p3A9cbetaTCR
(Plasmid #69600)

• Purpose: (Empty Backbone) TCR beta chain transgene expression plasmid which has a unique ClaI site for the insertion of a TCR beta chain VDJ exon
• Depositing Lab: Francis R. Carbone
• Publication: Barnden et al Immunol Cell Biol. 1998 Feb;76(1):34-40.

Backbone

• Vector backbone: pBluescript KS
• Backbone manufacturer: Invitrogen
• Modifications to backbone: See comments
• Vector type: Mammalian Expression

Growth in Bacteria

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

Cloning Information

• Cloning method: Restriction Enzyme

Terms and Licenses

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

Depositor Comments

A 1.0 kb Sma-I/Not-I fragment containing the TCR b-chain enhancer element was excised from pKS913.Cb18.31 and cloned into the pKS
Bluescript plasmid (Stratagene), which had been modified to remove the Kpn-I and Cla-I restriction sites from the polylinker resulting in the pKS.bEnh plasmid. A Sal-I fragment, containing the B3 TCR b-chain enhancer, promoter and coding regions, was isolated from
pKS913.Cb18.31 and cloned into the Sal-I site of the pKS.bEnh
plasmid to give the construct pKS913.Cb18.31(Kpn-I)/Cla-I)). A
4.2 kb Eco-I/Cla-I fragment comprising the 5’ region and leader
sequence from the hybridoma 2B4 was isolated from the p3A9 b
shuttle vector. This fragment was cloned into the pGEM-T
plasmid (Promega) resulting in the pG3A9 plasmid. A
4.2 kb Apa-I/Cla-I fragment, containing the 2B4 5’ region and
leader sequence, was isolated from the pG3A9 plasmid. This
fragment was cloned into pKS319.Cb18.31(Kpn-I)/Cla-I)) after
removal of the B3 TCR b-chain promoter and the TCRBVDJ
coding regions by a combined Apa-I/Cla-I restriction enzyme digest, resulting in the p3A9cbTCR plasmid.