Difference between revisions of "Part:BBa K1475006:Design"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1475006 short</partinfo> | <partinfo>BBa_K1475006 short</partinfo> | ||
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===Design Notes=== | ===Design Notes=== | ||
− | + | This part was constructed first by adding promoter, RBS and terminator to the iGEM registry basic TetR with LVA rapid degradation tag [https://parts.igem.org/Part:BBa_C0040 Part:BBa_C0040] | |
+ | Forward primer (containing: XbaI restriction site, [https://parts.igem.org/Part:BBa_J23106 Promoter:BBa_J23106] and [https://parts.igem.org/Part:BBa_B0030 RBS:BBa_B0030]): | ||
+ | <br> | ||
+ | CGCTTCTAGAGTCCCTATCAGTGATAGAGATTGACATCCCTATCAGTGATAGAGATACTGAGCACTACTAGATTAAAGAGGAGAAATACTAGATGCGTAAAGGAGAAGAAG | ||
+ | <br> | ||
+ | Reverse primer (containing: suffix and [https://parts.igem.org/Part:BBa_B1002 terminator:BBa_B1002]): | ||
+ | <br> | ||
+ | CTGCAGCGGCCGCTACTAGTAGCGAAAAAACCCCGCCGAAGCGGGGTTTTTTGCGTTATTAAGCTACTAAAGCGTAGTTTTCGTC | ||
+ | <br> | ||
+ | Then this PCR product and the [https://parts.igem.org/Part:BBa_E0840 GFP generator:BBa_E0840] was assembled using standard assembly. | ||
+ | |||
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===Source=== | ===Source=== | ||
− | + | TetR: ''Escherichia coli'' | |
+ | <br> | ||
+ | GFP: ''Aequeora victoria'' (GFPmut3b) [3] | ||
+ | |||
===References=== | ===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 | ||
+ | <br> | ||
+ | 2. Tetsystems, 2008: Principles and Components Description. http://www.tetsystems.com/science-technology/principles-components | ||
+ | <br> | ||
+ | 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 | ||
+ | <br> | ||
+ | 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 |
Latest revision as of 19:37, 17 October 2014
GFP controlled by TetR+LVA and pTet.
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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