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Golden Gate assembly is one of the most widely used DNA assembly methods due to its robustness and modularity. However, despite its popularity, the need for BsaI-free parts, the introduction of scars between junctions, as well as the lack of a comprehensive study on the linkers hinders its more widespread use. Here, we first developed a novel sequencing scheme to test the efficiency and specificity of 96 linkers of 4-bp length and experimentally verified these linkers and their effects on Golden Gate assembly efficiency and specificity. We then used this sequencing data to generate 200 distinct linker sets that can be used by the community to perform efficient Golden Gate assemblies of different sizes and complexity. We also present a single-pot scarless Golden Gate assembly and BsaI removal scheme and its accompanying assembly design software to perform point mutations and Golden Gate assembly. This assembly scheme enables scarless assembly without compromising efficiency by choosing optimized linkers near assembly junctions.

Sugarcane, a tropical C4 grass in the genus Saccharum (Poaceae), accounts for nearly 80% of sugar produced worldwide and is also an important feedstock for biofuel production. Generating transgenic sugarcane with predictable and stable transgene expression is critical for crop improvement. In this study, we generated a highly expressed single copy locus as landing pad for transgene stacking. Transgenic sugarcane lines with stable integration of a single copy nptII expression cassette flanked by insulators supported higher transgene expression along with reduced line to line variation when compared to single copy events without insulators by NPTII ELISA analysis. Subsequently, the nptII selectable marker gene was efficiently excised from the sugarcane genome by the FLPe/FRT site-specific recombination system to create selectable marker free plants. This study provides valuable resources for future gene stacking using site-specific recombination or genome editing tools.

Data sets for material included in "A 13-year record indicates differences in the duration and depth of soil carbon accrual among potential bioenergy crops" by Kantola et al., 2025, in Global Change Biology Bioenergy. Data include soil organic carbon (SOC), carbon stable isotope ratios, annual belowground biomass, and annual post-harvest litter for four crops, maize/soybean, miscanthus, switchgrass, and prairie, between 2008 and 2021.

Overwintering ability is an important selection criterion for Miscanthus breeding in temperate regions. Insufficient overwintering ability of the currently leading Miscanthus biomass cultivar, M. ×giganteus (M×g) ‘1993–1780′, in regions where average annual minimum temperatures are −26.1°C (USDA hardiness zone 5) or lower poses a pressing need to develop new cultivars with superior cold tolerance. To facilitate breeding of Miscanthus, this study characterized phenotypic and genetic variation of overwintering ability in an M. sinensis germplasm panel consisting of 564 accessions, evaluated in field trials at three locations in North America and two in Asia. Genome‐wide association (GWA) and genomic prediction analyses were performed. The Korea/N China M. sinensis genetic group is a valuable gene pool for cold tolerance. The Yangtze‐Qinling, Southern Japan, and Northern Japan genetic groups were also potential sources of cold tolerance. A total of 73 marker–trait associations were detected for overwintering ability. Estimated breeding value for overwintering ability based on these 73 markers could explain 55% of the variation for first winter overwintering ability among M. sinensis. Average genomic prediction ability for overwintering ability across 50 fivefold cross‐validations was high (~0.73) after accounting for population structure. Common genomic regions for overwintering ability were detected by GWA analyses and a previous parallel QTL mapping study using three interconnected biparental F1 populations. One QTL on Miscanthus LG 8 encompassed five GWA hits and a known cold‐responsive gene, COR47. The other overwintering ability QTL on Miscanthus LG 11 contained two GWA hits and three known cold stress‐related genes, carboxylesterase 13 (CEX13), WRKY2 transcription factor, and cold shock domain (CSDP1). Miscanthus accessions collected from high latitude locations with cold winters had higher rates of overwintering, and more alleles for overwintering, than accessions collected from southern locations with mild winters.

Yarrowia lipolytica has been used to produce both citric acid and lipid-based bioproducts at high titers. In this study, we found that pH differentially affects citric acid and lipid production in Y. lipolytica W29, with citric acid production enhanced at more neutral pH’s and lipid production enhanced at more acid pH’s. To determine the mechanism governing this pH-dependent switch between citric acid and lipid production, we profiled gene expression at different pH’s and found that the relative expression of multiple transporters is increased at neutral pH. These results suggest that this pH-dependent switch is mediated at the level of citric acid transport rather than changes in the expression of the enzymes involved in citric acid and lipid metabolism. In further support of this mechanism, thermodynamic calculations suggest that citric acid secretion is more energetically favorable at neutral pH’s, assuming the fully protonated acid is the substrate for secretion. Collectively, these results provide new insights regarding citric acid and lipid production in Y. lipolytica and may offer new strategies for metabolic engineering and process design.

Enzymatic reduction of oxyanions such as sulfite (SO32−) requires the delivery of multiple electrons and protons, a feat accomplished by cofactors tailored for catalysis and electron transport. Replicating this strategy in protein scaffolds may expand the range of enzymes that can be designed de novo. Mirts et al. selected a scaffold protein containing a natural heme cofactor and then engineered a cavity suitable for binding a second cofactor—an iron-sulfur cluster (see the Perspective by Lancaster). The resulting designed enzyme was optimized through rational mutation into a catalyst with spectral characteristics and activity similar to that of natural sulfite reductases.

Conversion of corn fiber to ethanol in the dry grind process could increase ethanol yields, reduce downstream processing costs and improve overall process profitability. This work investigates the in-situ conversion of corn fiber into ethanol (cellulase addition during simultaneous saccharification and fermentation) during dry grind process. Addition of 30 FPU/g fiber cellulase resulted in 4.6% increase in ethanol yield compared to the conventional process. Use of excess cellulase (120 FPU/g fiber) resulted in incomplete fermentation and lower ethanol yield compared to the conventional process. Multiple factors including high concentrations of ethanol and phenolic compounds were responsible for yeast stress and incomplete fermentation in excess cellulase experiments.

Most native producers of ribosomally synthesized and post-translationally modified peptides (RiPPs) utilize N-terminal leader peptides to avoid potential cytotoxicity of mature products to the hosts. Unfortunately, the native machinery of leader peptide removal is often difficult to reconstitute in heterologous hosts. Here we devised a general method to produce bioactive lanthipeptides, a major class of RiPP molecules, in Escherichia coli colonies using synthetic biology principles, where leader peptide removal is programmed temporally by protease compartmentalization and inducible cell autolysis. We demonstrated the method for producing two lantibiotics, haloduracin and lacticin 481, and performed analog screening for haloduracin. This method enables facile, high throughput discovery, characterization, and engineering of RiPPs.

This dataset contains files and relevant metadata for real-world and synthetic LFR networks used in the manuscript "Well-Connectedness and Community Detection (2024) Park et al. presently under review at PLOS Complex Systems. The manuscript is an extended version of Park, M. et al. (2024). Identifying Well-Connected Communities in Real-World and Synthetic Networks. In Complex Networks & Their Applications XII. COMPLEX NETWORKS 2023. Studies in Computational Intelligence, vol 1142. Springer, Cham. https://doi.org/10.1007/978-3-031-53499-7_1. “The Overview of Real-World Networks image provides high-level information about the seven real-world networks. TSVs of the seven real-world networks are provided as [network-name]_cleaned to indicate that duplicated edges and self-loops were removed, where column 1 is source and column 2 is target. LFR datasets are contained within the zipped file. Real-world networks are labeled _cleaned_ to indicate that duplicate edges and self loops were removed. #LFR datasets for the Connectivity Modifier (CM) paper ### File organization Each directory `[network-name]_[resolution-value]_lfr` includes the following files: * `network.dat`: LFR network edge-list * `community.dat`: LFR ground-truth communities * `time_seed.dat`: time seed used in the LFR software * `statistics.dat`: statistics generated by the LFR software * `cmd.stat`: command used to run the LFR software as well as time and memory usage information

Curated networks and clustering output from the manuscript: Well-Connected Communities in Real-World Networks https://arxiv.org/abs/2303.02813

The data within consist of compressed output files in the form of edgelists (*.edgelist.gz) and nodelists (*.aux.parquet) from large citation network simulations using an agent-based model. The code and instructions are available at: <a href="https://github.com/illinois-or-research-analytics/SASCA">https://github.com/illinois-or-research-analytics/SASCA</a>. In addition, we provide a distribution of citation frequencies drawn from a random sample of PubMed journal articles (pooled_50k_pubmed_unique.csv) and a table of recencies- the frequency with which citations are made to the previous year, the year before that and so on (recency_probs_percent_stahl_filled.csv). A manuscript describing the SASCA-s simulator has been submitted for review and will be referenced in a future version of this data repository if it is accepted. The prefixes sj and er refer to the real world and Erdos-Renyi random graph respectively that were used to initiate simulations. These 'seed' networks are available from the Github site referenced above.

These codes implement the master equation microkinetic modeling (ME-MKM) calculations of Adams et al. (J. Phys. Chem. C 2025, 129, 15, 7285–7294), as well as the automatic derivatives for activation energies and reaction orders in their follow-up work (in review).

This is the data set for the article entitled "Pollinator seed mixes are phenologically dissimilar to prairie remnants," a manuscript pending publication in Restoration Ecology. This represents the core phenology data of prairie remnant and pollinator seed mixes that were used for the main analyses. Note that additional data associated with the manuscript are intended to be published as a supplement in the journal. * In this V2, a second tab was added to the Rest.Ecol.data.xlsx file. This new sheet listed original data source citations that match the RELIX data base, a sister project.

Purpose-grown perennial herbaceous species are nonfood crops specifically cultivated for bioenergy production and have the potential to secure bioenergy feedstock resources while enhancing ecosystem services. This study assessed soil greenhouse gas emissions (CO2 and N2O), nitrate (NO3-N) leaching reduction potential, evapotranspiration (ET), and water-use efficiency (WUE) of bioenergy switchgrass (Panicum virgatum L.) in comparison to corn (Zea mays L.). The study was conducted on field-scale plots in Urbana, IL, during the 2020–2022 growing seasons. Switchgrass was established in 2020 and urea-fertilized at 56 kg N ha−1 year−1. Corn management followed best management practices for the US Midwest, including no-till and 202 kg N ha−1 year−1 fertilization, applied as urea–ammonium nitrate (32%). Our results showed lower direct N2O emissions in switchgrass compared to corn. Although soil CO2 emissions did not differ significantly during the establishment year, emissions in subsequent years were over 50% higher in switchgrass than in corn, likely due to increased belowground biomass, which was over five times higher in switchgrass. Nitrate-N leaching decreased as the switchgrass stand matured, reaching 80% lower than in corn by the third year. Differences in ET and WUE between corn and switchgrass were not significant; however, results indicate a trend toward reduced WUE in switchgrass under drought, driven by lower aboveground biomass production. Our study demonstrates that switchgrass can be implemented at a commercial scale without negatively impacting the hydrological cycle, while potentially reducing N losses through nitrate-N leaching and soil N2O emissions, and enhancing belowground C storage.

Miscanthus x giganteus (Mxg) is a promising perennial crop for producing natural colorants, renewable fuels, and bioproducts. However, natural recalcitrance and high pretreatment cost are major barriers to their complete conversion. In this study, a green processing method has been investigated for efficient recovery of natural pigments (anthocyanins), fermentable sugars, and pure lignin from Mxg genotypes using choline chloride-based natural deep eutectic solvents (NADES) systems. Interestingly, choline chloride: lactic acid (ChCl: LA) NADES-processed biomass resulted in 67.8 ± 2.1 μg g−1 of anthocyanins from dry biomass. A maximum of 87.4%–94.1% glucose yield was achieved after enzymatic saccharification. The effective extraction of lignin with high purity with higher β-aryl ether (βO4) bonds from advanced crops is crucial for lignin valorization. Notably, highly pure lignin (≈93.4% ± 1.4%) is achieved after low-temperature NADES pretreatment while retaining lignin’s native structure. 31P nuclear magnetic resonance demonstrated that total phenolics for ChCl: LA-lignin resulted in 1.20 mmol g−1 hydroxyls. The relative monolignol composition of syringyl (S), guaiacyl (G), and p-hydroxyphenyl (H) is 19.0, 65.7, and 14.3%, respectively, as evidenced by heteronuclear single quantum coherence analysis. This study provides a novel approach for obtaining high-purity lignin for catalytic depolymerization for oligomers and bifunctional monoaromatics production and leverages current cellulosic biorefinery technologies.

4D-STEM datasets for solution-treated (CrCoNi)93Al4Ti2Nb MEA in [111], [112], and [114] zone. Data used for Ultramicroscopy article "Differentiating electron diffuse scattering via 4D-STEM spatial fluctuation and correlation analysis in complex FCC alloys". Experiment details can be found in the paper. Data-specific details are listed in the Readme file.

Data from the 2025 publication in the Wilson Journal of Ornithology with the same name.

This dataset contains all raw and processed data used to generate the figures in the main text and supplementary material of the paper "High dynamic-range quantum sensing of magnons and their dynamics using a superconducting qubit." The data can be used to reproduce the plots and validate the analysis. Accompanying Jupyter notebooks provide step-by-step analysis pipelines for figure generation. The dataset also includes drawings for the mechanical samples used to perform the experiment. In addition, the dataset provides ANSYS HFSS electromagnetic simulation files used to design and analyze the resonator structures and estimate field distributions.

Supplementary data and code associated with the Biogeosciences paper published by Cecilia Prada et al. "Soil and Biomass Carbon Storage is Much Higher in Central American than Andean Montane Forests". There are 16 files associated with this paper (1) AGB.csv providing the site, plot, treeID, mnemn, family, agb, and AGcarbon for each tree in the dataset. Column headings are described in the file AGB_metadata.csv (2) AGB_metadata.csv Metadata (column descriptions) for AGB.csv (3) CWD_D.csv Complete information on the downed coarse woody debris (CWD) measured in each plot (4) CWD_D_metadata.csv Metadata (column descriptions) for CWD_D.csv (5) CWD_S.csv Complete information on the standing coarse woody debris measured in each plot (6) CWD_S_metadata.csv Metadata (column descriptions) for CWD_S.csv (7) SoilC.csv Estimated soil carbon storage (Mg C) at each sampling location in each plot (8) SoilC_metadata.csv Metadata (column descriptions) for SoilC.csv (9) Table.csv Data source, soil carbon value (Mg C) and elevation from published data sources (10) Table_metadata.csv Metadata (column descriptions) for Table.csv (11) TableS1.csv Data source, above ground carbon value (Mg C) and elevation from published data sources (12) TableS1_metadata.csv Metadata (column descriptions) for TableS1.csv (13) RScript.R Annotated code for data analysis and figures (14)Full_dataset.csv Full set of environmental data and carbon data by plot (15) Full_dataset_metadata.csv Metadata (column descriptions) for Full_dataset.csv (16) Species list and species codes.csv Full family, genus and species names for the species codes (column mnemn in AGB.csv)

Chronic wasting disease (CWD) surveillance data from Illinois and Wisconsin, USA between the fiscal years 2003 and 2022 (calendar years 2002 and 2021). Data is reported at the township level as defined by the US Public Survey System. CWD cases, animals tested for CWD, and the apparent prevalence calculated from these values are given by township and fiscal year. Data has been anonymized by replacing original township names with identification numbers to maintain the privacy of landowners. Variables include Tests, Cases, and nonlinear transformations of Tests and Cases (inverse, square root, and log transformations).

This dataset contains the data used for the publication “Aboveground rather than belowground productivity drives variability in Miscanthus x giganteus net primary productivity”. This dataset contains Miscanthus x giganteus biomass, carbon, and nitrogen tissue data for aboveground and belowground plant parts collected in 2021 for three different sites in Iowa with three different nitrogen application rates. Data at the Iowa sites were collected via biometric hand harvesting, belowground excavations, and soil coring both in-clump and beside-clump. Data were collected at two collection timepoints to calculate the contributions of belowground parts to Miscanthus x giganteus net primary productivity. This dataset also includes Miscanthus x giganteus and Switchgrass soil coring and excavation data collected in 2012 at the University of Illinois Urbana Champaign Energy Farm.

Physiological and yield data from a three year field experiment of soybean exposed to elevated ozone stress and reduced soil moisture at the SoyFACE experiment.

This dataset contains both processed and raw data that were leveraged to conduct analysis presented fully in the report "Community Vulnerability Assessment for Electric Vehicle Travelers Responsive to Extreme Flooding" and partially in the under review paper "Vulnerability Assessment of Electric Vehicles and their Charging Station Network during Evacuations".

Data and code for the paper titled "Electric Vehicle Charging Stations at Risk from Hazardous Events and Power Outages: Analytics and Resilience Implications" published in Renewable and Sustainable Energy Reviews journal (https://doi.org/10.1016/j.rser.2025.116144).

Data accompanying the article "Physics of Unraveling and Micromechanics of Hagfish Threads". Abstract of the article: Hagfish slime is a unique biological material composed of mucus and protein threads that rapidly deploy into a cohesive network when deployed in seawater. The forces involved in thread deployment and interactions among mucus and threads are key to understanding how hagfish slime rapidly assembles into a cohesive, functional network. Despite extensive interest in its biophysical properties, the mechanical forces governing thread deployment and interaction remain poorly quantified. Here, we present the first direct in situ measurements of the micromechanical forces involved in hagfish slime formation, including mucus mechanical properties, skein peeling force, thread–mucus adhesion, and thread–thread cohesion. Using a custom glass-rod force sensing system, we show that thread deployment initiates when peeling forces exceed a threshold of approximately 6.8 nN. To understand the flow strength required for unraveling, we used a rheo-optic setup to impose controlled shear flow, enabling us to directly observe unraveling dynamics and determine the critical shear rate for unraveling of the skeins, which we then interpreted using an updated peeling-based force balance model. Our results reveal that thread–mucus adhesion dominates over thread–thread adhesion and that deployed threads contribute minimally to bulk shear rheology at constant flow rate. These findings clarify the physics underlying the rapid, flow-triggered assembly of hagfish slime and inform future designs of synthetic deployable fiber–gel systems.