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GeneCorner plasmid details

Last data update: 27 October 2020 04:27 CET

Plasmid name: pCAGGS-E-hA20-Ct-ZF1 (LMBP 5928)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p5928.gb (View with Genome Compiler)
p5928.txt
p5928.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human TNF alpha induced protein 3 cDNA (TNFAIP3, A20, OTUD7C, GeneID 7128); C-terminal fragment
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: pCAGGSEhA20
Further information: The plasmid was constructed by amplifying the C-terminal fragment of human A20 (encoding AA440-790 - without ZF1) from pCAGGSEhA20 by PCR, and cloning it as a NotI/XhoI fragment into the NotI/XhoI opened pCAGGSEhA20 vector.
pCAGGS-E-hA20-Ct-ZF1 is useful for highly efficient expression of the E-tagged C-terminal fragment of human A20 under the control of the AG promoter and the hCMV-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 LT726815.1.
The nucleotide sequence of the human A20 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number M59465.1.
EMBL Accession number: M59465.1, view at EMBL, GenBank, DDBJ
LT726815.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 03/02/2009
Sequence detail:
Primers used to amplify the C-terminal fragment of human A20:

                               440                 445 446
forward: 5' AAGTAGCGGCCGCG.ATG.GGA.GAA.GCC.TAT.GAG.CCC.TTG 3'
                 NotI      *** 

                          790             786
reverse: 5' CCGCTCGAG.TTA.GCC.ATA.CAT.CTG.CTT.G 3'
               XhoI   +++

***: start codon.
+++: termination codon.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: NcoI, NotI/XhoI, PvuI and XmnI.
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: -
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-hA20-Ct-ZF1 (LMBP 5928) 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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