pBMN TS-HiBiT / Lyt2
(Plasmid #179757)

• Purpose: Expresses human thymidylate synthase (TS) with C-terminal HiBiT tag in mammalian cells. This retroviral vector has an IRES-Lyt-2 downstream of the cloning sites
• Depositing Lab: James Inglese
• Publication: Ciulla et al Nat Commun. 2025 May 10;16(1):4342. doi: 10.1038/s41467-025-59658-6.

Backbone

• Vector backbone: pBMN
• Backbone manufacturer: Prof. G. Nolan, Stanford University, Stanford, CA
• Backbone size w/o insert (bp): 6352
• Total vector size (bp): 7329
• Vector type: Retroviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: human TS-HiBiT and IRES-Lyt-2
• Alt name: TS-HiBiT
• Alt name: TYMS
• Species: H. sapiens (human)
• Insert Size (bp): 600
• Mutation: See Depositor Comments Below
• GenBank ID: NM_000791
• Entrez Gene: TYMS (a.k.a. DKCD, HST422, TMS, TS)
• Promoter: LTR/MMLV
• Tag / Fusion Protein:
  • HiBiT (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamH1 (not destroyed)
• 3′ cloning site: Not1 (not destroyed)
• 5′ sequencing primer: pBMN 5’
• 3′ sequencing primer: IRES reverse

Terms and Licenses

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

Depositor Comments

The human TS-HiBiT fusion protein was constructed to have the HiBiT tag (VSGWRLFKKIS) sequence (Schwinn, MK et al. (2018) ACS Chem. Biol. 13(2): 467–474) located in the C-terminus of TS. The human TS-HiBiT construct was synthesized (Bio Basic) and cloned into the retroviral vector pBMN IRES-Lyt-2 (G. Nolan, Stanford University, Stanford, CA) which expresses the coding region of mouse CD8a (Lyt-2) using the restriction sites BamH1 / Not1. All constructs were sequenced to validate authenticity.
BamH1- GCCACC-ATG-TS-HiBiT-Stop –Not1

Please note: Plasmid contains a 75bp deletion in one of the LTRs. This deletion is not known to affect plasmid function.