Difference between revisions of "Part:BBa K142040:Design"
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<partinfo>BBa_K142040 short</partinfo> | <partinfo>BBa_K142040 short</partinfo> | ||
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The nucleotide sequence of RMF from Escherichia coli MG1655 was obtained from GenBank and used for primer design. Amplification from DH5-alpha genomic DNA template and addition of BioBrick standard prefix and suffix RMF was performed by PCR using the following primers: | The nucleotide sequence of RMF from Escherichia coli MG1655 was obtained from GenBank and used for primer design. Amplification from DH5-alpha genomic DNA template and addition of BioBrick standard prefix and suffix RMF was performed by PCR using the following primers: | ||
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forward primer | forward primer | ||
5'-cgcggaattcgcggccgcttctagatgaagagacaaaaacgagatcgcctgg | 5'-cgcggaattcgcggccgcttctagatgaagagacaaaaacgagatcgcctgg | ||
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reverse primer | reverse primer | ||
5'-cgcgctgcagcggccgctactagtattattaggccattactaccctgtccgc | 5'-cgcgctgcagcggccgctactagtattattaggccattactaccctgtccgc | ||
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The PCR product was subcloned into vector pSB1A7. | The PCR product was subcloned into vector pSB1A7. | ||
===References=== | ===References=== |
Revision as of 21:13, 16 October 2008
ribosome modulation factor (RMF)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 55
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
RMF was amplified from E. coli, therefore sequence optimization was unnecessary. The RMF gene does not contain any restriction sites for EcoRI, XbaI, SpeI or PstI and could be turned into a BioBrick without further modification.
Source
The nucleotide sequence of RMF from Escherichia coli MG1655 was obtained from GenBank and used for primer design. Amplification from DH5-alpha genomic DNA template and addition of BioBrick standard prefix and suffix RMF was performed by PCR using the following primers:
forward primer 5'-cgcggaattcgcggccgcttctagatgaagagacaaaaacgagatcgcctgg
reverse primer 5'-cgcgctgcagcggccgctactagtattattaggccattactaccctgtccgc
The PCR product was subcloned into vector pSB1A7.