Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012739 | STOP-eGFP-ROSA26TV | 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:
- STOP-eGFP-ROSA26TV
- Antibiotic Resistance:
- Ampicillin
- Length:
- 14367 bp
- Replication origin:
- ori
- Source/Author:
- Klaus Rajewsky
- Copy Number:
- High copy number
- Expression Method:
- Transient
STOP-eGFP-ROSA26TV vector 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.
STOP-eGFP-ROSA26TV vector Sequence
LOCUS STOP-eGFP-ROSA26 14367 bp DNA circular SYN 11-SEP-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS STOP-eGFP-ROSA26TV
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14367)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 14367)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..14367
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 5..1086
/label=Rosa26 left arm
/note="left homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
misc_feature 1093..1196
/label=Ad SA
/note="adenovirus major late transcript splice acceptor
(Friedrich and Soriano, 1991)"
protein_bind 1207..1240
/label=Cre recombinase binding site
/bound_moiety="Cre recombinase"
/note="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
polyA_signal complement(1247..1295)
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
CDS complement(1305..2105)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
intron 2919..2984
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 3114..3134
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 3559..3693
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind complement(3713..3746)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
protein_bind 3779..3826
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
misc_feature 3848..4427
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4428..5144
/label=EGFP
/note="enhanced GFP"
protein_bind 5172..5219
/label=FLP recombinase from the Saccharomyces
cerevisi
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FRT"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
primer_bind complement(5244..5260)
/label=SK primer
/note="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
polyA_signal 5279..5503
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
misc_feature 5596..6379
/label=Rosa26 right arm
/note="right homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
promoter 9897..10387
/note="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 10414..10983
/label=DTA
/note="diphteria toxin Fragment A"
intron 11060..11125
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
polyA_signal 11199..11423
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter complement(11522..11540)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(11561..11577)
/label=M13 rev
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 11585..11601
/label=lac repressor encoded by lacI binding site
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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(11609..11639)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(11654..11675)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(11963..12551)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(12725..13582)
/label=AmpR
/note="beta-lactamase"
promoter complement(13583..13687)
/label=AmpR promoter
rep_origin complement(13713..14168)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 14310..14326
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 14336..14354
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"