pVEGFR1_TEV(C)_TCS(Q'G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1
(Plasmid
#101103)
-
PurposeEncodes dCas9(C)-VP64 with SAM components fused to VEGFR1
-
Depositing Lab
-
Depositing OrganizationUniversity of Oxford
Located in United Kingdom of Great Britain and Northern Ireland -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 101103 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 101103-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 101103-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
-
Vector backbonepTMt_TCS(Q’G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1
-
Modifications to backboneA sequence containing the VEGFR1 (FLT1) leader peptide, extracellular domain and transmembrane domain were PCR amplified from plasmid pDONR223-FLT1 (gift from William Hahn & David Root, Addgene plasmid #23912) and used to replace the TMt in pTMt_TCS(Q’G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1. In addition, the C-terminal TEV fragment was amplified from full length TEV protease as previously described (Wehr et al., 2006) and cloned between the VEGFR1 transmembrane domain and the TEV cleavage site.
-
Vector typeMammalian Expression, CRISPR, Synthetic Biology
Growth in Bacteria
-
Bacterial Resistance(s)Ampicillin, 100 μg/mL
-
Growth Temperature37°C
-
Growth Strain(s)DH5alpha
-
Copy numberHigh Copy
Gene/Insert
-
Gene/Insert nameVEGFR1-dCas9(C)
-
Tag
/ Fusion Protein
- HA
Resource Information
-
A portion of this plasmid was derived from a plasmid made bypDONR223-FLT1 (gift from William Hahn & David Root, Addgene plasmid #23912); pTMt_TCS(Q’G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1.
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
DNA (Catalog # 101103-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 101103-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
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:
pVEGFR1_TEV(C)_TCS(Q'G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1 was a gift from Tudor Fulga (Addgene plasmid # 101103 ; http://n2t.net/addgene:101103 ; RRID:Addgene_101103) -
For your References section:
Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors. Baeumler TA, Ahmed AA, Fulga TA. Cell Rep. 2017 Sep 12;20(11):2639-2653. doi: 10.1016/j.celrep.2017.08.044. 10.1016/j.celrep.2017.08.044 PubMed 28903044
