pRc/CMV HA TrkB-T1
(Plasmid #39980)

• Depositing Lab: Yves-Alain Barde
• Publication: Bibel et al EMBO J. 1999 Feb 1;18(3):616-22.

Backbone

• Vector backbone: pRC/CMV AC7 vector, a derivative of the pRC/CMV vector
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5500
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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: TrkB.T1
• Alt name: NTrk2 variant 2
• Species: R. norvegicus (rat)
• Insert Size (bp): 1500
• Mutation: TrkB.T1 variant; endogenous signal sequence was replaced with BM40 and HA tag
• GenBank ID: NM_001163168.1
• Entrez Gene: Ntrk2 (a.k.a. RATTRKB1, TRKB1, Tkrb, trk-B, trkB)
• Promoter: CMV
• Tags / Fusion Proteins:
  • HA (N terminal on insert)
  • BM40 signal sequence (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (unknown if destroyed)
• 3′ cloning site: NotI (unknown if destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: SP6

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

The T1 isoform of trkB was cloned from a pCMV-5 trkB.T1 construct using HindIII and XbaI to fuse it to the HA-tagged N-terminus of trkB in the vector pB-KS followed by subcloning into pRC/CMV AC7 with NheI and NotI.

See Middlemas et al (Mol Cell Biol. 1991 Jan;11(1):143-53., https://www.ncbi.nlm.nih.gov/pubmed?term=1846020 ) to learn more about the TrkB.T1 variant.

Please note that the endogenous signal sequence of TrkB-T1 was replaced by the BM40 signal peptide sequence (MRAWIFFLLCLAGRALAAPLA). When compared to translated sequences for GenBank ID M55292.1 or NM_001163168.1, the TrkB-T1 protein sequence starts at amino acid #32