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GeneCorner plasmid details

Last data update: 21 October 2021 10:13 CEST

Plasmid name: pEFBOS-EmCASP-1 (LMBP 3920)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p3920.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse cysteinyl aspartate specific proteinase 1 cDNA (caspase-1, CASP-1, ICE, Casp1)
E-tag; N-terminal
Promoter: Human elongation factor 1α promoter (EF1α)
Escherichia coli lac operon promoter
Simian virus 40 early promoter (SV40 early)
Ribosome
binding site:
-
Terminator: Human granulocyte macrophage colony stimulating factor polyadenylation signal (GM-CSF polyA)
Selection marker: Ampicillin (amp)
Replicon: Bacteriophage M13 origin
Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pEFBOS-EmCASP-7; pGEM-mCASP-1
Further information: The plasmid was constructed as follows: the NotI/MluI fragment of pEFBOS-mCASP-7 was replaced by a NotI/MluI fragment, generated by PCR, with the coding sequence of mouse caspase-1.
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 the mouse CASP-1 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number BC008152.1.
Other name of the plasmid is pEFBOSEICE.
EMBL Accession number: BC008152.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 23/03/1999
Sequence detail:
Nucleotide sequence at the start of the gene:

                              |-->        E-tag
5'...AGGATCCAATTGAATTCCACC.ATG.GGT.GCG.CCG.GTG.CCG.TAT.CCG.GAC.
      BamHI                MET Gly Ala Pro Val Pro Tyr Pro Asp
           MfeI



                                |--> mouse caspase-1
                     -->|       | 2           5       
     CCG CTG GAA CCG CGT GCG GCC GCT GAC AAG ATC CTG AGG...3'
     Pro Leu Glu Pro Arg Ala Ala Ala Asp Lys Ile Leu Arg 
                         NotI


Forward primer: 5' GCG TCC GCG GCC GCT GAC AAG ATC CTG AGG GCA AAG 3'
                           NotI  

Reverse primer: 5' GCG ACG CGT TTA ATG TCC CGG GAA GAG GTA GAA 3'
                       MluI
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 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: 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
Other culture collection numbers: -

Refer in your Materials and Methods:

pEFBOS-EmCASP-1 (LMBP 3920) 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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