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:
- pLD02X
- 载体抗性:
- Ampicillin
- 载体长度:
- 9609 bp
- 载体类型:
- Load vector
- 复制子:
- ori
- 宿主:
- Yeast
- 载体来源:
- Mishra D, Rivera PM, Lin A, Del Vecchio D, Weiss R.
- 启动子:
- URA3
pLD02X 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。
pLD02X 载体序列
LOCUS 40924_27628 9609 bp DNA circular SYN 18-DEC-2018
DEFINITION Load vector pLD02X, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9609)
AUTHORS Mishra D, Rivera PM, Lin A, Del Vecchio D, Weiss R.
TITLE A load driver device for engineering modularity in biological
networks
JOURNAL Nat. Biotechnol. 32 (12), 1268-1275 (2014)
PUBMED 25419739
REFERENCE 2 (bases 1 to 9609)
AUTHORS Mishra D, Rivera-Ortiz PM, Lin A, Del Vecchio D, Weiss R.
TITLE Direct Submission
JOURNAL Submitted (02-SEP-2014) Synthetic Biology Center, MIT, 500
Technology Square, NE47, Cambridge, MA 02139, USA
REFERENCE 3 (bases 1 to 9609)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9609)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
Biotechnol."; date: "2014"; volume: "32"; issue: "12"; pages:
"1268-1275"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-SEP-2014) Synthetic Biology Center, MIT, 500 Technology Square,
NE47, Cambridge, MA 02139, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9609
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 34..223
/label=CYC1 terminator
/note="transcription terminator for CYC1"
rep_origin complement(466..1054)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1228..2085)
/label=AmpR
/note="beta-lactamase"
promoter complement(2086..2190)
/label=AmpR promoter
rep_origin complement(2217..3559)
/direction=LEFT
/label=2u ori
/note="yeast 2u plasmid origin of replication"
promoter 3823..4043
/label=URA3 promoter
CDS 4044..4844
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
rep_origin complement(4978..5433)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
terminator complement(5512..5677)
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
protein_bind 5918..5938
/label=attB4
/note="core recombination site for the Gateway(R) BP
reaction"
regulatory 5972..6704
/regulatory_class="promoter"
misc_feature 6388..6427
/label=SKN7 binding site
/note="SKN7 binding site"
misc_feature 6428..6467
/label=SKN7 binding site
/note="SKN7 binding site"
protein_bind 6716..6740
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
primer_bind 6886..6902
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(6907..6931)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature complement(6909..6930)
/label=attB2
/note="attB2"
terminator 6954..7141
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
misc_feature 7244..7265
/label=attB4
/note="attB4"
protein_bind 7245..7265
/gene="mutant version of attB"
/label=attB4
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
regulatory 7299..8031
/regulatory_class="promoter"
misc_feature 7715..7754
/label=SKN7 binding site
/note="SKN7 binding site"
misc_feature 7755..7794
/label=SKN7 binding site
/note="SKN7 binding site"
protein_bind 8043..8067
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 8043..8064
/label=attP1
/note="attP1"
primer_bind 8213..8229
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(8234..8258)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
terminator 8273..8520
/label=CYC1 terminator
/note="transcription terminator for CYC1"
CDS 8681..9487
/label=KanR
/note="aminoglycoside phosphotransferase"