Last data update: 30 July 2021 10:22 CEST
Plasmid name: pCDM8 (LMBP 2393)
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|Price category:||Cat. 3 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Promoter:||Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer
Phage T7 gene 10 promoter (T7g10)
|Terminator:||Simian virus 40 polyadenylation signal (SV40 polyA)|
|Selection marker:||Suppressor tRNA gene (supF; requires a p3 containing host)|
|Replicon:||Bacteriophage M13 origin
Escherichia coli plasmid pMB1 origin
Polyoma virus origin
Simian virus 40 bidirectional origin (SV40)
|Host range:||Escherichia coli; use strains with the p3 helper plasmid (kan resistant, amber codon in amp and tet)
Mammalian cells; SV40 permissive cells
|Further information:||pCDM8 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 human CMV-IE promoter and enhancer and the SV40 polyadenylation site.
The T7 promoter is used for in vitro transcription of the cDNA insert.
The small HIV (human immunodeficiency virus) fragment (29 nucleotides) between the human CMV-IE and the T7 promoter sequences has an enhancer function.
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 supF tRNA suppressor gene is used for selection in E. coli (MC1061[p3]).
The M13 origin is useful for synthesis of single-stranded DNA upon infection with the M13 helper phage.
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 nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727510.1.
|EMBL Accession number:||LT727510.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||21/11/1995|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: NotI, PstI, SspI and XhoI.
This plasmid has also been fully sequenced but the NGS sequence data still need to be implemented.
|History of deposit:||This plasmid was deposited by Dr B. Seed(1).
(1) Department of Molecular Biology, Massachussetts General Hospital, Boston, USA
|Plasmid reference:||Seed, Nature 329 (1987), 840-842 [PMID: 3313052]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 MC1061[p3]|
|Related host reference:||Invitrogen Instruction Manual
Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
|Cultivation medium:||LB-Lennox + ampicillin (50 μg/ml) + kanamycin (50 μg/ml) + tetracycline (10 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCDM8 (LMBP 2393) is available at BCCM/GeneCorner. This plasmid was deposited by Dr B. Seed and was published in Seed, 1987.|
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.