Difference between revisions of "Part:BBa K2717021:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | + | ||
| + | GFP, pchBA, emrR binding promoter, emrR gene and gdh gene were obtained from plasmids or genomes by PCR methods. At the same time, V5tag and His tag were added after gdh to facilitate post-purification of proteins and westen bolt verification experiments. Then GFP and pchBA was fused by overlap to form a stretch, and the vector pUCyder was linearized by restriction enzyme linearization, next the gene fragment was ligated to the linearized vector by infusion. | ||
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===Source=== | ===Source=== | ||
| − | + | GFP is derived from the plasmid PSB1C3. pchBA is derived from the official kit. emrR binding promoter, emrR gene and gdh gene are all from the E.coli K12 genome. | |
| + | |||
===References=== | ===References=== | ||
Latest revision as of 20:19, 17 October 2018
final2(GFP-hairpin-pchBA-terminator-emrr-RBS2000-GDH-v5 tag-his tag)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1070
Illegal NgoMIV site found at 1474
Illegal NgoMIV site found at 1599
Illegal NgoMIV site found at 1610
Illegal NgoMIV site found at 1891
Illegal NgoMIV site found at 2359
Illegal AgeI site found at 4584
Illegal AgeI site found at 5476 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 5458
Illegal BsaI.rc site found at 644
Design Notes
GFP, pchBA, emrR binding promoter, emrR gene and gdh gene were obtained from plasmids or genomes by PCR methods. At the same time, V5tag and His tag were added after gdh to facilitate post-purification of proteins and westen bolt verification experiments. Then GFP and pchBA was fused by overlap to form a stretch, and the vector pUCyder was linearized by restriction enzyme linearization, next the gene fragment was ligated to the linearized vector by infusion.
Source
GFP is derived from the plasmid PSB1C3. pchBA is derived from the official kit. emrR binding promoter, emrR gene and gdh gene are all from the E.coli K12 genome.