PurposeEncodes SARS-CoV-2 Spike with 21 aa C-terminal deletions for lentiviral pseudotyping
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||155130||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4750
- Total vector size (bp) 8314
Modifications to backboneCGCCACC added 3' to EcoRI site and 5' to S start codon
Vector typeMammalian Expression, Lentiviral
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Growth Strain(s)NEB Stable
Copy numberHigh Copy
Gene/Insert nameSARS-CoV-2 Spike-delta21
Alt nameCodon optimized SARS-CoV-2 Spike
SpeciesSevere acute respiratory syndrome coronavirus 2
Insert Size (bp)3762
MutationCodon optimized to H. sapiens using IDT codon optimization tool; deleted 21 amino acids by deletion from 3769-3831
Entrez GeneS (a.k.a. GU280_gp02, spike glycoprotein)
- Promoter CMV
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site HindIII (not destroyed)
- 5′ sequencing primer CGCATGGTGTGGTCTTTCTCC
- 3′ sequencing primer agtccaagctaggcccttttg (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
Based on a plasmid described previously in Crawford et al. 2020, Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization Assays (https://www.mdpi.com/1999-4915/12/5/513). This is a version of the Spike expression plasmid used to pseudotype lentivirus which provided better titers in this protocol.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:HDM-SARS2-Spike-delta21 was a gift from Jesse Bloom (Addgene plasmid # 155130 ; http://n2t.net/addgene:155130 ; RRID:Addgene_155130)
For your References section:Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization Assays. Crawford KHD, Eguia R, Dingens AS, Loes AN, Malone KD, Wolf CR, Chu HY, Tortorici MA, Veesler D, Murphy M, Pettie D, King NP, Balazs AB, Bloom JD. Viruses. 2020 May 6;12(5). pii: v12050513. doi: 10.3390/v12050513. 10.3390/v12050513 PubMed 32384820