Coding

Part:BBa_K2515000:Design

Designed by: Slavil Peykov   Group: iGEM17_Bulgaria   (2017-10-26)
Revision as of 20:30, 1 November 2017 by Slavil (Talk | contribs) (→‎Design Notes)

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KillerOrange CDS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE 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.

References