Difference between revisions of "Part:BBa K4368007"
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<partinfo>BBa_K4368007 short</partinfo> | <partinfo>BBa_K4368007 short</partinfo> | ||
| − | + | ==Description== | |
| + | ''RFP'' (<partinfo>BBa_E1010</partinfo>) is the gene coding red fluorescent protein which is a protein synthesized by the cnidaria ''Discosoma sp.'' 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 the cI protein of the phage lambda (<partinfo>BBa_R1051</partinfo>). | ||
| + | |||
| + | ==Characterization== | ||
| + | The expression cassette sequence was digested with EcoRI and PstI enzymes and subsequently ligated with a ampicillin resistant plasmid backbone (Amp). | ||
| + | Transforming bacteria were created with this plasmid and seeded on LB-Agar+Amp plates. After growth, colonies were selected based on their color (red) 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_RFP.png|350px|center|]] | ||
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| − | <span class='h3bb'>Sequence and Features</span> | + | ==<span class='h3bb'>Sequence and Features</span>== |
<partinfo>BBa_K4368007 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4368007 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
pcI + rbs + RFP + terminator
Description
RFP (BBa_E1010) is the gene coding red fluorescent protein which is a protein synthesized by the cnidaria Discosoma sp. 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 the cI protein of the phage lambda (BBa_R1051).
Characterization
The expression cassette sequence was digested with EcoRI and PstI enzymes and subsequently ligated with a ampicillin resistant plasmid backbone (Amp). Transforming bacteria were created with this plasmid and seeded on LB-Agar+Amp plates. After growth, colonies were selected based on their color (red) 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]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 633
Illegal AgeI site found at 745 - 1000COMPATIBLE WITH RFC[1000]
Contribution
- Group: UMA_MALAGA
- Author: Molina Calvo, Alonso