pFastBac TwinStrepII Prescission SNAPf TEV cloning vector with BioBrick polycistronic restriction sites (11-SNAP-V1)
(Plasmid #55216)

• Purpose: (Empty Backbone)
• Depositing Lab: Scott Gradia
• Publication: Gradia et al Methods Enzymol. 2017;592:1-26. doi: 10.1016/bs.mie.2017.03.008. Epub 2017 May 15.

Backbone

• Vector backbone: pFastBac
• Vector type: Insect Expression
• Tags / Fusion Proteins:
  • TwinStrepII (N terminal on backbone)
  • SNAPf (N terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: LicBac dual V1 F (5'-cctataactattccggattattcataccgtc-3')
• 3′ sequencing primer: LicBac dual V1 Rv (5'-caggttcagggggaggtgtg-3')

Terms and Licenses

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

Depositor Comments

This plasmid is a LIC-adapted pFastBac vector. It uses the same PCR primer tags as with most of our other vectors, so one PCR product can be inserted into many different vectors at once.

This vector has a TEV cleavable TwinStrepII-Prescission-Snapf tag on the N-terminus.

Add the following tags to your PCR primers:
LicV1 Forward Tag TACTTCCAATCCAATGCA(ATG)
LicV1 Reverse Tag TTATCCACTTCCAATGTTATTA

Linearize this plasmid with SspI and gel purify the product, then T4-treat with dGTP. For the PCR product, T4-treat with dCTP.Series 11 vectors have BioBrick restriction sites to facilitate subcloning reactions to make polycistronic expression vectors. NotI, PacI, AsiSI, and SbfI are the restriction enzyme sites that flank your open reading frame. For more information, please see our website: http://qb3.berkeley.edu/qb3/macrolab/