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:
- pCC-TM52
- 载体抗性:
- Ampicillin
- 载体长度:
- 4410 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Jordan BR, Dimas RP, Jiang X-L., Morcos F
pCC-TM52 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。
pCC-TM52 载体序列
LOCUS V015766 4410 bp DNA circular SYN 30-JUL-2019
DEFINITION Exported.
ACCESSION V015766
VERSION V015766
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 4410)
AUTHORS Jordan BR, Dimas RP, Jiang X-L., Morcos F, Chan CTY.
TITLE Engineering DNA recognition and allosteric response properties of
TetR family proteins by using a module-swapping strategy
JOURNAL Nucleic Acids Res. (2019) In press
REFERENCE 2 (bases 1 to 4410)
AUTHORS Jordan BR, Dimas RP, Jiang X-L., Morcos F, Chan CTY.
TITLE Direct Submission
JOURNAL Submitted (02-MAR-2018) Department of Biology, The University of
Texas at Tyler, 3900 University Blvd, BEP 133, Tyler, TX 75799, USA
REFERENCE 3 (bases 1 to 4410)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res. (2019) In press"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-MAR-2018) Department of Biology, The University of Texas at
Tyler, 3900 University Blvd, BEP 133, Tyler, TX 75799, USA"
FEATURES Location/Qualifiers
source 1..4410
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 59..916
/label="AmpR"
/note="beta-lactamase"
rep_origin 1093..1681
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator 1941..2027
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind complement(2330..2351)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
regulatory 2387..2428
/label="bidir terminator"
/note="bidir terminator"
/regulatory_class="terminator"
regulatory 2446..2589
/regulatory_class="promoter"
CDS 2610..3185
/codon_start=1
/transl_table=11
/product="TetR-MphR"
/label="TetR-MphR"
/protein_id="QBZ38513.1"
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLVRMMERGVEQVRHYLNAIPIGAGPQGLWEFLQVLVRSMNTRNDFSVNYLISW
YELQVPELRTLAIQRNRAVVEGIRKRLPPGAPAAAELLLHSVIAGATMQWAVDPDGELA
DHVLAQIAAILCLMFPEHDDFQLLQPHA"
terminator complement(3200..3294)
/label="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS complement(3335..4066)
/gene="ermC"
/label="rRNA adenine N-6-methyltransferase"
/note="rRNA adenine N-6-methyltransferase from
Staphylococcus aureus. Accession#: P02979"
promoter complement(4067..4171)
/label="AmpR promoter"
terminator 4325..4372
/label="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"