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_E0040
1
GFP
green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212
2004-09-29T11:00:00Z
2016-01-26T02:09:38Z
Released HQ 2013
GFP (mut3b) [note that this part does not have a barcode]
false
true
_11_1_
4206
61
7
In stock
false
true
jcbraff
annotation1934520
1
GFP protein
range1934520
1
1
720
BBa_R0040
1
p(tetR)
TetR repressible promoter
2003-01-31T12:00:00Z
2015-05-08T01:14:14Z
Lutz, R., Bujard, H., <em>Nucleic Acids Research</em> (1997) 25, 1203-1210.
Released HQ 2013
Sequence for pTet inverting regulator driven by the TetR protein.</P>
false
true
_1_
0
24
7
In stock
false
<P> <P>BBa_R0040 TetR-Regulated Promoter is based on a cI promoter. It has been modified to include two TetR binding sites and the BioBrick standard assembly head and tail restriction sites.<P>
true
June Rhee, Connie Tao, Ty Thomson, Louis Waldman
annotation1986784
1
BBa_R0040
range1986784
1
1
54
annotation1986787
1
-10
range1986787
1
43
48
annotation1986785
1
-35
range1986785
1
20
25
annotation1986783
1
TetR 1
range1986783
1
1
19
annotation1986786
1
TetR 2
range1986786
1
26
44
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
annotation2000
1
-35
range2000
1
20
25
annotation1999
1
lac O1
range1999
1
3
19
annotation7064
1
BBa_R0011
range7064
1
1
54
annotation2002
1
-10
range2002
1
43
48
annotation2001
1
lac O1
range2001
1
26
42
BBa_C0161
1
luxI
autoinducer synthetase for AHL (no LVA)
2004-05-26T11:00:00Z
2015-08-31T04:07:24Z
Released HQ 2013
same as C0061 except no LVA tag
false
true
_11_1_
0
61
7
In stock
false
true
jcbraff
annotation1891615
1
LuxI
range1891615
1
1
585
BBa_C0062
1
luxr
luxR repressor/activator, (no LVA?)
2003-01-31T12:00:00Z
2015-08-31T04:07:23Z
<em>V. fischeri</em> <genbank>AF170104</genbank>
Released HQ 2013
In complex with HSL, LuxR binds to the Lux promoter, activating transcription from Pr <bb_part>BBa_R0062</bb_part>, and repressing transcription from Pl <bb_part>BBa_R0063</bb_part>. <p>The lux cassette of V. fischeri contains a left and a right promoter. The right promoter gives weak constitutive expression of downstream genes.This expression is up-regulated by the action of the Lux activator, LuxR complexed to HSL. Two molecules of LuxR protein form a complex with two molecules the signalling compound homoserine lactone (HSL). This complex binds to a palindromic site on the promoter, increasing the rate of transcription.</p>
false
true
_1_
0
24
7
In stock
false
<P> <P>2 silent point mutants were introduced in the coding sequence to remove internal XbaI and PstI sites. Mutation sites were chosen to replace codons commonly used in <em>E. coli</em> with codons used at a similar frequency. <P>
true
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr
annotation2213986
1
Help:Barcodes
range2213986
1
757
781
annotation1765
1
A
range1765
1
492
492
annotation1762
1
prefix
range1762
1
1
2
annotation1764
1
T
range1764
1
174
174
annotation7039
1
BBa_C0062
range7039
1
1
756
annotation1766
1
luxR
range1766
1
1
750
BBa_K1499503
1
BBa_K1499503
quorum sensing machinery that activates GFP expression with extra terminator
2014-10-07T11:00:00Z
2015-05-08T01:10:46Z
BBa_K1499500
This is the same part as Part:BBa_K1499500, except that it has two more terminators between the luxR gene and the luxPR promoter.
false
false
_1879_
0
22124
9
It's complicated
false
After ligating the BBa_K1499500 construct together and into the BioBrick backbone, we transformed it into the NEB 5-alpha strain of E. coli. The colonies that fluoresced were most likely to have been successfully transformed, and thus these were sent off for sequencing. The sequencing data showed that our construct was correct (see below), so we were able to submit for BioBricking.
Lux sequencing data.jpg
Although our sequencing data was good, we realized that we had not induced the lac-repressible promoter, and thus the colonies should not have been fluorescing yet. After doing more research, we realized that the strain of E. coli we were using was not lactose deficient, thus the quorum sensing cascade was being activated by the natural presence of lactose. We ordered a lacI^q strain of E. coli, which are lac deficient, so that we could have complete control of GFP expression. However, when we performed the transformation of the construct into this strain, all of the colonies still fluoresced.
After doing further research, we discovered that the luxPR promoter (BBa_R0062) [2], will induce backward transcription if the luxR protein is present but the AHL molecule is not. In the lac deficient strain of E. coli, AHL is not produced without IPTG induction, but luxR expression is controlled by the constitutive ptet promoter. We hypothesize that since luxR was present in these cells without AHL, the backward transcription from luxPR hindered the function of the terminators that separated the luxR gene from the GFP gene. Thus GFP may have been expressed because the ptet promoter, which is constitutive, was able to affect its expression.
In order to bypass this issue, inserted two more terminators between the luxR gene and the luxPR promoter, in hopes that this would stop the backward transcription from having an effect on GFP expression, allowing us to work towards measuring the time delay created by our construct. The double-promoter did help reduce the GFP expression, though not completely.
false
Aryo Sorayya
component2411386
1
BBa_B0012
component2411405
1
BBa_B0012
component2411383
1
BBa_C0161
component2411384
1
BBa_B0010
component2411421
1
BBa_B0032
component2411390
1
BBa_R0040
component2411416
1
BBa_R0062
component2411424
1
BBa_E0040
component2411427
1
BBa_B0012
component2411396
1
BBa_B0034
component2411425
1
BBa_B0010
component2411375
1
BBa_R0011
component2411399
1
BBa_C0062
component2411403
1
BBa_B0010
component2411381
1
BBa_B0034
component2411411
1
BBa_B0012
component2411409
1
BBa_B0010
annotation2411403
1
BBa_B0010
range2411403
1
1681
1760
annotation2411411
1
BBa_B0012
range2411411
1
1906
1946
annotation2411425
1
BBa_B0010
range2411425
1
2765
2844
annotation2411381
1
BBa_B0034
range2411381
1
64
75
annotation2411386
1
BBa_B0012
range2411386
1
763
803
annotation2411383
1
BBa_C0161
range2411383
1
82
666
annotation2411405
1
BBa_B0012
range2411405
1
1769
1809
annotation2411390
1
BBa_R0040
range2411390
1
812
865
annotation2411396
1
BBa_B0034
range2411396
1
874
885
annotation2411384
1
BBa_B0010
range2411384
1
675
754
annotation2411424
1
BBa_E0040
range2411424
1
2037
2756
annotation2411375
1
BBa_R0011
range2411375
1
1
54
annotation2411409
1
BBa_B0010
range2411409
1
1818
1897
annotation2411416
1
BBa_R0062
range2411416
1
1955
2009
annotation2411399
1
BBa_C0062
range2411399
1
892
1647
annotation2411427
1
BBa_B0012
range2411427
1
2853
2893
annotation2411421
1
BBa_B0032
range2411421
1
2018
2030
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_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
annotation1686
1
T7 TE
range1686
1
8
27
annotation1690
1
polya
range1690
1
28
41
annotation1687
1
stop
range1687
1
34
34
annotation7020
1
BBa_B0012
range7020
1
1
41
BBa_B0032
1
BBa_B0032
RBS.3 (medium) -- derivative of BBa_0030
2003-01-31T12:00:00Z
2015-08-31T04:07:20Z
Released HQ 2013
Weak1 RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0030</bb_part>, <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0033</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 ("RBS-2" in figure 4-14 of thesis). <P>
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.
annotation1709
1
RBS-3\Weak
range1709
1
1
13
annotation1710
1
RBS
range1710
1
7
10
annotation7027
1
BBa_B0032
range7027
1
1
13
BBa_R0062
1
lux pR
Promoter (luxR & HSL regulated -- lux pR)
2003-01-31T12:00:00Z
2015-05-08T01:14:15Z
<em>V. fischeri</em>
Released HQ 2013
Promoter activated by LuxR in concert with HSL</p> <p>The lux cassette of V. fischeri contains a left and a right promoter. The right promoter gives weak constitutive expression of downstream genes.This expression is up-regulated by the action of the LuxR activator protein complexed with the autoinducer, 3-oxo-hexanoyl-HSL. Two molecules of LuxR protein form a complex with two molecules of the signalling compound homoserine lactone (HSL). This complex binds to a palindromic site on the promoter, increasing the rate of transcription.
false
true
_1_
0
24
7
In stock
false
<P> <P>This promoter is based on the <em>Vibrio fischeri </em>quorum sensing gene promoters. Two genes LuxI and LuxR and transcribed in opposite directions as shown below. The original sequence from which the parts <bb_part>BBa_R0062</bb_part> and <bb_part>BBa_R0063</bb_part> were derived is shown in the picture below. <p><img src="<bb_file>Image1.gif</bb_file>" width="614" height="362"><P>
true
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr
annotation2046
1
-35
range2046
1
20
25
annotation2045
1
LuxR/HSL
range2045
1
1
20
annotation2048
1
start
range2048
1
53
53
annotation7070
1
BBa_R0062
range7070
1
1
55
annotation2047
1
-10
range2047
1
42
47
BBa_B0010_sequence
1
ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctc
BBa_R0062_sequence
1
acctgtaggatcgtacaggtttacgcaagaaaatggtttgttatagtcgaataaa
BBa_B0034_sequence
1
aaagaggagaaa
BBa_R0040_sequence
1
tccctatcagtgatagagattgacatccctatcagtgatagagatactgagcac
BBa_C0161_sequence
1
atgactataatgataaaaaaatcggattttttggcaattccatcggaggagtataaaggtattctaagtcttcgttatcaagtgtttaagcaaagacttgagtgggacttagttgtagaaaataaccttgaatcagatgagtatgataactcaaatgcagaatatatttatgcttgtgatgatactgaaaatgtaagtggatgctggcgtttattacctacaacaggtgattatatgctgaaaagtgtttttcctgaattgcttggtcaacagagtgctcccaaagatcctaatatagtcgaattaagtcgttttgctgtaggtaaaaatagctcaaagataaataactctgctagtgaaattacaatgaaactatttgaagctatatataaacacgctgttagtcaaggtattacagaatatgtaacagtaacatcaacagcaatagagcgatttttaaagcgtattaaagttccttgtcatcgtattggagacaaagaaattcatgtattaggtgatactaaatcggttgtattgtctatgcctattaatgaacagtttaaaaaagcagtcttaaattaataa
BBa_K1499503_sequence
1
aattgtgagcggataacaattgacattgtgagcggataacaagatactgagcacatactagagaaagaggagaaatactagatgactataatgataaaaaaatcggattttttggcaattccatcggaggagtataaaggtattctaagtcttcgttatcaagtgtttaagcaaagacttgagtgggacttagttgtagaaaataaccttgaatcagatgagtatgataactcaaatgcagaatatatttatgcttgtgatgatactgaaaatgtaagtggatgctggcgtttattacctacaacaggtgattatatgctgaaaagtgtttttcctgaattgcttggtcaacagagtgctcccaaagatcctaatatagtcgaattaagtcgttttgctgtaggtaaaaatagctcaaagataaataactctgctagtgaaattacaatgaaactatttgaagctatatataaacacgctgttagtcaaggtattacagaatatgtaacagtaacatcaacagcaatagagcgatttttaaagcgtattaaagttccttgtcatcgtattggagacaaagaaattcatgtattaggtgatactaaatcggttgtattgtctatgcctattaatgaacagtttaaaaaagcagtcttaaattaataatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagtccctatcagtgatagagattgacatccctatcagtgatagagatactgagcactactagagaaagaggagaaatactagatgaaaaacataaatgccgacgacacatacagaataattaataaaattaaagcttgtagaagcaataatgatattaatcaatgcttatctgatatgactaaaatggtacattgtgaatattatttactcgcgatcatttatcctcattctatggttaaatctgatatttcaatcctagataattaccctaaaaaatggaggcaatattatgatgacgctaatttaataaaatatgatcctatagtagattattctaactccaatcattcaccaattaattggaatatatttgaaaacaatgctgtaaataaaaaatctccaaatgtaattaaagaagcgaaaacatcaggtcttatcactgggtttagtttccctattcatacggctaacaatggcttcggaatgcttagttttgcacattcagaaaaagacaactatatagatagtttatttttacatgcgtgtatgaacataccattaattgttccttctctagttgataattatcgaaaaataaatatagcaaataataaatcaaacaacgatttaaccaaaagagaaaaagaatgtttagcgtgggcatgcgaaggaaaaagctcttgggatatttcaaaaatattaggttgcagtgagcgtactgtcactttccatttaaccaatgcgcaaatgaaactcaatacaacaaaccgctgccaaagtatttctaaagcaattttaacaggagcaattgattgcccatactttaaaaattaataacactgatagtgctagtgtagatcactactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatatactagagacctgtaggatcgtacaggtttacgcaagaaaatggtttgttatagtcgaataaatactagagtcacacaggaaagtactagatgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataatactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata
BBa_B0032_sequence
1
tcacacaggaaag
BBa_E0040_sequence
1
atgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataa
BBa_B0012_sequence
1
tcacactggctcaccttcgggtgggcctttctgcgtttata
BBa_R0011_sequence
1
aattgtgagcggataacaattgacattgtgagcggataacaagatactgagcaca
BBa_C0062_sequence
1
atgaaaaacataaatgccgacgacacatacagaataattaataaaattaaagcttgtagaagcaataatgatattaatcaatgcttatctgatatgactaaaatggtacattgtgaatattatttactcgcgatcatttatcctcattctatggttaaatctgatatttcaatcctagataattaccctaaaaaatggaggcaatattatgatgacgctaatttaataaaatatgatcctatagtagattattctaactccaatcattcaccaattaattggaatatatttgaaaacaatgctgtaaataaaaaatctccaaatgtaattaaagaagcgaaaacatcaggtcttatcactgggtttagtttccctattcatacggctaacaatggcttcggaatgcttagttttgcacattcagaaaaagacaactatatagatagtttatttttacatgcgtgtatgaacataccattaattgttccttctctagttgataattatcgaaaaataaatatagcaaataataaatcaaacaacgatttaaccaaaagagaaaaagaatgtttagcgtgggcatgcgaaggaaaaagctcttgggatatttcaaaaatattaggttgcagtgagcgtactgtcactttccatttaaccaatgcgcaaatgaaactcaatacaacaaaccgctgccaaagtatttctaaagcaattttaacaggagcaattgattgcccatactttaaaaattaataacactgatagtgctagtgtagatcac
igem2sbol
1
iGEM to SBOL conversion
Conversion of the iGEM parts registry to SBOL2.1
James Alastair McLaughlin
Chris J. Myers
2017-03-06T15:00:00.000Z