pcDNA3 Flag TSC2
(Plasmid #14129)

• Depositing Lab: Brendan Manning
• Publication: Manning et al Mol Cell. 2002 Jul . 10(1):151-62.

Backbone

• Vector backbone: pcDNA3-Flag
• Backbone manufacturer: FLAG (MDYKDDDDK) was cloned between HindIII and BamHI of pcDNA3
• Backbone size w/o insert (bp): 5500
• 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

• Gene/Insert name: TSC2
• Alt name: tuberin
• Species: H. sapiens (human)
• Insert Size (bp): 5400
• GenBank ID: NM_001114382.3
• Entrez Gene: TSC2 (a.k.a. LAM, PPP1R160, TSC4)
• Tag / Fusion Protein:
  • Flag (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: SP6

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest
• A portion of this plasmid was derived from a plasmid made by: David J. Kwiatkowski
• Articles Citing this Plasmid:
  • 19 References

Terms and Licenses

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

Depositor Comments

The first 276 base pairs of TSC2 were PCR-amplified from pcDNA6-TSC2 and inserted between the BamHI and NotI sites in frame with the FLAG-coding sequence in pcDNA3-FLAG. The remainder of TSC2 was subcloned from pcDNA6-TSC2 and inserted between the NotI and XbaI sites of the above construct.

Diagnostic digests reveal an extra BamHI site in this plasmid. BamHI will not linearize the construct.