pAGH42
(Plasmid #107249)

• Purpose: Vipp1-mGFPmut3 (or vipp1-gfp) construct with spectinomycin resistance cassette downstream used to replace the native copy of Vipp1 in Synechocystis (sll0617).
• Depositing Lab: Erin O'Shea
• Publication: Gutu et al Mol Microbiol. 2018 Jan 22. doi: 10.1111/mmi.13912.

Backbone

• Vector backbone: pGEM-3ZfP
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 3170
• Total vector size (bp): 8170
• Modifications to backbone: cloned the insert by Gibson into BamHI/SphI linearized vector backbone
• Vector type: Bacterial Expression ; suicide vector for destined for double homologous recombination of the insert

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: nothing special
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: vipp1-gfp
• Alt name: sll0617 translationally fused to mGFPmut3 at C-terminus
• Species: Synechocystis sp. PCC 6803
• Promoter: native promoter of sll0617
• Tag / Fusion Protein:
  • mGFPmut3 (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7 forward
• 3′ sequencing primer: M13 reverse

Resource Information

• Supplemental Documents:
  • pAGH42a-Vipp1GFPspec.gb

Terms and Licenses

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

Depositor Comments

the insert is a composite of several fragments, all stitched together by Gibson cloning.