Part:BBa_K1475006:Design

GFP controlled by TetR+LVA and pTet.

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 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 1485
  

Design Notes

This part was constructed first by adding promoter, RBS and terminator to the iGEM registry basic TetR with LVA rapid degradation tag Part:BBa_C0040 Forward primer (containing: XbaI restriction site, Promoter:BBa_J23106 and RBS:BBa_B0030):
CGCTTCTAGAGTCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGATACTGAGCACTACTAGATTAAAGAGGAGAAATACTAGATGCGTAAAGGAGAAGAAG
Reverse primer (containing: suffix and terminator:BBa_B1002):
CTGCAGCGGCCGCTACTAGTAGCGAAAAAACCCCGCCGAAGCGGGGTTTTTTGCGTTATTAAGCTACTAAAGCGTAGTTTTCGTC
Then this PCR product and the GFP generator:BBa_E0840 was assembled using standard assembly.

Source

TetR: Escherichia coli
GFP: Aequeora victoria (GFPmut3b) [3]

References

1. C. Krafft, et al.: Interaction of Tet Repressor with Operator DNA and with Tetracycline Studied by Infrared and Raman Spectroscopy. Biophysical Journal, Volume 74, Issue 1, January 1998, Pages 63–71. http://www.sciencedirect.com/science/article/pii/S0006349598777677
2. Tetsystems, 2008: Principles and Components Description. http://www.tetsystems.com/science-technology/principles-components
3. Cormack, B.P., Valdivia, R.H., and S. Falkow. FACS-optimized mutants of green fluorescent protein (GFP). Gene 173: 33-38 (1996). http://www.sciencedirect.com/science/article/pii/0378111995006850
4. Aagaard, L., et al.: A Facile Lentiviral Vector System for Ekspression of Doxycycline-Inducible dhRNAs: Knockdown of the Pre-miRNA Processing Enzyme Drosha. Molecular Therapy, 2007. 15:5, p. 938-945. http://www.nature.com/mt/journal/v15/n5/full/6300118a.html