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:
- pCIB4
- 载体抗性:
- Ampicillin
- 载体长度:
- 8922 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 宿主:
- Yeast
- 载体来源:
- Dequard-Chablat M, Nguyen T-T., Contamine V, Hermann-Le Denmat S, Malagnac F.
- 启动子:
- URA3
pCIB4 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。
pCIB4 载体序列
LOCUS 40924_10731 8922 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pCIB4, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8922)
AUTHORS Dequard-Chablat M, Nguyen T-T., Contamine V, Hermann-Le Denmat S,
Malagnac F.
TITLE Efficient tools to target DNA to Podospora anserina
JOURNAL Fungal Genet Rep 59, 21-25 (2012)
REFERENCE 2 (bases 1 to 8922)
AUTHORS Dequard-Chablat M, Nguyen T-T., Contamine V, Hermann-Le Denmat S,
Malagnac F.
TITLE Direct Submission
JOURNAL Submitted (29-JUN-2012) IGM, UPS XI, CNRS-UMR 8621, 15 rue Georges
Clemenceau, Orsay, Ile de France 91405, France
REFERENCE 3 (bases 1 to 8922)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8922)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Fungal
Genet Rep 59, 21-25 (2012)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(29-JUN-2012) IGM, UPS XI, CNRS-UMR 8621, 15 rue Georges Clemenceau,
Orsay, Ile de France 91405, France"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8922
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 196..416
/label=URA3 promoter
CDS 417..1217
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
rep_origin complement(1351..1806)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 1951..1967
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 1977..1995
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 1995..2002
/label=modified AscI restriction site
/note="modified AscI restriction site"
5'UTR 2003..2963
/gene="Pa_4_5450"
gene 2003..2963
/gene="Pa_4_5450"
/label=Pa_4_5450
/note="from Podospora anserina chromosome 4"
misc_feature 2963..2970
/label=modified SwaI restriction site
/note="modified SwaI restriction site"
regulatory 2971..3306
/gene="GPD"
/gene_synonym="Pa_3_5110"
/regulatory_class="promoter"
gene 2971..3306
/gene="GPD"
/gene_synonym="Pa_3_5110"
/label=GPD
CDS 3313..3876
/label=NrsR
/note="nourseothricin acetyltransferase"
regulatory 3929..4318
/gene="AS1"
/gene_synonym="Pa_1_16650"
/regulatory_class="terminator"
gene 3929..4318
/gene="AS1"
/gene_synonym="Pa_1_16650"
/label=AS1
misc_feature 4319..4326
/label=modified PacI restriction site
/note="modified PacI restriction site"
3'UTR 4327..5285
/gene="Pa_4_5450"
gene 4327..5285
/gene="Pa_4_5450"
/label=Pa_4_5450
/note="from Podospora anserina chromosome 4"
misc_feature 5285..5292
/label=modified AscI restriction site
/note="modified AscI restriction site"
promoter complement(5320..5338)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5359..5375)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5383..5399)
/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(5407..5437)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5452..5473)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5761..6349)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6523..7380)
/label=AmpR
/note="beta-lactamase"
promoter complement(7381..7485)
/label=AmpR promoter
rep_origin complement(7512..8854)
/direction=LEFT
/label=2u ori
/note="yeast 2u plasmid origin of replication"