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:
- pUDE204
- 载体抗性:
- Ampicillin
- 载体长度:
- 11697 bp
- 载体类型:
- Expression vector
- 复制子:
- ori
- 载体来源:
- Kozak BU, van Rossum HM, Benjamin KR, Wu L, Daran JM, Pronk JT, van Maris AJ.
- 启动子:
- TPI1
pUDE204 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。
pUDE204 载体序列
LOCUS V002341 11697 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V002341
VERSION V002341
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11697)
AUTHORS Kozak BU, van Rossum HM, Benjamin KR, Wu L, Daran JM, Pronk JT, van
Maris AJ.
TITLE Replacement of the Saccharomyces cerevisiae acetyl-CoA synthetases
by alternative pathways for cytosolic acetyl-CoA synthesis
JOURNAL Metab. Eng. (2013) In press
PUBMED 24269999
REFERENCE 2 (bases 1 to 11697)
AUTHORS Kozak BU, van Rossum HM, Benjamin KR, Wu L, Daran J-M.G., Pronk JT,
van Maris AJA.
TITLE Direct Submission
JOURNAL Submitted (31-MAY-2013) Biotechnology, Delft University of
Technology, Julianalaan 67, Delft 2628BC, The Netherlands
REFERENCE 3 (bases 1 to 11697)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11697)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Assembly Method :: IDBA v. 0.20
Sequencing Technology :: Illumina
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
(2013) In press"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(31-MAY-2013) Biotechnology, Delft University of Technology,
Julianalaan 67, Delft 2628BC, The Netherlands"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11697
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(183..771)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(945..1802)
/label="AmpR"
/note="beta-lactamase"
promoter complement(1803..1907)
/label="AmpR promoter"
rep_origin complement(1934..3276)
/direction=LEFT
/label="2u ori"
/note="yeast 2u plasmid origin of replication"
promoter 3538..3753
/label="URA3 promoter"
CDS 3754..4554
/label="URA3"
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
rep_origin complement(4688..5143)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 5284..5300
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 5310..5328
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 5838..6357
/label="TPI1 promoter"
/note="strong constitutive promoter for yeast triose
phosphate isomerase"
CDS 6358..8637
/gene="pflB"
/label="Formate acetyltransferase 1"
/note="Formate acetyltransferase 1 from Escherichia coli
(strain K12). Accession#: P09373"
regulatory 8641..9141
/label="GDN2 terminator"
/note="GDN2 terminator"
/regulatory_class="terminator"
regulatory 9146..10146
/label="FBA1 promoter"
/note="FBA1 promoter"
/regulatory_class="promoter"
CDS 10147..10881
/gene="pflA"
/label="Pyruvate formate-lyase 1-activating enzyme"
/note="Pyruvate formate-lyase 1-activating enzyme from
Escherichia coli O6:H1 (strain CFT073 / ATCC 700928 /
UPEC). Accession#: P0A9N5"
regulatory 10885..11385
/label="PMA1 terminator"
/note="PMA1 terminator"
/regulatory_class="terminator"
primer_bind complement(11386..11402)
/label="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(11439..11457)
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(11478..11494)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 11502..11518
/label="lac operator"
/bound_moiety="lac repressor encoded by lacI"
/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(11526..11556)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(11571..11592)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."