BBa_B0034 1 BBa_B0034 RBS (Elowitz 1999) -- defines RBS efficiency 2003-01-31T12:00:00Z 2015-08-31T04:07:20Z Released HQ 2013 RBS based on Elowitz repressilator. false true _1_ 0 24 7 In stock false Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix. <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 for this part: <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. annotation23325 1 conserved range23325 1 5 8 BBa_K215251 1 BBa_K215251 Lpp + 5tmr OmpA + GS Linker + TEV site + NheI Site 2009-09-29T11:00:00Z 2015-05-08T01:11:30Z Harvard iGEM 2006: BBa_J36848. We added a GS Linker and TEV site. This is a fusion protein that displays your favorite gene inserted into the NheI site. See BBa_K215250 for 1tmr OmpA. false false _320_ 0 5567 9 Not in stock false Anything inserted into the NheI shouldn't have a start codon and gene should still be in frame. false Alex Leone annotation2027619 1 GS Linker - GGGSGGGSGGG range2027619 1 436 468 annotation2027620 1 TEV Site - ENLYFQG range2027620 1 469 489 annotation2027618 1 BBa_J36837 - 5tmr OmpA range2027618 1 94 435 annotation2027617 1 BBa_J36835 - Lpp Signaling Peptide range2027617 1 1 87 annotation2027643 1 NheI range2027643 1 490 495 BBa_K215201 1 BBa_K215201 Generic Surface Display Construct: IPTG Inducible Lpp + 5tmr OmpA + NheI Site 2009-09-29T11:00:00Z 2015-05-08T01:11:30Z The Lpp + 5tmr OmpA is from Havard iGEM 2006: BBa_J36850, which is from [cite]. This is a generic surface display construct that displays a protein with the OmpA surface display system as described in [cite]. To insert a gene, you will need to design a primer that removes the start codon and adds a XbaI site that keeps the rest of the gene in frame with OmpA. This assumes that the gene is a biobrick with the standard suffix following the gene. If you are creating the gene from scratch, biobrick it then follow this guide so that there is the standard suffix following the gene. For example, assume our gene (E0040) starts with: 5'-atgcgtaaaggagaagaacttt...-3' The design process would be: # Start with the XbaI site: 5'-TCTAGA-3' # Add ~20bp of the gene immediately after the XbaI site, starting _after_ the start codon (atg), and try to end on a G or C: 5'-TCTAGAcgtaaaggagaagaacttt-3' for E0040 from above. These ~20bp will determine the melting temperature for the primer. # Add 6-8 random nucleotides at the start. Try to balance out the primer to ~%50 g&c to a&t: 5'-cgggcTCTAGAcgtaaaggagaagaacttt-3' # Tweak the number of nucleotides until the melting point roughly matches that of Vr. Then to add the gene to the construct, again assuming the gene is a biobrick with standard suffix: # PCR with the designed forward primer and Vr # Then run the PCR product in a digest with XbaI and PstI, and digest this part (the display construct) with NheI and PstI. The XbaI site has a sticky end that binds with NheI. # Standard ligation and transformation. false false _320_ 0 5567 9 It's complicated true We took the Lpp Signaling Peptide and OmpA 5 trans-membrane region from Havard 2006, BBa_J368450, with two custom primers: * Lpp Forward: 5'-gcggccgctTCTAGAtgaaagctactaaactggtactggg-3' * OmpA Reverse with GS Linker, GGGSGGGSGGG, and a TEV site, ENLYFQG, and SpeI site: 5'-cggccgctACTAGTaGCTAGCaccctgaaaatacaggttttcACCACCACCAGAACCACCACCAGAACCACCACCgctgcctttgtacggcatacgacc-3' false Alex Leone component2027650 1 BBa_B0034 component2027656 1 BBa_K215251 component2027644 1 BBa_R0011 annotation2027656 1 BBa_K215251 range2027656 1 82 576 annotation2027644 1 BBa_R0011 range2027644 1 1 54 annotation2027650 1 BBa_B0034 range2027650 1 64 75 BBa_R0011 1 lacI+pL Promoter (lacI regulated, lambda pL hybrid) 2003-01-31T12:00:00Z 2015-05-08T01:14:14Z represillator of Elowitz and Leibler (2000) Released HQ 2013 Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) <p> The PLlac 0-1 promoter is a hybrid regulatory region consisting of the promoter P(L) of phage lambda with the cI binding sites replaced with lacO1. The hybrid design allows for strong promotion that can nevertheless be tightly repressed by LacI, the Lac inhibitor (i.e. repressor) (<bb_part>BBa_C0010</bb_part>) ([LUTZ97]). The activity of the promoter can be regulated over a >600-fold range by IPTG in E.Coli DH5-alpha-Z1 (same paper reference). false true _1_ 0 24 7 In stock false <P> <P>hybrid promoter design to create strong promoter that is, at the same time, highly repressible. note that the upstream operator installed in this hybrid is slightly different than the one in the original source (Lutz and Bujard, 1997). the most upstream operator region is slightly truncated in the represillator version, so that both operators in the hybrid are the same sequence. see references for details. also, the sequence has been truncated after the transcriptional start site.<P>LacI binds to this regulator. This part is incompatible with species containing active LacI coding regions. Lactose and IPTG disable the operation of LacI and increase transcription. This part is incompatible with environments containing lactose or lactose analogs. true Neelaksh Varshney, Grace Kenney, Daniel Shen, Samantha Sutton annotation1999 1 lac O1 range1999 1 3 19 annotation7064 1 BBa_R0011 range7064 1 1 54 annotation2000 1 -35 range2000 1 20 25 annotation2001 1 lac O1 range2001 1 26 42 annotation2002 1 -10 range2002 1 43 48 BBa_B0034_sequence 1 aaagaggagaaa BBa_K215201_sequence 1 aattgtgagcggataacaattgacattgtgagcggataacaagatactgagcacatactagagaaagaggagaaatactagatgaaagctactaaactggtactgggcgcggtaatcctgggttctactctgctggcaggttgctccagcaacgctaaaatcgatcagactagaaacccgtatgttggctttgaaatgggttacgactggttaggtcgtatgccgtacaaaggcagcgttgaaaacggtgcatacaaagctcagggcgttcaactgaccgctaaactgggttacccaatcactgacgacctggacatctacactcgtctgggtggcatggtatggcgtgcagacactaaatccaacgtttatggtaaaaaccacgacaccggcgtttctccggtcttcgctggcggtgttgagtacgcgatcactcctgaaatcgctacccgtctggaataccagtggaccaacaacatcggtgacgcacacaccatcggcactcgtccggacaacggtggtggttctggtggtggttctggtggtggtgaaaacctgtattttcagggtgctagc BBa_K215251_sequence 1 atgaaagctactaaactggtactgggcgcggtaatcctgggttctactctgctggcaggttgctccagcaacgctaaaatcgatcagactagaaacccgtatgttggctttgaaatgggttacgactggttaggtcgtatgccgtacaaaggcagcgttgaaaacggtgcatacaaagctcagggcgttcaactgaccgctaaactgggttacccaatcactgacgacctggacatctacactcgtctgggtggcatggtatggcgtgcagacactaaatccaacgtttatggtaaaaaccacgacaccggcgtttctccggtcttcgctggcggtgttgagtacgcgatcactcctgaaatcgctacccgtctggaataccagtggaccaacaacatcggtgacgcacacaccatcggcactcgtccggacaacggtggtggttctggtggtggttctggtggtggtgaaaacctgtattttcagggtgctagc BBa_R0011_sequence 1 aattgtgagcggataacaattgacattgtgagcggataacaagatactgagcaca 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