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:
- pInducer20
- 载体抗性:
- Chloramphenicol, Ampicillin
- 载体长度:
- 11808 bp
- 载体类型:
- Mammalian Expression, Lentiviral
- 复制子:
- ori
- 筛选标记:
- Neomycin (select with G418)
- 启动子:
- UbC
- 克隆方法:
- Restriction Enzyme
- 5'引物:
- LNCX_primer
pInducer20 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。
pInducer20 载体序列
LOCUS V007219 11808 bp DNA circular SYN 08-SEP-2021
DEFINITION Exported.
ACCESSION V007219
VERSION V007219
KEYWORDS pInducer20
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11808)
AUTHORS Meerbrey KL, Hu G, Kessler JD, Roarty K, Li MZ, Fang JE,
Herschkowitz JI, Burrows AE, Ciccia A, Sun T, Schmitt EM, Bernardi
RJ, Fu X, Bland CS, Cooper TA, Schiff R, Rosen JM, Westbrook TF,
Elledge SJ
TITLE The pINDUCER lentiviral toolkit for inducible RNA interference in
vitro and in vivo.
JOURNAL Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3665-70. doi:
10.1073/pnas.1019736108. Epub 2011 Feb 9.
PUBMED 21307310
REFERENCE 2 (bases 1 to 11808)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11808)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1073/pnas.1019736108"; journalName: "Proc Natl Acad Sci U S A";
date: "2011-03-1- 1"; volume: "108"; issue: "9"; pages: "3665-70"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11808
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(75..818)
/label="rtTA-Advanced"
/note="improved tetracycline-controlled transactivator"
primer_bind complement(1021..1040)
/label="hUBCpro-F"
/note="Human Ubiquitin C (UbC) promoter, forward primer"
misc_feature complement(1255..1811)
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS complement(2174..2200)
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
protein_bind 2253..2377
/label="attR2"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(2421..2723)
/label="ccdB"
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
CDS complement(3068..3724)
/label="CmR"
/note="chloramphenicol acetyltransferase"
promoter complement(3778..3808)
/label="lac UV5 promoter"
/note="E. coli lac promoter with an 'up' mutation"
protein_bind complement(3833..3957)
/label="attR1"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(4066..4090)
/label="LNCX"
/note="Human CMV promoter, forward primer"
promoter complement(4087..4125)
/label="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind complement(4158..4428)
/label="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
misc_feature complement(4483..4600)
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(4703..4744)
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
CDS complement(4893..4937)
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
misc_feature complement(5122..5355)
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature complement(5852..5977)
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
LTR complement(6024..6657)
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(6849..6868)
/label="EBV-rev"
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6918..6934)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6918..6934)
/label="M13 Reverse"
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6931..6953)
/label="M13/pUC Reverse"
/note="In lacZ gene"
protein_bind 6942..6958
/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(6966..6996)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(7011..7032)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7161..7180)
/label="SV40pro-F"
/note="SV40 promoter/origin, forward primer"
primer_bind complement(7272..7289)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(7443..8031)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8205..9062)
/label="AmpR"
/note="beta-lactamase"
promoter complement(9063..9167)
/label="AmpR promoter"
primer_bind 9235..9253
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
LTR complement(9372..9605)
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
misc_feature complement(9680..10268)
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(10448..11239)
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
misc_feature complement(11255..11805)
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"