BBa_K1428001
1
BBa_K1428001
Gene cluster for Expressing a purifiable Eut Bacterial Microcompartment
2014-11-02T12:00:00Z
2015-05-08T01:10:23Z
BBa_K311004, pET28a
The following features of the gene cluster are listed from upstream to downstream: A T7 promoter, a T7 RBS, His6-tagged EutS, a Plac promoter, an RBS (B0034), EutM, an RBS (B0034), EutN, a Plac promoter, an RBS (B0034), EutL, an RBS (B0034), EutK.
The whole gene cluster encodes five proteins, EutS, EutM, EutN, EutL, EutK, arranged in a specific order such that when induced with 0.2mM IPTG or allowed for leaky expression at 30C overnight, these 5 gene products will be expressed in a specific molar ratio so that they can assemble into the intact organelle of the bacterial microcompartment (BMC) of the Ethanolamine-utilization type (Eut). It was derived from BBa_K311004 and the pET28a vector in 2 cloning steps.
If the whole part is put into a bacterial, one can readily express out a whole organelle in the form of BMC for compartmentalization, which, when combined with our second Biobrick BBa_K1428000, can target any coding Biobrick inside the BMC for both in vivo and in vitro applications, for example, metabolic engineering to avoid toxic intermediate damage to the cell, bioremediation for pollutant collection, drug delivery systems by storing nucleic acids, peptides and proteins for replacement therapy, or even virology model studies as the whole thing resembles a viral capsid.
false
false
_1806_
0
20650
9
Not in stock
false
Careful considerations about the linker peptide between the His6-tag and EutS must be considered to avoid (1) out-frame fusion and (2) linker peptides which are hydrophobic and inflexible.
false
Lai Hei Ming
annotation2429972
1
His6-tag
range2429972
1
100
117
annotation2429980
1
CAP binding site
range2429980
1
1509
1530
annotation2429983
1
EutL
range2429983
1
1613
2272
annotation2429973
1
Thrombin site
range2429973
1
127
144
annotation2429979
1
EutN
range2429979
1
1106
1393
annotation2429977
1
RBS
range2429977
1
689
696
annotation2429981
1
Modified Plac Promoter
range2429981
1
1545
1575
annotation2429978
1
EutM
range2429978
1
703
993
annotation2429970
1
T7 RBS
range2429970
1
64
80
annotation2429982
1
RBS
range2429982
1
1599
1606
annotation2429974
1
EutS
range2429974
1
148
483
annotation2429976
1
Modified Plac Promoter
range2429976
1
635
665
annotation2429971
1
Start codon
range2429971
1
88
90
annotation2429969
1
lac operator
range2429969
1
20
44
annotation2429968
1
T7 Promoter
range2429968
1
1
19
annotation2429975
1
CAP binding site
range2429975
1
599
620
annotation2429984
1
EutK
range2429984
1
2285
2779
BBa_K1428001_sequence
1
taatacgactcactataggggaattgtgagcggataacaattccccactagaaataattttgtttaactttaagaaggagatataccatgggcagcagccatcatcatcatcatcacagcagcggcctggtgccgcgcggcagccatatgaataaagaacgcattattcaggaatttgtgccgggcaaacaggtcacgctggcgcatctcattgcgcaccctggtgaagaactggcgaaaaagatcggcgttcccgatgctggcgcgattggcattatgactttaacgccgggggaaacggcgatgatcgctggcgatttagcgatgaaggctgcggatgtacacatcggttttctcgatagatttagcggcgcgctggtgatttatggcaccgtcggcgcagtagaagaagcgctattacagacggtcagcgggctggggcgattattaaactttaccctgtgtgagttaacaaaaagttaagcggccgcctcgagcatcaccatcaccatcactgattctccttacgcatctgtgcggtatttcacaccgcatgtactagacccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcgtctagtagaaggaggagatctatggaagcattaggaatgattgaaacccggggcctggttgcgctgattgaggcctccgatgcgatggtaaaagccgcgcgcgtgaagctggtcggcgtgaagcagattggcggtggcctgtgtactgccatggtgcgtggcgatgtggcggcgtgcaaagccgcaaccgatgctggcgccgctgcggcgcagcgcattggcgagttggtctccgtacacgtgattccacgcccgcacggcgatctggaagaagtgttcccgatcagcttcaaaggcgacagcaacatttgatggtaggccggataagacgcgcagcgtcgccatccggcatgaatgcgcgccattgcctgatggtgcgcagattcaaggcctacgacccaacgggcaatccatggaggcggatatgaaactggcagtcgtcacaggacaaatcgtatgtaccgtccgccatcagggactggcgcatgacaaattgctgatggtggaaatgatcgatgcccagggcaaccccgacgggcagtgtgccgtcgctatcgacagtatcggggcgggaaccggagagtgggtactgctggtcagcggcagttccgcacgccaggcgcatcgcagcgaattatcaccggtcgatctgtgcgtcattggcatcgtcgatgaagtggtggctggcgggaaagtggttttccataaataggcggccgcctcgagcatcaccatcaccatcactgattctccttacgcatctgtgcggtatttcacaccgcatgtactagacccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcgtctagtagaaggaggagatctatgcctgcattagatttaattcgaccttcagtgactgccatgcgcgtgattgcctccgtgaatgacggctttgcgcgggaacttaaattaccgccacatatacgtagtctcggactcatcacggcagattctgatgatgtgacctatattgccgctgacgaagcgacaaaacaggcgatggtggaagtggtctatggccgctccctctatgccggggcggctcacgggccgtcgcctaccgctggtgaggtgttgattatgctgggcggaccgaacccggcggaagtgcgcgccggtctggatgcaatggtcgccagcattgaaaacggcgcggcgttccagtgggcgaacgatgcggaaaataccgcattcctggcgcatgtggtttcgcgtaccggctcgtacctctcgtctaccgcaggtatcgcgctgggcgatccaatggcctacctggtggcgccgccgctggaagcgacattcggcattgacgcggcgatgaaatccgctgacgttcagctggtgacttacgtgccgccgccgtcggaaaccaactattcggccgcgttcttaacgggaagccaggccgcgtgtaaagccgcctgcaacgcctttaccgacgccgtgctggatatcgcccgtaaccctgtccagcgtgcgtaacggaggctgccaatgatcaatgccctgggattactggaagtggacggaatggtcgccgccgtcgatgcggcggatgctatgctgaaagcggccaacgtgcgtttgctcagtcatcaagtgctcgaccctggcaggctcacgctggtggtggagggcgatctggcggcgtgtcgtgcggcgctggatgccggaagcgctgccgcgcagcgcacaggccgtgtgattagtcgcaaagaaattggtcgtccggaagaggacacccagtggctgattggcgggtttgcgcgtgcgacaacgccgactgagaaagcaccccaggtacccgcaacgcctgagttcgctgaggcgttattggcgctgctggcatcagtacgccagggaatgacggcgggagaggttgccgcgcattttggctggccgctggagcaggccagaaacgtgctggaacaactcttttctgacggagcgttgcgtaaacgcagtagtcgctatcgcatcaaaaattaa
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