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pBABe-HA-ER-IPpoI vector (V000980)

Price Information

Cat No. Plasmid Name Availability Add to cart
V000980 pBABe-HA-ER-IPpoI 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:
pBABe-HA-ER-IPpoI
Antibiotic Resistance:
Ampicillin
Length:
6600 bp
Type:
Mammalian Expression, Retroviral
Replication origin:
ori
Selection Marker:
Puromycin
Promoter:
SV40
Cloning Method:
Restriction Enzyme
5' Primer:
PBABE 5'

pBABe-HA-ER-IPpoI vector Vector Map

pBABe-HA-ER-IPpoI6600 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300LTRMMLV Psigag (truncated)HAEstrogen receptorSV40 promoterPuroRLTRL4440oriAmpRAmpR promoter

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.

pBABe-HA-ER-IPpoI vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V000980                 6600 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V000980
VERSION     V000980
KEYWORDS    pBABe-HA-ER-IPpoI
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6600)
  AUTHORS   Berkovich E, Monnat RJ Jr, Kastan MB
  TITLE     Roles of ATM and NBS1 in chromatin structure modulation and DNA
            double-strand break repair.
  JOURNAL   Nat Cell Biol. 2007 Jun;9(6):683-90. Epub 2007 May 7.
   PUBMED   17486112
REFERENCE   2  (bases 1 to 6600)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6600)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Cell
            Biol."; date: "2007-06"; volume: "9(6)"; pages: "683-90. Epub 2007
            May 7"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /label="LTR"
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     misc_feature    542..741
                     /label="MMLV Psi"
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             942..1358
                     /label="gag (truncated)"
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     CDS             1386..1412
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             1800..2162
                     /gene="ESR1"
                     /label="Estrogen receptor"
                     /note="Estrogen receptor from Macaca mulatta. Accession#:
                     P49886"
     promoter        2935..3264
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3274..3870
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             3972..4563
                     /label="LTR"
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     primer_bind     complement(4693..4710)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4864..5452)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5615..6472)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6473..6577)
                     /label="AmpR promoter"