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:
- pSUMO1
- 载体抗性:
- Streptomycin
- 载体长度:
- 7815 bp
- 载体类型:
- Bacterial Expression
- 复制子:
- CloDF13 ori
- 拷贝数:
- Low Copy
- 启动子:
- rbs
- 克隆方法:
- Restriction Enzyme
- 5'引物:
- GATGAGAAAGAGAATCTCAG
- 3'引物:
- TGCTAGTTATTGCTCAGCGG
pSUMO1 载体载体图谱
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。
pSUMO1 载体载体序列
LOCUS V001155 7815 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V001155
VERSION V001155
KEYWORDS pSUMO1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7815)
AUTHORS Weber AR, Schuermann D, Schar P
TITLE Versatile Recombinant SUMOylation System for the Production of
SUMO-Modified Protein.
JOURNAL PLoS One. 2014 Jul 9;9(7):e102157. doi:
10.1371/journal.pone.0102157. eCollection 2014.
PUBMED 25007328
REFERENCE 2 (bases 1 to 7815)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7815)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
date: "2014-07-9"; pages: "
10.1371/journal.pone.0102157. eCollection 2014"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7815
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 2..20
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 21..45
/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)."
RBS 60..82
/label="RBS"
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 101..118
/label="6xHis"
/note="6xHis affinity tag"
CDS 140..430
/gene="SUMO1"
/label="Small ubiquitin-related modifier 1"
/note="Small ubiquitin-related modifier 1 from Sus scrofa.
Accession#: A7WLH8"
promoter 579..597
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 598..622
/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)."
CDS 665..1138
/gene="ube2i"
/label="SUMO-conjugating enzyme UBC9"
/note="SUMO-conjugating enzyme UBC9 from Xenopus
tropicalis. Accession#: Q28CQ4"
CDS 1154..1198
/label="S-Tag"
/note="affinity and epitope tag derived from pancreatic
ribonuclease A"
terminator 1250..1297
/label="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 1382..1400
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1401..1425
/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)."
CDS 1495..3414
/gene="UBA2"
/label="SUMO-activating enzyme subunit 2"
/note="SUMO-activating enzyme subunit 2 from Homo sapiens.
Accession#: Q9UBT2"
CDS 3446..4483
/gene="SAE1"
/label="SUMO-activating enzyme subunit 1"
/note="SUMO-activating enzyme subunit 1 from Homo sapiens.
Accession#: Q9UBE0"
terminator 4514..4561
/label="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(4735..5523)
/label="SmR"
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
promoter complement(5524..5615)
/label="AmpR promoter"
rep_origin complement(5663..6401)
/direction=LEFT
/label="CloDF13 ori"
/note="Plasmids containing the CloDF13 (CDF) origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
protein_bind complement(6577..6598)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(6614..7693)
/label="lacI"
/note="lac repressor"
promoter complement(7694..7771)
/label="lacI promoter"