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PurposeDoxycycline-inducible all-in-one lentivirus vector to express H2BeGFP fusion protein. System for labelling slow-cycling cells in vivo.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 165494 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepRRL-cPPT-hPGKTMPrtTA-WPRE
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Backbone manufacturer-
- Backbone size w/o insert (bp) 8023
- Total vector size (bp) 9139
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Modifications to backbone-
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Vector typeLentiviral
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Selectable markersGFP selection by fluorescent-activated cell sorting (FACS).
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)NEB Stable
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Growth instructions-
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Copy numberUnknown
Gene/Insert
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Gene/Insert nameHuman histone H2B
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Alt name-
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SpeciesH. sapiens (human)
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Insert Size (bp)1116
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Mutation-
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GenBank IDX00088
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Entrez GeneH2BC21 (a.k.a. GL105, H2B, H2B-GL105, H2B.1, H2BE, H2BFQ, H2BGL105, H2BQ, HIST2H2BE)
- Promoter TetO-CMV
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Tag
/ Fusion Protein
- EGFP (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site Xba I (destroyed during cloning)
- 3′ cloning site Nhe I (not destroyed)
- 5′ sequencing primer TCGAGTAGGCGTGTACGGT
- 3′ sequencing primer TGAAGAATGTGCGAGACC (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byThe coding sequence of histone2B fused to eGFP (H2BeGFP) was amplified from the Addgene plasmid ID: 11680 deposited by Geoff Wahl.
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
ABOUT BACKBONE:
The lentiviral backbone was originally designed and constructed by Barde I et al. (Barde I et al. Mol Ther. 2006. 13(2): 382-90. PMID: 16275162).
ABOUT TEST DIGESTION:
Since Xba I restriction site is lost during cloning, we recommend checking the plasmid using the enzymes: Xho I and Nhe I. You should get two fragments around 1600 bp (insert+vector) and 7500 bp (vector).
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pSIN-TRE-H2BeGFP-rtTA2 was a gift from Hector G. Palmer (Addgene plasmid # 165494 ; http://n2t.net/addgene:165494 ; RRID:Addgene_165494) -
For your References section:
TET2 controls chemoresistant slow-cycling cancer cell survival and tumor recurrence. Puig I, Tenbaum SP, Chicote I, Arques O, Martinez-Quintanilla J, Cuesta-Borras E, Ramirez L, Gonzalo P, Soto A, Aguilar S, Eguizabal C, Caratu G, Prat A, Argiles G, Landolfi S, Casanovas O, Serra V, Villanueva A, Arroyo AG, Terracciano L, Nuciforo P, Seoane J, Recio JA, Vivancos A, Dienstmann R, Tabernero J, Palmer HG. J Clin Invest. 2018 Aug 31;128(9):3887-3905. doi: 10.1172/JCI96393. Epub 2018 Aug 6. 10.1172/JCI96393 PubMed 29944140