BBa_B0012 1 BBa_B0012 TE from coliphageT7 2003-01-31T12:00:00Z 2015-08-31T04:07:20Z Derived from the TE terminator of T7 bacteriophage between Genes 1.3 and 1.4 <genbank>V01146</genbank>. Released HQ 2013 Transcription terminator for the <i>E.coli</i> RNA polymerase. false false _1_ 0 24 7 In stock false <P> <P>Suggested by Sri Kosuri and Drew Endy as a high efficiency terminator. The 5' end cutoff was placed immediately after the TAA stop codon and the 3' end cutoff was placed just prior to the RBS of Gene 1.4 (before AAGGAG).<P> Use anywhere transcription should be stopped when the gene of interest is upstream of this terminator. false Reshma Shetty annotation1690 1 polya range1690 1 28 41 annotation7020 1 BBa_B0012 range7020 1 1 41 annotation1686 1 T7 TE range1686 1 8 27 annotation1687 1 stop range1687 1 34 34 BBa_K1526001 1 BBa_K1526001 Gsh1*: Mutated Glutamate-Cysteine Ligase(Gsh1) from Saccharomyces cerevisiae 2014-10-04T11:00:00Z 2015-05-08T01:10:50Z http://nar.oxfordjournals.org/content/14/11/4393.full.pdf Glutamate-Cysteine Ligase(previously known as Gamma glutamylcysteine synthetase) catalyzes the first step in glutathione (GSH) biosynthesis; L-glutamate + L-cysteine + ATP -> gamma-glutamyl cysteine + ADP + Pi Expression is induced by oxidants, cadmium, and mercury. Protein abundance increases in response to DNA replication stress. (http://biocyc.org/YEAST/NEW-IMAGE?type=GENE-IN-MAP-IN-PWY&object=YJL101C) Gsh1* can be used to overproduce cysteine to make gamma-glutamyl cysteine, which is the monomer that forms phytochelatins (n=10-20). In order to do this, glutamate-cysteine ligase is needed to catalyse the reaction. We can overexpress GSH1* using the pYodA(BBa K896008) promoter. false false _1908_ 0 21661 9 In stock false The original gene(Gsh1/GshA) was mutated to obtain Gsh1*. false Ruth Haley annotation2427850 1 Start Codon range2427850 1 25 27 annotation2427849 1 BBa_B0034 range2427849 1 7 18 annotation2427852 1 Stop range2427852 1 1579 1581 annotation2427851 1 Gsh1* range2427851 1 25 1581 BBa_K1526005 1 BBa_K1526005 LacRS + Gsh1* 2014-10-06T11:00:00Z 2015-05-08T01:10:50Z E coli LacI + Gsh1 false false _1908_ 0 12384 9 It's complicated false codon optimized for e coli false Cau?? Westmann component2427890 1 BBa_K1526009 component2427895 1 BBa_K1526001 annotation2427895 1 BBa_K1526001 range2427895 1 1567 3147 annotation2427890 1 BBa_K1526009 range2427890 1 1 1558 BBa_P0312 1 BBa_P0312 PoPS -> LacI [S0156] 2004-04-26T11:00:00Z 2015-05-08T01:14:10Z Released HQ 2013 -- No description -- false false _1_ 0 24 7 In stock false true Randy Rettberg component944617 1 BBa_C0012 component944606 1 BBa_B0033 component944634 1 BBa_B0012 component944624 1 BBa_B0010 annotation944624 1 BBa_B0010 range944624 1 1179 1258 annotation944617 1 BBa_C0012 range944617 1 18 1145 annotation944606 1 BBa_B0033 range944606 1 1 11 annotation944634 1 BBa_B0012 range944634 1 1267 1307 BBa_B0033 1 BBa_B0033 RBS.4 (weaker) -- derivative of BBa_0030 2003-01-31T12:00:00Z 2015-08-31T04:07:20Z Released HQ 2013 Weaker RBS based on Ron Weiss thesis. Strengths relative to <bb_part>BBa_B0030</bb_part>, <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0032</bb_part>. false true _41_44_48_46_1_ 0 24 7 In stock false Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix (&quot;RBS-3&quot; in figure 4-14 of 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 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. annotation7028 1 BBa_B0033 range7028 1 1 11 annotation1714 1 RBS range1714 1 7 10 annotation1713 1 RBS-4\Weaker range1713 1 1 11 BBa_R0010 1 LacI promoter (lacI regulated) 2003-01-31T12:00:00Z 2015-05-08T01:14:14Z The Plac insert was PCR'd from the MG1655 strain of E.coli K12. Released HQ 2013 Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) <p> The pLac regulatory region is a 243 base-pair sequence with standard BioBrick prefix and suffix sections on its ends. It contains two protein binding sites: CAP, which is generally present in E.coli and is assocciated with cell health and availability of glucose., and LacI, the Lac inhibitor <bb_part>BBa_C0010</bb_part> which binds in an dimerized cooperative manner to inhibit the transcription of the protein that follows. In the presence of lactose or IPTG, an analog of lactose, LacI is unable to correctly bind and inhibit transcription. This allows <bb_part>BBa_R0010</bb_part> to be used as a inverter or as a detector of lactose or IPTG. false true _1_ 0 24 7 In stock false <P> <P><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 this regulator. This part is incompatible with environments containing lactose or lactose analogs. true annotation1961227 1 start range1961227 1 173 173 annotation1961221 1 end of LacI coding region (inactive) range1961221 1 1 88 annotation1961225 1 -10 range1961225 1 161 166 annotation1961226 1 LacI binding site range1961226 1 166 200 annotation1961223 1 CAP binding site range1961223 1 89 126 annotation1961222 1 BBa_R0010 range1961222 1 1 200 annotation1961224 1 -35 range1961224 1 137 142 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_K1526009 1 BBa_K1526009 Lac repressor System (LacRS) 2014-10-06T11:00:00Z 2015-05-08T01:10:50Z E. coli Lac operon false false _1908_ 0 12384 9 It's complicated false Codon optimized to E. coli false Hanna Esser component2419778 1 BBa_J23100 component2419792 1 BBa_P0312 component2419793 1 BBa_R0010 annotation2419792 1 BBa_P0312 range2419792 1 44 1350 annotation2419793 1 BBa_R0010 range2419793 1 1359 1558 annotation2419778 1 BBa_J23100 range2419778 1 1 35 BBa_C0012 1 lacI lacI repressor from E. coli (+LVA) 2003-01-31T12:00:00Z 2015-08-31T04:07:23Z represillator of Elowitz and Leibler (2000) Released HQ 2013 Coding region for the LacI protein with an LVA degradation tail and without an RBS. LacI binds to the pLac regulator <bb_part>BBa_R0010</bb_part> and PLlac01 hybrid regulator <bb_part>BBa_R0011</bb_part> and inhibits transcription. IPTG (Isopropylthiogalactoside) binds to LacI and inhibits its operation, therefore promoting transcription.</P> <P>A rapid degredation tail (LVA) has been added to improve the High to Low performance of this part.</P> false false _1_ 0 24 7 In stock false References (unparsed) here: <p>Elowitz, M.B., Leibler, S. A synthetic oscillatory network of transcriptional regulators. <em>Nature</em> 403, 335-338 (2000). <a href="http://biobricks.ai.mit.edu/BB_References.htm#ELOW00">[ELOW00]</a><br> <br> </P> <P> References (unparsed) here: <p>Elowitz, M.B., Leibler, S. A synthetic oscillatory network of transcriptional regulators. <em>Nature</em> 403, 335-338 (2000). <a href="http://biobricks.ai.mit.edu/BB_References.htm#ELOW00">[ELOW00]</a><br> <br> </P> <P>Sequence taken from the repressilator of Elowitz and Leibler (2000). The obtained sequence was compared to the wild-type sequence for LacI obtained through a database search. The sequence had been modified from the wild-type in that wild-type GTG start was changed to an ATG start (note, actual ORF in E.coli has several GTG starts it would seem). The LVA tag has been added for quicker degradation.<P> Incompatible with systems containing LacI, lactose, or IPTG. true Grace Kenney, Daniel Shen, Neelaksh Varshney, Samantha Sutton annotation7031 1 BBa_C0012 range7031 1 1 1128 annotation2213988 1 Help:Barcodes range2213988 1 1129 1153 annotation1722 1 LVA range1722 1 1090 1128 annotation1723 1 lacI-LVA range1723 1 1 1128 BBa_B0010 1 BBa_B0010 T1 from E. coli rrnB 2003-11-19T12:00:00Z 2015-08-31T04:07:20Z Transcriptional terminator consisting of a 64 bp stem-loop. false false _1_ 0 24 7 In stock false true Randy Rettberg annotation4184 1 stem_loop range4184 1 12 55 annotation7018 1 BBa_B0010 range7018 1 1 80 BBa_J23100_sequence 1 ttgacggctagctcagtcctaggtacagtgctagc BBa_B0010_sequence 1 ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctc BBa_B0033_sequence 1 tcacacaggac BBa_R0010_sequence 1 caatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacaca BBa_K1526001_sequence 1 tactagaaagaggagaaatactagatgatccctgatgtgtcacaagccctggcctggctggaaaaacatccgcaggctctgaaaggcattcagcgtggacttgaacgcgaaactctccgcgtaaatgcggacggtacgctggcgacgaccggccatccggaagccttaggctctgcgcttacgcacaaatggatcacgaccgatttcgctgaggccttgctcgaatttattacaccggtggatggcgacattgaacatatgctgacattcatgcgcgacttgcatcggtatacagcgcgtaacatgggagacgaacggatgctcccgctgagcatgccgtgctatatcgcggaaggtcaggacattgaactggcccaatatggtaccagcaacacgggccgctttaagaccctgtatcgcgaagggctgaaaaaccgttacggtgcactgatgcagacaatttccggagtgcactataacttctcactgccaatggcgttctggcaggcgaaatgtggcgacatctctggcgctgacgcgaaagaaaaaatctccgcgggttatttccgcgtgattcgcaactactaccgtttcggctgggttattccgtatctttttggggccagcccggctatttgttccagttttctccaaggcaaaccaacttccctgccgtttgaaaaaacggaatgcggtatgtactacctcccgtacgcaaccagcttacgtctgagcgatctgggctataccaacaaatctcaatcaaatctgggcattacgtttaacgatctttacgaatatgtggcagggctgaaacaggcgattaaaaccccctcggaagaatatgccaaaattggaatcgagaaggacgggaagcgcctacagattaatagtaacgtcctacagatcgaaaacgagctgtacgccccgatccgtcctaaacgtgtttcccgctcgggcgaaagtccgtcagatgccttacttcgtggcggcattgagtatatcgaggttcgctccctggacatcaaccctttcagtcccattggggtggatgaacagcaggtgcgcttcctggacttgtttatggtctggtgtgcgttggcggacgcgcctgaaatgtcttcatcggagctggcgtgtacacgcgtgaactggaatcgcgtgatcctggaaggccgcaaacctggcctgaccctgggtattgggtgcgagacagcccaattcccactccctcaagtgggtaaagacctgttccgtgatttaaaacgcgtcgcgcaaaccctcgactctattaatggtggtgaggcgtatcagaaggtgtgcgacgaacttgtcgcttgttttgataacccggatcttacctttagtgcccgtattttgcgtagcatgatcgataccggaatcggtggcacaggtaaagcgtttgcagaagcatatcggaatttgctgcgcgaggagccgctggaaatcctgcgcgaggaagattttgtagcggaacgtgaagggagcgagcgccgtcagcaggagatggaagctgctgatacggaaccattcgccgtctggttagaaaaacacgcttga BBa_P0312_sequence 1 tcacacaggactactagatggtgaatgtgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaagacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggatatctcggtagtgggatacgacgataccgaagacagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcaggctgcaaacgacgaaaactacgctttagtagcttaataactctgatagtgctagtgtagatctctactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata BBa_C0012_sequence 1 atggtgaatgtgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaagacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggatatctcggtagtgggatacgacgataccgaagacagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcaggctgcaaacgacgaaaactacgctttagtagcttaataactctgatagtgctagtgtagatctc BBa_K1526009_sequence 1 ttgacggctagctcagtcctaggtacagtgctagctactagagtcacacaggactactagatggtgaatgtgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaagacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggatatctcggtagtgggatacgacgataccgaagacagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcaggctgcaaacgacgaaaactacgctttagtagcttaataactctgatagtgctagtgtagatctctactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagcaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacaca BBa_B0012_sequence 1 tcacactggctcaccttcgggtgggcctttctgcgtttata BBa_K1526005_sequence 1 ttgacggctagctcagtcctaggtacagtgctagctactagagtcacacaggactactagatggtgaatgtgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaagacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggatatctcggtagtgggatacgacgataccgaagacagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcaggctgcaaacgacgaaaactacgctttagtagcttaataactctgatagtgctagtgtagatctctactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagcaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacacatactagagtactagaaagaggagaaatactagatgatccctgatgtgtcacaagccctggcctggctggaaaaacatccgcaggctctgaaaggcattcagcgtggacttgaacgcgaaactctccgcgtaaatgcggacggtacgctggcgacgaccggccatccggaagccttaggctctgcgcttacgcacaaatggatcacgaccgatttcgctgaggccttgctcgaatttattacaccggtggatggcgacattgaacatatgctgacattcatgcgcgacttgcatcggtatacagcgcgtaacatgggagacgaacggatgctcccgctgagcatgccgtgctatatcgcggaaggtcaggacattgaactggcccaatatggtaccagcaacacgggccgctttaagaccctgtatcgcgaagggctgaaaaaccgttacggtgcactgatgcagacaatttccggagtgcactataacttctcactgccaatggcgttctggcaggcgaaatgtggcgacatctctggcgctgacgcgaaagaaaaaatctccgcgggttatttccgcgtgattcgcaactactaccgtttcggctgggttattccgtatctttttggggccagcccggctatttgttccagttttctccaaggcaaaccaacttccctgccgtttgaaaaaacggaatgcggtatgtactacctcccgtacgcaaccagcttacgtctgagcgatctgggctataccaacaaatctcaatcaaatctgggcattacgtttaacgatctttacgaatatgtggcagggctgaaacaggcgattaaaaccccctcggaagaatatgccaaaattggaatcgagaaggacgggaagcgcctacagattaatagtaacgtcctacagatcgaaaacgagctgtacgccccgatccgtcctaaacgtgtttcccgctcgggcgaaagtccgtcagatgccttacttcgtggcggcattgagtatatcgaggttcgctccctggacatcaaccctttcagtcccattggggtggatgaacagcaggtgcgcttcctggacttgtttatggtctggtgtgcgttggcggacgcgcctgaaatgtcttcatcggagctggcgtgtacacgcgtgaactggaatcgcgtgatcctggaaggccgcaaacctggcctgaccctgggtattgggtgcgagacagcccaattcccactccctcaagtgggtaaagacctgttccgtgatttaaaacgcgtcgcgcaaaccctcgactctattaatggtggtgaggcgtatcagaaggtgtgcgacgaacttgtcgcttgttttgataacccggatcttacctttagtgcccgtattttgcgtagcatgatcgataccggaatcggtggcacaggtaaagcgtttgcagaagcatatcggaatttgctgcgcgaggagccgctggaaatcctgcgcgaggaagattttgtagcggaacgtgaagggagcgagcgccgtcagcaggagatggaagctgctgatacggaaccattcgccgtctggttagaaaaacacgcttga 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