Difference between revisions of "Part:BBa K4368004"
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<partinfo>BBa_K4368004 short</partinfo> | <partinfo>BBa_K4368004 short</partinfo> | ||
| − | + | ==Description== | |
| − | + | ''GlgC16'' encodes for the ADP-glucose pyrophosphorylase gene of ''Escherichia coli'' (<partinfo>BBa_K118016</partinfo>). This enzyme is responsible for the synthesis of starch-like starch using glucose as a substrate. 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>). | |
| + | <partinfo>BBa_K118016</partinfo> works under the control of this promoter to build the glucose concentration-based gene regulatory circuit that integrates all our parts. | ||
| + | ==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:glgC.png|350px|center|]] | ||
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| − | <span class='h3bb'>Sequence and Features</span> | + | ==<span class='h3bb'>Sequence and Features</span>== |
<partinfo>BBa_K4368004 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4368004 SequenceAndFeatures</partinfo> | ||
| − | + | ||
| + | ==Contribution == | ||
| + | *'''Group:''' [https://2022.igem.wiki/uma-malaga/index.html UMA_MALAGA] | ||
| + | *'''Author:''' Jiménez Amores, Carmen | ||
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Latest revision as of 18:54, 7 October 2022
pcI + rbs + glgC16 + terminator
Description
GlgC16 encodes for the ADP-glucose pyrophosphorylase gene of Escherichia coli (BBa_K118016). This enzyme is responsible for the synthesis of starch-like starch using glucose as a substrate. 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). BBa_K118016 works under the control of this promoter to build the glucose concentration-based gene regulatory circuit that integrates all our parts.
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]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 272
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Contribution
- Group: UMA_MALAGA
- Author: Jiménez Amores, Carmen