Flavinkins (@Flavinkins)

@Parsifaler https://anandamide.substack.com/p/curious-kittens However, examining these sequences (that were found from the Pfizer vials) revealed that while there were plasmid backbones that were highly similar, they are not identical and there were key differences between these plasmids and those that have been found within these Pseudomonas assemblies. For one, None of these plasmids harbored a mammalian/eukaryotic promoter that is used to drive the expression of the Spike protein, which is expected as these are supposed to be used to generate mRNA for vaccine production. Secondly, A duplication is discovered within one of the Pseudomonas-related sequences, whereas a deletion is found in the other plasmid with the Spike protein, while substitutions within the backbone fragment are found within both of the P.aeruginosa-linked plasmid sequences. This indicates that, compared to these unknown primary backbone sequences, the P.aeruginosa assemblies were found to have undergone a substantial level of evolution that involved duplication, deletion, and substitutions within the backbone sequences for these plasmids--suggesting an ancient origin of the plasmids found within the Pseudomonas assemblies. Third, The C-terminal of the Spike protein inserts, when found, were found to be native--expected for an mRNA vaccine. One of the proteins found was an S1-only protein. The C-terminal membrane anchor coiled-coil sequence is not found in any of these plasmids. This rules out mRNA vaccination as the origin for these Pseudomonas-linked plasmid sequences. Finally, While many P.aeruginosa strains are resistant toward Neomycin and Kanamycin, Neo/Kan, not found within the chromosomes of these assemblies, does in fact additionally confer GmR (Gentamicin resistance) toward the P.aeroginosa strains. While it is possible that the plasmids may have derived from contamination within the BioProject, the supposed natural host for these plasmids, laboratory strains of E.coli, was nowhere to be seen within the BioProject PRJNA839565. The closest sequences found within the BioProject by BLAST just returned other strains of Pseudomonas, Sampled in China in 2019. https://www.ncbi.nlm.nih.gov/nucleotide/CP081287.1?report=genbank&log$=nucltop&blast_rank=1&RID=Z2K3722G01R https://archive.md/hm8zm https://archive.md/kkSkI https://archive.md/O9Kkr https://archive.md/LYema https://www.ncbi.nlm.nih.gov/nucleotide/JAMOHA010000033.1?report=genbank&log$=nucltop&blast_rank=1&RID=Z2M30VUK01R https://www.ncbi.nlm.nih.gov/nucleotide/JAMOGL010000063.1?report=genbank&log$=nucltop&blast_rank=2&RID=Z2M30VUK01R https://www.ncbi.nlm.nih.gov/nucleotide/JAMOGK010000062.1?report=genbank&log$=nucltop&blast_rank=3&RID=Z2M30VUK01R https://archive.md/GTo6k This also included contigs that have very low coverage, supposedly consistent with "contamination origin". This indicates that 1: the plasmids found have evolved substantially compared to their closest ancestors within the labs. 2: they are found in Pseudomonas spp., instead of laboratory origin E.coli. This is expected as mammalian expression vectors with the use of SV40 Ori and CMV promoter is conventionally maintained within the lab using AmpR, and one lab that deals with mammalian expression typically maintain their plasmids using only one antibiotic for convenience in the preparation of the medium necessary for culturing the bacterial host. https://www.ncbi.nlm.nih.gov/nuccore/JAMOGH010000091.1 https://archive.md/ERB08 While in the U.S., AmpR may have been avoided in vaccine preparation (mRNA production vectors instead of mammalian expression vectors) within the Moderna facilities, the same basic rule for non-bacterially-oriented vectors, one antibiotic resistance gene per lab, is maintained within these vaccine-derived Spike expression vectors (all vectors found within the vaccine vials used Neo/Kan and nothing else). This is distinct from that is seen in the Pseudomonas assemblies, where Eukaryotic-oriented Neo/Kan accompanied with AmpR in only one of the plasmid versions was found. This also indicates that both mutations (substitutions seen in the backbone), duplications/deletions, and recombinations have shaped the plasmid backbone found within these P.aeruginosa assemblies and that this did not happen within a lab (E.coli is not found within PRJNA839565).