Deinococcus radiodurans uracil-DNA glycosylase 2
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000COMPATIBLE WITH RFC
This part was isolated from the genomic DNA of Deinococcus radiodurans (1). First, genomic DNA was extracted from cells growing in an overnight liquid culture. The gene was simultaneously amplified from genomic DNA and added BioBrick ends using the following protocol:
Forward primer uracil-DNA glycosylase 2 (49 bp)
Reverse primer uracil-DNA glycosylase 2 (54 bp)
60 ng genomic DNA/reaction
1 uM primer mix
25 ul AmplitaQ gold in a total reaction volume of 50 uL
1 min of 95C denaturation and 45 sec of annealing, x34 cycles
Note 1: The stop codon was modified from the wild-type TGA to the registry's standard TAA using the reverse amplification primer.
Note 2: This coding part does not contain a promoter, RBS, transcriptional terminator, etc. These elements must be added for proper functionality.
Isolated directly from Deinococcus radiodurans genomic DNA, Chromosome 1 NCBI Genome
1. White, O et al. (1999) Genome sequence of the radioresistant bacterium Deinococcus radiodurans R1 Science 286(5444):1571-7. PMID: 10567266.
2. Sandigursky, M et al. (2004) Multiple uracil-DNA glycosylase activities in Deinococcus radiodurans. DNA Repair (Amst) 3(2):163-9. PMID: 14706350.