FOUR A DIVERGENT COLLECTION EPUB
Download this ebook at: ppti.info?book= [PDF] Four: A Divergent Collection [PDF] Four: A Divergent Collection. [EbooK Epub] Four: A Divergent Collection (Divergent Series Story) Ebook | READ ONLINE For download this book click button below. Fans of the Divergent trilogy by #1 New York Times bestselling author Veronica Roth will be thrilled by Four: A Divergent Collection, a companion volume that includes four pre-Divergent stories plus three additional scenes from Divergent, all told from Tobias's point of view.
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Four: A Divergent Collection (Divergent Series Prequel Stories) by Veronica Roth A collection of four pre-Divergent stories plus three additional scenes from. Download PDF Four: A Divergent Collection, PDF Download Four: A Divergent Collection, Download Four: A Divergent Collection, PDF Four: A. Divergent Series Ultimate Four-Book Collection: Divergent; Insurgent; Allegiant; Four (Divergent Series Read online, or download in secure EPUB format.
Here we present a general strategy to engineer aTF response to new inducer molecules, using the E.
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As new inducer targets, we chose gentiobiose, fucose, and synthetically derived lactitol and sucralose, four saccharides not metabolizable by E. These molecules represent targets with increasing apparent chemical difference from known LacI inducers Supplementary Fig.
Design, synthesis and assembly of variants To capture the effects of protein residues both proximal and distal to the ligand-binding pocket, we used three methods to create LacI variants: computational design, protein-wide single-amino-acid saturation mutagenesis and error-prone PCR.
Using an adaptation of the Rosetta software suite 23 , 24 , we computationally designed LacI variants to bind the three target ligands that are most dissimilar from the native inducer: fucose, lactitol and sucralose Online Methods and Supplementary Note.
Rosetta has been used to design proteins with new ligand-binding interactions 25 , though it does not account for allostery.
We generated DNA oligonucleotides encoding designed variants by microarray-based synthesis 26 , which specifies a pool of exact oligonucleotide sequences Online Methods.
Because of oligonucleotide length limitations, residues mutable during Rosetta design were confined to three segments of lacI encoding residues 73—, — and — , encompassing the majority of the ligand-binding pocket. We synthesized and cloned LacI libraries encoding each single segment mean of 4.
Substitution of aTF residues distal to the ligand interface can influence induction through long-range effects 19 , 27 — Thus, we created a variant library encoding all LacI single-amino-acid substitutions using microarray-synthesized DNA by tiling mutable sequences in windows of 36 residues, totaling 6, variants.
Finally, we amplified lacI codons 67— by error-prone PCR to generate a library with a mean of five mutations per gene Online Methods.
What is Kobo Super Points?
A screen to identify LacI variants with new ligand response Affinity-based screens can evaluate binding, but not allostery, so we developed an in vivo selection-screening method designed to capture aTF variants functional in both allosteric states: DNA-bound in the absence of inducer and allosterically activated by inducer Fig.
Into the genome of E. Subsequently we collected variants that activated transcription of the GFP gene in response to a target ligand by fluorescence-activated cell sorting FACS; Supplementary Fig. Your result was textbook Abnegation. Like she's sad about something. You can say whatever you want here. I chose food over a weapon. I threw myself in the path of the dog to save the little girl. I knew that after I made those choices, the test would end and I would receive Abnegation as a result.
And I don't know that I would have made different choices if my father hadn't coached me, hadn't controlled every part of my aptitude test from afar. So what was I expecting? What faction did I want? Any of them. Any of them but Abnegation. I don't care what she says-this isn't a safe room. There are no safe rooms, no safe truths, no safe secrets to tell.
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I can still feel the dog's teeth closing around my arm, tearing my skin. I nod to Tori and start toward the door, but just before I leave, her hand closes around my elbow. But you never will. I return to the cafeteria and sit down at the Abnegation table, among the people who barely know me. My father doesn't permit me to come to most community events.
He claims that I'll cause a disruption, that I'll do something to hurt his reputation. I don't care. I'm happier in my room, in the silent house, than surrounded by the deferential, apologetic Abnegation. The consequence of my constant absence, though, is that the other Abnegation are wary of me, convinced there's something wrong with me, that I'm ill or immoral or strange. Even those willing to nod at me in greeting don't quite meet my eyes.
I sit with my hands clenching my knees, watching the other tables, while the other students finish their aptitude tests. The Erudite table is covered in reading material, but they aren't all studying-they're just making a show of it, trading conversation instead of ideas, their eyes snapping back to the words every time they think someone's watching them. The Candor are talking loudly, as always. The Amity are laughing, smiling, pulling food from their pockets and passing it around.
The Dauntless are raucous and loud, slung over the tables and chairs, leaning on one another and poking one another and teasing.
I wanted any other faction. Any other faction but mine, where everyone has already decided that I am not worth their attention. Finally an Erudite woman enters the cafeteria and holds up a hand for silence.
The Abnegation and Erudite quiet down right away, but it takes her shouting "Quiet! The Choosing Ceremony will be tomorrow at the Hub. Plan to arrive at least ten minutes before it begins. You are dismissed.
I know the path my fellow Abnegation will take out of here, down the hallway and out the front doors to the bus stop. They could be there for over an hour letting other people get on in front of them. I don't think I can bear any more of this silence.
Instead of following them, I slip out a side door and into an alley next to the school. I've taken this route before, but usually I creep along slowly, not wanting to be seen or heard. Today all I want to do is run. I sprint to the end of the alley and into the empty street, leaping over a sinkhole in the pavement. My loose Abnegation jacket snaps in the wind, and I peel it from my shoulders, letting it trail behind me like a flag and then letting it go.
I push the sleeves of my shirt up to my elbows as I run, slowing to a jog when my body can no longer stand the sprint. It feels like the entire city is rushing past me in a blur, the buildings blending together.
I hear the slap of my shoes like the sound is separate from me. Finally I have to stop, my muscles burning. I'm in the factionless wasteland that lies between the Abnegation sector and Erudite headquarters, Candor headquarters, and our common places. At every faction meeting, our leaders, usually speaking through my father, urge us not to be afraid of the factionless, to treat them like human beings instead of broken, lost creatures.
But it never occurred to me to be afraid of them. I move to the sidewalk so I can look through the windows of the buildings.
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Most of the time all I see is old furniture, every room bare, bits of trash on the floor. When most of the city's residents left-as they must have, since our current population doesn't fill every building-they must not have left in a hurry, because the spaces they occupied are so clean. Nothing of interest remains.
When I pass one of the buildings on the corner, though, I see something inside. The room just beyond the window is as bare as any of the others I've walked by, but past the doorway inside I can see a single ember, a lit coal.
I frown and pause in front of the window to see if it will open. At first it won't budge, and then I wiggle it back and forth, and it springs upward.
I push my torso through first, and then my legs, toppling to the ground inside in a heap of limbs. My elbows sting as they scrape the floor.
The building smells like cooked food and smoke and sweat.
I inch toward the ember, listening for voices that will warn me of a factionless presence here, but there's only silence.Computational approaches can sample a much larger mutagenic space; for example, homology modeling-based redesign of PobR was used for 3,4-dihydroxybenzoate response 20 , and mechanistic insights were leveraged to introduce vanillin response to QacR The woman barely looked at me on my way out of the room.
Had a memorable face. Marie Lu.
Today all I want to do is run. To all appearances, my room is as sparse and clean as every other Abnegation room.
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