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Team:Gifu/Parts
From 2014.igem.org
Parts
<groupparts>iGEM014 Gifu</groupparts>Our team´s parts in part.igem.org
Histidine tag (8 AA) and RFP
This part codes for a RFP. A histidine tag is attached to a RFP that is coded by this part. So the RFP can be purified by immobilized metal ions affinity chromatography.
Histidine tag (8 AA) and RFP (without stop codon)
A Histidine tag is attached to a RFP that is coded by this part. So the RFP can be purified by immobilized metal ions affinity chromatography.
!!Caution!!
A stop codon in this part is removed. Therefore, this part doesn’t have an ability of usual protein coding parts. The combination of this part and the mRNA circularization device is capable of generating the RFP (+histidine tag) polymer. The part to generate it is registered ([http://parts.igem.org/Part:BBa_K1332011 BBa_K1332011]).
The 5´ side of the intron from td gene of T4 phage without stop codon
This part is a part of the mRNA circularization device (3’ side). A stop codon in this part is removed. The combination of this part and a terminator is the mRNA circularization device (3’ side) (BBa_K1332009).
The 5´ side of the intron from td gene of T4 phage
This part is a part of the mRNA circularization device (3’ side). The combination of this part and a terminator is the mRNA circularization device (3’ side) (BBa_K1332010). But there is a stop codon in this part, so a circular mRNA made by BBa_K1332010 (mRNA circularization device (3’ side); It consists of this part and double terminator.) doesn’t cause continuous translation.
The 3´ side of the intron from td gene of T4 phage
This part is a part of the mRNA circularization device (5’ side). The combination of this part, a promoter (lacI regulated) (BBa_R0010) and RBS (BBa_B0034) is the mRNA circularization device (5’ side) (BBa_K1332008).
mRNA circularization device (5´ side)
This part consists of a promoter (lacI regulated), the 3´ side of the intron in td gene of T4 phage and RBS. The protein coding sequence that is inserted between this device and mRNA circularization device (3’ side) can be circularized. If you circularized the protein coding sequence (Its a stop codon have been removed.) with mRNA circularization device (3’ side) for endless translation, you can get a circular mRNA that is translated semi-permanently.
mRNA circularization device (3´ side) (endless translation)
This part consists of the 5´ side of the intron in td gene of T4 phage and a double terminator. The protein coding sequence that is inserted between this device and mRNA circularization device (5’ side) can be circularized. If you circularized the protein coding sequence (It’s a stop codon have been removed.), you can get a circular mRNA that is translated semi-permanently.
mRNA ircularization device (3´ side)
This part consists of the 5´ side of the intron in td gene of T4 phage and a double terminator. The protein coding sequence that is inserted between this device and mRNA circularization device (5´ side) can be circularized. The 5´side of the intron in td gene of T4 phage contains a stop codon, so a circular mRNA made by this part doesn’t cause continuous translation.
Histidine tag (8 AA) and RFP semi-permanent generator
This generator is capable of synthesizing a RFP (+histidine tag) polymer. This generator consists of a circularization device (5´ side), histidine tag (8 AA) and RFP (without stop codon) and a circularization device (3´ side). A mRNA is circular, so translation continues semi-permanently. A synthesis of the RFP (+histidine tag) become possible by a simply transformation, but the coloration of RFP is weak.
TTHA0715 with sequence cut by thrombin semi-permanent generator
This generator is capable of synthesizing TTHA0715 (with amino acid sequence cut by thrombin). This generator consists of a circularization device (5´ side), TTHA0715 (with amino acid sequence cut by thrombin) and a circularization device (3´ side). A mRNA is circular, so translation continues semi-permanently. TTHA0715 is a cold shock protein and ABC transporter ATP-binding protein. The use of this device is to confirm the existence of the long-chain protein that is made by a circular mRNA. But this part didn’t work.
SmtA semi-permanent generator
This generator is capable of synthesizing SmtA (BBa_K519010). This generator consists of a mRNA circularization device (5´ side), SmtA (BBa_K519010) and a mRNA circularization device (3´ side). A mRNA is circular, so translation continues semi-permanently. SmtA is a metallothionein that can catch heavy metal ions such as Zn2+.
