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:
- pTHSSe_70
- 载体抗性:
- Kanamycin
- 载体长度:
- 7391 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Segall-Shapiro TH, Sontag ED, Voigt CA.
pTHSSe_70 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。
pTHSSe_70 载体序列
LOCUS V002649 7391 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V002649
VERSION V002649
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7391)
AUTHORS Segall-Shapiro TH, Sontag ED, Voigt CA.
TITLE Engineered promoters enable constant gene expression at any copy
number in bacteria
JOURNAL Nat. Biotechnol. (2018) In press
PUBMED 29553576
REFERENCE 2 (bases 1 to 7391)
AUTHORS Segall-Shapiro TH, Sontag ED, Voigt CA.
TITLE Direct Submission
JOURNAL Submitted (06-MAR-2018) Synthetic Biology Center, Department of
Biological Engineering, Massachusetts Institute of Technology, 77
Massachusetts Avenue, Cambridge, MA 02139, USA
REFERENCE 3 (bases 1 to 7391)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7391)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
Biotechnol. (2018) In press"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-MAR-2018) Synthetic Biology Center, Department of Biological
Engineering, Massachusetts Institute of Technology, 77 Massachusetts
Avenue, Cambridge, MA 02139, USA"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7391
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory complement(13..102)
/label="ECK120029600"
/note="ECK120029600"
/regulatory_class="terminator"
regulatory 107..175
/label="PT7A1w1"
/note="PT7A1w1"
/regulatory_class="promoter"
misc_RNA 177..227
/label="sTRSV HHRz"
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
regulatory 251..273
/label="RBSVioEc"
/note="RBSVioEc"
/regulatory_class="ribosome_binding_site"
CDS 274..849
/codon_start=1
/product="VioE"
/label="VioE"
/protein_id="AVR55197.1"
/translation="MENREPPLLPARWSSAYVSYWSPMLPDDQLTSGYCWFDYERDICR
IDGLFNPWSERDTGYRLWMSEVGNAASGRTWKQKVAYGRERTALGEQLCERPLDDETGP
FAELFLPRDVLRRLGARHIGRRVVLGREADGWRYQRPGKGPSTLYLDAASGTPLRMVTG
DEASRASLRDFPNVSEAEIPDAVFAAKR"
regulatory 850..873
/label="RBSVioAc"
/note="RBSVioAc"
/regulatory_class="ribosome_binding_site"
CDS 874..2127
/gene="vioA"
/label="Flavin-dependent L-tryptophan oxidase VioA"
/note="Flavin-dependent L-tryptophan oxidase VioA from
Chromobacterium violaceum (strain ATCC 12472 / DSM 30191 /
JCM 1249 / NBRC 12614 / NCIMB 9131 / NCTC 9757).
Accession#: Q9S3V1"
regulatory 2131..2154
/label="RBSVioBc"
/note="RBSVioBc"
/regulatory_class="ribosome_binding_site"
CDS 2155..5148
/gene="vioB"
/label="2-imino-3-(indol-3-yl)propanoate dimerase"
/note="2-imino-3-(indol-3-yl)propanoate dimerase from
Chromobacterium violaceum (strain ATCC 12472 / DSM 30191 /
JCM 1249 / NBRC 12614 / NCIMB 9131 / NCTC 9757).
Accession#: Q9S3V0"
terminator 5166..5237
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 5253..5280
/label="T7Te terminator"
/note="phage T7 early transcription terminator"
misc_feature 5299..5319
/label="BioBrick suffix"
/note="universal suffix for all parts"
terminator 5320..5377
/label="his operon terminator"
/note="This putative transcriptin terminator from the E.
coli his operon has a 2-bp deletion introduced during
synthesis. Its efficiency has not been determined."
rep_origin 5549..6137
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6338..7129)
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
terminator complement(7324..7367)
/label="bacterial terminator"
/note="putative bacterial transcription terminator"
misc_feature 7370..7391
/label="BioBrick prefix"
/note="BioBrick prefix for parts that do not start with
'ATG'"