Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000780 | pMuLE Lenti Dest iRFP | 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:
- pMuLE Lenti Dest iRFP
- Antibiotic Resistance:
- Chloramphenicol, Ampicillin
- Length:
- 9377 bp
- Type:
- Mammalian Expression, Lentiviral ; MuLE Gateway De
- Replication origin:
- ori
- Selection Marker:
- iRFP
- Copy Number:
- High Copy
- Promoter:
- mPGK
pMuLE Lenti Dest iRFP 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.
pMuLE Lenti Dest iRFP vector Sequence
LOCUS V000780 9377 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000780
VERSION V000780
KEYWORDS pMuLE Lenti Dest iRFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9377)
AUTHORS Albers J, Danzer C, Rechsteiner M, Lehmann H, Brandt LP, Hejhal T,
Catalano A, Busenhart P, Goncalves AF, Brandt S, Bode PK,
Bode-Lesniewska B, Wild PJ, Frew IJ
TITLE A versatile modular vector system for rapid combinatorial mammalian
genetics.
JOURNAL J Clin Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743.
PUBMED 25751063
REFERENCE 2 (bases 1 to 9377)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 9377)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Clin
Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9377
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 94..327
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 399..533
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 539..674
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(707..725)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(735..751)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 896..1351
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1380..1484
/label="AmpR promoter"
CDS 1485..2342
/label="AmpR"
/note="beta-lactamase"
rep_origin 2516..3104
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3258..3275
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 3392..3413
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3428..3458
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3466..3482
/label="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)."
primer_bind 3490..3506
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 3527..3545
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 3573..3800
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 3801..3981
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 4025..4150
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 4643..4876
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 5060..5104
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 5253..5294
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 5365..5482
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 5537..5661
/label="attR1"
/note="recombination site for the Gateway(R) LR reaction"
promoter 5686..5716
/label="lac UV5 promoter"
/note="E. coli lac promoter with an 'up' mutation"
CDS 5770..6426
/label="CmR"
/note="chloramphenicol acetyltransferase"
CDS 6771..7073
/label="ccdB"
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(7117..7241)
/label="attR2"
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 7272..7860
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
promoter 7874..8373
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 8417..9364
/label="iRFP713"
/note="phytochrome-based near-infrared fluorescent protein,
originally termed iRFP (Filonov et al., 2011)"