Difference between revisions of "Part:pSB1C3:Experience"
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The pSB1C3 backbone was utilized to create a new plasmid backbone specifically engineered for use with the Golden Gate assembly method. Flanking regions were added within the prefix and suffix that facilitate assembly through the use of Bsa1 overhangs. This makes for a scarless ligation into the vector. The resulting sequence is a standard pSB1C3 with a part of choice, as the overhangs are removed during the Bsa1 cleavage. Three varieties of the backbone were made. | The pSB1C3 backbone was utilized to create a new plasmid backbone specifically engineered for use with the Golden Gate assembly method. Flanking regions were added within the prefix and suffix that facilitate assembly through the use of Bsa1 overhangs. This makes for a scarless ligation into the vector. The resulting sequence is a standard pSB1C3 with a part of choice, as the overhangs are removed during the Bsa1 cleavage. Three varieties of the backbone were made. | ||
*pSB1C3 ProAV for promoters <a = href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K799024">BBa_K799024</a>. | *pSB1C3 ProAV for promoters <a = href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K799024">BBa_K799024</a>. | ||
| − | + | [[Part:BBa_K799024]] | |
*BBa_K799025: pSB1C3 orfAV for coding regions | *BBa_K799025: pSB1C3 orfAV for coding regions | ||
*BBa_K799026: pSB1C3 ttAV for terminators | *BBa_K799026: pSB1C3 ttAV for terminators | ||
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Revision as of 21:00, 2 October 2012
This experience page is provided so that any user may enter their experience using this part.
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how you used this part and how it worked out.
Applications of pSB1C3
User Reviews
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UNIPV-Pavia iGEM 2011 |
This cloning vector has been improved, cloning in it the strong pTet promoter, between E and X restriction sites; the mRFP coding sequence from BBa_J61002, has been placed between S and P restirction sites, in order to facilitate the cloning of coding sequences downstream pTet promoter in this high copy number plasmid. |
The 2011 Nevada iGEM Team attempted to use pSB1C3 as a template to amplify the chloramphenicol resistance cassette for use in a new part. When they attempted to design primers for this purpose, we were unable to identify a start codon. We subsequently contacted Dr. Knight and Austin Che about the problem. They re-examined the sequence and found that the sequence in the registry was missing two nucleotides which resulted in an apparent frameshift. Upon fixing the sequence annotation, we were able to design our amplification primers for the chloramphenicol cassette.
iGEM JHU Wetware 2012 The pSB1C3 backbone was utilized to create a new plasmid backbone specifically engineered for use with the Golden Gate assembly method. Flanking regions were added within the prefix and suffix that facilitate assembly through the use of Bsa1 overhangs. This makes for a scarless ligation into the vector. The resulting sequence is a standard pSB1C3 with a part of choice, as the overhangs are removed during the Bsa1 cleavage. Three varieties of the backbone were made.
- pSB1C3 ProAV for promoters <a = href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K799024">BBa_K799024</a>.
- BBa_K799025: pSB1C3 orfAV for coding regions
- BBa_K799026: pSB1C3 ttAV for terminators
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