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URL: http://www.addgene.org/31602
Proper Citation: RRID:Addgene_31602
Insert Name: BoNT/A LC
Bacterial Resistance: Ampicillin
Defining Citation: PMID:18940613
Vector Backbone Description: Backbone Size:0; Vector Backbone:pBN3; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: The author states that although the construct is from a toxic protein, it only contains one third of the biologically active protein,i.e., it is inactive and harmless per se, and it does not require BL4 level for its expression/manipulation. Toxic BoNT/A is made of two 'chains' (i.e. polypeptides) linked to each other by one disulfide bond. The light chain (LC) is 50KDa long, whereas the heavy chain (HC) is 100 KDa. The HC is necessary for the toxin to target and enter motorneurons. This construct is the LC of BoNTA, and it completely lacks the HC, so it is unable to intoxicate neurons. Protein sequence of BoNT/A fragment present in this plasmid (corresponds to YP_001386738.1) MSGLEVSFEELRTFGGHDAKFIDSLQENEFRLYYYNKFKDIASTLNKAKSIVGTTASLQYMKNVFKEKYLLSEDTSGKFSVDKLKFDKLYKMLTEIYTEDNFVKFFKVLNRKTYLNFDKAVFKINIVPKVNYTIYDGFNLRNTNLAANFNGQNTEINNMNFTKLKNFT Nucleotide sequence of BoNT/A fragment present in this plasmid based on Addgene's Sanger sequencing results: ATGAGTGGGTTAGAAGTAAGCTTTGAGGAACTTAGAACATTTGGGGGACATGATGCAAAGTTTATAGATAGTTTACAGGAAAACGAATTTCGTCTATATTATTATAATAAGTTTAAAGATATAGCAAGTACACTTAATAAAGCTAAATCAATAGTAGGTACTACTGCTTCATTACAGTATATGAAAAATGTTTTTAAAGAGAAATATCTCCTATCTGAAGATACATCTGGAAAATTTTCGGTAGATAAATTAAAATTTGATAAGTTATACAAAATGTTAACAGAGATTTACACAGAGGATAATTTTGTTAAGTTTTTTAAAGTACTTAACAGAAAAACATATTTGAATTTTGATAAAGCCGTATTTAAGATAAATATAGTACCTAAGGTAAATTACACAATATATGATGGATTTAATTTAAGAAATACAAATTTAGCAGCAAACTTTAATGGTCAAAATACAGAAATTAATAATATGAATTTTACTAAACTAAAAAATTTTACT
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Source: Addgene
