pLEX_970_puro_DEST_V5_TLOC1 (220aa)
(Plasmid #45287)

• Depositing Lab: William Hahn
• Publication: Hagerstrand et al Cancer Discov. 2013 Jun 13. [Epub ahead of print]

Backbone

• Vector backbone: pLEX_970
• Vector type: Mammalian Expression, Lentiviral
• 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: TLOC1
• Species: H. sapiens (human)
• Insert Size (bp): 660
• GenBank ID: KC005990
• Entrez Gene: SEC62 (a.k.a. Dtrp1, HTP1, TLOC1, TP-1)
• Tag / Fusion Protein:
  • V5 (C terminal on backbone)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: pLEX-fwd
• 3′ sequencing primer: pLEX-rev

Resource Information

• A portion of this plasmid was derived from a plasmid made by: CCSB Human Orfeome Collection

Terms and Licenses

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

Depositor Comments

The initial pDONR223 clone was obtained from the CCSB Human Orfeome Collection. Upon sequencing it was found that the construct contained a 660 bp long variant as compared to the expected 1200 bp long version.

To determine whether this shorter version is expressed in cancer cells, and to exclude the possibility of this being a cloning artifact, we cloned TLOC1 from T47D cells and found both forms to be expressed. We re-cloned both the full length and shorter splice variants by ligation of BP flanked PCR products into pDONR223 by a Gateway BP reaction.

TLOC1 was transferred into DEST vectors from TLOC1 containing pDONR223 vectors through a Gateway LR reaction.