His-ZZ-TEV-SNAPf DHC1_IC2C_LIC2_Tctex1_Robl1_LC8
(Plasmid #111903)

• Purpose: Full length human cytoplasmic dynein 1 complex, optimised for Sf9 expression. 8xHis-ZZ tag followed by TEV cleavage site and SNAPf tag on the N-terminus of dynein heavy chain 1.
• Depositing Lab: Andrew Carter
• Publication: His-ZZ-LTLT-SNAPf DHC1_IC2C_LIC2_Tctex1_Robl1_LC8 (unpublished)

Backbone

• Vector backbone: pACEBac1
• Backbone manufacturer: Geneva Biotech
• Backbone size w/o insert (bp): 3063
• Total vector size (bp): 26718
• Modifications to backbone: Vector and cloning strategy based on the biGBac system (Weissmann et al. 2014). The equivalent of pBig1 and pBig2 plasmids were generated according to Weissmann et al. with the exception that the pACEBac1 plasmid (Geneva Biotech) was used as a template. pBig1 equivalent plasmids contain a Gentamycin resistance marker, and pBig2 equivalent plasmids contain both Gentamycin and Chloramphenicol resistance genes.
• Vector type: Insect Expression

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Gentamicin, 25 & 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: His-ZZ-LTLT-SNAPf-DYNC1H1
• Alt name: Dynein heavy chain 1
• Alt name: DYNC1H1
• Species: H. sapiens (human)
• Insert Size (bp): 15169
• Mutation: Optimised for expression in Sf9 cells.
• Entrez Gene: DYNC1H1 (a.k.a. CDCBM13, CMT2O, DHC1, DHC1a, DNCH1, DNCL, DNECL, DYHC, Dnchc1, HL-3, SMALED1, p22)
• Tags / Fusion Proteins:
  • 8xHis (N terminal on insert)
  • TEV site (N terminal on insert)
  • ZZ-tag (N terminal on insert)
  • SNAPf-tag (N terminal on insert)

Gene/Insert 2

• Gene/Insert name: DYNC1I2
• Alt name: Cytoplasmic dynein 1 intermediate chain 2 isoform C
• Species: H. sapiens (human)
• Insert Size (bp): 1839
• Mutation: Optimised for expression in Sf9 cells.
• Entrez Gene: DYNC1I2 (a.k.a. DIC74, DNCI2, IC2, NEDMIBA)

Gene/Insert 3

• Gene/Insert name: DYNC1LI2
• Alt name: Cytoplasmic dynein 1 light intermediate chain 2
• Species: H. sapiens (human)
• Insert Size (bp): 1479
• Mutation: Optimised for expression in Sf9 cells.
• Entrez Gene: DYNC1LI2 (a.k.a. DNCLI2, LIC2)

Gene/Insert 4

• Gene/Insert name: DYNLT1
• Alt name: Dynein light chain Tctex-type 1
• Alt name: TCTEX1
• Species: H. sapiens (human)
• Insert Size (bp): 342
• Mutation: Optimised for expression in Sf9 cells.
• Entrez Gene: DYNLT1 (a.k.a. CW-1, TCTEL1, tctex-1)

Gene/Insert 5

• Gene/Insert name: DYNLRB1
• Alt name: Dynein light chain roadblock-type 1
• Alt name: ROBLD1
• Species: H. sapiens (human)
• Insert Size (bp): 291
• Mutation: Optimised for expression in Sf9 cells.
• Entrez Gene: DYNLRB1 (a.k.a. BITH, BLP, DNCL2A, DNLC2A, ROBLD1)

Gene/Insert 6

• Gene/Insert name: DYNLL1
• Alt name: Dynein light chain 1, cytoplasmic
• Alt name: LC8
• Species: H. sapiens (human)
• Insert Size (bp): 270
• Mutation: Optimised for expression in Sf9 cells.
• Entrez Gene: DYNLL1 (a.k.a. DLC1, DLC8, DNCL1, DNCLC1, LC8, LC8a, PIN, hdlc1)

Resource Information

• Supplemental Documents:
  • p2065_Hs Dyn FL WT_His-ZZ-LTLT-SNAPf DHC1_IC2C_LIC2_Tctex1_Robl1_LC8.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Please note that there are some discrepancies between Addgene's quality control sequence and the depositor's sequence. The depositor noted that these discrepancies do NOT affect plasmid function.

This plasmid replaces pDyn3 from Schlager et al. 2014.