2017
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<p>As the maximum activity was observed in pPMMVFLt12
construct, for the present study the strong promoter having the synthetic cis element
<p><b>
Construct Formation:</b></p>
14 KB (1,923 words) - 17:58, 29 October 2017
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...The detailed cloning workflow which led to the finished Evaluation Vector
construct with the pBS1C backbone.</figcaption></figure>
...d it into the pSB1C3 backbone (via EcoRI and PstI digest) and verified the
construct by sequencing. It has been submitted to the parts registry under <a target=
13 KB (2,057 words) - 00:02, 2 November 2017
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construct. Our dCas9 will target the region between the transcription start
a component of the natural CRISPR-system, but it is a synthetic construct
12 KB (1,959 words) - 12:52, 24 October 2017
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...signaling cascade for the biosensor with a CRE-luciferase construct as the reporter.
to construct stable cell lines. Based on these cell lines, we infected different
79 KB (5,538 words) - 08:25, 25 October 2017
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<li>
Construct KD Tree based on the normalized word vectors of the key words in the databa
<p>This is the final step to
construct our Recommendation System , overall we use collaborative filtering strategy
27 KB (3,894 words) - 23:43, 1 November 2017
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...unidirectional terminator (ECK120034435) between them. This construct can be regarded as an
...troduced one or two pairs of attB/P sites on both sides of ECK120034435 to construct
40 KB (3,961 words) - 23:39, 1 November 2017
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...proteins on the dual-expression vector (pETDuet-1, pCDFDuet-1) for further construct other circuit using for detection.</p>
...striction on the use of T3 + sfGFP Endonuclease PstI and EcoRI digested to construct the expressed product. The electrophoresis tank buffer was 0.5 x TBE and th
31 KB (3,572 words) - 04:17, 9 December 2017
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...proteins on the dual-expression vector (pETDuet-1, pCDFDuet-1) for further construct other circuit using for detection.</p>
...striction on the use of T3 + sfGFP Endonuclease PstI and EcoRI digested to construct the expressed product. The electrophoresis tank buffer was 0.5 x TBE and th
27 KB (3,288 words) - 19:29, 1 November 2017
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<div class="page-heading">MESA construct replication</div>
The first few weeks, we tried to replicate the MESA construct using Luciferase-GFP fusion protein as the output signal and dsRED as a tra
125 KB (17,334 words) - 16:58, 27 October 2017
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<li> MG1655 transformed with <i>cysPUWA</i> construct in LB media with varying concentrations of potassium chromate (20-160 uM)
<li> MG1655 transformed with <i>cysPUWA</i> construct in LB media with no potassium chromate </li>
11 KB (1,541 words) - 01:55, 2 November 2017
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...apoprotein. After the first time purification, we refolded the protein to construct the correct structure and then we activated the apo-HRP with hemin to produ
...nce we failed to express GFP and ER-alpha at the same time, we will try to construct the gene containing GFP and ER-alpha and express them together for the purp
19 KB (2,852 words) - 01:27, 2 November 2017
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...ntibiotics. Previous work shows that when colonies containing the reporter construct are grown in LB medium there is very little response to the addition of try
...gem.org/wiki/index.php?title=Part:BBa_K2330002"> BBa_K2330002</a> reporter construct)</p>
9 KB (1,404 words) - 03:44, 2 November 2017
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...FP</em> being inserted into a DVK-AF plasmid. <strong>A:</strong> Displays construct assembly, BsaI:R indicates the
...eft. A, E and F = MoClo fusion sites. <strong>B:</strong> Overview of each construct and DVK-AF once it has been cut by BsaI. <strong>C</strong>: After ligation
7 KB (1,210 words) - 01:03, 2 November 2017
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<div class="note-p">Initially we planned to
construct pBBR1-<i>rhlABC</i>-<i>fimS</i>-<i>lacI</i>-<i>lacO</i>-<i>BcsA</i> and pBa
<div class="note-p">We continued OE PCR, simultaneously, we went on to
construct pBad30-cm-<i>fimE</i>.</div>
17 KB (2,496 words) - 03:52, 2 November 2017
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All plasmids except pSB1C3-PtNTT2 were tagged with the same cMyc-GFP construct for imaging and Western Blots. cMyc was used as a linker to allow the targe
...-cMyc-GFP with a TAT signal peptide. No bands can be observed for the full construct, but a very weak band can be seen between 55 and 70 kDa for the fusion prot
69 KB (10,514 words) - 03:26, 2 November 2017
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After obtaining the E.coli, we designed a construct, known as a biobrick, to allow the bacteria to express the proteins lipase
...came the gene for plastic degradation. One construct had esterase, and one construct had lipase.
7 KB (1,019 words) - 19:41, 19 November 2017
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...tical-align:middle;">Cloned <i>rdhANP</i> into PSB1C3 and transformed the construct into DH5α</td></tr>
...tical-align:middle;">Cloned <i>rdhANP</i> into PSB1C3 and transformed the construct into DH5α</td></tr>
31 KB (4,040 words) - 07:51, 1 November 2017
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...giving it yellow color. Construct 1 is cloned at AatII and ApaI site, and construct 2 is cloned at SalI and BamHI site.</h3></section>
...e copy plasmid, pACYC177. Construct 1 was cloned at XhoI and XmaI site and construct 2 was cloned at BamHI and AatII site.</h3></section><br />
5 KB (751 words) - 04:13, 14 December 2017
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<li> pSB1C3-TraR construct from the 11 of September 2017 </li>
<li> pSB1C3-TraAC construct from the 11 of September 2017 </li>
37 KB (5,520 words) - 03:50, 2 November 2017
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...org/Team:UT-Knoxville/Project#PuPromoter">here</a> to read more about this construct. </li>
...uct, our Pu promoter showed ~4x GFP expression than our solely 0.1 mM IPTG construct. Thus, our regulatory sequence including xylR regulatory protein and Pu pr
23 KB (2,783 words) - 02:29, 2 November 2017
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...onstruct 1, Ptet- RBS1- T7 RNA polymerase –terminator 1-terminator 2 and construct 2, PT7-RBS2- Chromoprotein II- RBS3-tetR-terminator3. Different parts like
<h3>For cloning of construct 1 of module 2, Pmar-RBS4- ToxR-terminator 4 and construct 2, Pctx-RBS5- chromoprotein I- RBS6-TetR- terminator5. Different parts like
7 KB (1,022 words) - 04:12, 14 December 2017
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...rescence microscopy images of Low and High strength Anderson promoter-PduD construct associated mCherry expressed alone and co-expressed with Eut subunits.</cen
An added benefit of our SMNLK+PPK construct is that it would allow us to determine whether it actually worked via DAPI
8 KB (1,131 words) - 03:26, 2 November 2017
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<h2>Whole construct assembly design</h2>
Final construct looks like this (Figure 3):</p><br>
15 KB (1,912 words) - 02:32, 2 November 2017
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...>BBa_K2350020</a></b>). Not only this, we even grouped endolysin-holin’s
construct with Nrt-A and created a new part (<b><a href="http://parts.igem.org/Part:B
...xt-decoration:none;">BBa_I742157</a></b>) . Same with indigoidine, we also
construct this part on our special designed backbone pPIGBACK so that it can be expre
6 KB (931 words) - 19:49, 1 November 2017
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...spoke with Team Tokyo Tech, who inspired us to <b style="color:#ff7575;">construct a plasmid containing Melanin (black pigment) sequence</b> for maximizing li
...onths of research on increasing the oil production in algae, we started to construct plasmids which would be used in our project. However, some question arose d
16 KB (2,075 words) - 23:43, 1 November 2017
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.... Suddenly I came up with an interesting idea that what if the scaffold is
construct by <font color="#1E90FF"><b>protain</b></font>. Then I realized that scaffo
<p class="dummy-text">So now how do we
construct the scaffold. We knew that there was many protein self-assembly system but
23 KB (3,147 words) - 09:16, 29 August 2017
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...ctive protein degradation tag (and a pdt-less inducible mScarlet-I control construct), all under the control of the pTet promoter. These parts, in combination w
...he strong constitutive promoter J23100. We also included a tagless control construct (J23100 mScarlet with no pdt) as a comparison. In order to demonstrate that
30 KB (4,615 words) - 02:13, 2 November 2017
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...t molecules can be recruited. This way, it will be possible to enhance the construct with bioactive functionalities. </br>
4 KB (555 words) - 19:55, 27 June 2017
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<p id="homeP">These construct backbones are assembled via Gibson Assembly and transformed into <i>E. coli
...tios to a single tube and assemble the insert and vector into an a plasmid construct.
10 KB (1,326 words) - 03:47, 2 November 2017
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...<div class="figure-text"><strong>Figure 1. The STAR-chromoprotein
construct.</strong></div>
...<div class="figure-text"><strong>Figure 2. The STAR 3 - As - CP
construct</strong></div>
17 KB (1,418 words) - 09:26, 10 December 2017
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The full genetic construct is comprised of three parts: pλR LacI, one of the three tsPurple parts (ts
...G, the relative levels of degradation would be compared; theoretically the construct with no degradation tag would show no degradation, the DAS tag (which is a
11 KB (1,456 words) - 16:09, 16 December 2017
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...y xylulose present, giving the xylulose molecules a chance to activate our construct.
...osities of fluids would cause non-uniform mixing of the samples inside the construct.
7 KB (1,150 words) - 23:05, 1 November 2017
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<figcaption><strong>Figure 9.1:</strong> Sketch of our GFP-LOV-PTS1 construct</figcaption>
<figcaption><strong>Figure 9.2:</strong> Sketch of our PTS2-import construct</figcaption>
85 KB (11,201 words) - 02:57, 2 November 2017
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...In this work, our goal is to construct a biological odor sensor in an easily cultured eukaryotic cells (HEK293) by
...dor compound/receptor pairs (β-citronellol/OR1A1 and bourgeonal/OR1D2) to construct the iSmeller odor sensors and evaluate their sensitivity and specificity. S
22 KB (1,602 words) - 16:07, 25 October 2017
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...ttom of the tube. 10μl of MilliQ ultrapure water was added to each of the construct wells. The reconstituted plasmid DNA was stored at -4°C.</p><br>
...each construct (16 in total). Using P200 pipette tips, 2 colonies for each construct were picked from their respective plates. The test tubes were placed on a l
16 KB (2,686 words) - 03:06, 2 November 2017
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total of 9 total parts that needed to be stitched into a full construct. This led to the use of the Gibson Assembly
in order to increase the rate of success of creating our construct. </h4><br>
13 KB (1,605 words) - 02:04, 2 November 2017
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...que that is similar to blue/white screening. User can use this biobrick to construct their toehold switches that use pT7 as the promoter and mRFP as the reporte
20 KB (2,806 words) - 20:57, 1 November 2017
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...od of time before undergoing knock out by Cre recombinase. Knocking out of
construct will lead to inactivation of the gene of interest, providing a secure and s
<p>There are two scenarios of activity in Time Module
Construct:</p>
4 KB (681 words) - 05:02, 3 October 2017
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...we interviewed different experts to evaluate the safety assessment of our construct. The talks focused on the importance of incorporating deficiencies. After m
...ion that there are no major security vulnerabilities, since we planned our construct according to
6 KB (822 words) - 03:45, 2 November 2017
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<p> The strategy we chose to construct the vector is to fuse B0015 and AmpR together first.
...p>10, 11</sup> Thus, we used IDT service to synthesize these two genes and construct it on pPIGBACK, a vector which can express the carrying genes in <i>S. elon
33 KB (4,501 words) - 17:42, 1 November 2017
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...tion tent which acts to increase the humidity to promote fast germination. Construct with clear plastic and kebab sticks.
...ormed the second gateway reaction with pB2GW7 to produce our final plasmid construct. </p>
39 KB (5,765 words) - 03:46, 1 November 2017
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4. Construct the INP-mSA part by Gibson assembly. <br/>
3. Construct the BirA-pYC230 circuit by T4 DNA ligase and transform into EBY100 strain.
34 KB (3,489 words) - 13:59, 18 November 2017
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...<th style="background-color: #377b7e; color: #ffffff; font-weight: bold;">Construct</th>
...ckground fluorescence for both biological replicates. The positive control construct exhibited reproducible patterns of fluorescent protein expression over the
10 KB (1,509 words) - 02:40, 2 November 2017
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<h2>General Protein Construct</h2>
<h2>General RNA Construct</h2>
12 KB (1,662 words) - 02:15, 2 November 2017
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...he usage of our non-canonical amino acids 2-nitrophenylalanine we aimed to construct a purification column that can induce the elution of the target protein by
...oncept. This led to another version of the column we have not been able to construct yet. However, we instead created a 3D-model to show it.
104 KB (12,792 words) - 20:31, 14 December 2017
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...arts.igem.org/partsdb/part_info.cgi?part_name=BBa_K2404013"> 35S:Luc+:NosT construct </a>, and quantified the activity of the 35S promoter using a luciferase re
...ref="http://parts.igem.org/Part:BBa_K2404013:Experience">35S:Luc+:NosT</a> construct. The red regions show high levels of luciferase expression. This part can n
6 KB (922 words) - 17:02, 31 October 2017
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...to use a flow cytometer to thoroughly quantify the expression of the deGFP construct, using fluorescence-activated cell sorting (FACS) as this method was not ac
After the arrival of the DNA, we transformed the deGFP construct into <i>E. coli</i> Top10 cells before we ran the sample through the flow c
4 KB (614 words) - 03:32, 2 November 2017
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...rescent Protein (GFP) variant for the first time. We created an expression construct (J23100 promoter and B0034 RBS in front of deGFP) and characterised the deG
...the sarcosine oxidase enzyme for the first time. We created an expression construct (T7 promoter and B0034 RBS in front of SOx CDS) and demonstrated that SOx i
15 KB (2,035 words) - 18:30, 1 November 2017
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The Cre-Cas9 construct was successfully assembled together into a plasmid. To make the P<sub><i>FU
...switch worked. The results came back negative. Sequencing of the Cre-Cas9 construct showed mutations in the Cre-recombinase part and might be the reason to why
256 KB (35,572 words) - 17:24, 30 October 2017
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...from Development and Application of a Synthetically-Derived Lead Biosensor Construct for Use in Gram-Negative Bacteria, in the journal Sensors. The backbone, kn
...from Development and Application of a Synthetically-Derived Lead Biosensor Construct for Use in Gram-Negative Bacteria. It shows that the promoter expressed GFP
7 KB (1,112 words) - 21:21, 13 October 2017
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...ruct was inserted into the vectors via Gibson Assembly method. The ligated construct was then transformed in competent E.coli and plated on chloramphenicol (CAM
24 KB (3,933 words) - 12:37, 16 October 2017
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...sensor Construct for Use in Gram-Negative Bacteria" published in Cell. The construct was used with permission by Dr. Ashley Franks.
7 KB (1,209 words) - 16:28, 28 October 2017
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<td>Construct comprised of PtNTT2, lacO and Cas9</td>
<td>Construct for the integration of unnatural base pairs</td>
28 KB (4,174 words) - 00:37, 2 November 2017
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...genes hpaB and hpaC in E. coli. To simplify our assembly, we designed our construct to position hpaB and hpaC coding sequences in succession under the control
...vector containing a pMMB67EH ori and spectinomycin resistance marker. Our construct was assembled using golden gate cloning techniques. To do so, we first used
5 KB (681 words) - 05:01, 16 December 2017
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...of the pockets. We choose to start with only empty pockets of the Scaffold Construct.<br/>
<li><span>The binding of an inducer to the pocket of the Scaffold Construct.</span></li>
7 KB (1,169 words) - 22:19, 1 November 2017
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...Ba_J23117" style="color:#C1D35D">j23117</a>. Along with one promoter, each
construct also had one of the two RBS (
...parts.igem.org/Part:BBa_J364100" style="color:#C1D35D">j364100</a>). Every
construct had the same GFP (<a href="http://parts.igem.org/Part:BBa_E0040" style="col
19 KB (2,490 words) - 02:59, 2 November 2017
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...m.org/Part:BBa_K2475002">PrD-YFP</a>, and C is used as a shorthand for our
construct <a href="http://parts.igem.org/Part:BBa_K2475001">PrD-CFP</a>. The samples
<p>In order to determine if [psi-] C was fluorescing because of our
construct, we checked its fluorescence value after 12 hours of growth in YPD. A signi
11 KB (1,552 words) - 03:28, 2 November 2017
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In the near future we would like to finish our final construct for the dCas9 project and measure the Auxin concentration to determine if w
...re how much (ideally: functional) respective protein is produced with each construct.
7 KB (894 words) - 11:24, 1 November 2017
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Our construct
.../. Gene sequence was optimized to eliminate illegal restriction sites. The construct was synthezied by IDT.
3 KB (495 words) - 22:32, 14 December 2017
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<br><br> To have a construct work in <i>Agrobacterium</i> we needed to use a plasmid that could replicat
<i>ietAS</i> construct. The resulting ligation was then transformed into chemically competent <i>E
17 KB (2,525 words) - 03:58, 2 November 2017
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...he plcA gene is under the control of PlcR, its promoter is usually used to construct reporter system.</p>
...take the pDG1730-pAim-AimR-AimP-pAimX-GFP (abbreviated as pDG1730-Aim-GFP) construct as an example. The Aim sequence was synthesized. After that, we cloned the
20 KB (2,735 words) - 13:46, 1 November 2017
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...<b>Figure 3-12 CsgD and OmpR234 Expression </b> Our construct includes a strong promoter, two strong RBS, csgD, ompR234 and double termin
...gure 3-13). Sequencing results from Tri-I Biotech confirmed that our final construct is correct.
25 KB (2,679 words) - 13:33, 31 October 2017
