pMIGR1-FLAG-BCR-ABL-P210 vector (V013619)

Price Information

Cat No. Plasmid Name Availability Add to cart
V013619 pMIGR1-FLAG-BCR-ABL-P210 In stock, 1 week for quality controls

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

Basic Vector Information

Vector Name:
pMIGR1-FLAG-BCR-ABL-P210
Antibiotic Resistance:
Ampicillin
Length:
12564 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Retrovirus
Promoter:
MSCV
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pMIGR1-FLAG-BCR-ABL-P210 vector Vector Map

pMIGR1-FLAG-BCR-ABL-P21012564 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000FLAGTyrosine-protein kinase ABL1IRES2EGFP3' LTRlac promoterCAP binding siteoriAmpRAmpR promoter5' LTRMESV Psigag (truncated)

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.

pMIGR1-FLAG-BCR-ABL-P210 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V013619                12564 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013619
VERSION     V013619
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 12564)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             156..179
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             2961..6272
                     /gene="ABL1"
                     /label="Tyrosine-protein kinase ABL1"
                     /note="Tyrosine-protein kinase ABL1 from Homo sapiens.
                     Accession#: P00519"
     misc_feature    6284..6860
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             6861..7577
                     /label="EGFP"
                     /note="enhanced GFP"
     LTR             7682..8196
                     /label="3' LTR"
                     /note="3' long terminal repeat from murine embryonic stem
                     cell virus"
     promoter        complement(8382..8412)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(8427..8448)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(8736..9324)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(9498..10355)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(10356..10460)
                     /label="AmpR promoter"
     LTR             11288..11804
                     /label="5' LTR"
                     /note="5' long terminal repeat from murine embryonic stem
                     cell virus"
     misc_feature    11868..12209
                     /label="MESV Psi"
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     CDS             join(12276..12564,1..128)
                     /label="gag (truncated)"
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"