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| | ====Violacein loxPsym Integration==== | | ====Violacein loxPsym Integration==== |
| | + | We currently have 23/24 promoters and 3'UTRs from our Golden Gate Assembly-compatible library blunt-end ligated into the vector pTZ19u with SmaI. The promoters and 3'UTRs were PCR'd from yeast genomic DNA with primers containing designed BsaI overhangs. Following miniprep and digestion with BsaI, the promoters and 3'UTRs can be combined with our violacein ORFs. The last fragment, RPS2utr, has had the BsaI overhangs added, and will soon be ligated into the vector and transformed into ''E. coli''. |
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| | [[File:(26)prom utr miniprep digest 2 exposure 2 8 15 11.jpg|center|thumb|610px|Some of the Golden Gate Assembly-compatible promoter/3'UTR library following miniprep and digestion with BsaI (8/15/2011)]] | | [[File:(26)prom utr miniprep digest 2 exposure 2 8 15 11.jpg|center|thumb|610px|Some of the Golden Gate Assembly-compatible promoter/3'UTR library following miniprep and digestion with BsaI (8/15/2011)]] |
Yeast Toolkit Results
5 Violacein Expression Cassettes Ready for Vector Ligation and Yeast Transformation
Violacein loxPsym Integration
We currently have 23/24 promoters and 3'UTRs from our Golden Gate Assembly-compatible library blunt-end ligated into the vector pTZ19u with SmaI. The promoters and 3'UTRs were PCR'd from yeast genomic DNA with primers containing designed BsaI overhangs. Following miniprep and digestion with BsaI, the promoters and 3'UTRs can be combined with our violacein ORFs. The last fragment, RPS2utr, has had the BsaI overhangs added, and will soon be ligated into the vector and transformed into E. coli.
Some of the Golden Gate Assembly-compatible promoter/3'UTR library following miniprep and digestion with BsaI (8/15/2011)
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prom_utr_miniprep_digest_2_exposure_2_8_15_11.jpg)
