Last data update: 24 January 2024 16:39 CET
Plasmid name: pSPgHIL2f (LMBP 2317)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p2317.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human interleukin 2 genomic DNA (IL2, IL-2, TCGF, GeneID 3558); fragment |
| Promoter: | Escherichia coli lac operon promoter Phage SP6 promoter |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ) |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pSP65NN; pUCgHIL2f |
| Further information: | The plasmid was constructed as follows: 1) pUCgHIL2f was first cleaved with XbaI and secondly with MnlI (total digestion); 2) The labelled XbaI-MnlI fragment, containing the human interleukin 2 (hIL2) information, was isolated and XhoI linkers (5'-CCTCGAGG-3') were added; 3) After XhoI treatment, the XbaI-XhoI fragment was then inserted into the XbaI-XhoI opened pSP65NN vector. The hIL2 information inserted in this plasmid contains the first exon (coding partially for the protein signal sequence), the first intron, the second exon and part of the third exon (up to the XbaI site); note that the second intron of the hIL2 gene is not present; so, the second exon and part of the third exon are contiguous (this junction region is derived from the cDNA sequence). The sequence of the genomic hIL2 insert was obtained from 'GENBANK Release 55; Locus: HUMIL2'; it differs in one nucleotide (position 164) from the incompletely determined hIL2 sequence as described by Degrave et al. (EMBO J., 1983). 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. |
| EMBL Accession number: | J00264, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 03/10/1990 |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by V. Vande Vijver(1) and Prof. Dr E. Remaut(1). (1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | Degrave et al., EMBO J. 2 (1983), 2349-2353 [PMID: 6321163] |
| 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 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pSPgHIL2f (LMBP 2317) is available at BCCM/GeneCorner. This plasmid was deposited by V. Vande Vijver and Prof. Dr E. Remaut and was published in Degrave et al., 1983. |
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.