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GeneCorner plasmid details

Last data update: 27 November 2020 04:15 CET

Plasmid name: pCAGGS-EmCASP-1m1 (LMBP 3914)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p3914.gb (View with Genome Compiler)
p3914.txt
p3914.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse cysteinyl aspartate specific proteinase 1 cDNA (caspase-1, CASP-1, ICE, Casp1); p45-2DE mutant
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; pCAGGS-caspase-1 2D/E
Further information: The construction of the plasmid is described in Lamkanfi et al. (2004).
The mouse CASP-1 coding sequence contains point mutations (p45-2DE mutant), converting two Asp (D) codons of the prodomain to Glu (E) to destroy potential cleavage sites, and creating additional BalI (MscI) and KpnI restriction sites.
pCAGGS-EmCASP-1m1 is useful for highly efficient expression of mutated mouse caspase-1 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 LT727286.1.
The nucleotide sequence of the wild-type mouse CASP-1 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number BC008152.1.
Other name of the plasmid is pCAGGSEICEASP.
EMBL Accession number: BC008152.1, view at EMBL, GenBank, DDBJ
LT727286.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 24/10/2001
Sequence detail:
Nucleotide sequence of the mutated mouse CASP-1 insert:


                                           E-tag
                      ---------------------------------------------------
5' ... GAATTCCACC ATG.GGT.GCG.CCG.GTG.CCG.TAT.CCG.GAC.CCG.CTG.GAA.CCG.CGT.GCG.GCC.
       EcoRI      Met Gly Ala Pro Val Pro Tyr Pro Asp Pro Leu Glu Pro Arg Ala Ala 
                  ***                       BspMII                        NotI


        |--> mouse CASP-1
        | 2   3   4        100         103     105                 110
     GCT.GCT.GAC.AAG...TTT.GTT.GCT.ACA.GAA.GAA.TCT.AAA.GGT.GGC.CAT.CCT.TCA.TCC.TCA.
     Ala Ala Asp Lys   Phe Val Ala Thr Glu Glu Ser Lys Gly Gly His Pro Ser Ser Ser 
                                       --^           ^   ^
                                        E            BalI (MscI)

 
                          prodomain --> | --> p30
                             120     122|
     GAA.ACA.AAG.GAA.GAA.CAG.AAC.AAA.GAA.GAG.GGT.ACC.TTT.CCA.GGA.CTG.ACT. ... 3'
     Glu Thr Lys Glu Glu Gln Asn Lys Glu Glu Gly Thr Phe Pro Gly Leu Thr
                                     --^   ^   ^
                                      E  KpnI

***: start codon.
^: mutated nucleotide.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BalI, KpnI, NcoI and SphI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by P. Schotte(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: Lamkanfi et al., J. Biol. Chem. 279 (2004), 24785-24793 [PMID: 15039421]
Related plasmid reference: Van Criekinge et al., J. Biol. Chem. 271 (1996), 27245-27248 [PMID: 8910297]
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:

pCAGGS-EmCASP-1m1 (LMBP 3914) is available at BCCM/GeneCorner. This plasmid was deposited by P. Schotte and Prof. Dr R. Beyaert and was published in Lamkanfi et al., 2004.

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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