Difference between revisions of "Part:BBa K3352001"
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We obtained the amino acid sequence of phi29 DNA polymerase from Bacillus phage phi29. We proceeded to optimize the DNA sequence for expression in E. coli and removed the EcoRI cutting site. We attached a 6x his-tag upstream of the phi29 DNA Polymerase for purification purposes followed by a glycine-serine linker (GS linker) to form our ORF. | We obtained the amino acid sequence of phi29 DNA polymerase from Bacillus phage phi29. We proceeded to optimize the DNA sequence for expression in E. coli and removed the EcoRI cutting site. We attached a 6x his-tag upstream of the phi29 DNA Polymerase for purification purposes followed by a glycine-serine linker (GS linker) to form our ORF. | ||
Revision as of 15:50, 18 October 2020
Φ29 DNA Polymerase with His-Tag and GS linker Sequence
Construct Design
We obtained the amino acid sequence of phi29 DNA polymerase from Bacillus phage phi29. We proceeded to optimize the DNA sequence for expression in E. coli and removed the EcoRI cutting site. We attached a 6x his-tag upstream of the phi29 DNA Polymerase for purification purposes followed by a glycine-serine linker (GS linker) to form our ORF.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 595
Illegal SapI.rc site found at 993