Regulatory
Part:BBa_K3020001:Design
Designed by: WANG WENJIA Group: iGEM19_BIT (2019-09-06)
recA promoter-Respond to sos response and initiate expression of downstream repair proteins
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The recA promoter fragment was cloned from the genome of E.coli K12 MG1655 strain as a template.
Primers For PCR
| PCR amplification of upstream primers, addition of EcoRI restriction sites | 5’-AGGAgaattcCAGATGATCGGCGTACGCG-3’ |
| PCR amplification of downstream primers, addition of XbaI restriction sites | 5’-CATGtctagaTTTTACTCCTGTCATGCCGGG-3’ |
| PCR system | |
|---|---|
| enzyme | 2xPCR Mastermix（Tiangen） |
| Upstream and downstream primers | 2μL |
| Genomic template | 2μL |
| ddH2O | to 50μL |
PCR Time Protocol
| Temperature(°C) | Time(sec) | Number of Cycles |
|---|---|---|
| 94 | 180 | -- |
| 94 | 30 | 30 |
| 56 | 30 | |
| 72 | 60 | |
| 72 | 300 | -- |
Source
The recA promoter fragment was cloned from the genome of E.coli K12 MG1655 strain as a template.Please note that the recA promoter sequences from different genomes are not consistent.
References
Norman A , Hestbjerg Hansen L , Sørensen, Søren J. Construction of a ColD cda Promoter-Based SOS-Green Fluorescent Protein Whole-Cell Biosensor with Higher Sensitivity toward Genotoxic Compounds than Constructs Based on recA, umuDC, or sulA Promoters[J]. Appl Environ Microbiol, 2005, 71(5):2338-2346.