Last data update: 24 January 2024 16:39 CET
Plasmid name: pSPT1hTNFR75 (LMBP 2582)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p2582.gb
(View with Genome Compiler) p2582.txt p2582.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human tumor necrosis factor receptor p75 cDNA (TNFR75, TNFR2, TNFRSF1B); with Kozak consensus sequence |
| Promoter: | Phage SP6 promoter Phage T7 gene 10 promoter (T7g10) |
| Ribosome binding site: |
- |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pSPT19 |
| Further information: | The plasmid was constructed by inserting the human TNFR75 coding sequence as an Asp718(KpnI)-HindIII fragment into the unique Asp718 (KpnI) and HindIII sites of pSPT19. The full-length human TNFR75 coding sequence was assembled from a partial cDNA clone using oligonucleotide synthesis and PCR. All non-coding sequences at both the 5' and 3' ends were removed. Unique HindIII (5') and Asp718 (KpnI) (3') restriction sites were introduced. In addition, the Kozak consensus sequence was inserted upstream of the ATG start codon. Due to the modifications of the non-coding sequences, the receptor can be well expressed whereas the natural sequence is expressed in low amounts. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727531.1. The nucleotide sequence of the human TNFR75 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number M32315.1. However, due to the PCR amplification, there is one base pair exchange in one codon that does not affect the amino acid sequence (silent mutation; W. Lesslauer, F. Hoffmann - La Roche LTD, Basel, Switserland: personal communication). This mutation was not experimentally verified by BCCM/GeneCorner. Other name of the plasmid is pSPT1. |
| EMBL Accession number: | M32315.1, view at EMBL, GenBank, DDBJ LT727531.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 07/10/1991 |
| Sequence detail: | Nucleotide sequence in front of the ATG start codon:
Kozak consensus sequence
<------------->
5' AAGCTTGCCGCCACC.ATG.GCG. 3'
HindIII ***
***: start codon. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: EcoNI/FspI, EcoRI and HindIII/KpnI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by F. Hoffmann - La Roche LTD(1). (1) Basel, Switserland |
| Plasmid reference: | - |
| Related plasmid reference: | Dembic et al., Cytokine 2 (1990), 231-237 [PMID: 1966549] |
| 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: |
pSPT1hTNFR75 (LMBP 2582) is available at BCCM/GeneCorner. This plasmid was deposited by F. Hoffmann - La Roche LTD. |
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.