Types | DnaRegion
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Roles | Protein_Domain
polypeptide_domain
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Sequences | BBa_K300002_sequence (Version 1)
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Description
-
Notes
It is identical to
BBa_K208001, but it lacks the stop codon in order to support protein fusions.
It has been designed as a head domain:
The Prefix sequence is 5'-GAATTCGCGGCCGCTTCTAG-3' (RFC10 Prefix)
The Suffix sequence is 5'-ACTAGTAGCGGCCGCTGCAG-3' (RFC23 Suffix)
For these reasons, a tail domain or an internal domain (compatible with RFC23) are ready to be assembled downstream to create protein fusions.
CONSTRUCTION METHOD:
BBa_K208001 (provided by iGEM HQ in pSB3K3 in RFC23 standard) was PCR-amplified/mutagenized with primers:
phaP10F 5'-GCTTCTAGATGATCCTCACCCCGGAACA-3'
phaPSR: 5'-GCTACTAGTGGCAGCCGTCGTCTTCTTTG-3'
in order to delete the stop codon.
The PCR product was ran on a 1% agarose gel, gel-extracted, cut with XbaI-SpeI and ligated with
pSB1AK3 (previously cut with XbaI-SpeI and dephosphorylated).
Positive clones were found through digestion screening/sequencing.
Source
Ralstonia eutropha genomic DNA.