Coding
Part:BBa_K4574060
Designed by: Isabel Aschenbrenner Group: iGEM23_Munich (2023-10-06)
MOR4649-LC
pcDNA3.4 vector containing anti-IL-23A light chain of MOR-4649 (guselkumab) monoclonal IgG1 antibody.
Design
This construct is comprised of the following components:
- XhoI (Part:BBa_K4574029): XhoI restriction site, allows for insert extraction
- Kozak (Part:BBa_K4574027): Consensus sequence which enhances the initiation of translation
- hVL-Leader (Part:BBa_K4574001): Human light chain lambda leader sequence for antibody secretion
- MOR4649-VL (Part:BBa_K4574004): Variable light chain locus targeting human interleukin-23 subunit α (IL-23A)
- hIG-CL (Part:BBa_K4574018): Human immunoglobulin light chain constant lambda region
- 2xStop (Part:BBa_K4574028): Double stop codon sequence ensuring enhanced translation termination
- SacI (Part:BBa_K4574030): SacI restriction site, allows for insert extraction
- pcDNA3.4 (Part:BBa_K4574000): pcDNA3.4-TOPO backbone for constitutive high-level transgene expression in mammalian systems
Usage and Biology
This construct has been used in mammalian cell culture (HEK293T, Exi293, RAMOS). Specifically, it has been co-expressed (together with its respective heavy chain Part:BBa_K4574061) using a high-yield Expi293 expression system (Gibco, a brand of Thermo Fisher Scientific, Braunschweig, Germany; catalog number: A14635) and purified in FPLC using Protein A agarose cartridges (ibidi, Goettingen, Germany; catalog number: 6-2022-001).
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 6725
Illegal SpeI site found at 6066
Illegal PstI site found at 307
Illegal PstI site found at 457
Illegal PstI site found at 2697 - 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 6066
Illegal PstI site found at 307
Illegal PstI site found at 457
Illegal PstI site found at 2697 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1647
Illegal BglII site found at 5829
Illegal XhoI site found at 1 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 6725
Illegal SpeI site found at 6066
Illegal PstI site found at 307
Illegal PstI site found at 457
Illegal PstI site found at 2697 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 6725
Illegal SpeI site found at 6066
Illegal PstI site found at 307
Illegal PstI site found at 457
Illegal PstI site found at 2697
Illegal NgoMIV site found at 553
Illegal NgoMIV site found at 1290
Illegal NgoMIV site found at 1807
Illegal NgoMIV site found at 3148
Illegal NgoMIV site found at 3433
Illegal AgeI site found at 647
Illegal AgeI site found at 748 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1530
Illegal BsaI.rc site found at 4961
Illegal SapI site found at 3878
Illegal SapI.rc site found at 2997
Illegal SapI.rc site found at 3207
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Categories
Parameters
None |