Part:BBa_K1957008:Design
HyaB N-terminally tagged subunit of NiFe Hydrogenase
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 282
Illegal BamHI site found at 1479 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1067
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1
Illegal BsaI.rc site found at 1766
Illegal SapI.rc site found at 37
Design 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
Source
The source of the part will be its sequence which was retrieved from GenBank.
References
Myers C.R. and Myers J.M. (1997). 'Replication of plasmids with the p15A origin in Shewanella putrefaciens MR-1', Applied Microbiology, 24, pp.221-225