Last data update: 24 January 2024 16:39 CET
Plasmid name: pTT3-hNTN1-bait (LMBP 11508)
New search | Print data sheet |
Price category: | Cat. 2 (cf. price list) |
Status: | GeneCorner non-core plasmid |
Depositor's sequence: | p11508.gb |
Sequence analysis results Genecorner: |
Sanger: ef32243336.fasta Sanger: ef32243358.fasta |
Cloned DNA: |
Human pregnancy specific beta-1-glycoprotein 1 cDNA (PSG1, B1G1, PBG1, PSBG1, PS-beta-G-1, GeneID 5669); signal sequence Human netrin 1 cDNA (NTN1, MRMV4, NTN1L, GeneID 9423); codon optimized sequence 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 the codon optimized human NTN1 coding sequence into the unc5bCD4d3+4-bio vector for Avidity-based Extracellular Interaction Screening (AVEXIS) assays. This vector expresses the human NTN1 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). Other name of the plasmid is NETRIN1-bio-AVEXIS. |
EMBL Accession number: | - |
Latest sequence update: | 09/09/2019 |
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 X. Gao(1) and Dr R. Meijers(1). (1) EMBL Hamburg, Germany |
Plasmid reference: | Liu et al., Neuron 97 (2018), 1261-1267 [PMID: 29503192] [DOI: 10.1016/j.neuron.2018.02.010] |
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 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-hNTN1-bait (LMBP 11508) is available at BCCM/GeneCorner. This plasmid was deposited by Dr X. Gao and Dr R. Meijers and was published in Liu et al., 2018. |
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.