MSCB-hDot1Lwt vector (V008318)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V008318	MSCB-hDot1Lwt	In stock, 1 week for quality controls	Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:: MSCB-hDot1Lwt

Antibiotic Resistance:: Ampicillin

Length:: 9994 bp

Type:: Retroviral

Replication origin:: ori

Promoter:: mPGK

MSCB-hDot1Lwt vector Map

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Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

MSCB-hDot1Lwt vector Sequence

LOCUS       V008318                 9994 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V008318
VERSION     V008318
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"
            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="Histone-lysine N-methyltransferase, H3 lysine-79
                     specific"
                     /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)"