Difference between revisions of "Part:BBa K3726068:Design"
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additionally, regarding to CDS_lv0_SigB we have removed an internal BsmbI site in the base 862 in order to make this pat MoClo compatible with the marburg collection. However, the Sapl and BbsI sites should be removed in order to make this part RFC1000 compatible.  
 
additionally, regarding to CDS_lv0_SigB we have removed an internal BsmbI site in the base 862 in order to make this pat MoClo compatible with the marburg collection. However, the Sapl and BbsI sites should be removed in order to make this part RFC1000 compatible.  
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===Source===
 
===Source===
Line 21: Line 20:
  
 
===References===
 
===References===
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D. Kaczmarzyk, J. Anfelt, A. Särnegrim and E. Hudson, "Overexpression of sigma factor SigB improves temperature and butanol tolerance of Synechocystis sp. PCC6803", 2021.

Revision as of 01:52, 20 October 2021


Lv.1 SigB - A


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1103
    Illegal PstI site found at 649
    Illegal PstI site found at 2005
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1103
    Illegal NheI site found at 1730
    Illegal PstI site found at 649
    Illegal PstI site found at 2005
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1103
    Illegal XhoI site found at 1139
    Illegal XhoI site found at 1376
    Illegal XhoI site found at 2158
    Illegal XhoI site found at 2278
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1103
    Illegal PstI site found at 649
    Illegal PstI site found at 2005
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1103
    Illegal PstI site found at 649
    Illegal PstI site found at 2005
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 863


Design Notes

The part is a MoClo Lv.1 part, assembled in the acceptor Lv1 entry vector “BBa_K3228069” P1_pANs_SpecR, which is a self replicating shuttle vector. This part has been assembled following the marburg collection standard, then the transcriptional unit is flanked by the part 5'Con3 “BBa_K2560067” that will allow the assembly of a Lv2 construct, and by the part “BBa_K3726109” 3CON5(H)_NS3(mod)-down (PCC 11801) that allows homologous recombination within the PCC 11801 Genome.

This design will allow us to test which “tolerance and secretion ” variant has better efficiency by its own account as a replicative vector, and then assemble them as an integrative Lv2 Moclo construct. To know more about this, visit our webpage: https://2021.igem.org/Team:MADRID_UCM/Design

additionally, regarding to CDS_lv0_SigB we have removed an internal BsmbI site in the base 862 in order to make this pat MoClo compatible with the marburg collection. However, the Sapl and BbsI sites should be removed in order to make this part RFC1000 compatible.

Source

This construct has been made by golden gate reaction.

References

D. Kaczmarzyk, J. Anfelt, A. Särnegrim and E. Hudson, "Overexpression of sigma factor SigB improves temperature and butanol tolerance of Synechocystis sp. PCC6803", 2021.