pVITRO-Fab_MTX-LUCID
(Plasmid #119047)

• Purpose: Plasmid encoding the mammalian expression of the Metotraxate LUCID based on antibodies (BRET semi-synthetic biosensor)
• Depositing Lab: Kai Johnsson
• Publication: Xue et al Angew Chem Int Ed Engl. 2017 Jun 12;56(25):7112-7116. doi: 10.1002/anie.201702403. Epub 2017 May 16.

Backbone

• Vector backbone: pVITRO1-Hygro
• Backbone manufacturer: Invivogen
• Backbone size w/o insert (bp): 6295
• Total vector size (bp): 9219
• Vector type: Mammalian Expression
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Hygromycin, 200 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert 1

• Gene/Insert name: Fab_MTX-LUCID_part1
• Alt name: SNAP_p30_nLuc_Fab MTX (VL)
• Species: Synthetic
• Insert Size (bp): 1983
• Promoter: EF-1α + CMV/SV40 enhancers
• Tags / Fusion Proteins:
  • Strep-tag (C terminal on insert)
  • His-tag (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: mAB-S3: cagGTGTTGTGAAagCC
• 3′ sequencing primer: mAB-S7: CCTTTTTTAGGTGTTGTGAAAaC

Gene/Insert 2

• Gene/Insert name: Fab_MTX-LUCID_part2
• Alt name: Fab MTX (VL)
• Species: Synthetic
• Insert Size (bp): 795
• Promoter: IRES
• Tag / Fusion Protein:
  • FLAG-tag

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: mAB-S8: gGGATTTTGGTCATGGCTAG
• 3′ sequencing primer: mAB-S9: TGGTGTACCCTGTGGATG

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • NanoLuc Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry