Last data update: 28 November 2020 04:24 CET
Plasmid name: pMcMIL4m (LMBP 2028)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Mouse interleukin 4 cDNA (IL4); mutated sequence|
|Terminator:||Phage fd terminator|
|Selection marker:||Chloramphenicol (cam)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli|
|Further information:||This phasmid was constructed by site-specific mutagenesis of pMaMIL4, where unique ApaI and DraII sites were created and the amino acid -1 (Ser) was mutated to Ala.
pMcMIL4m provides resistance to chloramphenicol, but is sensitive to ampicillin.
After mutagenesis, use mutS strains for primary transformation (e.g. sup(-) strain WK6mutS, sup(+) strain BMH71-18mutS); for segregation of possible mutants: sup(-) strains (e.g. WK6).
The nucleotide sequence of the 3' UTR region of the inserted mIL4 gene (from nucleotide position 4232 to 4358) has been experimentally verified by sequencing the non-coding strand in the orientation EcoRI-ScaI; the nucleotide sequence around the EcoRI site was not unambiguous (Marc Logghe: personal communication).
Since the nucleotide sequence of the 5' UTR region of the inserted mIL4 gene is not known, there is uncertainty as to the cutting frequency of the restriction enzymes indicated.
|EMBL Accession number:||-|
|Latest sequence update:||05/12/1994|
Nucleotide sequence around the start of the mature mIL4 gene: signal sequence | mature mIl4 gene | -5 -4 -3 -2 -1| 1 2 mIL4 wt: GAA TGT ACC AGG AGC|CAT ATC Glu Cys Thr Arg Ser|His Ile mutated mIL4: GAA TGT ACC AGG GCC|CAT ATC Glu Cys Thr Arg Ala|His Ile ApaI DraII
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by F. Hanssens(1) and Prof. Dr E. Remaut(1).
(1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|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 + chloramphenicol (25 μg/ml)|
|Other culture collection numbers:||-|
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.