The present technology provides for rapid, simple and cost effective technique and vector to assemble DNA molecules in a pre determined orientation of sense-intron / spacer antisense or its combinations to produce DNA constructs usefull in transformation of Eukaryotic organisms such as plants, yeast etc. the method employs single PCR reaction, single ligation and single bacterial transformation reaction to generate the vector containing the DNA molecules in desired order for assembling an inverted repeat interspersed with an intron. Using this approach, DNA constructa are assembled with reduced cost, efoort and time.