Basic Vector Information
- Vector Name:
- pLenti-EnCMV-MCS-P2A-mCherry-SV40-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8514 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- RSV
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pLenti-EnCMV-MCS-P2A-mCherry-SV40-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-P2A-mCherry-SV40-Puro vector Sequence
LOCUS 62056_13850 8514 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 8514)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8514
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 113..134
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 149..179
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 187..203
/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 211..227
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 248..266
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 294..520
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 521..701
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 748..873
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1366..1599
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1784..1828
/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 1977..2018
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 2095..2211
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 2234..2537
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2538..2741
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 2905..3612
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
promoter 3646..3975
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 3985..4581
/label=PuroR
/note="puromycin N-acetyltransferase"
misc_feature 4651..5239
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5326..5559
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5631..5765
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter complement(5948..5966)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5976..5992)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 6134..6589
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6615..6719
/label=AmpR promoter
CDS 6720..7577
/label=AmpR
/note="beta-lactamase"
rep_origin 7751..8339
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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