BBa_I746916
1
BBa_I746916
superfolder GFP coding sequence
2008-09-29T11:00:00Z
2015-08-31T04:08:05Z
Superfolder GFP was originally described by: Pedelacq et al (2006): "Engineering and characterization of a superfolder green fluorescent protein", Nature Biotech 24 (1) January 2006
This version was synthesised de novo (by Geneart).
This is the coding sequence of superfolder GFP (Pedelacq et al (2006): "Engineering and characterization of a superfolder green fluorescent protein", Nature Biotech 24 (1) January 2006).
It carries the following amino acid changes with respect to mut3 GFP (E0040), the currently most commonly used GFP in the registry:
S30R, Y39N, F64L, G65T, F99S, N105T, Y145F, M153T, V163A, I171V, A206V
Its in-vivo properties are considerably improved with respect to mut3 - it develops fluorescence about 3fold faster than mut3 GFP and reaches 4fold higher absolute fluorescence levels. Fluorescenct colonies can be identified with the naked eye even without UV or blue light illumination (that is to say the amount of blue light in normal daylight or lablight is sufficient).
Additionally it is more stable in vitro and refolds faster after in vitro denaturation with respect to mut3 GFP.
Note:
Superfolder GFP is available in constructs driven by the pBAD and T7 promoters: part numbers I746908 and I746909 respectively. Additionally 6-his tagged versions for protein purification exist: I746914 (pBAD driven) and I746915 (T7 driven).
false
false
_116_
0
2122
9
It's complicated
false
Codon optimisation before de novo synthesis was carried out for both, E.coli and Bacillus subtilis.
false
Stefan Milde
annotation1977535
1
stop
range1977535
1
715
720
annotation1977534
1
superfolder GFP coding region
range1977534
1
1
720
annotation1977533
1
start
range1977533
1
1
3
BBa_K1486000
1
BBa_K1486000
CpxR coding region
2014-08-11T11:00:00Z
2015-05-08T01:10:42Z
The part has been extracted from the E coli genome.
CpxR is a transciption factor that dimerizes and becomes active when phosphorylated by the transmembrane histidine kinase CpxA. CpxA in turn becomes active when the bacterial cell is submitted to different kinds of shock (pH, temperature, pressure, protein misfolding in the inter-membrane space...).
false
false
_1866_
0
21042
9
Not in stock
false
When making composite parts with CpxR that are supposed to dimerize, we had to take care about the positioning of the other elements regarding the parallel dimerization of CpxR.
false
iGem EPFL 2014
annotation2382021
1
CpxR
range2382021
1
1
702
annotation2382022
1
START
range2382022
1
1
3
annotation2382023
1
STOP
range2382023
1
699
702
BBa_K1486005
1
BBa_K1486005
Arabinose promoter + sfGFP-CpxR [Cterm]
2014-08-12T11:00:00Z
2015-05-08T01:10:42Z
The arabinose promoter and the sfGFP were obtained from the registry, while the CpxR was obtained from the E coli genome.
This part is composed of an arabinose-inducible promoter for superfolded GFP attached by a linker to CpxR (on the C terminus of CpxR).
false
false
_1866_
0
21042
9
It's complicated
false
We had to test both attachments, N and C terminus of the CpxR.
false
iGem EPFL 2014
component2382088
1
BBa_K1486000
component2382093
1
BBa_I746916
component2382089
1
BBa_K1486003
component2382084
1
BBa_B0034
component2382082
1
BBa_I0500
annotation2382084
1
BBa_B0034
range2382084
1
1211
1222
annotation2382089
1
BBa_K1486003
range2382089
1
1925
1948
annotation2382082
1
BBa_I0500
range2382082
1
1
1210
annotation2382088
1
BBa_K1486000
range2382088
1
1223
1924
annotation2382093
1
BBa_I746916
range2382093
1
1949
2668
BBa_K1486003
1
BBa_K1486003
Flexible linker 2x (GGGS)
2014-08-11T11:00:00Z
2015-05-08T01:10:42Z
Synthetic.
Flexible linker used to fuse two proteins.
false
false
_1866_
0
21042
9
Not in stock
false
Designed considering the flexibility and separation needed between our fused proteins.
false
iGem EPFL 2014
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_I0500
1
pBad/araC
Inducible pBad/araC promoter
2003-12-05T12:00:00Z
2015-08-31T04:07:29Z
-- No description --
false
true
_1_
0
24
7
In stock
false
true
Sri Kosuri
annotation1498842
1
PC
range1498842
1
1043
1114
annotation1498841
1
Pbad
range1498841
1
1080
1210
annotation1498843
1
AraC
range1498843
1
1
1079
BBa_B0034_sequence
1
aaagaggagaaa
BBa_K1486003_sequence
1
ggtggtggttctggtggtggttct
BBa_K1486005_sequence
1
ttatgacaacttgacggctacatcattcactttttcttcacaaccggcacggaactcgctcgggctggccccggtgcattttttaaatacccgcgagaaatagagttgatcgtcaaaaccaacattgcgaccgacggtggcgataggcatccgggtggtgctcaaaagcagcttcgcctggctgatacgttggtcctcgcgccagcttaagacgctaatccctaactgctggcggaaaagatgtgacagacgcgacggcgacaagcaaacatgctgtgcgacgctggcgatatcaaaattgctgtctgccaggtgatcgctgatgtactgacaagcctcgcgtacccgattatccatcggtggatggagcgactcgttaatcgcttccatgcgccgcagtaacaattgctcaagcagatttatcgccagcagctccgaatagcgcccttccccttgcccggcgttaatgatttgcccaaacaggtcgctgaaatgcggctggtgcgcttcatccgggcgaaagaaccccgtattggcaaatattgacggccagttaagccattcatgccagtaggcgcgcggacgaaagtaaacccactggtgataccattcgcgagcctccggatgacgaccgtagtgatgaatctctcctggcgggaacagcaaaatatcacccggtcggcaaacaaattctcgtccctgatttttcaccaccccctgaccgcgaatggtgagattgagaatataacctttcattcccagcggtcggtcgataaaaaaatcgagataaccgttggcctcaatcggcgttaaacccgccaccagatgggcattaaacgagtatcccggcagcaggggatcattttgcgcttcagccatacttttcatactcccgccattcagagaagaaaccaattgtccatattgcatcagacattgccgtcactgcgtcttttactggctcttctcgctaaccaaaccggtaaccccgcttattaaaagcattctgtaacaaagcgggaccaaagccatgacaaaaacgcgtaacaaaagtgtctataatcacggcagaaaagtccacattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccatacccgtttttttgggctagcaaagaggagaaaatgaataaaatcctgttagttgatgatgaccgagagctgacttccctattaaaggagctgctcgagatggaaggcttcaacgtgattgttgcccacgatggggaacaggcgcttgatcttctggacgacagcattgatttacttttgcttgacgtaatgatgccgaagaaaaatggtatcgacacattaaaagcacttcgccagacacaccagacgcctgtcattatgttgacggcgcgcggcagtgaacttgatcgcgttctcggccttgagctgggcgcagatgactatctcccgaaaccgtttaatgatcgtgagctggtggcacgtattcgcgcgatcctgcgccgttcgcactggagcgagcaacagcaaaacaacgacaacggttcaccgacactggaagttgatgccttagtgctgaatccaggccgtcaggaagccagcttcgacgggcaaacgctggagttaaccggtactgagtttaccctgctctatttgctggcacagcatctgggtcaggtggtttcccgtgaacatttaagccaggaagtgttgggcaaacgcctgacgcctttcgaccgcgctattgatatgcacatttccaacctgcgtcgtaaactgccggatcgtaaagatggtcacccgtggtttaaaaccttgcgtggtcgcggctatctgatggtttctgcttcataataaggtggtggttctggtggtggttctatgcgtaaaggcgaagagctgttcactggtgtcgtccctattctggtggaactggatggtgatgtcaacggtcataagttttccgtgcgtggcgagggtgaaggtgacgcaactaatggtaaactgacgctgaagttcatctgtactactggtaaactgccggtaccttggccgactctggtaacgacgctgacttatggtgttcagtgctttgctcgttatccggaccatatgaagcagcatgacttcttcaagtccgccatgccggaaggctatgtgcaggaacgcacgatttcctttaaggatgacggcacgtacaaaacgcgtgcggaagtgaaatttgaaggcgataccctggtaaaccgcattgagctgaaaggcattgactttaaagaagacggcaatatcctgggccataagctggaatacaattttaacagccacaatgtttacatcaccgccgataaacaaaaaaatggcattaaagcgaattttaaaattcgccacaacgtggaggatggcagcgtgcagctggctgatcactaccagcaaaacactccaatcggtgatggtcctgttctgctgccagacaatcactatctgagcacgcaaagcgttctgtctaaagatccgaacgagaaacgcgatcatatggttctgctggagttcgtaaccgcagcgggcatcacgcatggtatggatgaactgtacaaatgatga
BBa_K1486000_sequence
1
atgaataaaatcctgttagttgatgatgaccgagagctgacttccctattaaaggagctgctcgagatggaaggcttcaacgtgattgttgcccacgatggggaacaggcgcttgatcttctggacgacagcattgatttacttttgcttgacgtaatgatgccgaagaaaaatggtatcgacacattaaaagcacttcgccagacacaccagacgcctgtcattatgttgacggcgcgcggcagtgaacttgatcgcgttctcggccttgagctgggcgcagatgactatctcccgaaaccgtttaatgatcgtgagctggtggcacgtattcgcgcgatcctgcgccgttcgcactggagcgagcaacagcaaaacaacgacaacggttcaccgacactggaagttgatgccttagtgctgaatccaggccgtcaggaagccagcttcgacgggcaaacgctggagttaaccggtactgagtttaccctgctctatttgctggcacagcatctgggtcaggtggtttcccgtgaacatttaagccaggaagtgttgggcaaacgcctgacgcctttcgaccgcgctattgatatgcacatttccaacctgcgtcgtaaactgccggatcgtaaagatggtcacccgtggtttaaaaccttgcgtggtcgcggctatctgatggtttctgcttcataataa
BBa_I746916_sequence
1
atgcgtaaaggcgaagagctgttcactggtgtcgtccctattctggtggaactggatggtgatgtcaacggtcataagttttccgtgcgtggcgagggtgaaggtgacgcaactaatggtaaactgacgctgaagttcatctgtactactggtaaactgccggtaccttggccgactctggtaacgacgctgacttatggtgttcagtgctttgctcgttatccggaccatatgaagcagcatgacttcttcaagtccgccatgccggaaggctatgtgcaggaacgcacgatttcctttaaggatgacggcacgtacaaaacgcgtgcggaagtgaaatttgaaggcgataccctggtaaaccgcattgagctgaaaggcattgactttaaagaagacggcaatatcctgggccataagctggaatacaattttaacagccacaatgtttacatcaccgccgataaacaaaaaaatggcattaaagcgaattttaaaattcgccacaacgtggaggatggcagcgtgcagctggctgatcactaccagcaaaacactccaatcggtgatggtcctgttctgctgccagacaatcactatctgagcacgcaaagcgttctgtctaaagatccgaacgagaaacgcgatcatatggttctgctggagttcgtaaccgcagcgggcatcacgcatggtatggatgaactgtacaaatgatga
BBa_I0500_sequence
1
ttatgacaacttgacggctacatcattcactttttcttcacaaccggcacggaactcgctcgggctggccccggtgcattttttaaatacccgcgagaaatagagttgatcgtcaaaaccaacattgcgaccgacggtggcgataggcatccgggtggtgctcaaaagcagcttcgcctggctgatacgttggtcctcgcgccagcttaagacgctaatccctaactgctggcggaaaagatgtgacagacgcgacggcgacaagcaaacatgctgtgcgacgctggcgatatcaaaattgctgtctgccaggtgatcgctgatgtactgacaagcctcgcgtacccgattatccatcggtggatggagcgactcgttaatcgcttccatgcgccgcagtaacaattgctcaagcagatttatcgccagcagctccgaatagcgcccttccccttgcccggcgttaatgatttgcccaaacaggtcgctgaaatgcggctggtgcgcttcatccgggcgaaagaaccccgtattggcaaatattgacggccagttaagccattcatgccagtaggcgcgcggacgaaagtaaacccactggtgataccattcgcgagcctccggatgacgaccgtagtgatgaatctctcctggcgggaacagcaaaatatcacccggtcggcaaacaaattctcgtccctgatttttcaccaccccctgaccgcgaatggtgagattgagaatataacctttcattcccagcggtcggtcgataaaaaaatcgagataaccgttggcctcaatcggcgttaaacccgccaccagatgggcattaaacgagtatcccggcagcaggggatcattttgcgcttcagccatacttttcatactcccgccattcagagaagaaaccaattgtccatattgcatcagacattgccgtcactgcgtcttttactggctcttctcgctaaccaaaccggtaaccccgcttattaaaagcattctgtaacaaagcgggaccaaagccatgacaaaaacgcgtaacaaaagtgtctataatcacggcagaaaagtccacattgattatttgcacggcgtcacactttgctatgccatagcatttttatccataagattagcggatcctacctgacgctttttatcgcaactctctactgtttctccatacccgtttttttgggctagc
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