Part:BBa_K5520004
pT7-LacO-His-CDA
This part contains BBa_K2406020, BBa_B0034, BBa_K5520002 and BBa_K731721. BBa_B0034 contains a natural ribosome binding site (RBS) to facilitate the expression of downstream genes. By inducing efficient expression of the CDA protein through the addition of IPTG, the protein can subsequently be purified using the His tag for further enzyme activity tests and product polymerization.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 149
Illegal NheI site found at 897 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 182
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 641
Usage and Biology
Plasmid construction PET-28a linearized vector (5371 bp) was amplified using PET-28a plasmid from the laboratory conserved as template and ZT-F and ZT-R as primers. Using CDA-F and CDA-R as primers, we amplified PET-28a-CDA as a vector using Gibson assembly. The T7 promoter、RBS、 lac operator, CDA gene, 6×His tag, kana antibiotic gene, and T7 terminator on the vector PET-28a were already inserted before experiments.
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