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GeneCorner plasmid details

Last data update: 20 October 2020 04:14 CEST

Plasmid name: pCAGGS-E-hABIN-1 (LMBP 5126)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p5126.gb (View with Genome Compiler)
p5126.txt
p5126.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human A20-binding inhibitor of NF-κB activation 1 cDNA (ABIN-1, TNIP1, NAF1)
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; pCAGGSEhA20
Further information: The plasmid was constructed by ligating the three following fragments: 1) the 2517 bp XhoI/PvuI fragment from pCAGGS; 2) the 2336 bp PvuI/NotI fragment from pCAGGSEhA20; 3) a 1919 bp NotI/XhoI PCR fragment, containing the human ABIN-1 coding sequence.
pCAGGS-E-hABIN-1 is useful for highly efficient expression of human ABIN-1 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 LT727094.1.
The nucleotide sequence of the human ABIN-1 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AJ011895.1, except at two positions: 1) the nucleotide at position 895 of the coding sequence is a 'G' in the EMBL record (i.e. P299A mutation according to the information available at BCCM/GeneCorner) but a 'C' in the plasmid sequence (wt sequence); 2) amino acid 9 (isoleucine) is encoded by the wt sequence 'ATC' in the EMBL record and by the silent mutation 'ATT' in the plasmid sequence, resulting in the loss of a DpnI and a Sau3AI site.
Other name of the plasmid is pCAGGS-E-hABIN1-FLCR.
EMBL Accession number: AJ011895.1, view at EMBL, GenBank, DDBJ
LT727094.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 29/10/2010
Sequence detail:
Primers used to amplify the hABIN-1 coding sequence:

forward: 5' ... CGGGATCCTAGCGGCCGCC.ATG.GAA.GGG.AGA.GGA.CCG ... 3'
                  BamHI   NotI      ***

reverse: 5' ... GCCGCTCGAG.TCA.CTG.AGG.CCC.CTC.ACG ... 3'
                    XhoI   +++


                                   7       9      11
WT according to NM_006058: 5' ... TAC.CGG.ATC.TAC.GAC ... 3'
                                  Tyr Arg Ile Tyr Asp
                                        DpnI
                                        Sau3AI

                                   7       9      11
silent mutant:             5' ... TAC.CGG.ATT.TAC.GAC ... 3'
                                  Tyr Arg Ile Tyr Asp
                                            ^
***: start codon.
+++: termination codon.
^  : mutated nucleotide.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: AvaI, DpnI, EcoRI, NotI/XhoI and PvuI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed by M. Haegman(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: Huang et al., Oncogene 27 (2008), 6131-6140 [PMID: 18622428]
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-hABIN-1 (LMBP 5126) 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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