HuEV-A
(Plasmid #68342)

• Purpose: (Empty Backbone) Gateway compatible human expression vector that adds a "flexible" 3XFLAG,V5,YFP N terminus tag
• Depositing Lab: Jef Boeke
• Publication: Mita et al Biol Proced Online. 2016 Aug 15;18:16. doi: 10.1186/s12575-016-0046-x. eCollection 2016.

Backbone

• Vector backbone: pCEP4
• Backbone manufacturer: Invitrogen
• Backbone size (bp): 10200
• Modifications to backbone: The HuEV-A vector was constructed from a one-step ligation reaction with two inserts and a vector fragment. The backbone for the HuEV-A vector was the previously described pCEP4-Puro plasmid (5). The first insert was a DNA fragment containing the chloramphenicol resistance gene, ccdB gene, and att-R2 site, and was constructed by joining a NotI/KpnI segment of the Gateway Reading frame A cloning vector (Life Technologies, Cat #11828-029) containing the chloramphenicol resistance gene and the att-R2 site to a second synthetic DNA fragment (UEV) containing the complementary att-R1 site along with a series of epitope tags, recombination sites, and the Venus YFP sequence with BsiWI (compatible with BsrGI) and EagI (compatible with NotI) termini. The pCEP Puro backbone vector was digested with KpnI and BsrGI, and the inserts were ligated into the backbone using T4 DNA ligase in ligase buffer (Enzymatics). Vectors were transformed into E. coli and selected on ampicillin/chloramphenicol plates.
• Vector type: Mammalian Expression, Cre/Lox
• Promoter: TET-CMV
• Selectable markers: Puromycin
• Tag / Fusion Protein:
  • 3xFLAG, V5 and YFP (N terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Ampicillin, 25 & 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): ccdB Survival
• Copy number: High Copy

Cloning Information

• Cloning method: Gateway Cloning

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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