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Pcmv pe2 tagrfp bleor

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PCMV-PE2-tagRFP-BleoR is a plasmid vector that contains a red fluorescent protein (tagRFP) and a blasticidin resistance gene (BleoR). The plasmid is driven by a cytomegalovirus (CMV) promoter.

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Lab products found in correlation

Nebuilder hifi dna assembly master mix, by New England Biolabs (2 mentions) Pcdna3.1 zeo, by Thermo Fisher Scientific (2 mentions) Plv312, by Addgene (2 mentions) Lenti grna puro, by Addgene (2 mentions) Ecori hf, by New England Biolabs (2 mentions)

Spelling variants of this product

PCMV-PE2 (17 citations)

2 protocols using pcmv pe2 tagrfp bleor

1

Construction of pCMV-PE2-tagRFP-BleoR Plasmid

Cited 1 time
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pCMV-PE2 (Addgene #132775, a gift from David Liu1 (link)) was digested with EcoRI-HF (NEB). After digestion, the plasmid was dephosphorylated by rSAP (NEB), followed by gel extraction. tagRFP was PCR amplified from pLV312.3 (Addgene #119944)19 (link). tagRFP was Gibson assembled into pCMV-PE2 using 50 ng backbone and 2:1 ratio of tagRFP amplicon. Assembly was performed with NEBuilder HiFi DNA Assembly Master Mix (NEB). Assembled plasmids were transformed into NEB Stable Competent E. coli, resulting in pCMV-PE2-tagRFP. P2A-BleoR (Zeocin resistance) was PCR amplified from an in-house plasmid that originated from pcDNA™3.1/Zeo (Invitrogen). pCMV-PE2-tagRFP was PCR amplified to get linearized plasmid, with amplicon end downstream of tagRFP. Gibson assembly was performed as described previously, resulting in pCMV-PE2-tagRFP-BleoR (Addgene #192508). Lenti-p3-eGFP plasmid was produced by replacing EF1a-PuroR (MluI-HF and ApaI) on Lenti-gRNA-puro (Addgene #84752, a gift from Hyongbum Kim20 (link)) with p3-eGFP sequence.
Mathis N., Allam A., Kissling L., Marquart K.F., Schmidheini L., Solari C., Balázs Z., Krauthammer M, & Schwank G. (2023). Predicting prime editing efficiency and product purity by deep learning. Nature biotechnology.
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2

Construction of pCMV-PE2-tagRFP-BleoR Plasmid

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
pCMV-PE2 (Addgene #132775, a gift from David Liu1 (link)) was digested with EcoRI-HF (NEB). After digestion, the plasmid was dephosphorylated by rSAP (NEB), followed by gel extraction. tagRFP was PCR amplified from pLV312.3 (Addgene #119944)19 (link). tagRFP was Gibson assembled into pCMV-PE2 using 50 ng backbone and 2:1 ratio of tagRFP amplicon. Assembly was performed with NEBuilder HiFi DNA Assembly Master Mix (NEB). Assembled plasmids were transformed into NEB Stable Competent E. coli, resulting in pCMV-PE2-tagRFP. P2A-BleoR (Zeocin resistance) was PCR amplified from an in-house plasmid that originated from pcDNA™3.1/Zeo (Invitrogen). pCMV-PE2-tagRFP was PCR amplified to get linearized plasmid, with amplicon end downstream of tagRFP. Gibson assembly was performed as described previously, resulting in pCMV-PE2-tagRFP-BleoR (Addgene #192508). Lenti-p3-eGFP plasmid was produced by replacing EF1a-PuroR (MluI-HF and ApaI) on Lenti-gRNA-puro (Addgene #84752, a gift from Hyongbum Kim20 (link)) with p3-eGFP sequence.
Mathis N., Allam A., Kissling L., Marquart K.F., Schmidheini L., Solari C., Balázs Z., Krauthammer M, & Schwank G. (2023). Predicting prime editing efficiency and product purity by deep learning. Nature biotechnology.
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