H2B-RFP in pENTR1A (w507-1)
(Plasmid #22525)

• Depositing Lab: Eric Campeau
• Publication: shMDC1 plasmids (unpublished)

Backbone

• Vector backbone: pENTR1A
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 2289
• Vector type: Entry vector

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: TOP10F', 37oC
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Human Histone H2B -Heteractis crispa red fluorescent protein
• Alt name: H2B-HcRed
• Species: H. sapiens (human)
• Insert Size (bp): 1082
• Entrez Gene: H2BC21 (a.k.a. GL105, H2B, H2B-GL105, H2B.1, H2BE, H2BFQ, H2BGL105, H2BQ, HIST2H2BE)
• Tag / Fusion Protein:
  • HcRed (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Sal I (not destroyed)
• 3′ cloning site: Not I (not destroyed)
• 5′ sequencing primer: ENTRforw
• 3′ sequencing primer: ENTRrev

Resource Information

• Addgene Notes:
  • Addgene diagnostic digest
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

H2B-HcRed is of unknown origin.

Please note that H2B-DsRed runs at a higher molecular weight than it is supposed to (~ 2kb instead of ~1 kb) on an agarose gel. Both the original vector and the Entry vector containing the H2B-RFP show the same migration pattern. Depositor confirmed plasmid sequence with ENTRforw and ENTRrev. In addition, depositor expressed the H2B-RFP and confirmed its location to mitotic chromosomes.