Composite
Part:BBa_K4482012:Design
Designed by: Matteo Da Ros Group: iGEM22_NYC_Empire_State (2022-09-30)
HIRMAb-HC-NT3
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1917
Illegal EcoRI site found at 2010
Illegal XbaI site found at 1771
Illegal SpeI site found at 262
Illegal SpeI site found at 2026
Illegal PstI site found at 1354 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1917
Illegal EcoRI site found at 2010
Illegal SpeI site found at 262
Illegal SpeI site found at 2026
Illegal PstI site found at 1354 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1917
Illegal EcoRI site found at 2010
Illegal BamHI site found at 1415
Illegal XhoI site found at 16 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1917
Illegal EcoRI site found at 2010
Illegal XbaI site found at 1771
Illegal SpeI site found at 262
Illegal SpeI site found at 2026
Illegal PstI site found at 1354 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1917
Illegal EcoRI site found at 2010
Illegal XbaI site found at 1771
Illegal SpeI site found at 262
Illegal SpeI site found at 2026
Illegal PstI site found at 1354 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1077
Illegal BsaI.rc site found at 39
Illegal SapI.rc site found at 387
Design Notes
The sequences for HIRMAb-HC and NT-3 were ordered from Twist Bioscience as gene fragments and combined together with Gibson Assembly into a mammalian expression vector.
Source
The coding sequence for NT-3 was obtained from NCBI database (NP_001096124) and it was codon optimized for its synthesis by Twist Bioscience. The protein sequence for HIRMAb-HC was obtained from US patent US-8858950-B2 and back translated to obtain a DNA coding sequence for the protein, this DNA coding sequence was codon optimized for synthesis by Twist Bioscience and for expression in mammalian cells.