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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pTT3-DrEfnb3b-bait (LMBP 11596)

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Price category: Cat. 2 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: not available
Sequence
analysis results
Genecorner:

Sanger: ef32240430.fasta

Sanger: ef32240549.fasta

Cloned DNA: Danio rerio ephrin-B3b cDNA (efnb3b, GeneID 114378); extracellular domain (ECD)
Rattus norvegicus Cd4 molecule cDNA (Cd4, GeneID 24932); fragment
Biotinylation sequence (bio-tag); C-terminal
Promoter: Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: Rabbit β-globin polyadenylation signal (β-globin polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Epstein-Barr virus origin of replication (oriP)
Host range: Escherichia coli
Mammalian cells
Parental clone: unc5bCD4d3+4-bio
Further information: The plasmid was constructed by cloning the D. rerio Efnb3b extracellular domain coding sequence into the unc5bCD4d3+4-bio vector for Avidity-based Extracellular Interaction Screening (AVEXIS) assays.
This vector expresses the D. rerio Efnb3b as a bait protein, linked to a tag containing domains 3 and 4 of the rat CD4 (Cd4d3+4) protein, followed by a biotinylation sequence that is a substrate for the E.coli BirA enzyme and can therefore be monobiotinylated on a specific lysine residue. The Cd4d3+4 tag is recognised by the OX68 monoclonal antibody and is used to quantify the expression of the bait proteins by ELISA. The bait proteins are therefore monomeric and monobiotinylated.
The unc5bCD4d3+4-bio vector was derived from pTT3, obtained from Dr Yves Durocher (National Research Council of Canada, Biotechnology Research Institute).
The nucleotide sequence of the D. rerio Efnb3b cDNA corresponds with the EMBL Nucleotide Sequence Database accession number KM655629.1.
Other name of the plasmid is Efnb3b-bio-AVEXIS.
EMBL Accession number: KM655629.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 14/02/2020
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 Dr U. Irion(1). It was constructed by C. Söllner(1).
(1) Max Planck Institute for Developmental Biology, Tübingen, Germany
Plasmid reference: Gao et al., Cell Rep. 12 (2015), 694-708 [PMID: 26190107] [DOI: 10.1016/j.celrep.2015.06.047]
Related plasmid reference: Bushell et al., Genome Res. 18 (2008), 622-630 [PMID: 18296487] [DOI: 10.1101/gr.7187808]
Söllner et al., Genome Biol. 10 (2009), R99 [PMID: 19765300] [DOI: 10.1186/gb-2009-10-9-r99]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH5αT1R
Host reference: -
Related host reference: Killmann et al., J. Bacteriol. 178 (1996), 6913-6920 [PMID: 8955314] [DOI: 10.1128/jb.178.23.6913-6920.1996]
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:

pTT3-DrEfnb3b-bait (LMBP 11596) is available at BCCM/GeneCorner. This plasmid was deposited by Dr U. Irion and was published in Gao et al., 2015.

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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