Part:BBa_K3268002
Strong expression of eforRed in E. coli
To make a composite module that can highly express any inserted protein-coding genes, we had designed and constructed this strongly expressed eforRed in E. coli as a proof of our concept.
1. The eforRed coding region was PCR amplified from BBa_K592012 using high-fidelity enzyme KOD-Plus.
2. The strong promoter along with BBa_B0034 RBS and BBa_B0015 transcriptional terminator on pSB1C3 plasmid backbone were PCR amplified from BBa_J364007 using high-fidelity enzyme KOD-Plus.
3. In order to make the strong promoter (along with BBa_B0034 RBS and BBa_B0015 transcriptional terminator on pSB1C3 plasmid backbone) useful to highly express inserted protein-coding genes, we had designed two PCR primers with HindII and BamHI sites respectively to generate a composite expression module without the original GFP coding sequence.
4. Any inserted protein-coding genes not containing HindII and BamHI sites in their coding sequences can then be amplified and cloned into our composite expression module improved from BBa_J364007.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 2047
Illegal PstI site found at 14 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 2047
Illegal NheI site found at 2075
Illegal NheI site found at 2098
Illegal PstI site found at 14
Illegal NotI site found at 7
Illegal NotI site found at 2053 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 2047
Illegal BamHI site found at 2124
Illegal XhoI site found at 1031
Illegal XhoI site found at 1923 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 2047
Illegal PstI site found at 14 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 2047
Illegal XbaI site found at 2062
Illegal PstI site found at 14 - 1000COMPATIBLE WITH RFC[1000]
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