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:
- pTcFlagN-CO-2.0
- 载体抗性:
- Chloramphenicol
- 载体长度:
- 7428 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Diaz-Olmos Y, Batista M, Ludwig A, Marchini FK.
pTcFlagN-CO-2.0 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。
pTcFlagN-CO-2.0 载体序列
LOCUS 62056_21015 7428 bp DNA circular SYN 06-OCT-2020
DEFINITION Cloning vector pTcFlagN-CO 2.0, complete sequence.
ACCESSION MT180300
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7428)
AUTHORS Diaz-Olmos Y, Batista M, Ludwig A, Marchini FK.
TITLE Characterising ISWI chromatin remodeler in Trypanosoma cruzi
JOURNAL Mem. Inst. Oswaldo Cruz 115, e190457 (2020)
PUBMED 32428081
REFERENCE 2 (bases 1 to 7428)
AUTHORS Diaz-Olmos Y, Batista M, Ludwig A, Marchini FK.
TITLE Direct Submission
JOURNAL Submitted (09-MAR-2020) LACTAS, Fiocruz, Professor Algacyr Munhoz
Mader, Curitiba 81350010, Brasil
REFERENCE 3 (bases 1 to 7428)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mem. Inst.
Oswaldo Cruz 115, e190457 (2020)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(09-MAR-2020) LACTAS, Fiocruz, Professor Algacyr Munhoz Mader,
Curitiba 81350010, Brasil"
FEATURES Location/Qualifiers
source 1..7428
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 665..1281
/note="Trypanosoma cruzi 18S ribosomal RNA promoter region"
/regulatory_class="promoter"
misc_feature 1294..1798
/note="Trypanosoma cruzi 3' intergenic region of the
TcCLB.511469.50 CDS"
protein_bind 1816..1940
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 1965..1995
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 2049..2705
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 3050..3352
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(3396..3520)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
CDS 3534..3599
/label=3xFLAG
/note="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
misc_feature 3617..4037
/note="Trypanosoma cruzi 3' intergenic region of the
TcCLB.504069.70 CDS"
CDS 4047..4841
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
misc_feature 4851..5168
/note="Trypanosoma cruzi 3' intergenic region of the
TcCLB.511145.40 CDS"
promoter complement(5240..5258)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5279..5295)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5303..5319)
/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(5327..5357)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5372..5393)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5681..6269)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6443..7300)
/label=AmpR
/note="beta-lactamase"
promoter complement(7301..7405)
/label=AmpR promoter