Basic Vector Information
- Vector Name:
- pLenti CMV GFP Zeo (637-7)
- Antibiotic Resistance:
- Bleomycin
- Length:
- 6474 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Zeocin
- Copy Number:
- High Copy
- Promoter:
- RSV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMVforw
pLenti CMV GFP Zeo (637-7) 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 CMV GFP Zeo (637-7) vector Sequence
LOCUS pLenti_CMV_GFP_Z 6474 bp DNA circular SYN 13-MAY-2021
DEFINITION 3rd gen lentiviral eGFP expression vector, CMV promoter, Zeo.
ACCESSION .
VERSION .
KEYWORDS pLenti CMV GFP Zeo (637-7).
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6474)
AUTHORS Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO,
Campisi J, Yaswen P, Cooper PK, Kaufman PD.
TITLE A versatile viral system for expression and depletion of proteins in
mammalian cells.
JOURNAL PLoS One. 2009 Aug 6;4(8):e6529.
PUBMED 19657394
REFERENCE 2 (bases 1 to 6474)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6474)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
date: "2009-08-6"; volume: "4(8)"; pages: "e6529"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6474
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 71..659
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 813..830
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 947..968
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 983..1013
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1021..1037
/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 1045..1061
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 1082..1100
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 1128..1354
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 1355..1535
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1582..1707
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 2200..2433
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2618..2662
/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 2811..2852
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 2929..3045
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 3068..3371
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3372..3575
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3572..3596
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 3671..4387
/label=EGFP
/note="enhanced GFP"
misc_feature 4406..4994
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
promoter 5050..5379
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 5391..5438
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 5457..5828
/label=BleoR
/note="antibiotic-binding protein"
LTR 5924..6157
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6229..6363
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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