Difference between revisions of "Part:BBa K4368006"
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<partinfo>BBa_K4368006 short</partinfo> | <partinfo>BBa_K4368006 short</partinfo> | ||
| − | ''GFP'' is the gene coding green fluorescent protein which is a protein synthesized by the jellyfish Aequorea victoria. In addition, this part includes the composition used by the team, which includes a strong rbs (<partinfo>BBa_B0030</partinfo>), a double terminator (<partinfo>BBa_B0015</partinfo>) as well as a promoter inducible by glucose concentration (<partinfo> | + | ==Description== |
| + | ''GFP'' (<partinfo>BBa_E0040</partinfo>) is the gene coding green fluorescent protein which is a protein synthesized by the jellyfish ''Aequorea victoria''. In addition, this part includes the composition used by the team, which includes a strong rbs (<partinfo>BBa_B0030</partinfo>), a double terminator (<partinfo>BBa_B0015</partinfo>) as well as a promoter inducible by glucose concentration (<partinfo>BBa_K118011</partinfo>). | ||
| + | |||
| + | ==Characterization== | ||
| + | The expression cassette sequence was digested with EcoRI and PstI enzymes and subsequently ligated with a chloramphenicol resistant plasmid backbone (Cm). | ||
| + | Transforming bacteria were created with this plasmid and seeded on LB-Agar+Cm plates. After growth, colonies were selected based on their color (white) and DNA extraction was performed using the Promega PureYield Plasmid Miniprep System kit. | ||
| + | The resulting DNA is used for further digestion with EcoRI and PstI. The digests are then run on a 0.75% agarose gel at 90 mV voltage and constant amperage. BioRad brand RedSafe is used as an intercalating developing agent. | ||
| + | |||
| + | ===Enzyme digestion=== | ||
| + | [[File:UMA_GFP.png|350px|center|]] | ||
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| − | <span class='h3bb'>Sequence and Features</span> | + | ==<span class='h3bb'>Sequence and Features</span>== |
<partinfo>BBa_K4368006 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4368006 SequenceAndFeatures</partinfo> | ||
| + | ==Contribution == | ||
| + | *'''Group:''' [https://2022.igem.wiki/uma-malaga/index.html UMA_MALAGA] | ||
| + | *'''Author:''' Molina Calvo, Alonso | ||
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Latest revision as of 18:53, 7 October 2022
pcstA + rbs + GFP + terminator
Description
GFP (BBa_E0040) is the gene coding green fluorescent protein which is a protein synthesized by the jellyfish Aequorea victoria. In addition, this part includes the composition used by the team, which includes a strong rbs (BBa_B0030), a double terminator (BBa_B0015) as well as a promoter inducible by glucose concentration (BBa_K118011).
Characterization
The expression cassette sequence was digested with EcoRI and PstI enzymes and subsequently ligated with a chloramphenicol resistant plasmid backbone (Cm). Transforming bacteria were created with this plasmid and seeded on LB-Agar+Cm plates. After growth, colonies were selected based on their color (white) and DNA extraction was performed using the Promega PureYield Plasmid Miniprep System kit. The resulting DNA is used for further digestion with EcoRI and PstI. The digests are then run on a 0.75% agarose gel at 90 mV voltage and constant amperage. BioRad brand RedSafe is used as an intercalating developing agent.
Enzyme digestion
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 804
Contribution
- Group: UMA_MALAGA
- Author: Molina Calvo, Alonso