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Team:Johns Hopkins/Parts
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==Parts== | ==Parts== | ||
| - | ===BBa_K363001=== | + | ===BBa_K363001-BBa_K363007=== |
| - | ''' | + | '''Calcium dependent response element binding sites for the Crz1 activator.''' |
| - | This part is a binding site for the Crz1 transcriptional activator. When placed upstream of a eukaryotic promoter sequence, this sequence results in the calcium dependant transcription of downstream coding sequences given that Crz1 and the upstream signal transducing phosphotase calcineurin are constitutively expressed. The signal cascade occurs when calcineurin binds to cytosolic calcium, activating its phosphotase activity. The active calcineurin dephosphorylates cytoplasmic Crz1, resulting in translocation of Crz1 to the nucleus where it binds to this part and promotes the assembly of general transcription factors at the promoter sequence. | + | This part is a binding site for the Crz1 transcriptional activator. When placed upstream of a eukaryotic promoter sequence (~700bp in the case of FKS2), this sequence results in the calcium dependant transcription of downstream coding sequences given that Crz1 and the upstream signal transducing phosphotase calcineurin are constitutively expressed. The signal cascade occurs when calcineurin binds to cytosolic calcium, activating its phosphotase activity. The active calcineurin dephosphorylates cytoplasmic Crz1, resulting in translocation of Crz1 to the nucleus where it binds to this part and promotes the assembly of general transcription factors at the promoter sequence. |
| - | === | + | ====BBa_K363001==== |
| - | ''' | + | '''The consensus calcium dependent response element binding sites for the Crz1 activator.''' |
| + | This part was derived from calcium dependent response element (CDRE) binding sequence of the FKS2 gene. The consensus sequence for Crz1 binding as described by Yoshimoto et al (2002) was mutated at non conserved residues to generate this part. This UAS is the consensus sequence for Crz1 binding, and is expected to have higher binding than the UAS characterized in BBa_K363006, as it is the consensus sequence and occurs with ~4 times higher frequency than the characterized sequence in the yeast genome. | ||
| - | + | ====BBa_K363006==== | |
| + | '''A characterized calcium dependent response element binding sites for the Crz1 activator.''' | ||
| + | This part was characterized as part of a larger sequence (see design notes for this part in the registry), as it occurs naturally as an activator of the yeast gene FKS2, which is codes for an enzyme involved in spore wall synthesis. We found that applying a voltage of 8V for 40-80 seconds provided maximal induction using our apparatus. This apparatus was, roughly speaking, a cylindrical coaxial electrode that was about .75cm across. We used SC media. For full details, see the 2010 Johns Hopkins Team wiki. | ||
| - | ==== | + | ====BBa_K363008 and BBa_K363008==== |
| - | + | '''The voltage-gated calcium channels of ''S. cerevisiae''.''' | |
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===Registry Parts=== | ===Registry Parts=== | ||
<groupparts>iGEM010 Johns_Hopkins</groupparts> | <groupparts>iGEM010 Johns_Hopkins</groupparts> | ||
Revision as of 18:04, 27 October 2010
Contents |
Parts
BBa_K363001-BBa_K363007
Calcium dependent response element binding sites for the Crz1 activator.
This part is a binding site for the Crz1 transcriptional activator. When placed upstream of a eukaryotic promoter sequence (~700bp in the case of FKS2), this sequence results in the calcium dependant transcription of downstream coding sequences given that Crz1 and the upstream signal transducing phosphotase calcineurin are constitutively expressed. The signal cascade occurs when calcineurin binds to cytosolic calcium, activating its phosphotase activity. The active calcineurin dephosphorylates cytoplasmic Crz1, resulting in translocation of Crz1 to the nucleus where it binds to this part and promotes the assembly of general transcription factors at the promoter sequence.
BBa_K363001
The consensus calcium dependent response element binding sites for the Crz1 activator. This part was derived from calcium dependent response element (CDRE) binding sequence of the FKS2 gene. The consensus sequence for Crz1 binding as described by Yoshimoto et al (2002) was mutated at non conserved residues to generate this part. This UAS is the consensus sequence for Crz1 binding, and is expected to have higher binding than the UAS characterized in BBa_K363006, as it is the consensus sequence and occurs with ~4 times higher frequency than the characterized sequence in the yeast genome.
BBa_K363006
A characterized calcium dependent response element binding sites for the Crz1 activator. This part was characterized as part of a larger sequence (see design notes for this part in the registry), as it occurs naturally as an activator of the yeast gene FKS2, which is codes for an enzyme involved in spore wall synthesis. We found that applying a voltage of 8V for 40-80 seconds provided maximal induction using our apparatus. This apparatus was, roughly speaking, a cylindrical coaxial electrode that was about .75cm across. We used SC media. For full details, see the 2010 Johns Hopkins Team wiki.
BBa_K363008 and BBa_K363008
The voltage-gated calcium channels of S. cerevisiae.
Registry Parts
<groupparts>iGEM010 Johns_Hopkins</groupparts>
