Difference between revisions of "Part:BBa K3440016"
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<partinfo>BBa_K3440016 short</partinfo> | <partinfo>BBa_K3440016 short</partinfo> | ||
| − | Pconst(BBa_J23100)-RBS(BBa_B0034)-Mtrb(BBa_K3146001)-Myc(BBa_K823036) | + | Pconst(BBa_J23100) - RBS(BBa_B0034) - Mtrb(BBa_K3146001) - Myc(BBa_K823036) |
===Usage and Biology=== | ===Usage and Biology=== | ||
| − | This part expresses mtrb constitutively. Mtrb is responsible for electricity production in Shewanella oneidensis. In our project, we created this part as an intermediate to build BBa_K3440019 and BBa_K3440021. | + | This part expresses mtrb constitutively. Mtrb is responsible for electricity production in Shewanella oneidensis (UniProtKB - Q8CVD4). In our project, we created this part as an intermediate to build BBa_K3440019 and BBa_K3440021. |
===Characterization=== | ===Characterization=== | ||
Due to the pandemics, we haven’t been able to use biobricks to create the iGEM Stockholm 2020 parts. Those parts were ordered as gene blocks from Integrated DNA Technologies Inc.. | Due to the pandemics, we haven’t been able to use biobricks to create the iGEM Stockholm 2020 parts. Those parts were ordered as gene blocks from Integrated DNA Technologies Inc.. | ||
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| + | After heat shock transformation of the pSB1C3 plasmid containing the BBa_K3440016, we picked colonies from plates and PCR amplified them with primers VF and VR2. | ||
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| + | We ran gels of the product at 180V and for 30 mins (Figure 2). We obtained the expected size for the bands (2499bp) for cf3. | ||
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| + | [[File:T--Stockholm--gelheart.png|thumb|center|500px|Figure 2: Colony PCR Gel for BBa_K3440016 (cf)]] | ||
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| + | We therefore prepared plasmid preparations and glycerol stocks of those and sent some of them for sequencing to Microsynth AG. The sequence obtained did not correspond to mtrb. The part had not been insert into the plasmid. We therefore highly recommend other teams to always send their samples for sequencing, even when the bands on gels have the right size | ||
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| + | No further characterisation was done since this was only an intermediate part used to build others. | ||
Latest revision as of 23:31, 27 October 2020
Mtrb under constitutive promoter
Pconst(BBa_J23100) - RBS(BBa_B0034) - Mtrb(BBa_K3146001) - Myc(BBa_K823036)
Usage and Biology
This part expresses mtrb constitutively. Mtrb is responsible for electricity production in Shewanella oneidensis (UniProtKB - Q8CVD4). In our project, we created this part as an intermediate to build BBa_K3440019 and BBa_K3440021.
Characterization
Due to the pandemics, we haven’t been able to use biobricks to create the iGEM Stockholm 2020 parts. Those parts were ordered as gene blocks from Integrated DNA Technologies Inc..
After heat shock transformation of the pSB1C3 plasmid containing the BBa_K3440016, we picked colonies from plates and PCR amplified them with primers VF and VR2.
We ran gels of the product at 180V and for 30 mins (Figure 2). We obtained the expected size for the bands (2499bp) for cf3.
We therefore prepared plasmid preparations and glycerol stocks of those and sent some of them for sequencing to Microsynth AG. The sequence obtained did not correspond to mtrb. The part had not been insert into the plasmid. We therefore highly recommend other teams to always send their samples for sequencing, even when the bands on gels have the right size
No further characterisation was done since this was only an intermediate part used to build others.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2184
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]