pSP64TS Activin pro Vg1 Flag (DM#171)
(Plasmid #15010)

• Depositing Lab: Douglas Melton
• Publication: Reilly et al Cell. 1996 Sep 6. 86(5):743-54.

Backbone

• Vector backbone: pSP64TS
• Backbone size w/o insert (bp): 3000
• Vector type: Sp6 expression for use in Xenopus

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: activin-Vg1
• Alt name: AVgFLAG
• Species: X. laevis (frog)
• Insert Size (bp): 1200
• Tag / Fusion Protein:
  • Flag / HMK

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: Sp6

Terms and Licenses

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

Depositor Comments

Chimeric activin-Vg1 mRNA, encoding the pro-region of activin βB fused to the mature region of Vg1, resulting in the processing and secretion of mature Vg1.

The activin βB-Vg1 hybrid was constructed by PCR amplification of a fragment of a Xenopus activin βB gene encoding the N-terminal pro-region, the tetrabasic cleavage site and the first four amino acids of the C-terminal mature region. This fragment was ligated, in frame, to a PCR amplified fragment of the Vg1 gene encoding the C-terminal mature region lacking the first four amino acids. The ligation product was reamplified, digested at terminal restriction sites and cloned into pSP64TS.

The FLAG epitope (DYKDDDK) and a protein kinase recognition site (RRASV) were inserted three amino acids downstream of the tetrabasic cleavage site.

See Author's Map for more information.