pPtGE35
(Plasmid #107999)

• Purpose: Expresses TevCas9 in Phaeodactylum tricornutum / Encodes elements required for conjugation
• Depositing Lab: David Edgell
• Publication: Slattery et al ACS Synth Biol. 2018 Jan 8. doi: 10.1021/acssynbio.7b00191.

Backbone

• Vector backbone: pKS diaCas9_sgRNA
• Backbone manufacturer: Per Winge
• Backbone size w/o insert (bp): 8577
• Total vector size (bp): 12925
• Modifications to backbone: Addition of yeast Cen6-ArsH4-His3, origin of transfer, P. tricornutum ShBle cassette, and I-TevI nuclease doamin
• Vector type: CRISPR, Synthetic Biology ; Episomal vector for Phaeodactylum tricornutum
• Selectable markers: Zeocin, HIS3

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: OriT
• Alt name: Origin of Transfer
• Insert Size (bp): 771

Gene/Insert 2

• Gene/Insert name: 40SRPS8 Promoter
• Species: Phaeodactylum tricornutum
• Insert Size (bp): 600

Gene/Insert 3

• Gene/Insert name: ShBle
• Alt name: Zeocin resistance gene
• Species: Streptoalloteichus hindustanus
• Insert Size (bp): 375

Gene/Insert 4

• Gene/Insert name: 40SRPS8 Terminator
• Species: Phaeodactylum tricornutum
• Insert Size (bp): 500

Gene/Insert 5

• Gene/Insert name: Cen6-ArsH4-His3
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 1349

Gene/Insert 6

• Gene/Insert name: I-TevI nuclease and partial linker domain
• Alt name: diaTev169(G2S)2
• Species: Enterobacteria phage T4T
• Insert Size (bp): 528
• Tag / Fusion Protein:
  • Cas9 (C terminal on insert)

Resource Information

• Supplemental Documents:
  • pPtGE35.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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