Composite

Part:BBa_K3020000:Design

Designed by: WANG WENJIA   Group: iGEM19_BIT   (2019-09-04)
Revision as of 08:35, 8 September 2019 by WANGWENJIA (Talk | contribs) (→‎Design Notes)


Green fluorescent protein reporter gene under oxidative damage-sensitive initiation


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 812
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The recA promoter does not have a strict sequence range on the NCBI website. Therefore, when using the PCR to clone the target fragment, we designed the primer and completely cloned the recA repair protein base ATG upstream of 390 bp.

Source

  The recA promoter fragment was cloned from the genome of E.coli MG1655 strain as a template, and ligated upstream of the reporter gene by enzyme cleavage and other molecular cloning methods, and the target fragment was cloned into the standard vector pSB1A2.

  The eGFP gene sequence used in this experiment was obtained from NCBI, sequence ID: 20473140. Due to the preference of codons, the level of foreign gene expression was affected, and the gene sequence was given to Jin Weizhi (Su State) Bio Co., Ltd. performs codon optimization for E. coli hosts.

References