Difference between revisions of "Part:BBa K1463560"

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[[Image:reverse-rbs-rfp.jpg]]
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[[Image:reverse-rbs-rfp.jpg|thumb|500px|center|'''Fig. 1:''' Red fluorescence of colonies containing [https://parts.igem.org/Part:BBa_K1463560 BBa_K1463560] or [https://parts.igem.org/Part:BBa_I742130 BBa_I742130] reversed RBSs "downstream" of the reversed rfp gene [https://parts.igem.org/Part:BBa_K1463520 BBa_K1463520] transcribed from the PhiC31 integrase switch [https://parts.igem.org/Part:BBa_K1463000 BBa_K1463000]. The image was scanned using a GE Healthcare Typhoon FLA9500 using the 532 nm laser and LPG filter.]]
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Red fluorescence of colonies containing BBa_I742130 or K1463560 downstream of rfp gene transcribed from the PhiC31 integrase switch [https://parts.igem.org/Part:BBa_K1463000 K1463000]
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It was designed using the Salis RBS calculator software using the RBS sequence NNNNNNNNNNCTCTAGTA (10 Ns followed by a reversed scar) upstream of the RFP open reading frame.
 
It was designed using the Salis RBS calculator software using the RBS sequence NNNNNNNNNNCTCTAGTA (10 Ns followed by a reversed scar) upstream of the RFP open reading frame.
  
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[[File:Salis-rev-rbs.jpg]]
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[[Image:Salis-rev-rbs.jpg|thumb|812px|center|Fig. 2: Salis RBS calculator output for reverse RBS design]]
  
 
As can be seen, the designed RBS gives the start codon at position 39 a calculated translation initiation rate of "12204.43". This is 25 fold higher than the calculated translation initiation rate of [https://parts.igem.org/Part:BBa_I742130 BBa_I742130].
 
As can be seen, the designed RBS gives the start codon at position 39 a calculated translation initiation rate of "12204.43". This is 25 fold higher than the calculated translation initiation rate of [https://parts.igem.org/Part:BBa_I742130 BBa_I742130].
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<span class='h3bb'>Sequence and Features</span>
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===Sequence and Features===
 
<partinfo>BBa_K1463560 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1463560 SequenceAndFeatures</partinfo>
  

Latest revision as of 22:04, 16 October 2014

Reverse RBS

Reverse ribosome binding site designed using the Salis ribosome binding site calculator to have a strength of 12,204 when placed downstream of the reversed rfp part BBa_K1463520

Usage and Biology

This has been used to drive expression of the reversed RFP part BBa_K1463520 in our phiC31 integrase-based inversion switch BBa_K1463000.

It gave a higher level of fluorescence than a previous design for a reverse RBS BBa_I742130


Fig. 1: Red fluorescence of colonies containing BBa_K1463560 or BBa_I742130 reversed RBSs "downstream" of the reversed rfp gene BBa_K1463520 transcribed from the PhiC31 integrase switch BBa_K1463000. The image was scanned using a GE Healthcare Typhoon FLA9500 using the 532 nm laser and LPG filter.


It was designed using the Salis RBS calculator software using the RBS sequence NNNNNNNNNNCTCTAGTA (10 Ns followed by a reversed scar) upstream of the RFP open reading frame.

Fig. 2: Salis RBS calculator output for reverse RBS design

As can be seen, the designed RBS gives the start codon at position 39 a calculated translation initiation rate of "12204.43". This is 25 fold higher than the calculated translation initiation rate of BBa_I742130.


Sequence and Features


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]