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LMBP plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pMcMIL4m (LMBP 2028)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: pmcmil4m.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse interleukin 4 cDNA (IL4); mutated sequence
Promoter: -
Ribosome
binding site:
-
Terminator: Phage fd terminator
Selection marker: Chloramphenicol (cam)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Parental clone: pMaMIL4
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
Sequence detail:
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.
Class: Recombinant plasmid
Type: Plasmid
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
Plasmid reference: -
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)
Cultivation temperature: 37°C
Biosafety level: L1
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.

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