Difference between revisions of "Part:BBa K3409011"
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Contains the elements necessary to express the fourth gene from the Microcin PDI gene cluster (mcpD: a homolog of hlyD, coding for part of the secretion system). The sequence is organized as follows: a promoter (BBa_J23118), an RBS (BBa_J61118), the CDS for McpD (BBa_K3409006). Will constitutively generate McpD. For appropriate secretion, it must be followed by BBa_K3409012, which contains the CDS for McpB. | Contains the elements necessary to express the fourth gene from the Microcin PDI gene cluster (mcpD: a homolog of hlyD, coding for part of the secretion system). The sequence is organized as follows: a promoter (BBa_J23118), an RBS (BBa_J61118), the CDS for McpD (BBa_K3409006). Will constitutively generate McpD. For appropriate secretion, it must be followed by BBa_K3409012, which contains the CDS for McpB. | ||
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| + | ===Characterization=== | ||
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| + | NOTE: In this section: BB1 = part BBa_K3409010 - BB2 = part BBa_K3409011 – BB3 = part BBa_K3409012 | ||
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| + | PCR AMPLIFICATION OF PARTS | ||
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| + | [[File: PCR_BB1_BB2_BB3.png|border|700px]] | ||
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| + | Once received, the synthetized fragments are resuspended and amplified via polymerase chain reaction (PCR) to obtain a higher quantity of DNA. Every PCR sample is verified by gel electrophoresis to analyze the PCR results. First, the appropriate bands for BioBrick 1 (1.5 kb) , BioBrick 2 (1.4 kb) and BioBrick 3 (2.4 kb) amplified with the primers for Hifi Assembly method, are present at their expected size (Figure 1A). | ||
| + | Then, the appropriate bands for BioBrick 1 (1.5 kb), BioBrick 2 (1.4 kb) and BioBrick 3 (2.4 kb) amplified with the primers for BioBrick Assembly method, are present at their expected size on Figure 1B. | ||
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Revision as of 21:02, 25 October 2020
Microcin PDI cluster gene for McpD
Contains the elements necessary to express the fourth gene from the Microcin PDI gene cluster (mcpD: a homolog of hlyD, coding for part of the secretion system). The sequence is organized as follows: a promoter (BBa_J23118), an RBS (BBa_J61118), the CDS for McpD (BBa_K3409006). Will constitutively generate McpD. For appropriate secretion, it must be followed by BBa_K3409012, which contains the CDS for McpB.
Characterization
NOTE: In this section: BB1 = part BBa_K3409010 - BB2 = part BBa_K3409011 – BB3 = part BBa_K3409012
PCR AMPLIFICATION OF PARTS
Once received, the synthetized fragments are resuspended and amplified via polymerase chain reaction (PCR) to obtain a higher quantity of DNA. Every PCR sample is verified by gel electrophoresis to analyze the PCR results. First, the appropriate bands for BioBrick 1 (1.5 kb) , BioBrick 2 (1.4 kb) and BioBrick 3 (2.4 kb) amplified with the primers for Hifi Assembly method, are present at their expected size (Figure 1A). Then, the appropriate bands for BioBrick 1 (1.5 kb), BioBrick 2 (1.4 kb) and BioBrick 3 (2.4 kb) amplified with the primers for BioBrick Assembly method, are present at their expected size on Figure 1B.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]