Last data update: 24 January 2024 16:39 CET
Plasmid name: pGEMIL6gIL2PHOA (LMBP 2324)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p2324.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human interleukin 2 genomic DNA (IL2, IL-2, TCGF, GeneID 3558); mutated sequence Escherichia coli alkaline phosphatase A gene (phoA) |
| Promoter: | Human interleukin 6 promoter (IL6) Phage T7 gene 10 promoter (T7g10) 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 |
| Parental clone: | pGEMIL6gIL2f3; pSPgHIL2PHOA |
| Further information: | The plasmid was constructed by ligating the HindIII (filled in with Klenow DNA polymerase) - XbaI fragment from pSPgHIL2PHOA into the HindII-XbaI (nucleotide position 147) opened pGEMIL6gIL2f3. This construct codes for shortened (140 amino acids instead of 153) human interleukin 2 (hIL2), signal sequence included, fused to Escherichia 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 part was taken from Genbank (Release 55; Locus: HUMIL2). It differs in one nucleotide (position 2015) from the incompletely determined hIL2 sequence as described by Degrave et. al., 1983. The nucleotide sequence of the genomic hIL6 DNA corresponds with the EMBL Nucleotide Sequence Database accession number AC073072.11. However, the C nucleotide at promoter position -174 (polymorphism mutant) was changed into a G nucleotide (wild type). |
| EMBL Accession number: | AC073072.11, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 03/05/2004 |
| Sequence detail: | Nucleotide sequence at the fusion between hIL2 and phoA:
----------- exon 4 of hIL2 -----------> ------- phoA ------>
120 14
5' ... ACA.GCA.ACC.ATT.GTA.GAA.TTT.CTG.AAT.CGG.CCT.CAG.GGC.GAT.ATT.ACT ... 3'
Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Pro Gln Gly Asp Ile Thr
Punctuation indicates reading frame. |
| 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 G. Haegeman(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 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pGEMIL6gIL2PHOA (LMBP 2324) is available at BCCM/GeneCorner. This plasmid was deposited by V. Vande Vijver and Prof. Dr G. Haegeman. |
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.