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GeneCorner plasmid details

Last data update: 23 October 2020 04:27 CEST

Plasmid name: pCAGGS-E-mMyD88 (LMBP 6654)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p6654.gb (View with Genome Compiler)
p6654.txt
p6654.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse myeloid differentiation primary response gene 88 cDNA (Myd88, GeneID 17874)
E-tag; N-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-mMyD88
Further information: The plasmid was constructed as follows: 1) the 3' end of mouse Myd88 cDNA was PCR amplified from pCAGGS-E-mMyD88, reinserting the termination codon and reconstructing the unique BglII site; 2) this PCR fragment was ligated as an MscI/BglII fragment into the MscI/BglII opened pCAGGS-E-mMyD88 vector. The latter vector contains mouse MyD88 cDNA that was picked up from the mouse macrophage cell line Mf4/4 and fused in frame to the E-tag of a pCAGGS-based expression plasmid; however, the termination codon seemed to be missing.
This plasmid is useful for highly efficient expression of mouse MyD88 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 lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726837.1.
The nucleotide sequence of the mouse Myd88 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number U84409.1.
Name mentioned in Maelfait et al. (2008) is pCAGGS-E-MyD88.
Other name of the plasmid is pCAGGS-E-mMyD88-2.
EMBL Accession number: U84409.1, view at EMBL, GenBank, DDBJ
LT726837.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 11/03/2009
Sequence detail:
Primers used to amplify and reconstruct the 3' end of mouse Myd88:

Forward: 5' - CAG.GTG.GCC.AGA.GTG.GAA.AG 
                   MscI

Reverse: 5' - ATCTAGATCT.TCA.GGG.CAG.GGA.CAA.AGC.CTT.GG
                  BglII  +++

+++: termination codon.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, BglII, BstZI, EcoRV and NcoI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr S. Janssens(1) (2) and 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: -
Related plasmid reference: Janssens et al., Curr. Biol. 12 (2002), 467-471 [PMID: 11909531]
Janssens et al., Trends Biochem. Sci. 27 (2002), 474-482 [PMID: 12217523]
Janssens et al., FEBS Lett. 548 (2003), 103-107 [PMID: 12885415]
Maelfait et al., J. Exp. Med. 205 (2008), 1967-1973 [PMID: 18725521] [DOI: 10.1084/jem.20071632]
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: LMBP 05645

Refer in your Materials and Methods:

pCAGGS-E-mMyD88 (LMBP 6654) is available at BCCM/GeneCorner. This plasmid was deposited by Dr S. Janssens and 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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