Difference between revisions of "Part:BBa J119137"
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This part (also called '''pClone Green''') 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 promoter driving GFP 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 incorporate the BD18 bicistronic translational junction (see Part:BBa_J119024) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org. | This part (also called '''pClone Green''') 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 promoter driving GFP 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 incorporate the BD18 bicistronic translational junction (see Part:BBa_J119024) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org. | ||
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Revision as of 13:31, 19 August 2013
pClone Red - Device for Testing New Promoters via Golden Gate Assembly
This part (also called pClone Green) 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 promoter driving GFP 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 incorporate the BD18 bicistronic translational junction (see Part:BBa_J119024) engineered by Vivek Mutalik and The BIOFAB Team at biofab.org.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1499
Illegal AgeI site found at 1611 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 837
Illegal BsaI.rc site found at 760