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:
- pCVD036
- 载体抗性:
- Ampicillin
- 载体长度:
- 3526 bp
- 载体类型:
- Broad host range vector
- 复制子:
- ori
- 载体来源:
- Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
pCVD036 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。
pCVD036 载体序列
LOCUS 40924_49192 3526 bp DNA circular SYN 18-DEC-2018
DEFINITION Broad host range vector vector pCVD036, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3526)
AUTHORS Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B,
Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
TITLE Broad-host-range vector system for synthetic biology and
biotechnology in cyanobacteria
JOURNAL Nucleic Acids Res. (2014) In press
PUBMED 25074377
REFERENCE 2 (bases 1 to 3526)
AUTHORS Taton A, Unglaub F, Wright N, Zeng WY, Paz Yepes J, Brahamsha B,
Palenik B, Peterson T, Haerizadeh F, Golden SS, Golden JW.
TITLE Direct Submission
JOURNAL Submitted (17-JUN-2014) Division of Biological Sciences, University
of California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
REFERENCE 3 (bases 1 to 3526)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3526)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res. (2014) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(17-JUN-2014) Division of Biological Sciences, University of
California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3526
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 293..309
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 320..338
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 389..394
/label=EcoRV
/note="EcoRV"
misc_feature 392..412
/note="C2G; GC-adaptor for seamless assembly, flanking
antibiotic resistance markers, functional devices or custom
sequences; synthetic"
misc_feature 413..418
/label=AgeI
/note="AgeI"
regulatory 428..651
/note="PphoA_S7942; upstream region of phoA up to 25nt
upstream of the start codon in Synechococcus elongatus
PCC7942"
/regulatory_class="promoter"
regulatory 658..698
/note="RBS_LT; RBS and flanking sequences found in Life
Tech pENTR-SD/D Topo, start downstream of the att site and
includes the T7 gene enhancer, RBS, TOPO binding site and
directional TOPO overhang; Life Technologies; synthetic"
/regulatory_class="ribosome_binding_site"
RBS 669..689
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 699..1412
/label=yeGFP
/note="yeast-enhanced green fluorescent protein"
protein_bind complement(1435..1459)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
terminator 1469..1496
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
misc_feature 1505..1510
/label=XbaI
/note="XbaI"
misc_feature 1511..1531
/note="G5C5; GC-adaptor for seamless assembly, flanking
functional devices, custom sequences or cyanobacterial
replicons; synthetic"
misc_feature 1529..1534
/label=EcoRV
/note="EcoRV"
rep_origin complement(1779..2367)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2541..3398)
/label=AmpR
/note="beta-lactamase"
promoter complement(3399..3503)
/label=AmpR promoter