Part:BBa_K3726069:Design
Lv.1 SigB - B
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 850
Illegal PstI site found at 1752 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 850
Illegal NheI site found at 431
Illegal NheI site found at 454
Illegal NheI site found at 1477
Illegal PstI site found at 1752 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 850
Illegal XhoI site found at 886
Illegal XhoI site found at 1123
Illegal XhoI site found at 1905
Illegal XhoI site found at 2025 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 850
Illegal PstI site found at 1752 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 850
Illegal PstI site found at 1752 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 610
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 part MoClo compatible with the marburg collection. However, the Sapl 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.