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:
- pTRIPZ
- 载体抗性:
- Ampicillin
- 载体长度:
- 13320 bp
- 载体类型:
- Viral Expression & Packaging Vectors
- 复制子:
- ori
- 载体来源:
- Open Biosystems
- 拷贝数:
- High copy number
- 启动子:
- UbC
pTRIPZ 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。
pTRIPZ 载体序列
LOCUS V010351 13320 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V010351
VERSION V010351
KEYWORDS pTRIPZ
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13320)
AUTHORS Open Biosystems
TITLE Direct Submission
REFERENCE 2 (bases 1 to 13320)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
Excision of rtTA3 using Cre recombinase or BamHI yields a vector
that can be used for Tet-repressible expression.
FEATURES Location/Qualifiers
source 1..13320
/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
/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 1914..1955
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2063..2180
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(2248..2619)
/label="BleoR"
/note="antibiotic-binding protein"
promoter complement(2639..2686)
/label="EM7 promoter"
/note="synthetic bacterial promoter"
protein_bind 2722..2740
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2757..2775
/gene="tetO"
/label="tetracycline repressor TetR binding site"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2793..2811
/gene="tetO"
/label="tetracycline repressor TetR binding site"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2829..2847
/gene="tetO"
/label="tetracycline repressor TetR binding site"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2864..2882
/gene="tetO"
/label="tetracycline repressor TetR binding site"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2900..2918
/gene="tetO"
/label="tetracycline repressor TetR binding site"
/bound_moiety="tetracycline repressor TetR"
/note="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
promoter 2930..2968
/label="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 3005..3697
/label="TurboRFP"
/note="red fluorescent protein from Entacmaea quadricolor"
ncRNA 3713..3806
/label="5' miR-30a"
/note="sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
ncRNA 3832..3944
/label="3' miR-30a"
/note="sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
promoter complement(4039..4057)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 4070..5281
/label="UbC promoter"
/note="human ubiquitin C promoter"
protein_bind 5294..5327
/label="Cre recombinase binding site"
/bound_moiety="Cre recombinase"
/note="loxP511"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GTATACAT). loxP511 sites are compatible with each
other, but incompatible with wild-type loxP sites."
CDS 5334..6038
/label="rtTA3"
/note="reverse tetracycline transactivator 3"
protein_bind complement(6042..6075)
/label="loxP511"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are
compatible with each other, but incompatible with wild-type
loxP sites."
misc_feature 6098..6680
/label="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 6679..7272
/label="PuroR"
/note="puromycin N-acetyltransferase"
misc_feature 7291..7879
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 8086..8319
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 8426..8650
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 8696..9124
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 9138..9467
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 9516..10545
/label="HygR (inactivated)"
/note="HygR (inactivated)"
/note="The hygromycin resistance gene was inactivated by
destroying an EcoRI site within the coding sequence."
polyA_signal 10675..10796
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(10893..10923)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(10938..10959)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(11247..11835)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(12009..12866)
/label="AmpR"
/note="beta-lactamase"
promoter complement(12867..12971)
/label="AmpR promoter"
polyA_signal 13019..13153
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"