Difference between revisions of "Part:BBa K2717020: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 20:21, 17 October 2018
final1(GFP-TGATG-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 1023
Illegal NgoMIV site found at 1427
Illegal NgoMIV site found at 1552
Illegal NgoMIV site found at 1563
Illegal NgoMIV site found at 1844
Illegal NgoMIV site found at 2312
Illegal AgeI site found at 4537
Illegal AgeI site found at 5429 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 5411
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.