Last data update: 24 January 2024 16:39 CET
Plasmid name: pCDM9 (LMBP 4364)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
pcdm9.gb
(View with Genome Compiler) pcdm9.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human interferon β gene (IFNB, IFNB1); stuffer fragment |
| Promoter: | Phage T7 gene 10 promoter (T7g10) Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer |
| 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: | pCDM8; pSP64GSHIFNBNT; pACYC177 |
| Further information: | The plasmid was constructed as follows: First, the HindIII-NotI fragment of pSP64GSHIFNBNT (containing the human IFNβ gene) was inserted between the HindIII and NotI sites of pCDM8. This intermediate construction was DraIII opened, blunted with T4 polymerase (exonuclease) and ligated to the AsuII-BamHI fragment (sites filled in with Klenow DNA polymerase) from pACYC177, containing the ampicillin resistance gene. The inserted hIFNβ gene only has a stuffer function. It makes a SfiI and a NotI site easily available for cloning. pCDM9 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 CMV promoter 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 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 and SV40 origins of replication enables the vector to replicate episomally in mammalian cells expressing large T-antigen (e.g. WOB, COS cells). The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727197.1. The nucleotide sequence of the hIFNβ cDNA corresponds with the EMBL Nucleotide Sequence Database accession number V00546.1. |
| EMBL Accession number: | V00546.1, view at EMBL, GenBank, DDBJ LT727197.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 27/11/1995 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: NotI, PstI, SfiI and XhoI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by S. Dewaele(1)(2) and Prof. Dr R. Contreras(1)(2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) 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(λ) |
| Host reference: | Mertens et al., Gene 164 (1995), 9-15 [PMID: 7590329] |
| Related host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pCDM9 (LMBP 4364) 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.