T7 and cmv promoters

Manufactured by Promega

T7 and CMV promoters are commonly used genetic regulatory sequences found in various expression systems. The T7 promoter is derived from the bacteriophage T7 and is recognized by the T7 RNA polymerase, enabling high-level transcription of target genes. The CMV promoter, originating from the human cytomegalovirus, is a strong eukaryotic promoter commonly used for transgene expression in mammalian cell lines. Both promoters are widely utilized in molecular biology and biotechnology applications to drive the expression of recombinant proteins and other genetic constructs.

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Lab products found in correlation

Pcdh ef1 mcs t2a copgfp, by System Biosciences (1 mentions) E coli krx, by Promega (1 mentions) Quikchange 2 site directed mutagenesis kit, by Agilent Technologies (1 mentions) Pet28a, by Promega (1 mentions)

2 protocols using t7 and cmv promoters

Site-Directed Mutagenesis and Protein Expression

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CBR mutants/mutant libraries were constructed using QuikChange II Site-Directed Mutagenesis Kit (Agilent Technologies 200523) according to the manufacturer protocol. Oligonucleotides were from IDT. Mutagenesis reactions were used to transform E. coli KRX (Promega). Individual colonies were picked for plasmid preparation and DNA sequence verification. All plasmids for bacterial expression and transient mammalian cell expression were in a pF4Ag backbone (T7 and CMV promoters; Promega). For purification from bacterial overexpression, sequences were sub-cloned to pF6HisNK (Promega). CBR2opt was assembled synthetically as a mammalian codon optimized version of CBR2 expressing the identical enzyme sequence as CBR2 (Gene Dynamics). For stable cell line generation, Luc2 and CBR2opt were sub-cloned from their pF4Ag backbones into lentiviral vector pCDH-EF1-MCS-T2A-copGFP (System Biosciences).

Hall M.P., Woodroofe C.C., Wood M.G., Que I., van’t Root M., Ridwan Y., Shi C., Kirkland T.A., Encell L.P., Wood K.V., Löwik C, & Mezzanotte L. (2018). Click beetle luciferase mutant and near infrared naphthyl-luciferins for improved bioluminescence imaging. Nature Communications, 9, 132.

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Bacterial and Mammalian Plasmid Constructs

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Plasmids for bacterial expression and transient mammalian cell expression were in a pcDNA3.1(-) backbone (T7 and CMV promoters; Promega). For purification from bacterial overexpression, Smad3 cDNA sequences (RefSeq NM_016769.4) were sub-cloned to pET28a (Promega). To construct plasmids for dual reporter luciferase assays, Foxp3 promoter sequences (CAGAbox) were amplified and cloned into the pGL3 basic vector to become pGL3-CAGA.

Tang S., Zhang J., Lou F., Zhou H., Cai X., Wang Z., Sun L., Sun Y., Li X., Fan L., Li Y., Jin X., Deng S., Yin Q., Bai J., Wang H, & Wang H. (2024). A lncRNA Dleu2-encoded peptide relieves autoimmunity by facilitating Smad3-mediated Treg induction. EMBO Reports, 25(3), 1208-1232.

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