Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Nancy Teng
Date created: 2016-10-12 11:00:00
Date modified: 2016-10-13 11:48:20
HyaB N-terminally tagged subunit of NiFe Hydrogenase
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_K1957008_sequence (Version 1) |
Description
One of the three subunits that make up NiFe Hydrogenase in Shewanella oneidensis. Specifically this is the HyaB subunit, which is the membrane-bound subunit. This subunit has a strep-II tag on its CN-terminal for downstream protein assays. This gene encodes one out of the three subunits that can be used to make the entire NiFe hydrogenase construct.Notes
This is one of of the three NiFe Hydrogenase subunits. The entire NiFe construct can be made by ligating the remaining units into the pBAD expression vector via golden gate cloning. For the entire NiFe hydrogenase cluster the following BioBricks need to be ligated with Bba_K1957008: Bba_K1957005 with Bba_K1957006.BBa_J04450, which contained pSB1C3, would not conjugate properly into S.oneidensis, most likely due to conflicts in the origin of replication. The conflict could result in low copy numbers of pSB1C3 in S.oneidensis resulting in little antibiotic resistance. pSB1C3 contains pUC19 derived pMB1 origin of replication. In a paper by Myers and Myers (1997) they observed that pMB1 did not replicate in Shewanella oneidensis MR-1, formerly known as Shewanella putrefaciens MR-1. To overcome this issue, the three subunits were ligated into the pBAD expression vector using Golden Gate cloning