Difference between revisions of "Part:BBa K1949020:Design"
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fwd:5’-aaatctagatgcatcgaattctcgctgaaaaatcg-3’ | fwd:5’-aaatctagatgcatcgaattctcgctgaaaaatcg-3’ | ||
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rev:5’-cccctgcagcggccgctactagtattattattccttaaagtcattgaaatcatcatccgtc-3’ | rev:5’-cccctgcagcggccgctactagtattattattccttaaagtcattgaaatcatcatccgtc-3’ | ||
yafN gene fragment amplified by PCR contains XbaI site at N-terminal and SpeI and PstI site at C-terminal. Both terminals were cut by respective restriction enzymes and the fragment used for ligation with iGEM plasmid vectors. | yafN gene fragment amplified by PCR contains XbaI site at N-terminal and SpeI and PstI site at C-terminal. Both terminals were cut by respective restriction enzymes and the fragment used for ligation with iGEM plasmid vectors. | ||
| − | Material and method | + | |
| + | ===Material and method=== | ||
Revision as of 07:59, 11 October 2016
yafN
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 78
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
yafN(BBa_K1949020) was gained by PCR using E. coli MG1655 genomic DNA and the following primers.
fwd:5’-aaatctagatgcatcgaattctcgctgaaaaatcg-3’
rev:5’-cccctgcagcggccgctactagtattattattccttaaagtcattgaaatcatcatccgtc-3’
yafN gene fragment amplified by PCR contains XbaI site at N-terminal and SpeI and PstI site at C-terminal. Both terminals were cut by respective restriction enzymes and the fragment used for ligation with iGEM plasmid vectors.
Material and method
Source
From E.coli MG1655.
Material and method
References
[1] Zhang Y, Yamaguchi Y, Inoue M. Characterization of YafO, an Escherichia coli Toxin. J Biol Chem. 2009 Sep; 284(38): 25522–25531.
[2] Dalsgaard M, Jørgensen M, Gerdes K. Three new RelE-homologous mRNA interferases of Escherichia coli differentially induced by environmental stresses. Mol Microbiol. 2010 Jan; 75(2): 333–348.
[3] Singletary LA, Gibson JL, Tanner EJ, McKenzie GJ, Lee PL, Gonzalez C, et all. An SOS-Regulated Type 2 Toxin-Antitoxin System. J Bacteriol. 2009 Dec;191(24):7456-65.