10xHis-TEV-ATG5_ATG7_ATG10_ATG12_ATG16L1-TEV-mGFP-StrepII (Human autophagy E3(mGFP)-like enzyme)
(Plasmid #169077)

• Purpose: Insect codon optimized ATG genes (excluded tags): 10xHis-TEV-ATG5, ATG7, ATG10, ATG12 and ATG16L1-GFP-TEV-StrepII
• Depositing Lab: Sascha Martens
• Publication: Fracchiolla et al J Cell Biol. 2020 Jul 6;219(7). pii: 151802. doi: 10.1083/jcb.201912098.

Backbone

• Vector backbone: pGBdest
• Backbone manufacturer: Vienna Biocenter Core Facilities GmbH, Vienna, Austria (Protech)
• Backbone size w/o insert (bp): 4958
• Total vector size (bp): 12834
• Vector type: Bacterial Expression, Insect Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin and Gentamicin, 50 & 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: ATG5
• Alt name: ASP; APG5; APG5L; hAPG5; SCAR25; APG5-LIKE
• Species: H. sapiens (human)
• Insert Size (bp): 894
• GenBank ID: 9474 NC_000006.12
• Entrez Gene: ATG5 (a.k.a. APG5, APG5-LIKE, APG5L, ASP, SCAR25, hAPG5)
• Promoter: Polyhedrin promoter
• Tag / Fusion Protein:
  • HIS10 Tag, TEV cleavage site (N terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gateway Cloning
• 5′ sequencing primer: GGCGGTGAAAACCTGTATTTC
• 3′ sequencing primer: TTAGTCCGTAGGTTGGGGAAT

Gene/Insert 2

• Gene/Insert name: ATG7
• Alt name: GSA7; APG7L; APG7-LIKE
• Species: H. sapiens (human)
• Insert Size (bp): 2112
• GenBank ID: 10533 NC_000003.12
• Entrez Gene: ATG7 (a.k.a. APG7-LIKE, APG7L, GSA7, SCAR31)
• Promoter: Polyhedrin promoter

Cloning Information for Gene/Insert 2

• Cloning method: Gateway Cloning
• 5′ sequencing primer: ATGGCAGCGGCTACTGGTGAC
• 3′ sequencing primer: AGCGACGATGAGACTATTTAA

Gene/Insert 3

• Gene/Insert name: ATG10
• Alt name: APG10; APG10L; pp12616
• Species: H. sapiens (human)
• Insert Size (bp): 663
• GenBank ID: 83734 NC_000005.10
• Entrez Gene: ATG10 (a.k.a. APG10, APG10L, pp12616)
• Promoter: Polyhedrin promoter

Cloning Information for Gene/Insert 3

• Cloning method: Gateway Cloning
• 5′ sequencing primer: ATGGAGGAGGACGAGTTCATC
• 3′ sequencing primer: TTAGGGCACGTTTCTTTCGTC

Gene/Insert 4

• Gene/Insert name: ATG12
• Alt name: APG12; FBR93; APG12L; HAPG12
• Species: H. sapiens (human)
• Insert Size (bp): 423
• GenBank ID: 9140 NC_000005.10
• Entrez Gene: ATG12 (a.k.a. APG12, APG12L, FBR93, HAPG12)
• Promoter: Polyhedrin promoter

Cloning Information for Gene/Insert 4

• Cloning method: Gateway Cloning
• 5′ sequencing primer: ATGGCCGAAGAGCCCCAGAGC
• 3′ sequencing primer: TTAGCCCCACGCTTGTGATTT

Gene/Insert 5

• Gene/Insert name: ATG16
• Alt name: IBD10; WDR30; APG16L; ATG16A; ATG16L
• Species: H. sapiens (human)
• Insert Size (bp): 1881
• GenBank ID: 55054 NC_000002.12
• Entrez Gene: ATG16L1 (a.k.a. APG16L, ATG16A, ATG16L, IBD10, WDR30)
• Promoter: Polyhedrin promoter
• Tag / Fusion Protein:
  • GFP tag, GGlinker, TEV cleavage site. StrepII (C terminal on insert)

Cloning Information for Gene/Insert 5

• Cloning method: Gateway Cloning
• 5′ sequencing primer: ATGAGCAGCGGTTTGAGGGCG
• 3′ sequencing primer: GCGGTGCTCTGGGCTCAATAC

Resource Information

• Supplemental Documents:
  • 183_S_0_1385_1ATG-7_10_12_5_16-GFP_pGBdest.gb
• A portion of this plasmid was derived from a plasmid made by: Vienna Biocenter Core Facilities GmbH, Vienna, Austria (Protech)
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Internal construct reference: SMC-1100