Composite

Part:BBa_K3409011

Designed by: Albane Mabro   Group: iGEM20_Ionis_Paris   (2020-10-17)
Revision as of 21:02, 25 October 2020 by Albane Mabro (Talk | contribs)


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

PCR BB1 BB2 BB3.png

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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