pSB4A5 JQ929581
(Plasmid #38168)

• Depositing Lab: Drew Endy
• Publication: Bonnet et al Proc Natl Acad Sci U S A. 2012 Jun 5;109(23):8884-9. Epub 2012 May 21.

Backbone

• Vector backbone: pSB4A5
• Backbone size w/o insert (bp): 4485
• Vector type: Synthetic Biology ; BioBrick cloning vector

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: mKate2
• Species: Synthetic
• Insert Size (bp): 850

Gene/Insert 2

• Gene/Insert name: Superfolder GFP
• Species: Synthetic
• Insert Size (bp): 812

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: ccdB-Fwd
• 3′ sequencing primer: EGFP-C

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This construct has attB and attP sites (BP).

The DNA data register in BP and LR states consist of a constitutive promoter (BBa_J23119) flanked by BP or LR recombination sites positioned in opposite orientation, resulting in DNA inversion when recombined (Figs 1 & 2 and Figs S1E and
S1F). A Rrnp T1 terminator (BBa_J61048) was added in reverse orientation upstream of
the promoter to prevent transcriptional read-through in the opposite orientation, so that in each state, only one fluorescent protein is visibly expressed. On each side of the recombination target, we cloned superfolder GFP and mKate2 under translational
control of measured strong RBSs (BIOFAB pilot C-dog project http://biofab.org/data).