pCyCEN_Lisp2mCherry_hsp70_GFP
(Plasmid #137169)

• Purpose: Dual fluorescent reporter for liver stage expression in Plasmodium cynomolgi
• Depositing Lab: Clemens Kocken
• Publication: Voorberg-van der Wel et al Commun Biol. 2020 Jan 3;3:7. doi: 10.1038/s42003-019-0737-3. eCollection 2020.

Backbone

• Vector backbone: PUC19
• Modifications to backbone: Please note that there is another backbone present in the construct as well: pCR-BluntII
• Vector type: Unspecified
• Selectable markers: Human dhfr as resistance marker for Plasmodium

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: mCherry
• GenBank ID:
• Promoter: P. cynomolgi lisp2
• Tag / Fusion Protein:
  • T2A (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: N/A
• 3′ sequencing primer: N/A

Gene/Insert 2

• Gene/Insert name: Nanoluc
• Promoter: P. cynomolgi lisp2

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: N/A
• 3′ sequencing primer: N/A

Gene/Insert 3

• Gene/Insert name: dihydrofolate reductase
• Alt name: dhfr
• Species: H. sapiens (human)
• Promoter: P. cynomolgi hsp70
• Tag / Fusion Protein:
  • T2A (C terminal on insert)

Cloning Information for Gene/Insert 3

• Cloning method: Unknown
• 5′ sequencing primer: N/A
• 3′ sequencing primer: N/A

Gene/Insert 4

• Gene/Insert name: Green Fluorescent Protein
• Alt name: GFP
• Promoter: P. cynomolgi lisp2

Cloning Information for Gene/Insert 4

• Cloning method: Unknown
• 5′ sequencing primer: N/A
• 3′ sequencing primer: N/A

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • NanoLuc Luciferase Limited Use Label License
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The Addgene NGS result is not a full plasmid sequence and does not completely confirm the A/T rich Plasmodium cynomolgi strain M centromere sequence. A NotI restriction digest should be used to verify the plasmid, as an intact plasmid will yield 2 bands that add up to the full plasmid size of approximately 16 kb.