GFP-eSIBR
(Plasmid #71664)

• Purpose: EGFP coding region followed by enhanced synthetic inhibitory BIC/miR-155 RNA (eSIBR) backbone
• Depositing Lab: Philip Washbourne
• Publication: Fowler et al Nucleic Acids Res. 2015 Nov 17. pii: gkv1246.

Backbone

• Vector backbone: pcDNA3
• Backbone size w/o insert (bp): 5352
• Total vector size (bp): 6245
• Modifications to backbone: The single native BbsI site in pcDNA3 was mutated to allow cloning of amiRNAs into the eSIBR cassette

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: EGFP coding sequence followed by eSIBR cassette
• Insert Size (bp): 893
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: M13 reverse

Resource Information

• Supplemental Documents:
  • Annotated GenBank file
• A portion of this plasmid was derived from a plasmid made by: David Turner, University of Michigan

Terms and Licenses

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

Depositor Comments

The original SIBR cassette and instructions for cloning into and chaining SIBR amiRNAs are described in:
Chung, Kwan-Ho, et al. "Polycistronic RNA polymerase II expression vectors for RNA interference based on BIC/miR-155." Nucleic Acids Research 34.7 (2006): e53-e53.