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Team:Kyoto/GSDD/Parts
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We designed following three vectors and registered intermediate products of Timer vector and constitutive LacI generator optimized in yeast. | 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). | 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 pSB1A2. | 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 pSB1A2. | ||
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Revision as of 21:41, 17 October 2009
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 pSB1A2.
