Basic Vector Information
MSCB-hDot1Lwt 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 ℃.
MSCB-hDot1Lwt vector Sequence
LOCUS MSCB-hDot1Lwt. 9994 bp DNA circular SYN 13-MAY-2021
DEFINITION For packaging hDot1Lwt to retrovirus..
ACCESSION .
VERSION .
KEYWORDS MSCB-hDot1Lwt.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9994)
AUTHORS Okada Y, Feng Q, Lin Y, Jiang Q, Li Y, Coffield VM, Su L, Xu G,
Zhang Y
TITLE hDOT1L links histone methylation to leukemogenesis.
JOURNAL Cell. 2005 Apr 22;121(2):167-78.
PUBMED 15851025
REFERENCE 2 (bases 1 to 9994)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 9994)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell.";
date: "2005-04-22"; volume: "121(2)"; pages: "167-78"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9994
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1..417
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
CDS 428..454
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 455..5062
/gene="DOT1L"
/label=DOT1L
/note="Histone-lysine N-methyltransferase, H3 lysine-79
specific from Homo sapiens. Accession#: Q8TEK3"
promoter 5344..5843
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
LTR 5914..6428
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind 6590..6606
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(6614..6644)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6659..6680)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6797..6814)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6968..7556)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7730..8587)
/label=AmpR
/note="beta-lactamase"
promoter complement(8588..8692)
/label=AmpR promoter
enhancer 8849..9152
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 9153..9356
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 9595..9794
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
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