pSC-hPAX8_tdTomato-PGK-PuroTK
(Plasmid #206004)

• Purpose: plasmid to insert tdTomato reporter at the 3' end of the human PAX8 gene
• Depositing Lab: Darrell Kotton
• Depositing Organization: Boston University Medical Campus
  Located in the United States of America
• Publication: Undeutsch et al Stem Cell Reports. 2024 Dec 10;19(12):1690-1705. doi: 10.1016/j.stemcr.2024.10.004. Epub 2024 Nov 7.

Backbone

• Vector backbone: pSC-pTTR-tdTomato-T2A-TTR-puroTK-sameori 2
• Backbone manufacturer: Darrell Kotton
• Backbone size w/o insert (bp): 4415
• Total vector size (bp): 10052
• Vector type: Mammalian Expression
• Selectable markers: Puromycin ; thymidine kinase

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin and Kanamycin, 100 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: PAX8 5' homology arm, P2A-tdTomato
• Species: H. sapiens (human)
• Insert Size (bp): 614
• Entrez Gene: PAX8 (a.k.a. PAX-8)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: SalI, NdeI (not destroyed)
• 3′ cloning site: NdeI, PmeI (not destroyed)
• 5′ sequencing primer: GATATGTCGACAAAGACGAGGGCAATGAGACATG, TCTCCATATGGTGAGCAAGGGCGAG
• 3′ sequencing primer: ATAACATATGTTATAGCTAGCCAGATGGTCAAAGGCCGTGGCAG, ATTTGTTTAAACGGGCCCTCTAGACTCGACGCGGCCGCTTTACAGCTCGTCCATGCCGTACAGGAA

Gene/Insert 2

• Gene/Insert name: lox-P - PGK - puroTK - loxP
• Species: Synthetic
• Promoter: PGK

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: n/a

Gene/Insert 3

• Gene/Insert name: PAX8 3' homology arm
• Species: H. sapiens (human)
• Entrez Gene: PAX8 (a.k.a. PAX-8)

Cloning Information for Gene/Insert 3

• Cloning method: Restriction Enzyme
• 5′ cloning site: AvrII (not destroyed)
• 3′ cloning site: PsiI (not destroyed)
• 5′ sequencing primer: CTTATCCTAGGTTGCCATGGGGACAGTGGGAGCG
• 3′ sequencing primer: CTTGTTATAAGGTGGAATTTCTGAGGGAC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Hendrik Julian Undeutsch

Terms and Licenses

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

Depositor Comments

Please note that the depositor's annotated reference file has a minor difference in the tdTomato sequence compared to the Addgene verified sequence. We BLASTed our sequence result for tdTomato and confirmed it matches NCBI ID AAV52169.1, and do not predict it will alter tdTomato function. Please refer to the the Addgene sequence for the most accurate result.