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the His6-AV-Ubiquitin-semapeptide (nucleotide sequence) fusion protein encoding gene was amplified using forward primer (Primer 3) and T7 reverse primer (Universal primer) and pET24- Arg 34 GLP-l (9-37) without 6His Tag single copy (recombinant plasmid generated from combination of Primer 1 & Primer 2) as a template