Part:BBa_J119141
Device for Testing New Promoters via BsgI Golden Gate Assembly
This part will allow users to clone and test new promoters without gel purification or other preparation of DNA - it is ideal for teaching labs. It is a destination vector for Golden Gate Assembly (GGA) using BsgI and Ligase. A new promoter can be derived from synthetic oligos, PCR, or a plasmid clones. The new promoter must be flanked by BsgI sites that produce the 2 nt overhangs required for assembly. The left site must be 5' CTGCAC 3' and the right site must be 5' GTGCAG 3'. Successful GGA assembly replaces the reverve promoter driving RFP expression with the new promoter. The new promoter will drive RFP expression in the forward direction or GFP expression if it has reverse promoter function. The part incorporation of J23100 promoter was designed by Warsaw 2010.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal suffix found in sequence at 1589
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 772
Illegal NheI site found at 795
Illegal NheI site found at 1624
Illegal NheI site found at 1647
Illegal SpeI site found at 1590
Illegal PstI site found at 1604
Illegal NotI site found at 1597 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal suffix found in sequence at 1590
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 1590
Illegal PstI site found at 1604
Illegal AgeI site found at 1462
Illegal AgeI site found at 1574 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 72
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