BBa_K1162009
1
BBa_K1162009
10x-Histidine (10x-His) Tag with Met (ATG) For N-terminal purification
2013-09-14T11:00:00Z
2015-05-08T01:09:32Z
E. coli
This part uses assembly standard 23 (RFC 23-silver fusion) to allow for protein fusions. This 10x His Tag is similar to BBa_K844000 but has the double stop codon (TAA TAA) removed unlike BBa_K844000. (http://parts.igem.org/wiki/index.php?title=Part:BBa_K844000).
Another characteristic of this part is the presence of a Met (ATG) for direct His Tag and protein expression. Since the presence of the Met (ATG) this His Tag should be used for protein purification at the N-terminal. The user can place a promoter-rbs system of their choice in front of this His Tag for direct expression. The lack of any stop codon in this His Tag allows the user to add additional features downstream of the gene e.g. secretion tag or GFP (note, the gene itself must not contain any stop codons if the His Tag is to be used like this).
Related parts
BBa_K844000 (http://parts.igem.org/wiki/index.php?title=Part:BBa_K844000), this link also provides an extensive review of existing Histidine Tags in the Parts Registry.
false
false
_1474_
0
9404
9
It's complicated
true
RFC 23 and contains an atg (met). No stop codon.
false
Kathleen Miller
annotation2343739
1
start
range2343739
1
1
3
BBa_K208010
1
BBa_K208010
Lac Promoter (BBa_R0010) and RBS (B0034)
2009-10-11T11:00:00Z
2015-05-08T01:11:24Z
synthetic
Released HQ 2013
lac + rbs
false
false
_310_
0
3473
9
In stock
true
none
false
Elisabeth Linton
annotation2034625
1
BBa_B0034
range2034625
1
209
220
annotation2034626
1
BBa_R0010
range2034626
1
1
200
BBa_K1162010
1
BBa_K1162010
promoter+rbs and N-terminal 10x His Tag
2013-09-14T11:00:00Z
2015-05-08T01:09:32Z
E. coli
This promoter 10x His Tag system allows the user to place the protein they wish to be purified directly onto the back end of this construct. The user must be aware that the gene they were to express must be RFC 23 otherwise the expression system will be out of frame. Also the gene of interest should contain a stop codon at the end.
false
false
_1474_
0
9404
9
It's complicated
false
None.
false
Kathleen Miller
component2343748
1
BBa_K1162009
component2343746
1
BBa_K208010
annotation2343748
1
BBa_K1162009
range2343748
1
227
259
annotation2343746
1
BBa_K208010
range2343746
1
1
220
BBa_K208000
1
BBa_K208000
GFP
2009-10-07T11:00:00Z
2015-05-08T01:11:24Z
plasmid
GFP
false
false
_310_
0
2425
303
It's complicated
true
Mutate PstI sites
false
Elisabeth Linton
annotation2062216
1
Start
range2062216
1
1
3
annotation2062217
1
GFP Coding Region
range2062217
1
1
720
BBa_K1162013
1
BBa_K1162013
promoter+rbs+N terminal His Tag and GFP
2013-09-14T11:00:00Z
2015-05-08T01:09:32Z
E. coli.
This part demonstrates that the N-terminal 10x His Tag functions as desired. It contains a promoter and rbs (BBa_K208010) with a GFP fusion (BBa_K208000). His Tag and GFP are RFC 23 compatible which allows for protein fusion with the His Tag.
false
false
_1474_
0
9404
9
It's complicated
false
RFC 23 when making the N terminal His Tag and GFP fusion.
false
Kathleen Miller
component2343758
1
BBa_K208000
component2343755
1
BBa_K1162010
annotation2343758
1
BBa_K208000
range2343758
1
266
985
annotation2343755
1
BBa_K1162010
range2343755
1
1
259
BBa_K1162009_sequence
1
atgcatcatcaccatcaccaccatcatcaccat
BBa_K1162010_sequence
1
caatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacacatactagagaaagaggagaaatactagatgcatcatcaccatcaccaccatcatcaccat
BBa_K1162013_sequence
1
caatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacacatactagagaaagaggagaaatactagatgcatcatcaccatcaccaccatcatcaccattactagatggctagcaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgctacatacggaaagcttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttctcttatggtgttcaatgcttttcccgttatccggatcatatgaaacggcatgactttttcaagagtgccatgcccgaaggttatgtacaggaacgcactatatctttcaaagatgacgggaactacaagacgcgtgctgaagtcaagtttgaaggtgatacccttgttaatcgtatcgagttaaaaggtattgattttaaagaagatggaaacattctcggacacaaactcgagtacaactataactcacacaatgtatacatcacggcagacaaacaaaagaatggaatcaaagctaacttcaaaattcgccacaacattgaagatggatccgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcgacacaatctgccctttcgaaagatcccaacgaaaagcgtgaccacatggtccttcttgagtttgtaactgctgctgggattacacatggcatggatgagctctacaaataa
BBa_K208000_sequence
1
atggctagcaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggtgatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgctacatacggaaagcttacccttaaatttatttgcactactggaaaactacctgttccatggccaacacttgtcactactttctcttatggtgttcaatgcttttcccgttatccggatcatatgaaacggcatgactttttcaagagtgccatgcccgaaggttatgtacaggaacgcactatatctttcaaagatgacgggaactacaagacgcgtgctgaagtcaagtttgaaggtgatacccttgttaatcgtatcgagttaaaaggtattgattttaaagaagatggaaacattctcggacacaaactcgagtacaactataactcacacaatgtatacatcacggcagacaaacaaaagaatggaatcaaagctaacttcaaaattcgccacaacattgaagatggatccgttcaactagcagaccattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattacctgtcgacacaatctgccctttcgaaagatcccaacgaaaagcgtgaccacatggtccttcttgagtttgtaactgctgctgggattacacatggcatggatgagctctacaaataa
BBa_K208010_sequence
1
caatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaatttcacacatactagagaaagaggagaaa
igem2sbol
1
iGEM to SBOL conversion
Conversion of the iGEM parts registry to SBOL2.1
Chris J. Myers
James Alastair McLaughlin
2017-03-06T15:00:00.000Z