Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V010490 | pBABE-Puro | In stock, instant shipping |
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'
- Growth Strain(s):
- Top10
pBABE-Puro vector 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