LARG mCherry sspB in pcDNA3.1
(Plasmid #90459)

• Purpose: RhoA mediated cell migration
• Depositing Lab: Narasimhan Gautam
• Publication: O'Neill et al Dev Cell. 2018 Jun 15. pii: S1534-5807(18)30422-2. doi: 10.1016/j.devcel.2018.05.029.

Backbone

• Vector backbone: pcDNA3.1
• Backbone manufacturer: invitrogen
• Backbone size w/o insert (bp): 5428
• Total vector size (bp): 7687
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: LARG DH-PH (aa 766–1138)
• Alt name: Rho guanine nucleotide exchange factor 12 (ARHGEF12)
• Species: H. sapiens (human)
• Insert Size (bp): 1119
• GenBank ID: XM_017017422.1
• Entrez Gene: ARHGEF12 (a.k.a. LARG, PRO2792)
• Promoter: CMV

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (not destroyed)
• 3′ cloning site: BamH1 (not destroyed)
• 5′ sequencing primer: T7 primer
• 3′ sequencing primer: BGH reverse primer

Gene/Insert 2

• Gene/Insert name: mCherry
• Insert Size (bp): 710
• Promoter: CMV

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamH1 (destroyed during cloning)
• 3′ cloning site: BspE1 (destroyed during cloning)
• 5′ sequencing primer: T7 primer
• 3′ sequencing primer: BGH reverse primer

Gene/Insert 3

• Gene/Insert name: sspB
• Species: bacterial
• Insert Size (bp): 342
• Mutation: R73Q

Cloning Information for Gene/Insert 3

• Cloning method: Restriction Enzyme
• 5′ cloning site: BspE1 (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: T7 primer
• 3′ sequencing primer: BGH reverse primer

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

LARG sequence contains an Y830F variant that does not affect plasmid function. The sspB sequence was obtained from pLL7.0: mTiam1(64-437) tgRFPt-SSPBR73Q (Plasmid #60418).