Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000664 | lentiSAM v2 (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:
- lentiSAM v2 (Puro)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13713 bp
- Type:
- Mammalian Expression, Lentiviral, CRISPR
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- High Copy
- Promoter:
- U6
- Cloning Method:
- Restriction Enzyme
lentiSAM v2 (Puro) 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.
lentiSAM v2 (Puro) vector Sequence
LOCUS V000664 13713 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V000664
VERSION V000664
KEYWORDS lentiSAM v2 (Puro)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13713)
TITLE Lenti CRISPR Activate
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 13713)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 13713)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13713
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 89..418
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 466..513
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 532..903
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 1036..1169
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(1206..1222)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1230..1246)
/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)."
promoter complement(1254..1284)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(1299..1320)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1437..1454)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(1608..2196)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2370..3227)
/label="AmpR"
/note="beta-lactamase"
promoter complement(3228..3332)
/label="AmpR promoter"
primer_bind complement(3407..3426)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 3598..3977
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 3978..4180
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 4195..4375
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 4422..4547
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5040..5273
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 5458..5502
/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 5651..5692
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 5800..5917
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 5967..6207
/label="U6 promoter"
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 6253..6271
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 6323..6341
/label="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
promoter 6428..6639
/label="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 6656..6703
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label="nucleoplasmin NLS"
/translation="KRPAATKKAGQAKKKK"
CDS 6704..10804
/label="dCas9"
/note="catalytically dead mutant of the Cas9 endonuclease
from the Streptococcus pyogenes Type II CRISPR/Cas system"
CDS 10859..10879
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 10904..11053
/label="VP64"
/note="tetrameric repeat of the minimal activation domain
of herpes simplex virus VP16 (Beerli et al., 1998)"
CDS 11075..11131
/label="P2A"
/note="2A peptide from porcine teschovirus-1 polyprotein"
CDS 11132..11725
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 11765..12353
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(12356..12372)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(12357..12373)
/label="pBluescriptKS"
/note="For pBluescript vector"
LTR 12878..13058
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 13090..13314
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 13360..13713
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"