pENTR TERRA
(Plasmid #111383)

• Purpose: TERRA with approximately 130 telomere hexanucleotide repeats of TTAGGG and TTGGGG variant sequences
• Depositing Lab: Klaus Holzmann
• Publication: Kreilmeier et al Genes (Basel). 2016 Aug 16;7(8). pii: genes7080046. doi: 10.3390/genes7080046.

Backbone

• Vector backbone: pENTR/D-hH1
• Backbone manufacturer: Invitrogen
• Modifications to backbone: From the pENTR-hH1 plasmid with inserted telomere fragment, the H1 promoter was deleted for use with Gateway expression vectors with built-in expression cassettes using CMV promoters, like pAD/CMV and pLenti4/V5 (Invitrogen). To this end, the hH1 promoter was cut out with SacI and BamHI (Fermentas) and the remaining plasmid fragment was treated with Klenow fragment (Invitrogen) to generate blunt ends, ligated and transformed in competent E. coli as described before.
• Vector type: Gateway Entry Cloning

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: TERRA
• Species: H. sapiens (human)
• Insert Size (bp): 868

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: unknown

Resource Information

• Supplemental Documents:
  • pENTR TERRA.gb

Terms and Licenses

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

Depositor Comments

Depositing Scientist Sequence may not contain all correct information about the telomere sequence repeats such as canonical and variant repeat positions, and precise information about insert length. Further, the Addgene Partial NGS Result does not include the entire region of telomere repeats. Note that the BamHI site was destroyed during cloning.