Advances in nucleic acid based molecular diagnosis techniques, provided the basis for efficient, accurate and rapid detection of biological/ pathological agents. However, these techniques suffer from major limitations including lack of the genomic sequence of high-risk pathogens to calibrate techniques as positive controls, false positive results and restrictions on access to existing commercial tests. However with the advent of synthetic biology, it is possible to construct appropriate positive controls. Artificial genetic constructs are the synthetic constructs that contain genetic materials from highly dangerous bacteria or viruses, which can be used as positive controls in molecular techniques such as PCR. In this review, we introduce positive controls and simulator positive controls firstly and discuss about artificial genetic constructs, procedure of design and synthesis secondly. Finally, we discussed about single and chimeric artificial genetic constructs as two main categories of these positive control.