Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V018107 | pLVX-EGFP-IRES-puro | In stock, 1 week for quality controls |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pLVX-EGFP-IRES-puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8911 bp
- Type:
- Mammalian Expression, Lentiviral
- Source/Author:
- Robert Sobol
- Promoter:
- CMV
- Cloning Method:
- Gateway Cloning
- 5' Primer:
- M13 forward for pENTR vector
- 3' Primer:
- M13 reverse for pENTR vector
- Growth Strain(s):
- NEB Stable
- Growth Temperature:
- 37℃
pLVX-EGFP-IRES-puro 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.
pLVX-EGFP-IRES-puro vector Sequence
LOCUS Exported 8911 bp DNA circular SYN 14-APR-2025
DEFINITION mammalian expression of EGFP with puromycin selection.
ACCESSION .
VERSION .
KEYWORDS pLVX-EGFP-IRES-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8911)
AUTHORS Fang Q, Inanc B, Schamus S, Wang XH, Wei L, Brown AR, Svilar D,
Sugrue KF, Goellner EM, Zeng X, Yates NA, Lan L, Vens C, Sobol RW
TITLE HSP90 regulates DNA repair via the interaction between XRCC1 and DNA
polymerase beta.
JOURNAL Nat Commun. 2014 Nov 26;5:5513. doi: 10.1038/ncomms6513.
PUBMED 25423885
REFERENCE 2 (bases 1 to 8911)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun."; date: "2014-11-26"; volume: "5:5513. doi"; pages: "
10.1038/ncomms6513"
FEATURES Location/Qualifiers
source 1..8911
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2028..2143
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
enhancer 2201..2504
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2505..2708
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2659..2679
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2705..2729
/label=LNCX
/note="Human CMV promoter, forward primer"
protein_bind 2824..2848
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 2869..3588
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 2869..3585
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2914..2935)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3175..3194)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3522..3543
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind complement(3605..3629)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 3661..4247
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(3828..3845)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 4055..4074
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 4267..4866
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(4267..4286)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4763..4783
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 4880..5468
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4933..4953)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5351..5362)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5675..6308
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(6436..6452)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6436..6452)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6449..6471)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6460..6476
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(6484..6514)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6529..6550
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6667..6684)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6838..7426)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6918..6937)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7597..8457)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 8220..8239
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8458..8562)
/gene="bla"
/label=AmpR promoter
polyA_signal 8610..8744
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8647..8666)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8701..8720
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"