GREAT AT SMALL THINGS

0

GeneCorner plasmid details

Last data update: 29 October 2020 04:15 CET

Plasmid name: pCAGGS-E-mCASP-1-4DE (LMBP 6093)

Add to cart

New search

Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p6093.gb (View with Genome Compiler)
p6093.txt
p6093.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse cysteinyl aspartate specific proteinase 1 cDNA (caspase-1, CASP-1, ICE, Casp1); mutated sequence
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-EmCASP-1m1
Further information: The plasmid was constructed by introducing two additional D/E mutations in the coding sequence of mouse CASP-1 of pCAGGS-EmCASP-1m1.
pCAGGS-E-mCASP-1-4DE contains mouse CASP-1 cDNA in which the four cleavage sites were mutated (D103E, D122E, D296E and D314E), rendering an uncleavable mouse CASP-1 mutant. The plasmid contains additional silent mutations near D103E and D122E, creating a BalI (MscI) and a KpnI restriction site.
pCAGGS-E-mCASP-1-4DE is useful for highly efficient expression of the mouse CASP-1-4DE mutant 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 LT726821.1.
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-mcasp-1 4DE WT.
EMBL Accession number: BC008152.1, view at EMBL, GenBank, DDBJ
LT726821.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 20/01/2014
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 
                  ***                                                     NotI

     |--> mouse CASP-1
     | 2           5      100         103     105 106 107         110
  GCT.GCT.GAC.AAG.ATC ... GCT.ACA.GAA.GAA.TCT.AAA.GGT.GGC.CAT.CCT.TCA.TCC.TCA.GAA.
  Ala Ala Asp Lys Ile     Ala Thr Glu Glu Ser Lys Gly Gly His Pro Ser Ser Ser Glu
                                      --^           ^   ^
                                       E            BalI (MscI)

                       prodomain --> | --> p30
      115                 120     122|123 124 125                 130     295 296   
  ACA.AAG.GAA.GAA.CAG.AAC.AAA.GAA.GAG.GGT.ACC.TTT.CCA.GGA.CTG.ACT.GGG ... AAA.GAA.
  Thr Lys Glu Glu Gln Asn Lys Glu Glu Gly Thr Phe Pro Gly Leu Thr Gly     Lys Glu
                                  --^   ^   ^                                 --^
                                   E  KpnI                                     E  

              300                 305                 310             314 315
  TCA.GTA.AGA.GAC.TCT.GAA.GAG.GAT.TTC.TTA.ACG.GAT.GCA.ATT.TTT.GAA.GAT.GAA.GGC ... 3'
  Ser Val Arg Asp Ser Glu Glu Asp Phe Leu Thr Asp Ala Ile Phe Glu Asp Glu Gly
                                                                      --^
                                                                       E
***: Start codon.
^: Mutated nucleotide.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglII/NotI, HindIII, NcoI and PvuI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr K. Kersse(1) (2) and Prof. Dr P. Vandenabeele(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:

pCAGGS-E-mCASP-1-4DE (LMBP 6093) is available at BCCM/GeneCorner. This plasmid was deposited by Dr K. Kersse and 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.

New search