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:
- pCDH-EF1a-GaussiaSP-MCS-IRES-zeo
- 载体抗性:
- Ampicillin
- 载体长度:
- 7504 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Kulemzin SV, Gorchakov AA, Chikaev NA, Volkova OY, Kuznetsova VV, Taranin AV.
- 启动子:
- RSV
pCDH-EF1a-GaussiaSP-MCS-IRES-zeo 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。
pCDH-EF1a-GaussiaSP-MCS-IRES-zeo 载体序列
LOCUS V008631 7504 bp DNA circular SYN 17-DEC-2018
DEFINITION Exported.
ACCESSION V008631
VERSION V008631
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7504)
AUTHORS Kulemzin SV, Gorchakov AA, Chikaev NA, Volkova OY, Kuznetsova VV,
Taranin AV.
TITLE Modular lentiviral vector system for optimization of chimeric
antigen receptor design
JOURNAL Russ. J. Bioorganic Chem. (2016) In press
REFERENCE 2 (bases 1 to 7504)
AUTHORS Kulemzin SV, Gorchakov AA, Chikaev NA, Volkova OY, Kuznetsova VV,
Taranin AV.
TITLE Direct Submission
JOURNAL Submitted (19-AUG-2016) Immunogenetics, Institute of Molecular and
Cellular Biology, Lavrentyeva 8/2, Novosibirsk 630090, Russia
REFERENCE 3 (bases 1 to 7504)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7504)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Russ. J.
Bioorganic Chem. (2016) In press"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(19-AUG-2016) Immunogenetics, Institute of Molecular and Cellular
Biology, Lavrentyeva 8/2, Novosibirsk 630090, Russia"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7504
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 6..232
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 233..413
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 460..585
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1078..1311
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1496..1540
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 1689..1730
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 1807..1923
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1966..2177
/label="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 2190..2458
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
regulatory 2484..2493
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
misc_feature 2490..2540
/label="luciferase signal peptide from Gaussia princeps"
/note="luciferase signal peptide from Gaussia princeps"
misc_feature 2597..3183
/label="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 3202..3573
/label="BleoR"
/note="antibiotic-binding protein"
misc_feature 3583..4171
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4245..4478
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4550..4684
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 4711..4846
/label="SV40 ori"
/note="SV40 origin of replication"
primer_bind complement(4879..4895)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4903..4919)
/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)."
promoter complement(4927..4957)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(4972..4993)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5281..5869)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6043..6900)
/label="AmpR"
/note="beta-lactamase"
promoter complement(6901..7005)
/label="AmpR promoter"
primer_bind 7479..7495
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"