质粒编号
质粒名称
规格
价格
V014991
FUW_tetO_GFP_hOKMS
5 μg
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:
- FUW_tetO_GFP_hOKMS
- 载体抗性:
- Ampicillin
- 载体长度:
- 14810 bp
- 载体类型:
- Protein expression
- 复制子:
- ori
- 宿主:
- Mammalian cells, Lentivirus
- 启动子:
- SV40
- 5'引物:
- SV40pro-F：TATTTATGCAGAGGCCGAGG
- 3'引物:
- WPRE-R：CATAGCGTAAAAGGAGCAACA
- 培养温度:
- 37℃
FUW_tetO_GFP_hOKMS 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。
FUW_tetO_GFP_hOKMS 载体序列
LOCUS V014991 14810 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V014991
VERSION V014991
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 14810)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..14810
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 7..386
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 387..589
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 604..784
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 831..956
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1449..1682
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1867..1911
/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 2060..2101
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2209..2326
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 2385..2655
/label="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
CDS 2869..3585
/label="EGFP"
/note="enhanced GFP"
misc_feature 3700..4273
/label="IRES"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4280..5359
/label="hOct4"
/note="Homo sapiens Oct-4 gene. Encodes a transcription
factor containing a POU homeodomain that plays a key role
in embryonic development and stem cell pluripotency.
Aberrant expression in adult tissues is associated with
tumorigenesis."
CDS 5423..6832
/label="hKLF4"
/note="Homo sapiens Kruppel-like factor 4 (Klf4) gene.
Belongs to the relatively large family of SP1-like
transcription factors and is involved in the regulation of
proliferation, differentiation, apoptosis and somatic cell
reprogramming."
CDS 6899..8215
/label="c-Myc"
/note="human c-Myc proto-oncogene"
CDS 8285..9235
/label="hSOX2"
/note="Homo sapiens transcription factor SOX-2 gene.
Belongs to the SRY-related HMG-box (SOX) family of
transcription factors, which is involved in the regulation
of embryonic development and in the determination of cell
fate"
misc_feature 9271..9859
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 10384..10564
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 10596..10820
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 10866..11294
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 11308..11637
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 11685..11732
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 11751..12122
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 12255..12388
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(12473..12503)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(12518..12539)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(12827..13415)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(13589..14446)
/label="AmpR"
/note="beta-lactamase"
promoter complement(14447..14551)
/label="AmpR promoter"