pBABE-Puro vector (V010490)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V010490	pBABE-Puro	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:: pBABE-Puro

Antibiotic Resistance:: Ampicillin

Length:: 5169 bp

Type:: Viral Expression & Packaging Vectors

Replication origin:: ori

Source/Author:: Morgenstern JP, Land H.

Selection Marker:: Puromycin

Copy Number:: High copy number

Cloning Method:: Restriction Enzyme

5' Primer:: pBABE 5'

3' Primer:: pBABE 3'

pBABE-Puro vector Map

Copy Sequence View Map

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-Puro vector Sequence

LOCUS       pBABE-Puro.        5169 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Retroviral expression vector derived from the Moloney murine 
            leukemia virus. Contains a puromycin resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    pBABE-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5169)
  AUTHORS   Morgenstern JP, Land H.
  TITLE     Advanced mammalian gene transfer: high titre retroviral vectors with
            multiple drug selection markers and a complementary helper-free 
            packaging cell line.
  JOURNAL   Nucleic Acids Res. 1990;18:3587-96.
  PUBMED    2194165
REFERENCE   2  (bases 1 to 5169)
  AUTHORS   Cell Biolabs
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5169)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "1990"; volume: "18"; pages: "3587-96"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     Strong expression is driven by the viral promoter in the 5' LTR. The
            commercially provided sequence was modified to include the splice 
            donor mutation.
FEATURES             Location/Qualifiers
     source          1..5169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     misc_feature    541
                     /label=splice donor mutation
                     /note="splice donor mutation"
                     /note="T-to-C mutation that blocks RNA splicing"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             934..1350
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     misc_feature    1355..1402
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     promoter        1413..1742
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             1752..2348
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     LTR             2450..3043
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     rep_origin      complement(3415..4003)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4184..5041)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5042..5146)
                     /label=AmpR promoter