pFastBac1 hBRM
(Plasmid #1956)

• Depositing Lab: Robert Kingston
• Publication: Phelan et al Mol Cell 1999 Feb;3(2):247-53.

Backbone

• Vector backbone: pFastBac1
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4775
• Vector type: Insect Expression
• Selectable markers: Gentamicin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: BRM
• Alt name: BAF190b
• Species: H. sapiens (human)
• Entrez Gene: SMARCA2 (a.k.a. BAF190, BIS, BRM, NCBRS, SNF2, SNF2L2, SNF2LA, SWI2, Sth1p, hBRM, hSNF2a)
• Tag / Fusion Protein:
  • flag (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: Polyhedrin forward

Terms and Licenses

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

Depositor Comments

A C-terminal Flag tag was added to hBRM by PCR using a 3' primer designed to incorporate the FLAG sequence (DYKDDDDK) before the stop codon, followed by EcoRI and KpnI restriction sites. The PCR product was digested with HpaI and KpnI, and inserted into pCG/hBrm. pCG/FL-hBrm was digested with EcoRI, and the resulting fragment was cloned into the EcoRI site of pFastBac1. (kingston #1234)