pLPC-C-SF-TAP-DEST-TLK1-WT
(Plasmid #218829)

• Purpose: Expresses C-terminal Strep-FLAG-tagged wild type human TLK1 variant 5 in a retrovirus vector.
• Depositing Lab: Travis Stracker
• Publication: Identification of de novo mutations in TLK1 and MDM1 in a patient with a neurodevelopmental disorder and immunodeficiency (unpublished)

Backbone

• Vector backbone: pLPC-C-SF-TAP DEST: modified pLPC-N-Myc (Plasmid #12540) backbone with pDEST/C-SF-TAP cassette.
• Backbone size w/o insert (bp): 5855
• Total vector size (bp): 8162
• Modifications to backbone: The Gateway DEST cloning cassette and Strep-FLAG affinity tags were cloned from the pDEST/C-SF-TAP vector (https://doi.org/10.1002/pmic.200700038) and ligated into pLPC-N-Myc (Addgene #12540) using EcoRI and Xho1. TLK1 was cloned into the modified vector using a Gateway LR cloning reaction from a pENTR-TLK1 vector obtained from the Human ORFeome Version 1.1 library.
• Vector type: Mammalian Expression
• Selectable markers: Puromycin

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: TLK1
• Species: H. sapiens (human)
• Insert Size (bp): 2157
• GenBank ID: NM_001136554.2
• Entrez Gene: TLK1 (a.k.a. PKU-beta)
• Promoter: CMV
• Tag / Fusion Protein:
  • FLAG-STREP (N terminal on backbone)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: T7 Forward
• 3′ sequencing primer: SP6 Reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: This plasmid was constructed in part with a Human ORFeome Version 1.1 library plasmid, pLPC-N-Myc (Plasmid #12540) and pDEST/C-SF-TAP (https://doi.org/10.1002/pmic.200700038), a kind gift from Christian Gloeckner.

Terms and Licenses

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