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Revision as of 23:26, 31 October 2017
Composite Parts
The middle part of the full three part construct is one of three tsPurple sequences. Each sequence is composed of BBa_B0034 (RBS) and BBa_K1033906 (tsPurple- a purple chromoprotein also known as TinselPurple). When assembled with p-lambda-r-LacI and pLac-ClpXP-CI, the tsPurple constructs are expressed by p-lambda-r-LacI. The terminator for tsPurple is located at the beginning of the pLac-ClpXP-CI sequence, before the promoter for ClpXP-CI. When the construct is fully assembled (and induced with IPTG) ClpXP is expressed, which degrades tsPurple when there is a degradation tag attached at the end of the tsPurple construct. BBa_K2353000, the tsPurple construct without any degradation tag, shows the baseline expression of tsPurple as a basis for comparison. BBa_K2353001 has degradation tag DAS (BBa_M0052) which should cause moderately fast degradation of tsPurple, while BBa_K2353002 has the degradation tag LAA (BBa_M0050), which should cause high rates of degradation due to the high affinity of ClpXP for LAA.
Gel results from a digest of all three tsPurple parts. From top to bottom: BBa_K2353000, BBa_K2353002, BBa_K2353001