Last data update: 24 January 2024 16:39 CET
Plasmid name: pTT3-DrJam2b-bait (LMBP 11802)
New search | Print data sheet |
Price category: | Cat. 2 (cf. price list) |
Status: | GeneCorner non-core plasmid |
Depositor's sequence: | not available |
Sequence analysis results Genecorner: |
Sanger: ef32241116.fasta Sanger: ef32241669.fasta |
Cloned DNA: |
Danio rerio junctional adhesion molecule 2b cDNA (jam2b, sc:d0813, GeneID 100005301); 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 Jam2b extracellular domain coding sequence into the unc5bCD4d3+4-bio vector for Avidity-based Extracellular Interaction Screening (AVEXIS) assays. This vector expresses the D. rerio Jam2b 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 Jam2b cDNA corresponds with the EMBL Nucleotide Sequence Database accession number CU458870.1. Other name of the plasmid is jam2b-bio-AVEXIS. |
EMBL Accession number: | CU458870.1, view at EMBL, GenBank, DDBJ |
Latest sequence update: | 02/04/2020 |
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 G. Wright(1). It was constructed by Dr C. Söllner(1). (1) Cell Surface Signalling Laboratory, Wellcome Trust Sanger Institute, Cambridge, UK |
Plasmid reference: | Söllner et al., Genome Biol. 10 (2009), R99 [PMID: 19765300] [DOI: 10.1186/gb-2009-10-9-r99] |
Related plasmid reference: | Martin et al., Mol. Cell. Proteomics 9 (2010), 2654-2665 [PMID: 20802085] [DOI: 10.1074/mcp.M110.004119] Bushell et al., Genome Res. 18 (2008), 622-630 [PMID: 18296487] [DOI: 10.1101/gr.7187808] Gao et al., Cell Rep. 12 (2015), 694-708 [PMID: 26190107] [DOI: 10.1016/j.celrep.2015.06.047] |
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-DrJam2b-bait (LMBP 11802) is available at BCCM/GeneCorner. This plasmid was deposited by Dr G. Wright and was published in Söllner et al., 2009. |
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.