Part:BBa_I715020:Design

Carboxyl Half of GFP (aka GFP2)

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 174
  

Design Notes

Half-GFP program in the PTC-100™ Programmable Thermal Controller:

1. Step 1: 95° C 5 minutes
2. Step 2: 95° C 1 minute
3. Step 3: 47.24° C 1 minute
4. Step 4: 72° C 30 seconds
5. Repeat steps 2-4 29X

Primers:

• GFP2 Forward 5’GCATGAATTCGCGGCCGCTTCTAGATAAGAATGGAATCAAAGTTAA 3’
• GFP2 Reverse 5’GCATCTGCAGCGGCCGCTACTAGTTTATTATTTGTATAGTTCATCCATG 3’

Source

[http://www.bio.davidson.edu/Courses/Immunology/Students/spring2006/Acker/karen_project.html summary web page]