Basic Vector Information
- Vector Name:
- pLenti-EnCMV-MCS-dTomato-PGK-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9318 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- mPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLenti-EnCMV-MCS-dTomato-PGK-Puro vector Vector Map
Plasmid Resuspension 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 ℃.
pLenti-EnCMV-MCS-dTomato-PGK-Puro vector Sequence
LOCUS 62056_13845 9318 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9318)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9318
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 5..185
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 257..391
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 418..553
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(574..592)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(602..618)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 760..1215
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1241..1345
/label=AmpR promoter
CDS 1346..2203
/label=AmpR
/note="beta-lactamase"
rep_origin 2377..2965
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 3253..3274
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3289..3319
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3327..3343
/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 3351..3367
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 3388..3406
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 3434..3660
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
misc_feature 3888..4013
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 4506..4739
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 4924..4968
/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 5117..5158
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 5235..5351
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 5374..5677
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5678..5881
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 5977..7404
/label=tdTomato
/note="tandem dimeric (pseudo-monomeric) derivative of
DsRed (Shaner et al., 2004)"
promoter 7420..7919
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 7940..8536
/label=PuroR
/note="puromycin N-acetyltransferase"
misc_feature 8546..9134
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
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