Cloning and expression plasmid for one gene of interest
pUNO plasmids feature a wide choice of native and fusion genes. The gene of interest is under the control of of the strong and ubiquitous mammalian promoter, EF1α/HTLV.
pUNO plasmids can be used for in vitro or in vivo transfection experiments.
Invivogen also provides pUNO-mcs plamids, designed to serve as control and/or cloning vectors.
• hEF1/HTLV prom is a composite promoter comprising the Elongation Factor-1α (EF-1α) core promoter and the R segment and part of the U5 sequence (R-U5’) of the Human T-Cell Leukemia Virus (HTLV) Type 1 Long Terminal Repeat. The EF-1α promoter exhibits a strong activity, higher than viral promoters and, on the contrary to the CMV promoter, yields persistent expression of the transgene in vivo. The R-U5’ has been coupled to the EF-1α core promoter to enhance stability of DNA and RNA.
• MCS or Gene:
- The multiple cloning site contains the following restriction sites:
pUNO: 5’ - SgrAI, SalI, BamHI, Eco47III, PstI, NheI - 3’
pUNO1: 5’ - Sal I, SgrA I, BamH I, Eco47 III, Nco I, Nhe I - 3’
Each restriction site is compatible with many other enzymes, increasing the cloning options.
• SV40 pan: The Simian Virus 40 late polyadenylation signal enables efficient cleavage and polyadenylation reactions resulting in high levels of steady-state mRNA.
• ori is a minimal E. coli origin of replication with the same activity as the longer Ori.
• SV40 prom: The Simian Virus 40 promoter allows the expression of the blasticidin resistance gene in mammalian cells.
• CMV enh/prom: The human cytomegalovirus immediate-early gene 1 promoter/enhancer was originally isolated from the Towne strain and was found to be stronger than any other viral promoters.
• Resistance gene:
- Bsr (blasticidin resistance gene): The bsr gene from Bacillus cereus encodes a deaminase that confers resistance to the antibiotic Blasticidin S. The bsr gene is driven by the SV40 promoter in tandem with the bacterial EM7 promoter to allow blasticidin selection in both mammalian cells and E. coli bacteria.
- hph: Resistance to Hygromycin B is conferred by the hph gene from E. coli which encodes a phosphotransferase. The hph gene is driven by the CMV enhancer/promoter in tandem with the bacterial EM7 promoter allowing selection in both mammalian cells and E. coli.
• βGlo pAn: The human beta-globin 3’UTR and polyadenylation sequence allows efficient arrest of the transgene transcription.
- 20 µg of lyophilized DNA.
- 4 pouches of the relevant E. coli Fast-Media® (2 TB and 2 Agar)