BBa_K511000 1 BBa_K511000 Hef1a-LacOid Promoter MammoBlock 2011-09-24T11:00:00Z 2015-05-08T01:12:31Z This part is a synthetic promoter formed from the human elongation factor 1a promoter sequence and a LacOid operator that has been computationally shown to bind to the LacI repressor protein more tightly than the wildtype LacO operator. This part encodes a promoter with low-level, constitutive activity that can be repressed by variants of the LacI transcriptional repressor. As shown in Figures 1 and 2 below, cotransfection of a construct containing a yellow fluorescent protein (EYFP) driven by the Hef1a-LacO promoter with a construct containing a LacI-KRAB transcriptional repressor driven by the TRE-Tight promoter proves that it is possible to repress Hef1a-LacO expression with the LacI-KRAB LacI variant, which includes a Kruppel-associated box motif previously shown to silence chromatin and thereby inhibit transcription. false false _674_ 0 6719 9 It's complicated true When sequencing constructs that include this part, it is important to realize that the Hef1a-LacOid promoter is roughly a kilobase long; therefore, promoter-gene pairs will require either internal sequencing primers or sequencing from the gene side of the construct to be useful. false Grant Robinson annotation2139540 1 Hef1-alpha Promoter range2139540 1 1 1275 annotation2139541 1 Lac O1 range2139541 1 201 221 annotation2139542 1 Lac Oid range2139542 1 272 291 BBa_J85600 1 BBa_J85600 attB1 recombination site 2011-09-05T11:00:00Z 2015-05-08T01:08:30Z lambda phage attB1 recombination site Used for creating mammoblock (RFC65) composite promoter-gene pairs using recombination-based cloning. false false _406_ 0 106 406 Not in stock false None false Ron Weiss annotation2125966 1 attB1 range2125966 1 2 22 BBa_K511904 1 BBa_K511904 Repressible LacI Repressor Generator (Hef1a-LacO-LacI) MammoBlock Device 2011-09-27T11:00:00Z 2015-05-08T01:12:31Z Consult constituent components. This MammoBlock composite device produces the LacI repressor protein at a low, constitutive level that can be repressed by LacI variants. As a result, this device is expected to either limits its own production constant level or to oscillate; its behavior remains to be interrogated in future experiments. false false _674_ 0 6719 9 Not in stock false In order to adhere to the recombination-based MammoBlock standard, and because shipping Biosafety Level 2 mammalian expression vectors to the Registry of Standard Biological Parts is not currently possible, we are providing the sequence resulting from Gateway recombination of the constituent MammoBlock parts here without redistributing the physical DNA plasmids to the Registry. In future years, we hope to make use of improvements in the Registry's accommodations to provide our complete expression vectors. false Grant Robinson component2146354 1 BBa_K511000 component2146356 1 BBa_J85600 component2146359 1 BBa_K511204 annotation2146359 1 BBa_K511204 range2146359 1 1298 2404 annotation2146356 1 BBa_J85600 range2146356 1 1276 1297 annotation2146354 1 BBa_K511000 range2146354 1 1 1275 BBa_K511204 1 BBa_K511204 LacI Repressor MammoBlock 2011-09-26T11:00:00Z 2015-05-08T01:12:31Z This protein is a mammalian codon-optimized version of the LacI repressor originally derived from Escherichia coli. This part encodes a repressor protein that binds tightly to the lactose operator originating from Escherichia coli. This protein has been codon-optimized for mammalian expression and is capable of exerting moderate repression on promoters with internal lactose operator sites. Figures 1 and 2 below show typical levels of repression mediated by the LacI repressor. false false _674_ 0 6719 9 It's complicated false Not applicable. false Grant Robinson annotation2143781 1 Nuclear Localization Sequence range2143781 1 1080 1101 annotation2143755 1 LacI Repressor range2143755 1 1 1101 BBa_J85600_sequence 1 ccaagtttgtacaaaaaagcag BBa_K511000_sequence 1 tggctccggtgcccgtcagtgggcagagcgcacatcgcccacagtccccgagaagttggggggaggggtcggcaattgaaccggtgcctagagaaggtggcgcggggtaaactgggaaagtgatgtcgtgtactggctccgcctttttcccgagggtgggggagaaccgtatataagtgcagtagtcgccgtgaagctagaattgtgagcggataacaattccacagtcgaccctaggttgtgtcgcgagtgttggatcccagctgacaccaattgtgagcgctcacaattgctagccgttctttttcgcaacgggtttgccgccagaacacaggtaagtgccgtgtgtggttcccgcgggcctggcctctttacgggttatggcccttgcgtgccttgaattacttccacctggctgcagtacgtgattcttgatcccgagcttcgggttggaagtgggtgggagagttcgaggccttgcgcttaaggagccccttcgcctcgtgcttgagttgaggcctggcctgggcgctggggccgccgcgtgcgaatctggtggcaccttcgcgcctgtctcgctgctttcgataagtctctagccatttaaaatttttgatgacctgctgcgacgctttttttctggcaagatagtcttgtaaatgcgggccaagatctgcacactggtatttcggtttttggggccgcgggcggcgacggggcccgtgcgtcccagcgcacatgttcggcgaggcggggcctgcgagcgcggccaccgagaatcggacgggggtagtctcaagctggccggcctgctctggtgcctggcctcgcgccgccgtgtatcgccccgccctgggcggcaaggctggcccggtcggcaccagttgcgtgagcggaaagatggccgcttcccggccctgctgcagggagctcaaaatggaggacgcggcgctcgggagagcgggcgggtgagtcacccacacaaaggaaaagggcctttccgtcctcagccgtcgcttcatgtgactccacggagtaccgggcgccgtccaggcacctcgattagttctcgagcttttggagtacgtcgtctttaggttggggggaggggttttatgcgatggagtttccccacactgagtgggtggagactgaagttaggccagcttggcacttgatgtaattctccttggaatttgccctttttgagtttggatcttggttcattctcaagcctcagacagtggttcaaagtttttttcttccatttcaggt BBa_K511904_sequence 1 tggctccggtgcccgtcagtgggcagagcgcacatcgcccacagtccccgagaagttggggggaggggtcggcaattgaaccggtgcctagagaaggtggcgcggggtaaactgggaaagtgatgtcgtgtactggctccgcctttttcccgagggtgggggagaaccgtatataagtgcagtagtcgccgtgaagctagaattgtgagcggataacaattccacagtcgaccctaggttgtgtcgcgagtgttggatcccagctgacaccaattgtgagcgctcacaattgctagccgttctttttcgcaacgggtttgccgccagaacacaggtaagtgccgtgtgtggttcccgcgggcctggcctctttacgggttatggcccttgcgtgccttgaattacttccacctggctgcagtacgtgattcttgatcccgagcttcgggttggaagtgggtgggagagttcgaggccttgcgcttaaggagccccttcgcctcgtgcttgagttgaggcctggcctgggcgctggggccgccgcgtgcgaatctggtggcaccttcgcgcctgtctcgctgctttcgataagtctctagccatttaaaatttttgatgacctgctgcgacgctttttttctggcaagatagtcttgtaaatgcgggccaagatctgcacactggtatttcggtttttggggccgcgggcggcgacggggcccgtgcgtcccagcgcacatgttcggcgaggcggggcctgcgagcgcggccaccgagaatcggacgggggtagtctcaagctggccggcctgctctggtgcctggcctcgcgccgccgtgtatcgccccgccctgggcggcaaggctggcccggtcggcaccagttgcgtgagcggaaagatggccgcttcccggccctgctgcagggagctcaaaatggaggacgcggcgctcgggagagcgggcgggtgagtcacccacacaaaggaaaagggcctttccgtcctcagccgtcgcttcatgtgactccacggagtaccgggcgccgtccaggcacctcgattagttctcgagcttttggagtacgtcgtctttaggttggggggaggggttttatgcgatggagtttccccacactgagtgggtggagactgaagttaggccagcttggcacttgatgtaattctccttggaatttgccctttttgagtttggatcttggttcattctcaagcctcagacagtggttcaaagtttttttcttccatttcaggtccaagtttgtacaaaaaagcagatgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaagacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggatatctcggtagtgggatacgacgataccgaagacagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagccaaaaaagaagagaaaggtctaatag BBa_K511204_sequence 1 atgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaagacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggatatctcggtagtgggatacgacgataccgaagacagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagccaaaaaagaagagaaaggtctaatag 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