pIVT-desNb_EGFP-BleoR-mRuby2
(Plasmid #252240)

• Purpose: Template for IVTR of destabilized antibody against EGFP or EBFP with PuroR-mRuby2 as effector
• Depositing Lab: Kate Galloway
• Publication: Engineering STRAIGHT-IN single and dual lines in the male iPS11 parental line for programmable DNA integration (unpublished)

Backbone

• Vector backbone: pBR322/pUC
• Vector type: Mammalian Expression, Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: T7-desNb-EGFP-BleoR-mRuby2
• Species: Synthetic
• Insert Size (bp): 2000
• Mutation: N/A
• Promoter: T7

Cloning Information

• Cloning method: Gibson Cloning

Terms and Licenses

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

Depositor Comments

Due to the difficulties of maintaining and sequencing long homopolymers, Addgene cannot guarantee the exact length of polyA sequence. The depositor noted that they have also observed small deletions, but it still sufficiently confers mRNA stability. The enzymes or proteins expressed in these IVT reactions are only required for a very brief period of time, so long-term expression is neither intended nor necessary. If needed, researchers can PCR-amplify the transgene and add a longer polyA sequence.