S oybean vein necrosis virus (Orthotospovirus glycininecrovenae, SVNV) is an ambisense ssRNA virus in the genus Orthotospovirus first identified in Tennessee in 2008 (1). SVNV consists of three segments: S, M, and L. These encode a nucleocapsid protein (N), nonstructural proteins (NSs and NSm), glycoproteins (GN and GC), and an RNA-dependent RNA polymerase (RdRp) (2). The complete sequence of the SVNV17_Auburn_AL isolate was obtained using RNA-Seq and RACE. In 2023, soybean samples exhibiting symptoms of SVNV were collected. Total RNA was extracted from symptomatic leaves using the previous methodology (3), followed by ribosomal RNA depletion using the Illumina Ribo-Zero Plus rRNA Depletion Kit (Illumina, Cat: 20037135). Libraries were prepared with the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina and sequenced on an Illumina NovaSeq 6000 (150 bp PE, ~47 million reads). Quality control was conducted using FastQC (4), and adapter sequences were removed using BBDuk (https://sourceforge.net/projects/bbmap/). Processed reads were mapped to the SVNV-TN genome (GCA_004789395.1) using Bowtie (5). Variants (depth >80, Phred > 100) were called using BCFtools (6) and FreeBayes tool (7), and the average depth was calculated using SAMtools (6). Identified variants were visualized using IGV v2.3.57 (8). The consensus assembly was generated with BCFtools (6). For all tools, the default parameters were used. Missing terminal nucleotides were filled using RACE: 5′ ends with the Invitrogen 5′ RACE System (ThermoFisher, Cat: 18374058) using segment-specific primers (Table 1), and 3′ ends using E. coli poly(A) polymerase (NEB) followed by SuperScript III (Invitrogen) synthesis. PCR amplification used Phusion (ThermoFisher, F530S), cloned using the CloneJET PCR Cloning Kit, and 12 colonies were Sanger sequenced using an Applied Biosystems 3730xl sequencer. The genome of the SVNV17_Auburn_AL comprises 16,563 bases (2,602 bp [S], 4,948 bp [M], and 9,013 bp [L]) with a GC content of 35% and an average depth of 1,669×. The leaders are 58, 57, and 185 bases, while the trailers are 70, 91, and 30 bases for the S, M, and L segments, respectively. The first six bases (AGAGCA) at the 5′ ends are identical across all three segments and are complementary to the 3′ ends, forming a panhandle similar to other orthotospoviruses (9). Genome comparison between the SVNV17_Auburn_AL isolate and the TN strain revealed 43, 97, and 138 SNPs/indels in the S, M, and L segments (Fig. 1). To determine the impact on the protein level, the ORFs’ sequence was translated with ExPASy (10) and aligned to the TN strain with Clustal