Difference between revisions of "Part:BBa K3726068:Design"
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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 | 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. | + | 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=== | ===Source=== | ||
Revision as of 14:00, 20 October 2021
Lv.1 SigB - A
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1103
Illegal PstI site found at 649
Illegal PstI site found at 2005 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1103
Illegal PstI site found at 649
Illegal PstI site found at 2005 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1103
Illegal PstI site found at 649
Illegal PstI site found at 2005 - 1000INCOMPATIBLE 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.