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:
- pMK33-CTAP
- 载体抗性:
- Ampicillin
- 载体长度:
- 8793 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Veraksa A, Bauer A, Artavanis-Tsakonas S.
- 启动子:
- MT
pMK33-CTAP 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。
pMK33-CTAP 载体序列
LOCUS 40924_30985 8793 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMK33-CTAP, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8793)
AUTHORS Veraksa A, Bauer A, Artavanis-Tsakonas S.
TITLE Analyzing protein complexes in Drosophila with tandem affinity
purification-mass spectrometry
JOURNAL Dev. Dyn. 232 (3), 827-834 (2005)
PUBMED 15704125
REFERENCE 2 (bases 1 to 8793)
AUTHORS Veraksa A, Bauer A, Artavanis-Tsakonas S.
TITLE Direct Submission
JOURNAL Submitted (17-AUG-2004) MGH Cancer Center, Harvard Medical School,
Bldg 149, 13th Street, Charlestown, MA 02129, USA
REFERENCE 3 (bases 1 to 8793)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8793)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Dev. Dyn.";
date: "2005"; volume: "232"; issue: "3"; pages: "827-834"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(17-AUG-2004) MGH Cancer Center, Harvard Medical School, Bldg 149,
13th Street, Charlestown, MA 02129, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8793
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..8
/label=NotI site
/note="NotI site"
rep_origin complement(865..1453)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1627..2484)
/label=AmpR
/note="beta-lactamase"
promoter complement(2485..2589)
/label=AmpR promoter
promoter 4071..4350
/label=copia promoter
/note="strong promoter from the Drosophila transposable
element copia (Sinclair et al., 1986)"
CDS 4396..5418
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
intron 5784..5849
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 5979..5999
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 6424..6558
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 6559..6985
/label=MT promoter
/note="Drosophila metallothionein promoter"
misc_feature 6992..7015
/note="polylinker; unique cloning sites for XhoI, BamHI,
EcoRV, SpeI"
CDS 7022..7099
/label=CBP
/note="calmodulin-binding peptide"
CDS 7127..7147
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
CDS 7187..7360
/label=ProtA
/note="IgG-binding unit of Staphylococcus aureus protein A"
CDS 7364..7534
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
EAKKLNGAQAPK"