chassis_bacteriophage_t7
1
//chassis/bacteriophage/t7
BBa_K145014
1
T7
T7 RNA Polymerase with UmuD derived tag
Benjamien Moeyaert
2008-07-23T11:00:00Z
2015-05-08T01:10:28Z
false
false
_257_
0
2939
9
Released HQ 2013
In stock
true
T7 RNA Polymerase with N-terminal UmuD degradation tag.
This part is different from the simple composition of the tag and the polymerase, as there cannot be a stop codon in the scar.
Part composed of the T7 RNA Polymerase and the N-terminal degradation tag, but with no scar (no stop codon)
true
annotation1968241
1
tag
range1968241
1
1
87
annotation1968240
1
stop
range1968240
1
2737
2739
annotation1968238
1
start
range1968238
1
1
3
annotation1968242
1
cds
range1968242
1
88
2736
annotation1968239
1
stop
range1968239
1
2740
2742
BBa_R0085
1
BBa_R0085
T7 Consensus Promoter Sequence
Barry Canton
2005-02-21T12:00:00Z
2015-05-08T01:14:15Z
false
false
_11_6_
0
135
7
Released HQ 2013
In stock
false
The T7 promoter should only produce PoPS when the T7 polymerase is also being expressed.
Sequence obtained from Sri Kosuri
false
annotation1721520
1
Transcription Start Site
range1721520
1
18
18
annotation1721522
1
Initiation Region
range1721522
1
12
23
annotation1721521
1
Polymerase Binding Region
range1721521
1
1
11
BBa_K921002
1
BBa_K921002
T7 RNAP + IPTG->PoPs (Mutant III)
Eric Pederson
2012-09-30T11:00:00Z
2015-05-08T01:13:46Z
false
false
_1186_
0
12713
9
In stock
false
This is a mutant T7 promoter that includes a lacO sequence. This is repressible by the LacI protein and is induced in the presence of IPTG. The promoter sits in the 5' region of the gene of interest and initiates transcription when the cell expresses T7 RNA polymerase. Be sure you are using a cell strain that includes this gene (DE3).
There are three portions to the T7 promoter: the recognition site, the melting box (or TATA box) and the initation site. Mutations in these sequences caused differential behavior from the wild type.
The part consists of a sequences that resemble a T7 promoter and the lacO sequence found in E. coli.
false
annotation2205317
1
LacO
range2205317
1
19
47
annotation2205318
1
Initiation
range2205318
1
18
20
annotation2205316
1
T7 Promoter
range2205316
1
1
20
BBa_R0186
1
BBa_R0186
T7 promoter (lacI repressible)
Bartholomew Canton
2008-04-07T11:00:00Z
2015-05-08T01:14:15Z
false
false
_11_
0
135
84
Not in stock
false
A lac repressible T7 promoter. This promoter is similar to those found in the pET vectors with T7lac promoters. I mutated the -9 base to an A, which should make it less than 3% of the strength of consensus. The base numbering is as used by Studier and coworkers (JMB 1991, 219, 45-59). See BBa_R0183 for more information.
The -9 mutation from consensus was included to weaken the consensus promoter as described by Imburgio and co-workers (see BBa_R0183 for more information)
This sequence is based on the T7lac promoter described by Dubendorff and Studier (JMB 1991, 219, 45-59). The only modification is the mutation at -9.
false
annotation1962094
1
LacI binding site
range1962094
1
20
44
annotation1962095
1
T7 promoter
range1962095
1
1
20
BBa_R0180
1
BBa_R0180
T7 RNAP promoter
Barry Canton
2005-09-25T11:00:00Z
2015-05-08T01:14:15Z
false
false
_11_6_
0
135
6
Not in stock
false
T7 promoter with an A->T mutation at the -16 position of the consensus sequence
false
BBa_G00113
1
T7
T7 promoter sequencing primer, 20-mer
Reshma Shetty
2008-10-16T11:00:00Z
2015-08-31T04:07:28Z
false
true
_41_
0
126
162
It's complicated
false
This primer anneals to the T7 Promoter region of any vector containing the T7 Promoter.
<biblio>
#Wallace pmid=6282692
</biblio>
T7 promoter.
false
BBa_K113010
1
BBa_K113010
overlapping T7 promoter
Wu Jingjing, Wang Jinyu
2008-10-25T11:00:00Z
2015-05-08T01:09:20Z
false
false
_254_
0
2711
9
It's complicated
false
Two T7 promoters are reversely overlapping.
It is difficult to make this short DNA by PCR as both sides are T7 promoters, which are very similar. It is highly possible that they form loops before they can be synthesized by PCR. Thus we order two single stranded DNA and then anneal them together by touchdown PCR.
The sequence of T7 promoter is TAATACGACTCACTATA.
false
annotation1989277
1
T7 promoter
range1989277
1
4
21
annotation1989307
1
T7 promoter
range1989307
1
20
37
BBa_S03652
1
BBa_S03652
E0041:S03650
Heather Keller
2007-03-05T12:00:00Z
2015-05-08T01:14:26Z
false
false
_10_
0
571
10
In stock
false
false
component1920514
1
BBa_B0101
component1920515
1
BBa_B0010
component1920517
1
BBa_B0012
component1920512
1
BBa_E0041
annotation1920517
1
BBa_B0012
range1920517
1
891
931
annotation1920512
1
BBa_E0041
range1920512
1
1
728
annotation1920515
1
BBa_B0010
range1920515
1
803
882
annotation1920514
1
BBa_B0101
range1920514
1
737
794
BBa_K921000
1
BBa_K921000
T7 RNAP + IPTG->PoPs (Mutant I)
Eric Pederson
2012-09-30T11:00:00Z
2015-05-08T01:13:46Z
false
false
_1186_
0
12713
9
In stock
true
This is a mutant T7 promoter that includes a lacO sequence. This is repressible by the LacI protein and is induced in the presence of IPTG. The promoter sits in the 5' region of the gene of interest and initiates transcription when the cell expresses T7 RNA polymerase. Be sure you are using a cell strain that includes this gene (DE3).
There are three portions to the T7 promoter: the recognition site, the melting box (or TATA box) and the initation site. Mutations in these sequences caused differential behavior from the wild type.
The part consists of a sequences that resemble a T7 promoter and the lacO sequence found in E. coli.
false
annotation2205290
1
LacO
range2205290
1
19
46
annotation2205291
1
Initiation
range2205291
1
19
20
annotation2205289
1
T7 promoter
range2205289
1
1
19
BBa_R0184
1
BBa_R0184
T7 promoter (lacI repressible)
Bartholomew Canton
2007-04-25T11:00:00Z
2015-05-08T01:14:15Z
false
false
_11_
0
135
84
Not in stock
false
A mutant version of the T7lac promoter designed by Dubendorff and Studier. The promoter is based on the strong ??10 T7 promoter but incorporates an A->C mutation at -16. The LacI operator site is a partially symmetric 25bp sequence.
The -16 mutation was included to weaken the consensus promoter as described by Imburgio and co-workers (see BBa_R0183 for more information)
This sequence is based on the T7lac promoter described by Dubendorff and Studier (JMB 1991, 219, 45-59). The only modification is the mutation at -16.
false
annotation1932678
1
BBa_R0183 fragment
range1932678
1
1
20
annotation1932679
1
lacI binding site
range1932679
1
20
44
annotation1932680
1
Center of binding site symmetry
range1932680
1
31
31
BBa_K1833009
1
BBa_K1833009
T7 promoter -> 434 cI N terminus with P22 repressor specificity
Konstantin Borisov
2015-09-17T11:00:00Z
2015-09-17T11:13:57Z
false
false
_2259_
27388
27388
9
false
This part produces the DNA binding domain of the phage 434 cI repressor, with mutations that change the specificity of the DNA to which the protein binds. This protein domain was used as in "A two-hybrid system based on chimeric operator recognition for studying protein homo/heterodimerization in Escherichia coli" (G. Di Lallo, L. Castagnoli, P. Ghelardini and L. Paolozzi; Microbiology (2001), 147, 1651???1656). For more information on the uses of this part, visit 2015.igem.org/Team:Pitt/Protease/Project.
This part was designed with a T7 promoter so the protein could be overexpressed.
"A two-hybrid system based on chimeric operator recognition for studying protein homo/heterodimerization in Escherichia coli" (G. Di Lallo, L. Castagnoli, P. Ghelardini and L. Paolozzi; Microbiology (2001), 147, 1651???1656) described the use of this protein domain.
false
annotation2469007
1
P22-like 434 cI N-terminus
range2469007
1
33
317
annotation2469011
1
T7 terminator
range2469011
1
318
358
annotation2469004
1
pT7
range2469004
1
1
20
annotation2469005
1
rbs
range2469005
1
24
29
BBa_R0181
1
BBa_R0181
T7 RNAP promoter
Barry Canton
2005-09-25T11:00:00Z
2015-05-08T01:14:15Z
false
false
_11_6_
0
135
6
Not in stock
false
T7 promoter with an T->G mutation at the -17 position of the consensus sequence
false
BBa_R0183
1
BBa_R0183
T7 RNAP promoter
Barry Canton
2005-09-25T11:00:00Z
2015-05-08T01:14:15Z
false
false
_11_6_
0
135
6
Not in stock
false
T7 promoter with an A->C mutation at the -16 position of the consensus sequence
false
BBa_K1614000
1
BBa_K1614000
T7 promoter for expression of functional RNA
Stefan Holderbach
2015-09-13T11:00:00Z
2015-09-14T12:29:29Z
false
false
_2031_
12380
16422
9
false
Test
None.
T7 phage minimal promoter (consensus)
false
annotation2454728
1
T7 promoter
range2454728
1
1
17
BBa_K731700
1
BBa_K731700
Platform for terminators analysis under the control of T7 promoter
Giacomo Giacomelli, Anna Depetris
2012-08-13T11:00:00Z
2015-05-08T01:13:06Z
false
false
_977_
0
12063
9
It's complicated
true
We used this part to test terminators: the two subsequent proteins allow to quantify the termination efficiency as the ratio between Venus's and Cherry's fluorescence.
The vector is pET21b, that contains lacI, t7 promoter and ampicillin resistance gene.
The two fluorescent proteins have partially overlapping emission spectra, thus the fluorimetric measures should be taken with these parameters, to avoid interference between the two:
Venus excitation: 485nm, Venus emission: 528nm
Cherry excitation: 528nm, Cherry emission: 615nm
We inserted the prefix-suffix linker between the two proteins, to allow terminator characterization.
Using and modifying this construct is important to remember that the two proteins have strong N-terminal homology, especially designing primers.
For any future improvement, maybe it would be better two use two different fluorescence proteins, because these two has partially overlapping emission spectra and different folding kinetics.
Using and modifying this construct is important to remember that the two proteins have strong N-terminal homology, especially designing primers.
For any future improvement, maybe it would be better two use two different fluorescence proteins, because these two has partially overlapping emission spectra and different folding kinetics.
It's made from a Roberta Lentini's construct (from Mansy lab) that consist of pET21b with the two proteins and a 20bp linker between them.
That construct was mutated two times to eliminate illegal restriction sites; prefix-suffix linker was added by insertion PCR.
false
annotation2179558
1
ampicillin resistance gene
range2179558
1
1711
2370
annotation2179561
1
lacO
range2179561
1
6050
6072
annotation2179562
1
mCHERRY
range2179562
1
6119
6829
annotation2179560
1
T7 PROMOTER
range2179560
1
6031
6050
annotation2179557
1
T7 TERMINATOR
range2179557
1
843
890
annotation2179559
1
lacI
range2179559
1
4562
5653
annotation2179556
1
A206K VENUS
range2179556
1
36
752
BBa_Z0253
1
BBa_Z0253
T7 weak binding promoter
Sriram Kosuri
2006-05-15T11:00:00Z
2015-05-08T01:14:55Z
false
false
_11_10_
0
64
10
Not in stock
false
-- No description --
false
annotation1862691
1
ag->GA from concensus
range1862691
1
31
32
annotation1862684
1
ag->TA from concensus
range1862684
1
27
28
BBa_S03650
1
BBa_S03650
B0100:B0015
Heather Keller
2007-03-05T12:00:00Z
2015-05-08T01:14:26Z
false
false
_10_
0
571
10
Released HQ 2013
In stock
false
false
component1920491
1
BBa_B0012
component1920489
1
BBa_B0010
component1920488
1
BBa_B0101
annotation1920491
1
BBa_B0012
range1920491
1
155
195
annotation1920488
1
BBa_B0101
range1920488
1
1
58
annotation1920489
1
BBa_B0010
range1920489
1
67
146
BBa_K113012
1
BBa_K113012
weaken overlapping T7 promoter
Wu Jingjing, Wang Jinyu
2008-10-25T11:00:00Z
2015-05-08T01:09:20Z
false
false
_254_
0
2711
9
It's complicated
false
Two T7 promoters are reversely overlapping in two base pairs while a point mutation occurs at the 12th base pair, from A-T to T-A.
It is difficult to make this short DNA by PCR as both sides are T7 promoters, which are very similar. It is highly possible that they form loops before they can be synthesized by PCR. Thus we order two single stranded DNA and then anneal them together by touchdown PCR.
The sequence of T7 promoter is TAATACGACTCACTATA (from 5' to 3').
false
annotation1989402
1
T7 promoter
range1989402
1
20
37
annotation1989400
1
T7 promoter
range1989400
1
4
21
BBa_K921001
1
BBa_K921001
T7 RNAP + IPTG->PoPs (Mutant II)
Eric Pederson
2012-09-30T11:00:00Z
2015-05-08T01:13:46Z
false
false
_1186_
0
12713
9
It's complicated
false
This is a mutant T7 promoter that includes a lacO sequence. This is repressible by the LacI protein and is induced in the presence of IPTG. The promoter sits in the 5' region of the gene of interest and initiates transcription when the cell expresses T7 RNA polymerase. Be sure you are using a cell strain that includes this gene (DE3).
There are three portions to the T7 promoter: the recognition site, the melting box (or TATA box) and the initation site. Mutations in these sequences caused differential behavior from the wild type.
The part consists of a sequences that resemble a T7 promoter and the lacO sequence found in E. coli.
false
annotation2205296
1
T7 Promoter
range2205296
1
1
20
annotation2205298
1
Initiation
range2205298
1
18
20
annotation2205297
1
LacO
range2205297
1
22
48
BBa_R0182
1
BBa_R0182
T7 RNAP promoter
Barry Canton
2005-09-25T11:00:00Z
2015-05-08T01:14:15Z
false
false
_11_6_
0
135
6
Not in stock
false
T7 promoter with an A->T mutation at the -10 position of the consensus sequence
false
BBa_J64997
1
BBa_J64997
T7 consensus -10 and rest
Sai Duriseti
2007-03-25T11:00:00Z
2015-05-08T01:08:18Z
false
false
_98_
0
1428
98
Not in stock
false
false
annotation1921746
1
T7-10
range1921746
1
1
19
BBa_I719005
1
pT7
T7 Promoter
Imperial 2007
2007-10-23T11:00:00Z
2015-08-31T04:07:53Z
false
false
_128_
0
2097
9
Released HQ 2013
In stock
true
Just a T7 Promoter
None
---
true
BBa_R0187
1
BBa_R0187
T7 promoter (lacI repressible)
Bartholomew Canton
2008-04-07T11:00:00Z
2015-05-08T01:14:15Z
false
false
_11_
0
135
84
Not in stock
false
A lac repressible T7 promoter. This promoter is similar to those found in the pET vectors with T7lac promoters. I mutated the -9 base to an G, which should make it less than 3% of the strength of consensus. The base numbering is as used by Studier and coworkers (JMB 1991, 219, 45-59). See BBa_R0183 for more information.
The -9 mutation from consensus was included to weaken the consensus promoter as described by Imburgio and co-workers (see BBa_R0183 for more information)
This sequence is based on the T7lac promoter described by Dubendorff and Studier (JMB 1991, 219, 45-59). The only modification is the mutation at -9.
false
annotation1962097
1
T7 promoter
range1962097
1
1
20
annotation1962096
1
LacI binding site
range1962096
1
20
44
BBa_K113011
1
BBa_K113011
more overlapping T7 promoter
Wu Jingjing, Wang Jinyu
2008-10-25T11:00:00Z
2015-05-08T01:09:20Z
false
false
_254_
0
2711
9
It's complicated
false
Two T7 promoters are reversely overlapping in five base pairs, with the third one mutated from A to G.
It is difficult to make this short DNA by PCR as both sides are T7 promoters, which are very similar. It is highly possible that they form loops before they can be synthesized by PCR. Thus we order two single stranded DNA and then anneal them together by touchdown PCR.
The sequence of T7 promoter is TAATACGACTCACTATA (from 5' to 3').
false
annotation1989380
1
T7 promoter
range1989380
1
17
34
annotation1989359
1
T7 promoter
range1989359
1
4
21
BBa_I712074
1
BBa_I712074
T7 promoter (strong promoter from T7 bacteriophage)
Rok Gaber
2007-10-21T11:00:00Z
2015-08-31T04:07:46Z
false
false
_130_
0
1856
9
In stock
false
T7 promoter is very specific promoter which is transcribed only by specific T7 RNA polymerase. Usually this promoter is used in expression systems where T7 promoter is cotransfected with T7 RNA polymerase. That ensures strong transcription of desired genes.
T7 bacteriophage
true
BBa_K093001
1
T7Gene6
T7 Gene 6 exonuclease
John Heil, Shira Davis, Danielle Nash
2008-10-25T11:00:00Z
2015-05-08T01:08:40Z
false
false
_247_
0
3630
9
It's complicated
false
This is the T7 gene 6 ORF, that codes for the T7 gene 6 5'-3' exonuclease. For properties of T7 gene 6 exonuclease see: Kerr and Sadowski "Gene 6 Exonuclease of Bacteriophage T7" The Journal of Biological Chemistry Vol. 247 No. 1 January 10, pp 311-318, 1972.
We needed to change the stop codon to the standard stop codon TAA. We later realized that we should have added tandem TAA. The Tandem TAA stop codon is present in the composite parts containing this ORF. The ORF alone has the single TAA stop codon.
T7 genomic DNA
false
BBa_R0185
1
BBa_R0185
T7 promoter (lacI repressible)
Bartholomew Canton
2008-04-07T11:00:00Z
2015-05-08T01:14:15Z
false
false
_11_
0
135
84
Not in stock
false
A lac repressible T7 promoter. This promoter is similar to those found in the pET vectors with T7lac promoters. I mutated the -10 base to a G, which should make it about 4% of the strength of consensus. The base numbering is as used by Studier and coworkers (JMB 1991, 219, 45-59). See BBa_R0183 for more information.
The -10 mutation from consensus was included to weaken the consensus promoter as described by Imburgio and co-workers (see BBa_R0183 for more information)
This sequence is based on the T7lac promoter described by Dubendorff and Studier (JMB 1991, 219, 45-59). The only modification is the mutation at -10.
false
annotation1962092
1
T7 promoter sequence
range1962092
1
1
20
annotation1962093
1
LacI binding site
range1962093
1
20
44
BBa_Z0252
1
BBa_Z0252
T7 weak binding and processivity
T7.2
2005-04-04T11:00:00Z
2015-05-08T01:14:55Z
false
false
_10_
0
250
10
Not in stock
false
Promoter for T7 RNA polymerase
T7
false
annotation1476344
1
tga -> acg mutation from consensus
range1476344
1
10
12
annotation1476343
1
23bp conserved region
range1476343
1
6
28
annotation1476345
1
t -> a mutation from consensus
range1476345
1
21
21
BBa_J34814
1
BBa_J34814
T7 Promoter
Alexandra Choutko
2006-10-25T11:00:00Z
2015-08-31T04:08:47Z
false
false
_62_
0
1085
62
Not in stock
false
to be used qith t7 polymerase BBa_J34811
a
a
false
BBa_K1833000
1
BBa_K1833000
pT7-eGFP
Konstantin Borisov
2015-09-12T11:00:00Z
2015-09-18T11:04:36Z
false
false
_2259_
27388
27388
9
false
This part produces GFP in the presence of T7 RNA polymerase. It contains a T7 promoter, a strong RBS, the coding region for GFP, and a double terminator. The GFP is a mutant known as GFPmut3b, and the original citation is as follows:
Cormack, B.P., Valdivia, R.H., and S. Falkow. FACS-optimized mutants of green fluorescent protein (GFP). Gene 173: 33-38 (1996). (http://www.sciencedirect.com/science/article/pii/0378111995006850) It has a maximum excitation at 501 nm and maximum emission at 511 nm. For more information, see part BBa_E0040
The part containing GFP was cut with EcoRI and XbaI, and ligated with the annealed oligos to form the completed part. Note that due to the choice of technique, a Biobrick scar site remains between the promoter and RBS.
The RBS-GFP-terminator was obtained from as part BBa_E0840 from the 2015 Distribution (Plate 2, Well 24D). The T7 promoter was obtained as two short oligonucleotides annealed to form sticky ends. The part containing GFP was then cut with EcoRI and XbaI, and ligated with the annealed oligos to form the completed part. Note that due to the choice of technique, a Biobrick scar site remains between the promoter and RBS.
false
component2460130
1
BBa_E0840
component2460119
1
BBa_K1833999
annotation2460130
1
BBa_E0840
range2460130
1
30
907
annotation2460119
1
BBa_K1833999
range2460119
1
1
29
BBa_S03653
1
BBa_S03653
E0042:S03650
Heather Keller
2007-03-05T12:00:00Z
2015-05-08T01:14:26Z
false
false
_10_
0
571
10
Released HQ 2013
In stock
false
false
component1920500
1
BBa_E0042
component1920502
1
BBa_B0101
component1920503
1
BBa_B0010
component1920505
1
BBa_B0012
annotation1920500
1
BBa_E0042
range1920500
1
1
722
annotation1920503
1
BBa_B0010
range1920503
1
797
876
annotation1920502
1
BBa_B0101
range1920502
1
731
788
annotation1920505
1
BBa_B0012
range1920505
1
885
925
BBa_K1833999
1
BBa_K1833999
T7 promoter as annealed oligos
Konstantin Borisov
2015-09-14T11:00:00Z
2015-09-16T07:58:33Z
false
false
_2259_
27388
27388
9
false
This is a part used as a building block for T7 driven expression. However, this part uses a special method with the following synthesized nucleotides:
pT7sense: aattcgcggccgcttctagataatacgactcactatagggagaa
pT7antisense: ctagttctccctatagtgagtcgtattatctagaagcggccgcg
The following method can be used to insert a T7 promoter before a desired Biobrick part:
1. Cut the desired plasmid (which should contain an RBS and cds at minimum) with EcoRI and XbaI. Purify the plasmid with a method of your choice (both gel purification and PCR purification kits from Qiagen serve to remove the short undesired oligonucleotide from the digestion).
2. Be sure that your oligos have 5' phosphorylation (treat unphosphorylated oligos with T4 Polynucleotide Kinase). Then, anneal the two oligos by slowly cooling from 98C to room temperature.
3. Finally, ligate the cut plasmid with the annealed oligos by using a 1:10 molar ratio of plasmid to insert.
4. Transform the ligation reaction into E. coli, and plate.
5. Pick colonies, and verify the success of the cloning with sequencing or PCR. (Doing a diagnostic digest is not recommended as the size of the original plasmid and the desired plasmid are very similar.)
6. Add a composite part to the iGEM registry by using this part with the original protein coding part.
When ligating with a cut plasmid, this leaves a Biobrick scar between the T7 promoter and original Biobrick part.
Synthesized oligonucleotides as described on main page.
false
annotation2460074
1
pT7
range2460074
1
1
23
annotation2460116
1
Oligo scar
range2460116
1
24
29
BBa_Z0251
1
BBa_Z0251
T7 strong promoter
T7.2
2005-04-04T11:00:00Z
2015-05-08T01:14:55Z
false
false
_10_
0
250
10
Not in stock
false
Promoter for T7 RNA polymerase
bacteriophage T7
false
annotation1476333
1
consensus sequence
range1476333
1
6
28
BBa_S03651
1
BBa_S03651
S00159:P1010
Heather Keller
2007-03-05T12:00:00Z
2015-06-15T12:38:52Z
true
false
_10_
4206
571
10
It's complicated
false
false
component1920361
1
BBa_P1010
component1920355
1
BBa_S00159
annotation1920361
1
BBa_P1010
range1920361
1
185
859
annotation1920355
1
BBa_S00159
range1920355
1
1
176