Last data update: 24 January 2024 16:39 CET
Plasmid name: pMxCATlinkDD (LMBP 3908)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3908.gb
(View with Genome Compiler) p3908.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Chloramphenicol acetyltransferase gene (cat) Human tumor necrosis factor receptor p55 cDNA (TNFR55, TNFR1, TNFRSF1A); C-terminal fragment of the intracellular domain (death domain) |
| Promoter: | Mouse Mx promoter Escherichia coli lac operon promoter Phage SP6 promoter |
| Ribosome binding site: |
- |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Mammalian cells |
| Parental clone: | pMxCAT; pMxCAThTNFDD |
| Further information: | The plasmid was constructed as follows: a C-terminal fragment of the intracellular domain of the hTNFR55 cDNA, called death domain (DD) and starting with codon 326, was recovered from pMxhTNFR55i and introduced into pMxCAT, in frame at the 3' end of the chloramphenicol acetyltransferase coding sequence and preceded by an oligonucleotide coding for a flexible spacer peptide Gly-Gly-Ser. An additional initiation codon was introduced between the spacer and the hTNFR fragment. In this intermediate plasmid pMxCAThTNFDD, an oligonucleotide was inserted, coding for an extra 15 amino acids linker (Gly4Ser)3, located between the Gly-Gly-Ser linker and the cDNA sequence of the hTNFR55 death domain. When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727283.1. Other name of the plasmid is pMxCATDD. |
| EMBL Accession number: | LT727283.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 15/01/1999 |
| Sequence detail: | Nucleotide sequence around the fusion between the chloramphenicol acetyltransferase gene and the C-terminal part of the intracellular domain (death domain) of the human tumor necrosis factor receptor p55:
cat -->|
| spacer
5' … GGC.GGG.GCG.GGC.GGA.TCC.GGT.GGA.GGC.GGT.TCA.GGC.GGA.GGT.GGC.TCT.
Gly Gly Ala Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
| |--> human TNF receptor p55
| |--> death domain
| |326
GGC.GGT.GGC.GGA.TCC.ATG.GAC.CCC.GCG.ACG.CTG.TAC. … 3'
Gly Gly Gly Gly Ser Met Asp Pro Tyr Thr Leu Tyr |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglI, EcoNI, HindIII, NcoI, ScaI and XmnI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr G. Haegeman(1) and constructed by Dr V. Vandevoorde(1). (1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | Vandevoorde et al., J. Cell Biol. 137 (1997), 1627-1638 [PMID: 9199176] |
| Related plasmid reference: | Boone et al., FEBS Lett. 441 (1998), 275-280 [PMID: 9883899] |
| 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: |
pMxCATlinkDD (LMBP 3908) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr G. Haegeman and constructed by Dr V. Vandevoorde and was published in Vandevoorde et al., 1997. |
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.