GST-ENLYFQC-PNT(46-138)-His6
(Plasmid #42919)

• Depositing Lab: Barbara Imperiali
• Publication: Lukovic et al Angew Chem Int Ed Engl. 2009;48(37):6828-31. doi: 10.1002/anie.200902374.

Backbone

• Vector backbone: pGEX-4T-2
• Backbone manufacturer: Amersham
• Backbone size w/o insert (bp): 4970
• Total vector size (bp): 5300
• Vector type: Bacterial Expression

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: Ets-1 PNT domain (46-138)
• Alt name: Ets-1
• Alt name: C-ets-1
• Alt name: v-ets erythroblastosis virus E26 oncogene homolog 1
• Species: M. musculus (mouse)
• Insert Size (bp): 300
• Mutation: Contains the PNT domain (Serine 46 through Lysine 138)
• GenBank ID:
• Entrez Gene: Ets1 (a.k.a. D230050P06, Ets-1, Tpl1, p54, vs)
• Promoter: Ptac
• Tags / Fusion Proteins:
  • GST (N terminal on backbone)
  • TEV protease cleavage site (N terminal on insert)
  • 6xHis (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: pGEX5
• 3′ sequencing primer: pGEX3

Terms and Licenses

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

Depositor Comments

The gene fragment encoding residues 46-138 of Ets-1 (including the PNT domain) was amplified from an Ets-1 plasmid using primers to insert 5’ EcoR1 and 3’ Not1 restriction sites for incorporation into a pGEX-4T-2 vector. The primers also encoded an amino-terminal TEV protease cleavage site (ENLYFQ//C) and a carboxy-terminal hexahistidine tag. For this amplification the following PCR primers were used:

5’- GCC GGA ATT CGT GAA AAC CTG TAT TTT CAG TGC TCC CAA GCC TTG AAA
GCT -3’

5’- GCC CCC TTT TGC GGC CGC CTA GTG ATG GTG ATG GTG ATG ACC TTT CAC
ATC CTC TTT CTG -3’

The PCR-amplified fragments were digested with Not1 and EcoR1 and ligated to Not1/EcoR1- digested and CIP-treated pGEX-4T-2.

The protein coding sequence of the Ets-1 PNT domain is as follows:
SQALK ATFSGFTKEQ QRLGIPKDPR QWTETHVRDW VMWAVNEFSL KGVDFQKFCM NGAALCALGK DCFLELAPDF VGDILWEHLE ILQKEDVK

Alternate plasmid names: GST-TEV-PNT(46-138)-His6, GST-PNT, pGEX-PNT, GST-PNT-pGEX, GST-ENLYFQC-Ets-1(46-138)-His6, GST-TEV-Ets-1(46-138)-His6, GST-Ets-1(46-138), pGEX-Ets-1(46-138), GST-Ets-1(46-138)-pGEX