Part:BBa_K3726070:Design
Lv.1 AcrbV2 - A
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 2033
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1634
Illegal NheI site found at 1762
Illegal PstI site found at 2033 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1573
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 2033
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 2033
Illegal NgoMIV site found at 3169
Illegal NgoMIV site found at 4685
Illegal AgeI site found at 109
Illegal AgeI site found at 1408 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 1390
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 two connectors that will allow the assembly of a Lv2 construct. In its upstream region the connector is 5'Con2 K2560066, and in its downstream region 3'Con2 K2560071
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
For last, regarding to P_pBAD (BBa_K3726076), the Sapl site should be removed in order to make this part RFC1000 compatible. however, this part is MoClo compatible with the marburg collection.
Source
This construct has been made by golden gate reaction.
References
"Enhancing Tolerance to Short-Chain Alcohols by Engineering the Escherichia coli AcrB Efflux Pump to Secrete the Non-native Substrate n-Butanol", ACS Publications, 2021. [Online]. Available: https://pubs.acs.org/doi/10.1021/sb400065q.