BBa_K1173502
1
BBa_K1173502
<i>ompC</i> w/GFP + (<i>BglII+BamHI</i>)restriction sites
2013-09-19T11:00:00Z
2015-05-08T01:09:35Z
Synthesised by IDT (gBlocks). Based on a variety of BioBricks.
ompF is a specific promoter which is only activated when phosphorylated OmpR (OmpR-P) is present. OmpR is phosphorylated by the protein Cph8 (form by two proteins, Cph1 and EnvZ), which is usually associated with osmotic regulation. Cph8 is also responsive to red light; exposure to red light halts the phosphorylation of OmpR. The lack of OmpR-P means the ompF promoter is not actiavted, so the genes which follow it are not transcribed. There is a GFP after ompF, which has the nucleotide sequence from BBa_E0040. The GFP will be produced in most generally used competant cells, as Cph8 is present in their outer membranes for osmotic regulation. To control the levels of GFP produced, we recommend the use of EnvZ deficient cells and introducing the genes which code for Cph8 on a LCN plasmid. This would allow control of GFP production using only red light.
However, the novel concept of this part is that you may replace GFP with a gene of your choice. The GFP is flanked by restriction sites for BglII and BamHI. This allows you to "cut out" the GFP without interfering with the standard 3A assembly (EcoRI, XbaI, SpeI and PstI) and replace it with any gene, providing it also has the BglII and BamHI restriction sites. These can easily be added using traditional cloning using PCR primers. This will theoretically allow any gene to be controlled by the ompF promoter, and therefore exposure to red light.
false
false
_1486_
0
18234
9
It's complicated
false
You MUST be concious of the BglII and BamHI cut sites within the construct. These enzymes are used for several assembly techniques, so these must not be applied when assembling the part with others.
false
paul james
component2366891
1
BBa_M31985
component2366888
1
BBa_R0084
component2366894
1
BBa_K150008
component2366893
1
BBa_E0040
component2366890
1
BBa_B0034
component2366901
1
BBa_B0015
annotation2366893
1
BBa_E0040
range2366893
1
149
868
annotation2366891
1
BBa_M31985
range2366891
1
137
142
annotation2366890
1
BBa_B0034
range2366890
1
117
128
annotation2366901
1
BBa_B0015
range2366901
1
891
1019
annotation2366888
1
BBa_R0084
range2366888
1
1
108
annotation2366894
1
BBa_K150008
range2366894
1
877
882
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_B0015
1
BBa_B0015
double terminator (B0010-B0012)
2003-07-16T11:00:00Z
2015-08-31T04:07:20Z
Released HQ 2013
Double terminator consisting of BBa_B0010 and BBa_B0012
false
true
_1_
0
24
7
In stock
false
true
Reshma Shetty
component1916612
1
BBa_B0012
component1916610
1
BBa_B0010
annotation1916610
1
BBa_B0010
range1916610
1
1
80
annotation1916612
1
BBa_B0012
range1916612
1
89
129
BBa_M31985
1
BglII
BglII restriction enzyme site
2007-02-28T12:00:00Z
2015-05-08T01:14:01Z
none
BglII restriction enzyme site
false
false
_102_
0
1373
102
Not in stock
false
none
false
Tiffany Guo
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_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_R0084
1
OmpR
Promoter (OmpR, positive)
2004-01-27T12:00:00Z
2015-05-08T01:14:15Z
NC_000913 E. coli K12
Released HQ 2013
Positively regulated, OmpR-controlled promoter. This promoter is taken from the upstream region of ompF. Phosphorylated OmpR binds to the three operator sites and activates transcription.
false
false
_1_
0
24
7
In stock
false
The promoter includes three OmpR binding sites: F1, F2, and F3. The promoter starts at -107 and goes up to the transcriptional start site at +1. A distant fourth binding site, F4, has been deleted. It is involved in negative regulation of ompF (Head, Tardy and Kenney, 1998), so its deletion is intended to make the promoter solely activating. The efficacy of this promoter versus the ompC upstream region (BBa_R0082) or the truncated ompC upstream region (BBa_R0083) is currently unknown.
true
Stephen Lee, Roshan Kumar, Joe Levine, Ziyan Chu (Polkadorks, IAP 2004)
annotation301313
1
F3 OmpR
range301313
1
51
68
annotation301309
1
F2 OmpR
range301309
1
31
48
annotation301317
1
-35
range301317
1
73
78
annotation301308
1
F1 OmpR
range301308
1
11
28
annotation301318
1
-10
range301318
1
88
93
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
annotation1687
1
stop
range1687
1
34
34
annotation1690
1
polya
range1690
1
28
41
annotation1686
1
T7 TE
range1686
1
8
27
annotation7020
1
BBa_B0012
range7020
1
1
41
BBa_K150008
1
BamHI
BamHI restriction enzyme site
2008-10-20T11:00:00Z
2015-05-08T01:10:46Z
-
BamHI restriction site
false
true
_234_
0
3299
9
Not in stock
false
-
false
Pascal Kraemer
BBa_B0010_sequence
1
ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctc
BBa_K1173502_sequence
1
cttaaattttacttttggttacatattttttctttttgaaaccaaatctttatctttgtagcactttcacggtagcgaaacgttagtttgaatggaaagatgcctgcatactagagaaagaggagaaatactagagagatcttactagatgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataatactagagggatcctactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata
BBa_B0034_sequence
1
aaagaggagaaa
BBa_K150008_sequence
1
ggatcc
BBa_E0040_sequence
1
atgcgtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacggaaaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttcggttatggtgttcaatgctttgcgagatacccagatcatatgaaacagcatgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttcaaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgttaatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaattggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatggaatcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtccttcttgagtttgtaacagctgctgggattacacatggcatggatgaactatacaaataataa
BBa_M31985_sequence
1
agatct
BBa_R0084_sequence
1
cttaaattttacttttggttacatattttttctttttgaaaccaaatctttatctttgtagcactttcacggtagcgaaacgttagtttgaatggaaagatgcctgca
BBa_B0012_sequence
1
tcacactggctcaccttcgggtgggcctttctgcgtttata
BBa_B0015_sequence
1
ccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata
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