Difference between revisions of "Part:BBa K1957006"

Revision as of 18:14, 13 October 2016

HyaC subunit of NiFe Hydrogenase

One of the three subunits that make up NiFe Hydrogenase in Shewanella oneidensis. Specifically this is the HyaC subunit, which is the outer periplasmic subunit. This gene encodes one out of the three subunits that can be used to make the entire NiFe hydrogenase construct.

Figure 1: PCR Colony of NiFe Hydrogenase subunits. Lanes 14 to 16 have HyaC gene insert, which is 754bp. Ladder is Hyperladder Kb, sizes shown in bp

Restriction site incompatibility, [1000], is with regards to MoClo or Golden Gate cloning. This restriction site was used in the described Golden Gate cloning experiments in order to assemble the gene cluster into a single expression vector. The Bsal restriction enzyme sites flank the coding sequence, and digestion with Bsal should not disrupt the protein coding sequence. For more details on our Golden Gate cloning strategy and instructions on how to assemble the HyaCBA gene cluster into a single expression vector please see the NRP-UEA 2016 iGEM teams wiki.

Figure 2: Screenshot of sequencing data of colony aligned with HyaC using Benchling. DNA sequence of screenshot starts at 355bp

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 176
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1
Illegal BsaI.rc site found at 690

@@ Line 5: / Line 5: @@
 One of the three subunits that make up NiFe Hydrogenase in ''Shewanella oneidensis''. Specifically this is the HyaC subunit, which is the outer periplasmic subunit. This gene encodes one out of the three subunits that can be used to make the entire NiFe hydrogenase construct.
-[[File:T--NRP-UEA-Norwich--Gel_PCR_Colony_NiFe_.jpg|thumb|none|Figure 1: PCR Colony of NiFe Hydrogenase subunits. Lanes 14 to 16 have HyaC gene insert, which is 754bp. Ladder is Hyperladder Kb, sizes shown in bp]]
+[[File:T--NRP-UEA-Norwich--Gel_PCR_Colony_NiFe_.jpg|thumb|none|'''Figure 1: PCR Colony of NiFe Hydrogenase subunits'''. Lanes 14 to 16 have HyaC gene insert, which is 754bp. Ladder is Hyperladder Kb, sizes shown in bp]]
-Restriction site incompatibility, [1000], is with regards to MoClo or Golden Gate cloning. This restriction site was used in the described Golden Gate cloning experiments in order to assemble the gene cluster into a single expression vector. The Bsal restriction enzyme sites flank the coding sequence, and digestion with Bsal should not disrupt the protein coding sequence. For more details on our Golden Gate cloning strategy and instructions on how to assemble the HyaCBA gene cluster into a single expression vector please see the NRP-UEA 2016 iGEM teams wiki.
+Restriction site incompatibility, [1000], is with regards to MoClo or Golden Gate cloning. This restriction site was used in the described Golden Gate cloning experiments in order to assemble the gene cluster into a single expression vector. The ''Bsal'' restriction enzyme sites flank the coding sequence, and digestion with ''Bsal'' should not disrupt the protein coding sequence. For more details on our Golden Gate cloning strategy and instructions on how to assemble the HyaCBA gene cluster into a single expression vector please see the NRP-UEA 2016 iGEM teams wiki.
-[[File:T--NRP-UEA-Norwich--Sequencing_data_605_.jpg|thumb|none|Figure 2: Screenshot of sequencing data of colony aligned with HyaC using Benchling. DNA sequence of screenshot starts at 355bp]]
+[[File:T--NRP-UEA-Norwich--Sequencing_data_605_.jpg|thumb|none|'''Figure 2: Screenshot of sequencing data of colony aligned with HyaC using Benchling'''. DNA sequence of screenshot starts at 355bp]]
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