Last data update: 24 January 2024 16:39 CET
Plasmid name: pCDM92 (LMBP 3980)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3980.gb
(View with Genome Compiler) p3980.txt p3980.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human interferon β gene (IFNB, IFNB1); stuffer fragment |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Phage T7 gene 10 promoter (T7g10) |
| Ribosome binding site: |
- |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) Suppressor tRNA gene (supF; requires a p3 containing host) |
| Replicon: | Escherichia coli plasmid pMB1 origin Polyoma virus origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pCDM9 |
| Further information: | The plasmid was constructed as follows: pCDM9 was partially digested with SfiI, treated with T4 DNA polymerase and ligated. Consequently, the SfiI site in the origin region of SV40 is destroyed and the SfiI downstream from the T7 promoter becomes unique. The inserted hIFNβ gene only has a stuffer function. It makes a SfiI and a NotI site easily available for cloning. pCDM92 is a shuttle vector designed for high-level transient expression of cloned cDNA in mammalian cells. The protein encoded by the cDNA can be expressed under control of the hCMV-IE promoter and enhancer and the SV40 polyadenylation site. The T7 promoter can be used for in vitro transcription of the cDNA insert. The small HIV (human immunodeficiency virus) fragment (29 nucleotides) between the CMV-IE and T7 promoter sequences has an enhancer function. The SspI fragment (153 bp) of the 3' UTR region of the SV40 small t-antigen is missing, but without effect on the function of the vector. The presence of the polyoma origin of replication enables the vector to replicate episomally in mammalian cells expressing polyoma large T-antigen (e.g. L929 cells). The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727298.1. The nucleotide sequence of the hIFNβ cDNA corresponds with the EMBL Nucleotide Sequence Database accession number V00546.1. Other name of the plasmid is pCDM9SfiΔ. |
| EMBL Accession number: | V00546.1, view at EMBL, GenBank, DDBJ LT727298.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 07/12/1999 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: HindII, NciI/SfiI, NotI, SspI and XhoI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by S. Dewaele(1) and Prof. Dr R. Contreras(1). (1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061 |
| Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pCDM92 (LMBP 3980) is available at BCCM/GeneCorner. This plasmid was deposited by S. Dewaele and Prof. Dr R. Contreras. |
Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.