pEMI91
(Plasmid #74888)

• Purpose: Shuffled 9I27 polyprotein with shuffled codons to allow facile sequencing and unique RE for the 9 cassettes for versatile cloning. See Depositor Comments for more information
• Depositing Lab: Piotr Marszalek
• Publication: Scholl et al Biomacromolecules. 2016 Jun 8.

Backbone

• Vector backbone: pET-15b
• Backbone size w/o insert (bp): 5441
• Total vector size (bp): 8207
• Vector type: Bacterial 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: Titin I27 x 9
• Alt name: 9I27
• Alt name: UniProtKB Q8WZ42
• Species: H. sapiens (human)
• Insert Size (bp): 2766
• Entrez Gene: TTN (a.k.a. CMD1G, CMH9, CMPD4, CMYO5, CMYP5, EOMFC, HMERF, LGMD2J, LGMDR10, MYLK5, SALMY, TMD)
• Promoter: T7 promoter
• Tags / Fusion Proteins:
  • His-tag (N terminal on insert)
  • Strep-tag II (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: BlpI (not destroyed)
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: T7 terminator

Resource Information

• A portion of this plasmid was derived from a plasmid made by: GenScript
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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

Depositor Comments

Each Titin I27 domain is separated by a 4 amino acid linker - two aa being "RS" and the other two corresponding to the specific restriction site