BBa_K215012 1 BBa_K215012 Secretion Signal and Streptavidin Binding Tags on GFP 2009-10-17T11:00:00Z 2015-05-08T01:11:30Z E0040 (GFP) was amplified using NheI K215002 drives the expression of [http://partsregistry.org/wiki/index.php?title=Part:BBa_K215001 BBa_K215001] by placing it under the control of a strong, IPTG-inducible promoter (R0011) and RBS (R0034). Any favorite protein (afp) can be inserted into a composite tag [http://partsregistry.org/wiki/index.php?title=Part:BBa_K215001 BBa_K215001]. The composite tag has an NheI site (compatible with XbaI and SpeI, but make sure the resulting product stays in frame, described below) which places the afp in between a series of tags which can be used with the [http://2009.igem.org/Team:Washington 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: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K215107 BBa_K215107]). In order for the secretion tag to signal the secretion of afp the BioBrick [http://partsregistry.org/wiki/index.php?title=Part:BBa_K215107 BBa_K215107] must be present as well. Cutting a BioBrick coding sequence at X and S and pasting into the NheI site of this construct would result in a stop codon at the resulting scar site. Therefore, to insert a gene into the K215001 of this construct, a PCR product must be generated that keeps the gene of interest in frame with the rest of the sequence. To do this: false false _320_ 0 5005 9 Not in stock false sdsdsd false Jeff Nivala annotation2052467 1 K215012 range2052467 1 1 115 annotation2052469 1 K215012 range2052469 1 833 1581 annotation2052468 1 E0040 range2052468 1 116 832 BBa_K215012_sequence 1 atggacgttgaagcatggctgggtgcccgtgtaccgctggttgaaacgggcagcgcacatcaccaccatcatcatggttccgccgaaaacctctactttcagggcggctctgctgctagacgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaaactagcggttctgccgagaatctgtacttccaaggtggctccgctcatcaccaccatcatcacggtagcgcaggcggcactgacacctttgacttctctggttactctaacaatcagcgtatcaacctgaacgagggttctttctctgacgttggtggtctgaagggcaacgtttctatcgcgcatggtgttaccatcgaaaacgcgatcggtggttctggtaacgacatcctgatcggtaatggcgcggataacatcctccagggtggtgcgggtgacgacgttctgtacggttctacgggtgcggacacgctgacgggtggcgctggtcgtgacattttcgtatatggtagcggtcaagactctaccgtttctgcatacgactggatcacggattttcagacgggtatcgacaaaatcgacctgtccgctttccgtaatgaaggtcagctgtcttttgttcaggaccagtttactggtaaaggccaggaggttatgctgcaatgggacgcggcgaactctacgaccaatctgtggctgcacgaagcgggtcactcttctgttgactttctcgttcgtatcgttggccagaccgcgcagagcgacattatcgtctaataaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 Chris J. Myers James Alastair McLaughlin 2017-03-06T15:00:00.000Z