pAAVS1-Blast-PECAM1-Neo
(Plasmid #221149)

• Purpose: Expression of platelet and endothelial cell adhesion molecule 1 (CD31)
• Depositing Lab: Paul Burridge
• Publication: Pinheiro et al Sci Rep. 2024 Mar 26;14(1):7123. doi: 10.1038/s41598-024-57686-8.

Backbone

• Vector backbone: pAAVS1-P-CAG-DEST (Plasmid #80490)
• Backbone manufacturer: Knut Woltjen
• Total vector size (bp): 14899
• Modifications to backbone: Gateway destination cassette replaced with a promoterless mScarlet ORF flanked by 5' AscI and NotI sites and a 3' Sbfl site. A neomycin phosphotransferase ORF PCR-amplified with primers containing a 5′ NotI and 3′ SbfI site was cloned via TOPO cloning into the pCR4Blunt II-TOPO (Invitrogen, K280002) and used to replace the mScarlet reading frame using the NotI and SbfI sites. The Puromycin resistance reading frame was removed and replaced with the Blasticidin ORF from a previously made AAVS1-Blast plasmid using PmII and SpeI restriction sites.
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Growth instructions: In mammalian cells, use Blasticidin for selection of integrated constructs and use Neomycin for selection of cells with the PECAM1 promoter active.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Platelet And Endothelial Cell Adhesion Molecule 1
• Alt name: PECAM1
• Alt name: CD31
• Species: H. sapiens (human)
• Entrez Gene: PECAM1 (a.k.a. CD31, CD31/EndoCAM, GPIIA', PECA1, PECAM-1, endoCAM)
• Promoter: PECAM1

Cloning Information

• Cloning method: TOPO Cloning
• 5′ sequencing primer: na

Resource Information

• Supplemental Documents:
  • pAAVS1-Blast-PECAM-Neo.gb
• A portion of this plasmid was derived from a plasmid made by: PECAM1-eGFP was a gift from Victoria Bautch (Addgene plasmid # 14688; https://www.addgene.org/14688/; RRID:Addgene_14688).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry