Difference between revisions of "Part:BBa K1957005:Design"

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===References===
 
===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

Latest revision as of 21:40, 8 October 2016


HyaA subunit of NiFe Hydrogenase


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 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 1151


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.

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