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Plasmid cloning vectors for gene therapy with phosphohistidine-based oligos

Artificial DNA primers: phosphoramidate-based oligonucleotide synthesis

cDNA:

• N-6-benzoyl-deoxyadenosine phosphoramidite - Notice the P-N bound indicating the linkage of a phosphonate diester to a benzoyl atom :)
• N-3-oxo-hexanoyl-L-homoserine lactone - A light-inducible phosphohistidine-bound AHL ? A(denosine)TP/cAMP dependent.
• Reminder:
  • NHC = (N heterocycle carbene) - Chemical compounds with a N chiral prodrug switch - highly enantioselective molecules
  • N = indicates a Nitrogen atom
  • P = a Phosphonate (inorganic state) - a compound for oligo synthesis using the nucleoside phosphoramidite method.
  • P-N = phosphoramidate (a Nitrogen atom bound to a Phosphonate ester in nucleophilic substitution).

deoxynucleotide synthesis:

• cDNA synthesis building block = deoxynucleoside + 3'-phosphonate = synthetic nucleotide analog (dATP) 1

oligodeoxyribonucleotide synthesis:

• short 50-200(?) bp restriction endonucleases enzymes for cDNA catalysis/plasmid vector cloning in E. coli
• building blocks for oligonucleotides assembly with phosphoramidites method 1
• see also Oligonucleotide Synthesis

M13K07 cloning in E. coli:

• amino-acid sequence signature (Myc epitope tag) = EQKLISEEDL

References