Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V017845 | V5-GFP-TurboID-KDEL_pLX208 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- V5-GFP-TurboID-KDEL_pLX208
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10081 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Hyg
- Promoter:
- CMV
- 5' Primer:
- CMV-F
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
V5-GFP-TurboID-KDEL_pLX208 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.
V5-GFP-TurboID-KDEL_pLX208 vector Sequence
LOCUS . 10081 bp DNA circular UNK 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION
VERSION
KEYWORDS .
SOURCE .
ORGANISM .
.
FEATURES Location/Qualifiers
misc_feature 20..608
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 680..913
/label="3' LTR (ΔU3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 991..1125
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 1152..1287
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(1308..1326)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin 1494..1949
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1975..2079
/label="AmpR promoter"
CDS 2080..2937
/label="AmpR"
/note="β-lactamase"
rep_origin 3111..3699
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 3987..4008
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4023..4053
/label="lac promoter"
/note="promoter for the E. coli lac operon"
promoter 4122..4140
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 4168..4394
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 4395..4575
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 4622..4747
/label="HIV-1 Ψ"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5240..5473
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 5658..5702
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
promoter 5877..6377
/label="hPGK promoter"
/note="human phosphoglycerate kinase 1 promoter"
CDS 6389..7411
/label="HygR"
/note="hygromycin B phosphotransferase"
misc_feature 7465..7582
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 7699..8002
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 8003..8206
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 8343..9059
/label="EGFP"
/note="the original enhanced GFP (Yang et al., 1996)"
CDS 9078..10034
/label="TurboID"
/note="mutant of the E. coli biotin ligase BioID engineered
for improved catalytic efficiency (Branon et al., 2018)"