Basic Vector Information
- Vector Name:
- pLX302
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 9573 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Source/Author:
- Yang X, Boehm JS, Yang X, Salehi-Ashtiani K, Hao T, Shen Y, Lubonja
- Copy Number:
- High copy number
- Promoter:
- hPGK
pLX302 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 ℃.
pLX302 vector Sequence
LOCUS pLX302. 9573 bp DNA circular SYN 01-JAN-1980
DEFINITION Lentiviral Gateway(R) destination vector with a puromycin resistance
marker and a V5 epitope tag, for strong expression and improved
plasmid yield and viral titer.
ACCESSION .
VERSION .
KEYWORDS pLX302.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9573)
AUTHORS Yang X, Boehm JS, Yang X, Salehi-Ashtiani K, Hao T, Shen Y, Lubonja
R, Thomas SR, Alkan O, Bhimdi T, Green TM, Johannessen CM, Silver
SJ, Nguyen C, Murray RR, Hieronymus H, Balcha D, Fan C, Lin C,
Ghamsari L, Vidal M, Hahn WC, Hill DE, Root DE.
TITLE A public genome-scale lentiviral expression library of human ORFs.
JOURNAL Nat. Methods 2011;8:659-61.
PUBMED 21706014
REFERENCE 2 (bases 1 to 9573)
AUTHORS Root Lab
TITLE Direct Submission
REFERENCE 3 (bases 1 to 9573)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
Methods"; date: "2011"; volume: "8"; pages: "659-61"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9573
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 65..368
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 369..572
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 595..719
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 744..774
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 828..1484
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 1829..2131
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(2175..2299)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
CDS 2301..2342
/label=V5 tag
/note="epitope tag from simian virus 5"
misc_feature 2343..2351
/label=stop codons
/note="stop codons"
misc_feature 2384..2972
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 3044..3277
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 3355..3489
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 3516..3651
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(3672..3690)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3700..3716)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 3858..4313
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4339..4443
/label=AmpR promoter
CDS 4444..5301
/label=AmpR
/note="beta-lactamase"
rep_origin 5475..6063
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 6351..6372
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6387..6417
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6425..6441
/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 6449..6465
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 6486..6504
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 6532..6758
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 6759..6939
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6986..7111
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7604..7837
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 8022..8066
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
promoter 8241..8741
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
CDS 8763..9359
/label=PuroR
/note="puromycin N-acetyltransferase"
misc_feature 9404..9521
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
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