质粒编号
质粒名称
规格
价格
V018126
Luciferase-pcDNA3
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。
Luciferase-pcDNA3 is a mammalian expression vector containing the firefly luciferase gene under a CMV promoter, used for gene expression studies and reporter assays, with neomycin resistance for selection.
- 载体名称:
- Luciferase-pcDNA3
- 载体抗性:
- Ampicillin
- 载体长度:
- 7040 bp
- 载体类型:
- Mammalian Expression Vectors
- 载体来源:
- William Kaelin
- 拷贝数:
- High copy number
- 培养温度:
- 37℃
Luciferase-pcDNA3 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。
Luciferase-pcDNA3 载体序列
LOCUS Exported 7040 bp DNA circular SYN 14-APR-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Luciferase-pcDNA3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7040)
AUTHORS Safran M, Kim WY, O'Connell F, Flippin L, Gunzler V, Horner JW,
Depinho RA, Kaelin WG
TITLE Mouse model for noninvasive imaging of HIF prolyl hydroxylase
activity: assessment of an oral agent that stimulates erythropoietin
production.
JOURNAL Proc Natl Acad Sci U S A. 2006 Jan 3. 103(1):105-10.
PUBMED 16373502
REFERENCE 2 (bases 1 to 7040)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
Acad Sci U S A. 2006 Jan 3. 103(1):105-10."
FEATURES Location/Qualifiers
source 1..7040
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(105..125)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 110..439
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 290..425
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 352..371
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 506..1300
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(560..579)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1170..1189
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 1474..1595
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1511..1530)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1565..1584
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(1644..1660)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1644..1660)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1657..1679)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1668..1684
/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(1692..1722)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1737..1758
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1875..1892)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2046..2634)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2126..2145)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2805..3665)
/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 3428..3447
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3666..3770)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3845..3864)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 4036..4415
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4416..4619
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4570..4590
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 4664..4683
/label=T7
/note="T7 promoter, forward primer"
promoter 4664..4682
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 4719..4728
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 4725..6377
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(4791..4809)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
promoter complement(6394..6412)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(6395..6412)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(6432..6449)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6438..6662
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin join(6708..7040,1..96)
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6795..6814)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7005..7026
/label=F1ori-F
/note="F1 origin, forward primer"