Pcaggs n ha vector

All the ORFs of the ASFV strain China/2018/AnhuiXCGQ (GenBank accession number MK128995.1), including 11 members belonging to the MGF110, were generated by gene synthesis (GENEWIZ, Suzhou, China) and cloned into the pCMV vector with a C-terminal FLAG tag (Beyotime Biotechnology, Shanghai, China). The ATF4-RLuc and ATF4-EGFP reporters were constructed as previously described (31 (link)). Briefly, a full-length human ATF4 mRNA 5′ leader and ATF4 initiation codon were subcloned into pRL-TK (Promega, WI) and pEGFP-C1 (Clontech, Mountain View, CA), respectively. cDNAs of PDIA3, TMED4, and PSMA4 were amplified from total mRNA of PK-15 cells by conventional RT-PCR and inserted into the KpnI and XhoI sites of the pCAGGS-N-HA vector (Addgene, MA) to obtain hemagglutinin (HA)-tagged constructs. All constructed plasmids were verified by sequencing.
The organelle markers, including pDsRed2-ER, pDsRed2-Golgi, pDsRed2-Mito, pDsRed2-LAMP1, and pDsRed2-Peroxi vectors that encode targeting sequences of the corresponding organelles, were purchased from Clontech.

All the ORFs of the ASFV strain China/2018/AnhuiXCGQ (GenBank accession number MK128995.1), including 11 members belonging to the MGF110, were generated by gene synthesis (GENEWIZ, Suzhou, China) and cloned into the pCMV vector with a C-terminal FLAG tag (Beyotime Biotechnology, Shanghai, China). The ATF4-RLuc and ATF4-EGFP reporters were constructed as previously described (31 (link)). Briefly, a full-length human ATF4 mRNA 5′ leader and ATF4 initiation codon were subcloned into pRL-TK (Promega, WI) and pEGFP-C1 (Clontech, Mountain View, CA), respectively. cDNAs of PDIA3, TMED4, and PSMA4 were amplified from total mRNA of PK-15 cells by conventional RT-PCR and inserted into the KpnI and XhoI sites of the pCAGGS-N-HA vector (Addgene, MA) to obtain hemagglutinin (HA)-tagged constructs. All constructed plasmids were verified by sequencing.
The organelle markers, including pDsRed2-ER, pDsRed2-Golgi, pDsRed2-Mito, pDsRed2-LAMP1, and pDsRed2-Peroxi vectors that encode targeting sequences of the corresponding organelles, were purchased from Clontech.