Part:BBa_J119138
Device for Testing New Promoters via 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 BsaI and Ligase. A new promoter can be derived from synthetic oligos, PCR, or a plasmid clone. The new promoter must be flanked by BsaI sites that produce the 4 nt overhangs required for assembly. The left site must be 5' CGAC 3' and the right site must be 5' GCGG 3'. Successful GGA assembly replaces the reverse GFP expression casette with the new promoter. A functional new promoter will drive RFP expression in the forward direction. The part incorporate the BD18 bicistronic translational junction (see Part:BBa_J119024) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 1614 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal SpeI site found at 1615
Illegal PstI site found at 1629
Illegal NotI site found at 7
Illegal NotI site found at 1622 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
- 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 1615 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 1
Illegal XbaI site found at 16
Illegal SpeI site found at 1615
Illegal PstI site found at 1629
Illegal AgeI site found at 1487
Illegal AgeI site found at 1599 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 105
Illegal BsaI site found at 837
Illegal BsaI.rc site found at 28
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