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GeneCorner plasmid details

Last data update: 29 October 2020 04:15 CET

Plasmid name: pCAGGSmIL1B (LMBP 3853)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p3853.gb (View with Genome Compiler)
p3853.txt
p3853.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse interleukin 1 beta cDNA (Il1b, Il-1b, IL-1beta, GeneID 16176)
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: The plasmid was constructed by inserting the mouse IL1B prosequence cDNA as an EcoRV PCR fragment into the unique BalI site of pCAGGS, in the sense orientation relative to the AG promoter.
pCAGGSmIL1B is useful for highly efficient expression of the mouse IL1B prosequence under the control of the AG promoter and the hCMV- IE enhancer in various mammalian cells.
Sequence analysis at GeneCorner revealed 4 silent and 1 non-silent (Gln -> Leu replacement at codon 132) differences. According to the depositor however, the plasmid is biologically active upon cotransfection with casp-1 in HEK293 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 LT727274.1.
The nucleotide sequence of the wt mouse IL1B prosequence corresponds with the Genbank accession number NM_008361.3.
Name mentioned in Maelfait et al. (2008) is pCAGGS-pro-IL-1β.
Other name of the plasmid is pcaggsmpIL1β.
EMBL Accession number: NM_008361.3, view at GenBank
LT727274.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 20/02/2006
Sequence detail:
PCR primers used to amplify the mIL1B cDNA:

Forward primer: 5' GCGGATATCAGCGCTCCACC.ATG.GCA.ACT.GTT.CCT.GAA.CTC.AAC 3'
                      EcoRV             ***

Reverse primer: 5' GCGGATATCAGCGC.TTA.GGA.AGA.CAC.AGA.TTC.CAT.GGT 3'
                      EcoRV       +++

***: start codon.
+++: termination codon.
Punctuation indicates reading frame.


Nucleotide sequence of the mouse IL1B coding sequence in pCAGGSmIL1B (LMBP 3853):

ATG GCA ACT GTT CCT GAA CTC AAC TGT GAA ATG CCA CCT TTT GAC AGT GAT GAG AAT GAC
MET Ala Thr Val Pro Glu Leu Asn Cys Glu MET Pro Pro Phe Asp Ser Asp Glu Asn Asp

CTG TTC TTT GAA GTT GAC GGA CCC CAA AAG ATG AAG GGC TGC TTC CAA ACC TTT GAC CTG
Leu Phe Phe Glu Val Asp Gly Pro Gln Lys MET Lys Gly Cys Phe Gln Thr Phe Asp Leu

         43                                  52
GGC TGT CCT GAT GAG AGC ATC CAG CTT CAA ATC TCG CAG CAG CAC ATC AAC AAG AGC TTC
Gly Cys Pro Asp Glu Ser Ile Gln Leu Gln Ile Ser Gln Gln His Ile Asn Lys Ser Phe
          *                                   *                                    

AGG CAG GCA GTA TCA CTC ATT GTG GCT GTG GAG AAG CTG TGG CAG CTA CCT GTG TCT TTC 
Arg Gln Ala Val Ser Leu Ile Val Ala Val Glu Lys Leu Trp Gln Leu Pro Val Ser Phe 

CCG TGG ACC TTC CAG GAT GAG GAC ATG AGC ACC TTC TTT TCC TTC ATC TTT GAA GAA GAG
Pro Trp Thr Phe Gln Asp Glu Asp MET Ser Thr Phe Phe Ser Phe Ile Phe Glu Glu Glu

CCC ATC CTC TGT GAC TCA TGG GAT GAT GAT GAT AAC CTG CTG GTG TGT GAC GTT CCC ATT 
Pro Ile Leu Cys Asp Ser Trp Asp Asp Asp Asp Asn Leu Leu Val Cys Asp Val Pro Ile

    122                                     132
AGA CAA CTG CAC TAC AGG CTC CGA GAT GAA CAA CTA AAA AGC CTC GTG CTG TCG GAC CCA
Arg Gln Leu His Tyr Arg Leu Arg Asp Glu Gln Leu Lys Ser Leu Val Leu Ser Asp Pro
      *                                      *

TAT GAG CTG AAA GCT CTC CAC CTC AAT GGA CAG AAT ATC AAC CAA CAA GTG ATA TTC TCC
Tyr Glu Leu Lys Ala Leu His Leu Asn Gly Gln Asn Ile Asn Gln Gln Val Ile Phe Ser

ATG AGC TTT GTA CAA GGA GAA CCA AGC AAC GAC AAA ATA CCT GTG GCC TTG GGC CTC AAA
MET Ser Phe Val Gln Gly Glu Pro Ser Asn Asp Lys Ile Pro Val Ala Leu Gly Leu Lys

GGA AAG AAT CTA TAC CTG TCC TGT GTA ATG AAA GAC GGC ACA CCC ACC CTG CAG CTG GAG
Gly Lys Asn Leu Tyr Leu Ser Cys Val MET Lys Asp Gly Thr Pro Thr Leu Gln Leu Glu

                                                    214
AGT GTG GAT CCC AAG CAA TAC CCA AAG AAG AAG ATG GAA AAA CGG TTT GTC TTC AAC AAG
Ser Val Asp Pro Lys Gln Tyr Pro Lys Lys Lys MET Glu Lys Arg Phe Val Phe Asn Lys
                                                      *

ATA GAA GTC AAG AGC AAA GTG GAG TTT GAG TCT GCA GAG TTC CCC AAC TGG TAC ATC AGC
Ile Glu Val Lys Ser Lys Val Glu Phe Glu Ser Ala Glu Phe Pro Asn Trp Tyr Ile Ser

ACC TCA CAA GCA GAG CAC AAG CCT GTC TTC CTG GGA AAC AAC AGT GGT CAG GAC ATA ATT
Thr Ser Gln Ala Glu His Lys Pro Val Phe Leu Gly Asn Asn Ser Gly Gln Asp Ile Ile

GAC TTC ACC ATG GAA TCT GTG TCT TCC TAA 
Asp Phe Thr MET Glu Ser Val Ser Ser +++

*: Difference with the EMBL nucleotide sequence (Accession number M15131; SV 1).
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, HindIII and PstI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by 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: -
Related plasmid reference: 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 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
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:

pCAGGSmIL1B (LMBP 3853) is available at BCCM/GeneCorner. This plasmid was deposited by 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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