Team:Kyoto/GSDD/Parts

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Parts

Parts Design

We designed following three vectors and registered intermediate products of Timer vector and constitutive LacI generator optimized in yeast.

Timer Vector for functional measurement experiment

This part is designed for our functional measurement of Timer vector. It consists of four part (see figure); both ends of repeated sequence of LacI binding site, constitutive eGFP generator (pTEF2-kozac-eGFP) with degradation promoted sequence (PEST), yeast centromeric sequence (CEN), yeast replication origin (ARS), constitutive uracil generator (pTEF2-kozac-URA3) with SV40 nuclear localization sequences and two ADH1 terminators (SV40-ADH1). As mentioned in Mechanism, Timer vector is a linear DNA and inserted into pSB1A2. When we use this, we can cut out linear Timer vector with a pair of restriction enzyme frequently used in iGEM, (EcoRI or XbaI)and(SpeI or PstI).

Constitutive LacI generator

This part is also designed for our experiment. It consists of constitutive LacI generator (pTEF2-kozac-LacI) with SV40 nuclear localization sequences and two ADH1 terminators (SV40-ADH1). This part is also inserted into BYP5005(national bioresource project).