BBa_K2009820 1 sfGFP11 sfGFP11 2016-09-13T11:00:00Z 2016-09-15T02:33:25Z chemical synthesing sequence which is designed by ???Protein tagging and detection with engineered self-assembling fragments of green fluorescent protein Ste??phanie Cabantous, Thomas C Terwilliger & Geoffrey S Waldo&#65292;2005??? sfGFP11 length: 48bp Derived from:synthesis from Sangon sfGFP11??????PSB1C3 is an expression plasmid which insert sfGFP11 into PSB1C3. before sfGFP11, we add a linker(gatggagggtctggtggcggatca) to achieve our goal.SfGFP11 is a part of GFP(from 214bp to 230bp), GFP has been mutated to improve its solubilityand self-associating activity. When it express, it will emit green fluorescenceslightly under the fluorescence microscope. We try to find anideal protein tag to be work both invivo and invitro and it can provide a sensitive measurable signalwhich don???t need external chemical reagents or substrates. Finally we find away to accomplish this goal?????? dividing GFP into sfGFP1-10and sfGFP11. Either the sfGFP1-10 or sfGFP11 will emit green fluorescence slightly under the fluorescencemicroscope. However, when sfGFP1-10 and sfGFP11 express insame cell, they will interact each other and emit more intense fluorescence thaneach of them. The split GFP system is simple and does not change fusion proteinsolubility. Usage and biology:The split GFP system has manypractical applications. Obtaining soluble, well-folded recombinant proteins fordownstream applications requires screening large numbers of protein variants (mutants,fragments, fusion tags, folding partners) and testing many expression orrefolding conditions.(Ste??phanieCabantous, Thomas C Terwilliger & Geoffrey S Waldo,2005) Part sequence agagaccacatggtccttcatgagtatgtaaatgctgctgggattaca (All the sequence has been testified by Sangon) false false _2476_ 32614 32614 9 true We give up PCR from the gene of GFP due to the fact that to achieve the PCR, GFP gene should be mutated too many times. It is more convinent to chenical synthese directly. false Yin Wu annotation2483449 1 sfGFP11 range2483449 1 25 72 annotation2483448 1 linker range2483448 1 1 24 BBa_K2009602 1 BBa_K2009602 sfGFP11 with promoter and RBS 2016-09-13T11:00:00Z 2016-09-17T07:37:40Z chenical synthesis and link it to BBa_K081005 sfGFP11 length: 112bp Derived from:synthesis from Sangon BBa_J23100: promoter: BBa_B0030: RBS sfGFP11??????PSB1C3 is an expression plasmid which insert sfGFP11 into PSB1C3. before sfGFP11, we add a linker(gatggagggtctggtggcggatca) to achieve our goal.SfGFP11 is a part of GFP(from 214bp to 230bp), GFP has been mutated to improve its solubilityand self-associating activity. When it express, it will emit green fluorescenceslightly under the fluorescence microscope. We try to find anideal protein tag to be work both invivo and invitro and it can provide a sensitive measurable signalwhich don???t need external chemical reagents or substrates. Finally we find away to accomplish this goal?????? dividing GFP into sfGFP1-10and sfGFP11. Either the sfGFP1-10 or sfGFP11 will emit green fluorescence slightly under the fluorescencemicroscope. However, when sfGFP1-10 and sfGFP11 express insame cell, they will interact each other and emit more intense fluorescence thaneach of them. The split GFP system is simple and does not change fusion proteinsolubility. Usage and biology:The split GFP system has manypractical applications. Obtaining soluble, well-folded recombinant proteins fordownstream applications requires screening large numbers of protein variants (mutants,fragments, fusion tags, folding partners) and testing many expression orrefolding conditions.(Ste??phanieCabantous, Thomas C Terwilliger & Geoffrey S Waldo,2005) Part sequence ttgacggctagctcagtcctaggtacagtgctagctactagagattaaagaggagaaatactagagagaccacatggtccttcatgagtatgtaaatgctgctgggattaca (All the sequence has been testified by Sangon) false false _2476_ 32614 32614 9 false If we PCR from GFP directly, we have to mutate it many times and it is more convenient to directly synthese it. false Yin Wu component2483338 1 BBa_K2009820 component2483337 1 BBa_K081005 annotation2483338 1 BBa_K2009820 range2483338 1 67 114 annotation2483337 1 BBa_K081005 range2483337 1 1 58 BBa_B0030 1 BBa_B0030 RBS.1 (strong) -- modified from R. Weiss 2003-01-31T12:00:00Z 2015-08-31T04:07:20Z Released HQ 2013 Strong RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0032</bb_part>, <bb_part>BBa_B0033</bb_part>. false true _44_46_ 0 24 7 In stock false Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix (&quot;orig&quot; in figure 4-14 of Ron Weiss thesis). <p>No secondary structures are formed in the given RBS region. Users should check for secondary structures induced in the RBS by upstream and downstream elements in the +50 to -50 region, as such structures will greatly affect the strength of the RBS. Contact info <a href="mailto:(bchow@media.mit.edu)">Brian Chow</a> true Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003. annotation7025 1 BBa_B0030 range7025 1 1 15 annotation1702 1 RBS range1702 1 8 12 annotation1701 1 RBS-1\Strong range1701 1 1 15 BBa_J23100 1 BBa_J23100 constitutive promoter family member 2006-08-03T11:00:00Z 2015-08-31T04:08:40Z Isolated from library of promoters Released HQ 2013 Replace later false true _52_ 0 483 95 In stock true N/A true John Anderson BBa_K081005 1 BBa_K081005 constitutive promoter family member and RBS 2008-10-17T11:00:00Z 2015-05-08T01:08:34Z Promoter: John Anderson. RBS: Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003. Released HQ 2013 Constitutive promoter (strong) with RBS (strong, efficiency=0.3) false true _227_ 0 2583 9 In stock true We used BioBrick Standard Assembly. true Lorenzo Pasotti, Paolo Magni component1981863 1 BBa_J23100 component1981865 1 BBa_B0030 annotation1981863 1 BBa_J23100 range1981863 1 1 35 annotation1981865 1 BBa_B0030 range1981865 1 44 58 BBa_J23100_sequence 1 ttgacggctagctcagtcctaggtacagtgctagc BBa_K081005_sequence 1 ttgacggctagctcagtcctaggtacagtgctagctactagagattaaagaggagaaa BBa_B0030_sequence 1 attaaagaggagaaa BBa_K2009820_sequence 1 gatggagggtctggtggcggatcaagagaccacatggtccttcatgagtatgtaaatgctgctgggattaca BBa_K2009602_sequence 1 ttgacggctagctcagtcctaggtacagtgctagctactagagattaaagaggagaaatactagagagagaccacatggtccttcatgagtatgtaaatgctgctgggattaca 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