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:
- pDM1
- 载体抗性:
- Tetracycline
- 载体长度:
- 10456 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori2
- 载体来源:
- Chan CT, Lee JW, Cameron DE, Bashor CJ, Collins JJ.
pDM1 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。
pDM1 载体序列
LOCUS 40924_14770 10456 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pDM1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10456)
AUTHORS Chan CT, Lee JW, Cameron DE, Bashor CJ, Collins JJ.
TITLE 'Deadman' and 'Passcode' microbial kill switches for bacterial
containment
JOURNAL Nat. Chem. Biol. 12 (2), 82-86 (2016)
PUBMED 26641934
REFERENCE 2 (bases 1 to 10456)
AUTHORS Chan CTY., Lee JW, Cameron DE, Bashor CJ, Collins JJ.
TITLE Direct Submission
JOURNAL Submitted (12-OCT-2015) Biological engineering, MIT, 45 Carleton St,
MIT Building E25-Rm302, Cambridge, MA 02142, USA
REFERENCE 3 (bases 1 to 10456)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10456)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem.
Biol."; date: "2016"; volume: "12"; issue: "2"; pages: "82-86"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(12-OCT-2015) Biological engineering, MIT, 45 Carleton St, MIT
Building E25-Rm302, Cambridge, MA 02142, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..10456
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(105..1073)
/label=sopB
/note="partitioning protein for the bacterial F plasmid"
CDS complement(1076..2248)
/label=sopA
/note="partitioning protein for the bacterial F plasmid"
misc_feature complement(2574..2824)
/label=incC
/note="incompatibility region of the bacterial F plasmid"
CDS complement(2830..3582)
/label=repE
/note="replication initiation protein for the bacterial F
plasmid"
rep_origin complement(3673..3892)
/direction=LEFT
/label=ori2
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
rep_origin complement(3968..4582)
/direction=LEFT
/label=oriV
/note="origin of replication for the bacterial F plasmid"
promoter 5440..5542
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 5543..6199
/label=CmR
/note="chloramphenicol acetyltransferase"
terminator complement(6337..6431)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(6494..7207)
/label=yeGFP
/note="yeast-enhanced green fluorescent protein"
RBS 7213..7221
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
regulatory complement(7249..7322)
/label=PLtetO promoter
/note="PLtetO promoter"
/regulatory_class="promoter"
promoter complement(7249..7322)
/label=PLtetO-1 promoter
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
misc_feature 7268
/label=transcription start site
/note="transcription start site"
protein_bind 7304..7322
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature complement(7365..7448)
/label=ssrA tag (mf-lon)
/note="ssrA tag (mf-lon)"
CDS complement(7449..8528)
/label=lacI
/note="lac repressor"
RBS 8534..8542
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
promoter complement(8564..8637)
/label=PLtetO-1 promoter
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
promoter 8669..8698
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
misc_feature 8704
/label=transcription start site
/note="transcription start site"
protein_bind 8706..8722
/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)."
RBS 8755..8763
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 8769..9389
/label=TetR
/note="tetracycline repressor TetR"
regulatory 9435..9509
/label=Ptrc-2 promoter
/note="Ptrc-2 promoter"
/regulatory_class="promoter"
promoter 9448..9477
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 9485..9501
/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)."
misc_feature 9485
/label=transcription start site
/note="transcription start site"
RBS 9530..9538
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 9544..10251
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
terminator 10314..10400
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"