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GeneCorner plasmid details

Last data update: 23 October 2020 04:27 CEST

Plasmid name: pCAGGS-EmCASP-1C285A (LMBP 4937)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p4937.gb (View with Genome Compiler)
p4937.txt
p4937.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse cysteinyl aspartate specific proteinase 1 cDNA (caspase-1, CASP-1, ICE, Casp1); C285A 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
Further information: pCAGGS-EmCASP-1C285A codes for the enzymatically inactive mutant of mouse caspase-1.
The mutation is described as C285A, according to the numbering of the codons in the human caspase-1 coding sequence, although the mutated mouse codon is located on position 284.
The plasmid is useful for highly efficient expression of mouse CASP-1C285A, fused to the N-terminal E-tag, 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 LT727059.1.
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 pCAGGS-E-mCASP1 C/A.
EMBL Accession number: BC008152.1, view at EMBL, GenBank, DDBJ
LT727059.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 15/05/2008
Sequence detail:
E-tag
                      ---------------------------------------------------
5' ... GAATTCCACC ATG.GGT.GCG.CCG.GTG.CCG.TAT.CCG.GAC.CCG.CTG.GAA.CCG.CGT.GCG.GCC.
                  Met Gly Ala Pro Val Pro Tyr Pro Asp Pro Leu Glu Pro Arg Ala Ala 
       EcoRI                                BspMII                        NotI


   |--> mouse CASP-1
   | 2   3   4                            280             284         
    GCT.GAC.AAG...GAC.AAG.CCC.AAG.GTG.ATC.ATT.ATT.CAG.GCA.GCC.CGT.GGA.GAG.AAA.CAA...
                  Asp Lys Pro Lys Val Ile Ile Ile Gln Ala Ala Arg Gly Glu Lys Gln 
                                                          **
                                                   Q   A   A   R   G   

*: mutated nucleotide.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglII/NotI, NcoI, SphI and XbaI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr P. Vandenabeele(1) (2). It was constructed by M. Lamkanfi(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: Lamkanfi et al., J. Biol. Chem. 279 (2004), 24785-24793 [PMID: 15039421]
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-1C285A (LMBP 4937) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr P. Vandenabeele .

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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