p11U1ADb
(Plasmid #27384)

• Depositing Lab: David Bartel
• Publication: Shechner et al Science. 2009 Nov 27. 326(5957):1271-5.

Backbone

• Vector backbone: pET11a
• Backbone size w/o insert (bp): 5600
• Vector type: Bacterial Expression

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: U1A A1-98 Y31H/Q36R
• Alt name: U1A
• Species: H. sapiens (human)
• Mutation: A1-98 Y31H/Q36R double mutant
• Entrez Gene: SNRPA (a.k.a. Mud1, U1-A, U1A)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7 terminator

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Adrian Ferré-D’Amaré and Jennifer Doudna

Terms and Licenses

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

Depositor Comments

The U1A A1-98 Y31H/Q36R double mutant was expressed from the plasmid
p11U1ADb. To construct this plasmid, the protein-coding sequence from a vector
provided by Adrian Ferré-D’Amaré and Jennifer Doudna was inserted into the pET11a plasmid, thereby generating a vector that expressed more efficiently than its
predecessor. For protein expression, a saturated culture of E. coli BL21(DE3) cells transformed with p11U1ADb was diluted 1:1000 into LB supplemented with 100 μg/mL ampicillin and grown in baffled flasks at 37º C with vigorous shaking. Protein
overexpression was induced by the addition of 0.5 mM IPTG when cultures reached an
OD of 0.7.