Composite
Part:BBa_K2804005:Design
Designed by: Byron Francisco Mosquera Group: iGEM18_Ecuador (2018-10-05)
CBD cipA fused to sfGFP under the control of LacI promoter
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 230
Illegal AgeI site found at 1493 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
After runing a PCR for adding the basepairs 5'-ccg-3'and 5'-aaaac-3 to the N-terminus and C-terminus of the gBlocks CBD cipA under the control of LacI promoter and sfGFP and then allow the enzymatic cleavage by EcoRI and PstI, a 3A assembly was achieved using psb1c3 as the cloning vector.
Source
Fusion assembly
References
New England Biolabs. Cleavage Close to the End of DNA Fragments (oligonucleotides). https://www.neb.com/-/media/nebus/files/chart-image/cleavage_olignucleotides_old.pdf?la=en