质粒编号
质粒名称
规格
价格
V018107
pLVX-EGFP-IRES-puro
5 μg (冻干粉)
我们所展示的质粒图谱主要是从文献和开放数据库中收集而来，主要是为了方便研究工作，其中一小部分质粒进行了质量控制，可供科学家使用。
所有的产品都严格仅供科学研究使用，不能应用于临床试验，包括人体摄入、注射或外用的药理学用途。
确保质粒的关键元件正确，但是我们并不能保证实验结果。页面展示的图谱序列为理论序列，可能与测序结果不一致，请自行比对后确定是否满足要求。(如果测过序，本页面一般会提供下载)
开放数据库中的大多数载体序列都没有被完全测序。如果实际序列与参考序列的相似度超过99%，则将其视为正确。
由于科学研究是在探索未知，具有很大的不确定性，在任何情况下，我们都不承担超出质粒本身的额外经济损失或责任。
- 载体名称:
- pLVX-EGFP-IRES-puro
- 载体抗性:
- Ampicillin
- 载体长度:
- 8911 bp
- 载体类型:
- Mammalian Expression, Lentiviral
- 载体来源:
- Robert Sobol
- 启动子:
- CMV
- 克隆方法:
- Gateway Cloning
- 5'引物:
- M13 forward for pENTR vector
- 3'引物:
- M13 reverse for pENTR vector
- 感受态:
- NEB Stable
- 培养温度:
- 37℃
pLVX-EGFP-IRES-puro 载体图谱
质粒操作方法
1. 发货形式：质粒干粉(常温运输，存于-20度，请务必先转化提质粒后使用)
2. 收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O溶解质粒;（质粒复测的浓度有时候与标称值差距较大，这可能是因为冻干质粒在管中的位置、复溶效率、测量偏差以及管壁的吸附导致，因此建议先转化提质粒后再使用）
3. 取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；
4. 加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min (30℃培养1-1.5小时);
5. 6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上;
6. 将平板倒置37℃培养14h，如果要求是30℃则培养20h; (菌落过多则将质粒稀释后再转化；没有菌落则加入10μl质粒转化；建议不要直接转表达感受态， 要先转克隆感受态，重提质粒后再导入表达感受态);
7. 挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
pLVX-EGFP-IRES-puro 载体序列
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"