Difference between revisions of "Part:BBa K2515000:Design"
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The 5' end nucleotide sequence was codon optimized (no effect on AA sequence) to avoid formation of stable secondary structures. After this modification one can easily design primers for this part without scars. | The 5' end nucleotide sequence was codon optimized (no effect on AA sequence) to avoid formation of stable secondary structures. After this modification one can easily design primers for this part without scars. | ||
| + | If you want to have this synthesized by IDT, order the following sequence: | ||
| + | <pre> | ||
| + | KillerOrange CDS | ||
| + | GAATTCGCGGCCGCTTCTAGATGGAATGTGGTCCTGCTTTATTTCAATCAGATATGACATT | ||
| + | CAAAATTTTTATCGACGGGGAAGTAAACGGTCAGAAATTTACGATCGTAGCAGATGGCAGC | ||
| + | AGTAAGTTCCCGCATGGAGACTTCAATGTGCACGCAGTCTGTGAAACTGGGAAATTGCCCA | ||
| + | TGTCATGGAAGCCCATTTGCCACTTGATCCAATGGGGCGAGCCGTTTTTCGCCCGCTATCC | ||
| + | TGATGGGATTTCCCACTTCGCTCAGGAATGCTTTCCAGAAGGACTGTCAATCGATCGCACC | ||
| + | GTCCGTTTTGAAAACGATGGGACTATGACATCACATCATACTTATGAGCTGTCCGATACGT | ||
| + | GTGTTGTCTCACGTATTACAGTGAATTGTGATGGCTTTCAACCAGATGGGCCGATCATGCG | ||
| + | CGATCAACTGGTCGATATTTTGCCTTCAGAAACTCATATGTTCCCTCATGGACCCAACGCA | ||
| + | GTCCGTCAACTTGCCTTCATTGGCTTCACTACCGCCGACGGTGGCCTTATGATGGGTCATC | ||
| + | TGGACAGTAAGATGACATTTAACGGCAGCCGCGCCATTGAGATTCCCGGACCTCATTTTGT | ||
| + | CACTATCATCACCAAACAAATGCGTGATACATCTGATAAGCGTGATCATGTTTGCCAACGT | ||
| + | GAAGTAGCACACGCACACTCCGTGCCGCGCATTACTTCGGCAATTGGCTCTGACCAGGATT | ||
| + | AATAAACTAGTAGCGGCCGCTGCAG</pre> | ||
| + | |||
| + | and then clone it in a pSB vector of your choice using the following primers: | ||
| + | |||
| + | C_BioBrick_Prefix-F: TATGAATTCGCGGCCGCTTCTAG | ||
| + | |||
| + | C_BioBrick_Suffix-R: TATCTGCAGCGGCCGCTACTAGTA | ||
| + | |||
| + | <i>Note: C means cloning - these primers have 3 extra bases at their 5' ends so the EcoRI and PstI can cut efficiently. <b>Do not use the regular BioBrick_Prefix-F and R pair instead - it won't work!</b></i><br /><br /> | ||
===Source=== | ===Source=== | ||
Latest revision as of 20:30, 1 November 2017
KillerOrange CDS
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The 5' end nucleotide sequence was codon optimized (no effect on AA sequence) to avoid formation of stable secondary structures. After this modification one can easily design primers for this part without scars.
If you want to have this synthesized by IDT, order the following sequence:
KillerOrange CDS GAATTCGCGGCCGCTTCTAGATGGAATGTGGTCCTGCTTTATTTCAATCAGATATGACATT CAAAATTTTTATCGACGGGGAAGTAAACGGTCAGAAATTTACGATCGTAGCAGATGGCAGC AGTAAGTTCCCGCATGGAGACTTCAATGTGCACGCAGTCTGTGAAACTGGGAAATTGCCCA TGTCATGGAAGCCCATTTGCCACTTGATCCAATGGGGCGAGCCGTTTTTCGCCCGCTATCC TGATGGGATTTCCCACTTCGCTCAGGAATGCTTTCCAGAAGGACTGTCAATCGATCGCACC GTCCGTTTTGAAAACGATGGGACTATGACATCACATCATACTTATGAGCTGTCCGATACGT GTGTTGTCTCACGTATTACAGTGAATTGTGATGGCTTTCAACCAGATGGGCCGATCATGCG CGATCAACTGGTCGATATTTTGCCTTCAGAAACTCATATGTTCCCTCATGGACCCAACGCA GTCCGTCAACTTGCCTTCATTGGCTTCACTACCGCCGACGGTGGCCTTATGATGGGTCATC TGGACAGTAAGATGACATTTAACGGCAGCCGCGCCATTGAGATTCCCGGACCTCATTTTGT CACTATCATCACCAAACAAATGCGTGATACATCTGATAAGCGTGATCATGTTTGCCAACGT GAAGTAGCACACGCACACTCCGTGCCGCGCATTACTTCGGCAATTGGCTCTGACCAGGATT AATAAACTAGTAGCGGCCGCTGCAG
and then clone it in a pSB vector of your choice using the following primers:
C_BioBrick_Prefix-F: TATGAATTCGCGGCCGCTTCTAG
C_BioBrick_Suffix-R: TATCTGCAGCGGCCGCTACTAGTA
Note: C means cloning - these primers have 3 extra bases at their 5' ends so the EcoRI and PstI can cut efficiently. Do not use the regular BioBrick_Prefix-F and R pair instead - it won't work!
Source
gBlock synthesis. Coding sequence comes from the following article: "Genetically Encoded Photosensitizer Activated by Blue and Green Light" PLoS One. 2015 Dec 17;10(12):e0145287. doi: 10.1371/journal.pone.0145287. eCollection 2015.