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Targeting Construct Development for Gene Editing Studies

This genetic construct contains multiple cloning sites and selectable marker genes for gene editing experiments. Fragments from various sources were cloned to create the final construct for targeted genetic manipulations.

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Targeting Construct Development for Gene Editing Studies

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  1. f1 origin A pNosTerFRT-neo (5500bp) Ampr Ori BglII XbaI NotI SacI SfiI FseIAvrII……NotI XbaI NotI DsaI SacI FRT FRT Nos-ter Tn5 pGK neo 0.3kb 1.8kb UPSTREAM MULTIPLE CLONING SITE NotI XbaI KnpI SacI MluINheIXhoIBglII HindIII….. BglII EcoRI PstI SmaIBamHISalIPstI B TARGETING CONSTRUCT SpeI/XhoI fragment from pST33 was cloned into pNosTerFRT-Neo cut with NheI/SalI GA Pc2ox1 (i) pGAPc2ox1FRT-neo EcoRI EcoRI EcoRVNotI 0.9kb BamHI/SacI fragment from pBin-mgfp5-ER was blunt end ligated into pNosTerFRT-Neo cut with SmaI mgfp5-ER (ii) pmgfp5-ERFRT-neo 0.8kb

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