pBig1a zz TEV YBBR POT1 MBP TEV TPP1 MBP TEV TIN2 ZZ TEV TRF2 (4comp2)
(Plasmid #185448)

• Purpose: Coexpresses human POT1 with a zz affinity tag, YBBR site, and TEV site, human TRF2 with a zz tag and TEV site, and human TIN2 and TPP1 each with an MBP affinity tag and TEV site in insect cells
• Depositing Lab: Ahmet Yildiz
• Publication: Jack et al Dev Cell. 2022 Jan 24;57(2):277-290.e9. doi: 10.1016/j.devcel.2021.12.017.

Backbone

• Vector backbone: pBig1a
• Backbone size w/o insert (bp): 6500
• Total vector size (bp): 17533
• Vector type: Insect Expression

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: TRF2
• Species: H. sapiens (human)
• Insert Size (bp): 1626
• Mutation: See Depositor Comments Below
• GenBank ID: NP_056265.2
• Entrez Gene: TERF2 (a.k.a. TRBF2, TRF2)
• Tag / Fusion Protein:
  • ZZ (N terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CACCGTCAGCCGGGGCTGAACTTTCGTTTTC
• 3′ sequencing primer: CCAGCAGAAGATGCTGCGCTTCCTGGAGAGC

Gene/Insert 2

• Gene/Insert name: POT1
• Species: H. sapiens (human)
• Insert Size (bp): 1907
• GenBank ID: NP_056265.2
• Entrez Gene: POT1 (a.k.a. CMM10, CRMCC3, GLM9, HPOT1, PFBMFT8, TPDS3)
• Tags / Fusion Proteins:
  • ZZ (N terminal on insert)
  • YBBR (N terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CTTGAATCTTCAGCCTGTGAAAGCGAAC
• 3′ sequencing primer: CACCCCTGAATCAACTTAAGGGTGG

Gene/Insert 3

• Gene/Insert name: TPP1
• Species: H. sapiens (human)
• Insert Size (bp): 1371
• GenBank ID: AAX82621.1
• Entrez Gene: TPP1 (a.k.a. CLN2, GIG1, LPIC, SCAR7, TPP-1)
• Tag / Fusion Protein:
  • MBP (N terminal on insert)

Cloning Information for Gene/Insert 3

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CTGGTCATTCTCAGAGATGC
• 3′ sequencing primer: GAAATGCGGGAGGACCAGGA

Gene/Insert 4

• Gene/Insert name: TIN2
• Species: H. sapiens (human)
• Insert Size (bp): 1059
• GenBank ID: NM_012461.3
• Entrez Gene: TINF2 (a.k.a. DKCA3, TIN2)
• Tag / Fusion Protein:
  • MBP (N terminal on insert)

Cloning Information for Gene/Insert 4

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GCT ACG CCC CTG GTG GCG GG
• 3′ sequencing primer: CTA AAG GGA AAC AGC ATG AC

Resource Information

• Supplemental Documents:
  • 4-comp-ybbr-pbig1a-zzyb-pot1-hism-tpp1-hism-tin2-zztrf2.gb

Terms and Licenses

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

Depositor Comments

Please note: Plasmid contains a G682C mutation in MBP-Tin2. This mutation is not expected to affect the plasmid function.