Difference between revisions of "Team:Austin UTexas/Improve"
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<p>This Phytobrick is an improved version of the Biobrick BBa_K864100, which is the coding sequence for super yellow fluorescent protein (sYFP2). We added overall usability and stability to this sequence by not only turning it into level 0 Phytobrick for use in Golden Gate Assembly, but we removed mutational hotspot from the sequence, making it more stable.<p> | <p>This Phytobrick is an improved version of the Biobrick BBa_K864100, which is the coding sequence for super yellow fluorescent protein (sYFP2). We added overall usability and stability to this sequence by not only turning it into level 0 Phytobrick for use in Golden Gate Assembly, but we removed mutational hotspot from the sequence, making it more stable.<p> | ||
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<p>We took BBa_E1010 and added restriction sites then inserted it into PhytoBrick universal acceptor, BBa_P10500, in order to create a phytobrick for use in Golden Gate Assemblies with Bsa1.</p> | <p>We took BBa_E1010 and added restriction sites then inserted it into PhytoBrick universal acceptor, BBa_P10500, in order to create a phytobrick for use in Golden Gate Assemblies with Bsa1.</p> | ||
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<img src="https://static.igem.org/mediawiki/2018/3/38/T--Austin_UTexas--RCPlevel0.PNG"> | <img src="https://static.igem.org/mediawiki/2018/3/38/T--Austin_UTexas--RCPlevel0.PNG"> | ||
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<p>For this improvement, we took BBa_E1010 (RCP) and added the synthetic, high copy number promoter, CP25, and an RBS sequence to our Phytobrick. This will increase the expression of the RCP. We also made the sequence golden gate compatible by inserting the sequence into the PhytoBrick Universal Acceptor BBa_P10500, for future use in Golden Gate Assemblies. By linking a Promoter/RBS with a coding sequence, this will increase the efficiency of the BsaI reaction in order to make transcriptional units in Golden Gate Assemblies.</p> | <p>For this improvement, we took BBa_E1010 (RCP) and added the synthetic, high copy number promoter, CP25, and an RBS sequence to our Phytobrick. This will increase the expression of the RCP. We also made the sequence golden gate compatible by inserting the sequence into the PhytoBrick Universal Acceptor BBa_P10500, for future use in Golden Gate Assemblies. By linking a Promoter/RBS with a coding sequence, this will increase the efficiency of the BsaI reaction in order to make transcriptional units in Golden Gate Assemblies.</p> | ||
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Revision as of 15:10, 12 October 2018
Improve
BBa_xxxxxxx
This Phytobrick is an improved version of the Biobrick BBa_K864100, which is the coding sequence for super yellow fluorescent protein (sYFP2). We added overall usability and stability to this sequence by not only turning it into level 0 Phytobrick for use in Golden Gate Assembly, but we removed mutational hotspot from the sequence, making it more stable.
Originally, BBa_K864100 had the IS10 sequence: 5' - gctnagc - 3', which resulted in an insertion in the region of: 5’ -acaggcgtagtaccg- 3’. In BBa_xxxxxxx (the improved part), the IS10 sequence has been removed and the area of the insertion sequence is now 5’ -actggggttgttcca- 3’, which reduces the palindromic nature of the sequence. Palindromes in the DNA cause potential hairpins, which prevents RNA polymerase from translating the DNA correctly. We also turned this part into a level 0 Phytobrick by assembling it with the Universal Acceptor, BBa_P10500. This in turn will allow researchers to use this very fluorescent protein gene in Golden Gate Assembly reactions using the enzyme BsaI.
BBa_rcppartp
We took BBa_E1010 and added restriction sites then inserted it into PhytoBrick universal acceptor, BBa_P10500, in order to create a phytobrick for use in Golden Gate Assemblies with Bsa1.
BBa_linked
For this improvement, we took BBa_E1010 (RCP) and added the synthetic, high copy number promoter, CP25, and an RBS sequence to our Phytobrick. This will increase the expression of the RCP. We also made the sequence golden gate compatible by inserting the sequence into the PhytoBrick Universal Acceptor BBa_P10500, for future use in Golden Gate Assemblies. By linking a Promoter/RBS with a coding sequence, this will increase the efficiency of the BsaI reaction in order to make transcriptional units in Golden Gate Assemblies.
