pGBW-m4137382
(Plasmid #149539)

• Purpose: Mammalian expression plasmid for SARS-CoV-2 surface glycoprotein (Spike) for VSV pseudotyping
• Depositing Labs: Ginkgo Bioworks, Benjie Chen
• Publication: Truncated SARS-CoV-2 Spike protein constructs for making VSV pseudo virus (unpublished)

Backbone

• Vector backbone: AmpR gene, with start and stop codons
• Backbone size w/o insert (bp): 860
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: SARS-CoV-2 S (surface glycoprotein)
• Alt name: Spike protein
• Alt name: S protein
• Species: H. sapiens (human); Severe acute respiratory syndrome coronavirus 2
• Insert Size (bp): 6562
• Mutation: H. sapiens recode;19-aa Ctrunc
• Entrez Gene: S (a.k.a. GU280_gp02, spike glycoprotein)
• Promoter: CMV

Cloning Information

• Cloning method: Unknown

Resource Information

• A portion of this plasmid was derived from a plasmid made by: GinkgoBioworks
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • OpenMTA
• Industry Terms:
  • OpenMTA

Depositor Comments

NCBI reference sequence: YP_009724390.1 . This plasmid contains an 18 or 19bp truncation in the CMV promoter and was synthesized for making pseudotyped VSV particles. It has been shown that truncations either help expression and /or capture by VSVdeltaG. See https://www.ncbi.nlm.nih.gov/pubmed/19057867 for original paper on VSV pseudotyping, and https://www.cell.com/cell/pdf/S0092-8674(20)30229-4.pdf on more recent efforts with SARS-CoV-2.

Please note: Addgene NGS finds that the AmpR resistance marker is in the opposite orientation in the plasmid relative to the depositor's sequence. As the plasmid was able to grow in media with Ampicillin, this is not expected to affect plasmid function.