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GeneCorner plasmid details

Last data update: 29 October 2020 04:15 CET

Plasmid name: pCAGGSE-mIL1B-HA (LMBP 4354)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p4354.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse interleukin 1 beta cDNA (Il1b, Il-1b, IL-1beta, GeneID 16176)
E-tag; N-terminal
Influenza HA epitope encoding the haemagglutinin tagging peptide; C-terminal
Promoter: Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pCAGGS-E
Further information: The plasmid was constructed by PCR amplifying the mouse IL1B cds, introducing a C-terminal HA epitope, and cloning it as a NotI/XmaI fragment into the NotI/XmaI opened pCAGGS-E vector.
The plasmid is useful for highly efficient expression of the E- and HA-tagged mouse IL1B prosequence under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacIq strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The mouse IL1B insert was sequenced by BCCM/GeneCorner. The nucleotide sequence of the mouse IL1B prosequence cDNA corresponds with the Genbank accession number NM_008361.3 except for codon 185 of mouse IL1B which encodes for histidin (H) instead of tyrosin (Y), and codon 266 which consists of TCT instead of the wild type TCC.
Other names of the plasmid are pCAGGSEmproIL1βHAtag and pCAGGSEmproIL1betaHAtag.
EMBL Accession number: NM_008361.3, view at GenBank
Latest sequence update: 09/10/2013
Sequence detail:
Primers used to amplify the HA-tagged mouse IL1B cds:

                                 <---         mIL1B         ...                  
Forward: 5' CGCAAGGAAGCGGCGGCCGC.GCA.ACT.GTT.CCT.GAA.CTC.AAC.TG
                     NotI 
                           <---         HA-epitope        --->
Reverse: 5' TCGCCCCGGG.TTA.AGC.GTA.ATC.TGG.AAC.ATC.GTA.TGG.GTA.
                XmaI   +++
            <---         mIL1B         ...
            GGA.AGA.CAC.AGA.TTC.CAT.GGT.GAA.G

+++: termination codon.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI/SacII, BglII/EcoRI, HincII and HindIII.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr R. Beyaert(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) 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 + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pCAGGSE-mIL1B-HA (LMBP 4354) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert .

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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