pJM601-TRA-2::FLAG
(Plasmid #27129)

• Depositing Lab: Ding Xue
• Publication: Mapes et al Proc Natl Acad Sci U S A. 2010 Oct 19. 107(42):18022-7.

Backbone

• Vector backbone: pPD95.79
• Backbone size w/o insert (bp): 2700
• Vector type: Worm Expression

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: TRA-2 FLAG
• Alt name: C15F1.3
• Species: C. elegans (nematode)
• Insert Size (bp): 13801
• Mutation: Replaced TRA-2 stop codon with KpnI and EcoRI and inserted 3 copies of the FLAG tag
• GenBank ID: S42187
• Entrez Gene: tra-2 (a.k.a. CELE_C15F1.3)
• Tag / Fusion Protein:
  • 3x FLAG (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: SpeI (not destroyed)
• 5′ sequencing primer: n/a

Terms and Licenses

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

Depositor Comments

Cloning of this plasmid was achieved in the following steps:

(1)the sequence 3' (3'UTR) of the tags was added by (EcoRI/SpeI)
(2) 1.2 kb XhoI/KpnI tra-2 pcr fragment into SalI/KpnI cut vector (destroying one SalI/XhoI site but adding another in the insert) immediately 5' of the tag
(3) 7 kb HindIII/SalI fragment into same that partially replaced fragment added in step 2 containing most of the TRA-2 coding sequence
(4) 5 kb Xho/AgeI that added the promoter and upstream gene in the operon as well as some Tra-2 coding sequence