Basic Vector Information
2XMyc-LRRK2-WT vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
2XMyc-LRRK2-WT vector Sequence
LOCUS 2XMyc-LRRK2-WT. 11985 bp DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS 2XMyc-LRRK2-WT. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11985) TITLE The Parkinson Disease-associated Leucine-rich Repeat Kinase 2 (LRRK2) Is a Dimer That Undergoes Intramolecular Autophosphorylation JOURNAL J Biol Chem. 2008 Jun 13;283(24):16906-14. PUBMED 18397888 REFERENCE 2 (bases 1 to 11985) TITLE Direct Submission REFERENCE 3 (bases 1 to 11985) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol Chem."; date: "2008-06-13"; volume: "283(24)"; pages: "16906-14" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..11985 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 68..371 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 372..575 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 621..639 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" regulatory 671..680 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 680..709 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 725..754 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" protein_bind 773..797 /label=attB1 /note="recombination site for the Gateway(R) BP reaction" CDS 812..8392 /gene="LRRK2" /label=LRRK2 /note="Leucine-rich repeat serine/threonine-protein kinase 2 from Homo sapiens. Accession#: Q5S007" protein_bind complement(8414..8438) /label=attB2 /note="recombination site for the Gateway(R) BP reaction" promoter complement(8488..8506) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 8780..8901 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(8908..9363) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 9390..9492 /label=AmpR promoter promoter 9496..9853 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 9888..10679 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" primer_bind complement(10870..10889) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 10914..10961 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 11290..11878 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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