Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Jeff Nivala
Date created: 2009-09-29 11:00:00
Date modified: 2015-05-08 01:11:29
Secretion Signal and Streptavidin Binding Tags
| Types | DnaRegion |
| Roles | Tag tag |
| Sequences | BBa_K215001_sequence (Version 1) |
Description
This part is the vector into which afp is inserted to tag it for purification using the IPP system. It contains an NheI restriction site that allows for the insertion of the desired protein coding sequence (NheI is compatible with XbaI and SpeI). This Target Vector part contains two tags: a secretion tag that causes the protein to be exported to the extracellular space, and a nanotag that causes the protein to bind to stretavidin (or some analog). For this part to function properly, it must be combined with a promoter and RBS, and used in E.coli strain UW5alpha, which has two other plasmids containing the other genes required for proper secretion and display of the protein to be purified on the outer membrane.Notes
NheI was chosen as the protein insertion site because it is compatible with digested XbaI and SpeI restriction sites. The TEV sites and HIS tags flanking the NheI site are present to separate the inserted protein from the added tags. GSA linkers were used to separate the various features of the part.Source
Nanotag (bases 1-48): From Lamla and Erdmann (http://www.ncbi.nlm.nih.gov/pubmed/14680960).Secretion tag (last 181 AA): Gene Bank ID Erwinia chrysanthemi M60395.1
| Sequence Annotation | Location | Component / Role(s) |
| nanotag GSA linker His tag GSA linker TEV protease site GSA linker NheI GSA linker TEV protease site GSA linker His tag GSA linker Secretion tag terminator (B0015) | 1,48 49,57 58,75 76,84 85,104 105,114 115,120 121,129 130,150 151,159 160,177 178,186 187,735 736,865 | feature/tag tag sequence_feature feature/misc feature/tag tag sequence_feature feature/misc feature/misc sequence_feature feature/misc sequence_feature feature/misc sequence_feature feature/misc sequence_feature sequence_feature feature/misc sequence_feature feature/misc feature/tag tag sequence_feature feature/misc tag feature/tag feature/stop stop_codon |