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. 2020 Feb 11;21(4):1206. doi: 10.3390/ijms21041206

Figure 2.

Figure 2

Constructs in pGL3prom, pRF and pHpRF. pGL3-ACLY contains ACLY 5′ untranslated region (UTR) inserted into pGL3prom upstream firefly luciferase (FL) cistron. The control dicistronic vector pRF contains the SV40 early promoter (prom), a chimeric intron, and the cDNAs encoding Renilla luciferase (RL) and FL separated by a short linker sequence. pR-ACLY-F construct is obtained by cloning the ACLY 5′ UTR between the two cistrons RL and FL. The ACLY 5′ UTR was also cloned between the two cistrons RL and FL of pHpRF obtaining the pHpR-ACLY-F construct. The pHpRF differs from pRF because it contains an inverted repeat upstream the first cistron, in order to form a stem–loop at the 5′ end of the transcript. pRF(-P) and pR-ACLY-F(-P) constructs are identical to their corresponding pRF constructs described above, except that they do not contain the SV40 early promoter and the chimeric intron.