BBa_K2144101
1
LT
Vector containing Hisx6 and Sortase tag LPETGG
2016-10-13T11:00:00Z
2016-10-14T01:15:18Z
The plasmid was constructed using the iGEM-supplied pSB1C3 and a synthetically produced sequence.
This vector is constructed to simplify the usage of the transpeptidase Sortase A for future iGEM teams. Following a BamHI restriction site we constructed a synthetic sequence consisting of three main elements:
1) A linker (GSSS) to allow for limited mobility of the attached peptides and facilitate correct folding
2) A Sortase A tag (LPETGG) that is used to conjugate the proteins to poly-glycines
3) A standard Hisx6 tag commonly used for Immobilized Metal Affinity Chromatography (IMAC)
This sequence follows a BamHI restriction site which can be used by teams to clone in whichever peptide they would like to conjugate using Sortase A.
false
false
_2614_
31510
31510
9
false
BamHI is a very convenient restriction enzyme for our application as its restriction site will code for a glycine and a serine, both of which are often used in linkers and tend to be unproblematic in such sequences.
false
Sigrun Stulz
annotation2509042
1
conserved
range2509042
1
213
216
annotation2509049
1
polya
range2509049
1
1056
1069
annotation2509043
1
ori
range2509043
1
254
842
annotation2509048
1
T7 TE
range2509048
1
1036
1055
annotation2509037
1
-35
range2509037
1
137
142
annotation2509051
1
CmR
range2509051
1
1140
1799
annotation2509052
1
Hisx6
range2509052
1
2077
2094
annotation2509053
1
Sortase A Tag
range2509053
1
2059
2076
annotation2509046
1
stem_loop
range2509046
1
952
995
annotation2509054
1
Stop
range2509054
1
2095
2097
annotation2509035
1
BBa_R0010
range2509035
1
1
200
annotation2509044
1
barcode
range2509044
1
908
932
annotation2509041
1
BBa_B0034
range2509041
1
209
220
annotation2509045
1
BBa_B0010
range2509045
1
941
1020
annotation2509038
1
-10
range2509038
1
161
166
annotation2509036
1
CAP
range2509036
1
89
126
annotation2509050
1
stop
range2509050
1
1062
1062
annotation2509047
1
BBa_B0012
range2509047
1
1029
1069
annotation2509040
1
start
range2509040
1
173
173
annotation2509039
1
LacI
range2509039
1
166
200
BBa_K2144101_sequence
1
gtccggcaaaaaagggcaaggtgtcaccaccctgccctttttctttaaaaccgaaaagattacttcgcgttatgcaggcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccacaggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagctcgaggcttggattctcaccaataaaaaacgcccggcggcaaccgagcgttctgaacaaatccagatggagttctgaggtcattactggatctatcaacaggagtccaagcgagctcgatatcaaattacgccccgccctgccactcatcgcagtactgttgtaattcattaagcattctgccgacatggaagccatcacaaacggcatgatgaacctgaatcgccagcggcatcagcaccttgtcgccttgcgtataatatttgcccatggtgaaaacgggggcgaagaagttgtccatattggccacgtttaaatcaaaactggtgaaactcacccagggattggctgagacgaaaaacatattctcaataaaccctttagggaaataggccaggttttcaccgtaacacgccacatcttgcgaatatatgtgtagaaactgccggaaatcgtcgtggtattcactccagagcgatgaaaacgtttcagtttgctcatggaaaacggtgtaacaagggtgaacactatcccatatcaccagctcaccgtctttcattgccatacgaaattccggatgagcattcatcaggcgggcaagaatgtgaataaaggccggataaaacttgtgcttatttttctttacggtctttaaaaaggccgtaatatccagctgaacggtctggttataggtacattgagcaactgactgaaatgcctcaaaatgttctttacgatgccattgggatatatcaacggtggtatatccagtgatttttttctccattttagcttccttagctcctgaaaatctcgataactcaaaaaatacgcccggtagtgatcttatttcattatggtgaaagttggaacctcttacgtgcccgatcaactcgagtgccacctgacgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggcagaatttcagataaaaaaaatccttagctttcgctaaggatgatttctggaattcgcgggatccggcggcggcggcagccttcctgaaactggcggccatcaccatcaccatcactgatactagtagcggccgctgca
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