Difference between revisions of "Part:BBa K4344041"

Latest revision as of 14:20, 12 October 2022

pEX-A258-HSV-UL19-6xHis

Synthesized plasmid by Eurofins contains modified UL19 sequence. Plasmid with additional His-tag for purification.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 1055
Illegal XbaI site found at 1043
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1055
Illegal NheI site found at 1049
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1055
Illegal XhoI site found at 3463
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1055
Illegal XbaI site found at 1043
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 1055
Illegal XbaI site found at 1043
Illegal NgoMIV site found at 307
Illegal NgoMIV site found at 454
Illegal NgoMIV site found at 814
1000
COMPATIBLE WITH RFC[1000]

The sequence of UL19 encoding for HSV-1 major capsid protein was extracted from the complete https://www.ncbi.nlm.nih.gov/nuccore/KF498959.1 human herpesvirus 1 genome. The sequence was human codon optimised using https://www.benchling.com/ Benchling. The sequence was screened for published antibody binding sites and functional structure elements using the https://www.uniprot.org/ UniProt database. We identified the amino acids 862 to 880 in UL19 as a published antibody binding site (Han et al., 2019). Since it is a published antibody site, we anticipate that the corresponding DNA sequence is highly conserved in the HSV genome. Therefore, we chose a sequence for cloning and expression including this structure element. For UL19 bases 2446 to 3444 were selected resulting in a fragment of 999 bp in size. The fragment was further modified to mask unwanted restriction sites and ease primer design while keeping the amino acid sequence unchanged. UL19 was modified at the 16-18TCC>AGT and 21G>A. A His-tag (5’-CATCACCATCACCATCAC-3’) and a TAA stop codon were added at the 3’ sequence end.

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 <partinfo>BBa_K4344041 short</partinfo>
-Placeholder
+Synthesized plasmid by Eurofins contains modified UL19 sequence.
+Plasmid with additional His-tag for purification.
 <!-- Add more about the biology of this part here
@@ Line 12: / Line 13: @@
 <partinfo>BBa_K4344041 SequenceAndFeatures</partinfo>
+The sequence of <i>UL19</i> encoding for HSV-1 major capsid protein was extracted from the complete https://www.ncbi.nlm.nih.gov/nuccore/KF498959.1 human herpesvirus 1 genome. The sequence was human codon optimised using https://www.benchling.com/ Benchling.
+The sequence was screened for published antibody binding sites and functional structure elements using the https://www.uniprot.org/ UniProt database. We identified the amino acids 862 to 880 in <i>UL19</i> as a published antibody binding site <span class="scientific-src">(Han <i>et al.</i>, 2019)</span>. Since it is a published antibody site, we anticipate that the corresponding DNA sequence is highly conserved in the HSV genome. Therefore, we chose a sequence for cloning and expression including this structure element. For <i>UL19</i> bases 2446 to 3444 were selected resulting in a fragment of 999 bp in size. The fragment was further modified to mask unwanted restriction sites and ease primer design while keeping the amino acid sequence unchanged. <i>UL19</i> was modified at the 16-18TCC>AGT and 21G>A. A His-tag  (5’-CATCACCATCACCATCAC-3’) and a TAA stop codon were added at the 3’ sequence end.
 <!-- Uncomment this to enable Functional Parameter display