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Part:BBa_K3927015
NLS-NucA
Gene encoding for Staphylococcus aureus endonuclease nucA with a nuclear localisation sequence. Improved part based on part BBa_K1159105.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 25
Illegal AgeI site found at 478 - 1000COMPATIBLE WITH RFC[1000]
Description
Usage
Design
Characterization
Characterization of original part BBa_K1159105
Sequence for BBa_K1159105 was ordered from IDT, and using Gibson Assembly it was inserted into the pGmFaHBD-H plasmid to form pGNucA-H.
pGNucA-H (BY4741) was cultured in YPD-HygB for 24 hours, and a CFU assay was carried out 5 serial dilutions to a cell count of 10-5 and then plated on either YPD-HygB agar as a negative control, as well as YPGR-H to induce the production of NucA. Culture that was induced showed only a 20% decrease in cell mortality, and it was decided that the part needed to be improved upon.
Characterization of this part (Improved version)
Gibson Assembly was used to insert a 7 amino acid NLS sequence at the N-terminus of pGNucA-H forming the plasmid pGNLSNucA-H. pGNLSNucA-H was transfomed into BY4741 forming the strain pGNLSNucA-H (BY4741). pGNLSNucA-H (BY4741) was in YPD-HygB for 24 hours, and a CFU assay was carried out 5 serial dilutions to a cell count of 10-5 and then plated on either YPD-HygB agar as a negative control, as well as YPGR-H to induce the production of NucA.
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