Last data update: 24 January 2024 16:39 CET
Plasmid name: pSP64mIL2m (LMBP 3382)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p3382.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse interleukin 2 cDNA (IL2); mutated sequence |
| Promoter: | Phage SP6 promoter Escherichia coli lac operon 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: | pSP64; pAT153MIL2F2 |
| Further information: | The plasmid was constructed as follows: 1) Via site-specific mutagenesis according to Morinaga et al., using the synthetic oligo 5'-GTCAACTGCGCACCCACTTC-3', an FspI (MstI) site was introduced just in front of the first codon (GCA) of the mature mIL2 gene in the vector pAT153MIL2F2; 2) then, the PstI fragment from the mutated pAT153MIL2F2 vector, containing the mIL2 gene, was inserted into the unique PstI site of pSP64. The length of the 3' untranslated region of the mIL2 gene is not known. 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 the wild-type mouse IL2 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number X01772.1. |
| EMBL Accession number: | X01772.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 21/12/1995 |
| Sequence detail: | Nucleotide sequence around the start of the mature mIL2 gene:
wild type mIL2 gene: AAC.AGC.GCA.CCC
^
mutated mIL2 gene: AAC.TGC.GCA.CCC
^
MstI
^: Start position of the mature mIL2 gene. |
| 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 J. Demolder(1) and Prof. Dr R. Contreras(1). (1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | Demolder et al., Gene 111 (1992), 207-213 [PMID: 1541399] |
| Related plasmid reference: | Morinaga et al., Biotechnology 2 (1984), 636-639 |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 DH5α |
| Host reference: | Focus 8 (1986), 9 |
| Related host reference: | Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660] Rodriguez-Quinones et al., Focus 15 (1993), 110-112 Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8] Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187] Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051] |
| 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: |
pSP64mIL2m (LMBP 3382) is available at BCCM/GeneCorner. This plasmid was deposited by J. Demolder and Prof. Dr R. Contreras and was published in Demolder et al., 1992. |
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.