Last data update: 24 January 2024 16:39 CET
Plasmid name: pTT3-hDSCAM-ig1-9-bait (LMBP 11513)
| New search | Print data sheet |
| Price category: | Cat. 3 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p11513.gb |
| Sequence analysis results Genecorner: |
NGS: c09-gc-dec2019.fasta |
| Cloned DNA: |
Human pregnancy specific beta-1-glycoprotein 1 cDNA (PSG1, B1G1, PBG1, PSBG1, PS-beta-G-1, GeneID 5669); signal sequence Human DS cell adhesion molecule cDNA (DSCAM, CHD2-42, CHD2-52, GeneID 1826); codon optimized fragment Tobacco etch virus (TEV) protease recognition site; C-terminal Histidine tag (His-tag); C-terminal 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 part of the codon optimized human DSCAM coding sequence into the unc5bCD4d3+4-bio vector for Avidity-based Extracellular Interaction Screening (AVEXIS) assays. This vector expresses a fragment of the human DSCAM, consisting of immunoglobulin domains Ig1 up to and including Ig9, as a bait protein, linked to a TEV recognition site, His-tag and 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). Other name of the plasmid is DSCAM_ig1-9-bio-AVEXIS. |
| EMBL Accession number: | - |
| Latest sequence update: | 22/01/2020 |
| Authenticity test: | This plasmid has been fully sequenced but the NGS sequence data still need to be implemented. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr X. Gao(1) and Dr R. Meijers(1). (1) EMBL Hamburg, Germany |
| Plasmid reference: | - |
| Related plasmid reference: | Söllner et al., Genome Biol. 10 (2009), R99 [PMID: 19765300] [DOI: 10.1186/gb-2009-10-9-r99] Gao et al., Cell Rep. 12 (2015), 694-708 [PMID: 26190107] [DOI: 10.1016/j.celrep.2015.06.047] Liu et al., Neuron 97 (2018), 1261-1267 [PMID: 29503192] [DOI: 10.1016/j.neuron.2018.02.010] |
| Restricted distribution: | - BCCM/MTA adapted by EMBL - Restricted to academic users - Acknowledgment of the depositor in any publication resulting from the use of the MATERIAL |
| 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-hDSCAM-ig1-9-bait (LMBP 11513) is available at BCCM/GeneCorner. This plasmid was deposited by Dr X. Gao and Dr R. Meijers. |
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.