Last data update: 24 January 2024 16:39 CET
Plasmid name: pSPgHIL2PHOA (LMBP 2320)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p2320.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human interleukin 2 genomic DNA (IL2, IL-2, TCGF, GeneID 3558) Escherichia coli alkaline phosphatase A gene (phoA); mature sequence |
| 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: | pSPgHIL2f; pPLcHIL2PHOA |
| Further information: | This plasmid was constructed by replacing the XbaI-HindII fragment of pSPgHIL2f by the XbaI-BstEII (filled in with Klenow DNA polymerase) fragment of pPLcHIL2PHOA. This construct codes for a shortened human interleukin 2 (140 amino acids instead of 153), signal sequence included, fused to E. coli alkaline phosphatase A (phoA). The hIL2 information of this plasmid contains the first exon (coding partially for the protein signal sequence), the first intron, the second and third exon (mutated) and part of the fourth exon (mutated). Note that the second and third intron of the hIL2 gene are not present. So, the second and third exon and part of the fourth exon are contiguous (this junction region is derived from the cDNA sequence). The sequence of the genomic hIL2 information was taken from Genbank (Release 55; Locus: HUMIL2). It differs in one nucleotide (nucleotide position 164) from the incompletely determined hIL2 sequence as described by Degrave et. al., 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: | 15/10/1990 |
| Sequence detail: | Nucleotide sequence at the fusion between hIL2 and phoA:
exon 4 of hIL2| | phoA
120 14
Leu Asn Arg Pro Gln Gly
5' CTG AAT CG | G CCT CAG GGC 3' |
| 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 R. Leemans(1) and Prof. Dr E. Remaut(1). (1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related 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: |
pSPgHIL2PHOA (LMBP 2320) is available at BCCM/GeneCorner. This plasmid was deposited by R. Leemans and Prof. Dr E. Remaut. |
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.