Difference between revisions of "Part:BBa K215001"
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Any favorite protein (afp) can be inserted into the NheI (compatable with XbaI and SpeI, but make sure the resulting product stays in frame) which places the afp in between a series of tags which can be used with the [http://2009.igem.org/Team:Washington/Project/Display University of Washington 2009 Ideal Protein Purification system (IPP)]. The series of tags are, respectively: a Nano-Tag (binds to streptavidin), a 6x His tag (for IMAC purification), a TEV cute site (for removal of N-terminal tags), the NheI site (insertion of afp), TEV, 6x His tag, and the 181 C-terminal amino acids of prtB (secretion tag recognized by the secretion system: [https://parts.igem.org/wiki/index.php?title=Part:BBa_K215107 BBa_K215107). To use in E. coli we recommend using the composite part [https://parts.igem.org/wiki/index.php?title=Part:BBa_K215001 BBa_K215002], which already has a strong promoter and RBS attached. In order for the secretion tag to function, this part needs to be used in conjunction with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K215001 BBa_K2150107]. | Any favorite protein (afp) can be inserted into the NheI (compatable with XbaI and SpeI, but make sure the resulting product stays in frame) which places the afp in between a series of tags which can be used with the [http://2009.igem.org/Team:Washington/Project/Display University of Washington 2009 Ideal Protein Purification system (IPP)]. The series of tags are, respectively: a Nano-Tag (binds to streptavidin), a 6x His tag (for IMAC purification), a TEV cute site (for removal of N-terminal tags), the NheI site (insertion of afp), TEV, 6x His tag, and the 181 C-terminal amino acids of prtB (secretion tag recognized by the secretion system: [https://parts.igem.org/wiki/index.php?title=Part:BBa_K215107 BBa_K215107). To use in E. coli we recommend using the composite part [https://parts.igem.org/wiki/index.php?title=Part:BBa_K215001 BBa_K215002], which already has a strong promoter and RBS attached. In order for the secretion tag to function, this part needs to be used in conjunction with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K215001 BBa_K2150107]. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K215001 SequenceAndFeatures</partinfo> | <partinfo>BBa_K215001 SequenceAndFeatures</partinfo> | ||
Revision as of 23:34, 18 October 2009
Secretion Signal and Streptavidin Binding Tags
Any favorite protein (afp) can be inserted into the NheI (compatable with XbaI and SpeI, but make sure the resulting product stays in frame) which places the afp in between a series of tags which can be used with the [http://2009.igem.org/Team:Washington/Project/Display University of Washington 2009 Ideal Protein Purification system (IPP)]. The series of tags are, respectively: a Nano-Tag (binds to streptavidin), a 6x His tag (for IMAC purification), a TEV cute site (for removal of N-terminal tags), the NheI site (insertion of afp), TEV, 6x His tag, and the 181 C-terminal amino acids of prtB (secretion tag recognized by the secretion system: BBa_K215107). To use in E. coli we recommend using the composite part [https://parts.igem.org/wiki/index.php?title=Part:BBa_K215001 BBa_K215002, which already has a strong promoter and RBS attached. In order for the secretion tag to function, this part needs to be used in conjunction with BBa_K2150107.
Usage and Biology
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 115
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]