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:
- pTNS2-ColE1
- 载体抗性:
- Ampicillin
- 载体长度:
- 10445 bp
- 载体类型:
- Tn7 transposase expression vector
- 复制子:
- R6K γ ori
- 载体来源:
- Lee KW, Periasamy S, Mukherjee M, Xie C, Kjelleberg S, Rice SA.
pTNS2-ColE1 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。
pTNS2-ColE1 载体序列
LOCUS V002577 10445 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V002577
VERSION V002577
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10445)
AUTHORS Lee KW, Periasamy S, Mukherjee M, Xie C, Kjelleberg S, Rice SA.
TITLE Biofilm development and enhanced stress resistance of a model,
mixed-species community biofilm
JOURNAL ISME J 8 (4), 894-907 (2014)
PUBMED 24152718
REFERENCE 2 (bases 1 to 10445)
AUTHORS Lee KWK., Periasamy S, Mukherjee M, Xie C, Kjelleberg S, Rice SA.
TITLE Direct Submission
JOURNAL Submitted (15-SEP-2014) Singapore Centre on Environmental Life
Sciences Engineering, Nanyang Technological University, 60 Nanyang
Drive, SBS-01N-27 637551, Singapore
REFERENCE 3 (bases 1 to 10445)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10445)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ISME J";
date: "2014"; volume: "8"; issue: "4"; pages: "894-907"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-SEP-2014) Singapore Centre on Environmental Life Sciences
Engineering, Nanyang Technological University, 60 Nanyang Drive,
SBS-01N-27 637551, Singapore"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10445
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(690..2216)
/codon_start=1
/gene="tnsD"
/product="TnsD"
/label="tnsD"
/note="Tn7 transposition site selector"
/protein_id="AIZ73053.1"
/translation="MRNFPVPYSNELIYSTIARAGVYQGIVSPKQLLDEVYGNRKVVAT
LGLPSHLGVIARHLHQTGRYAVQQLIYEHTLFPLYAPFVGKERRDEAIRLMEYQAQGAV
HLMLGVAASRVKSDNRFRYCPDCVALQLNRYGEAFWQRDWYLPALPYCPKHGALVFFDR
AVDDHRHQFWALGHTELLSDYPKDSLSQLTALAAYIAPLLDAPRAQELSPSLEQWTLFY
QRLAQDLGLTKSKHIRHDLVAERVRQTFSDEALEKLDLKLAENKDTCWLKSIFRKHRKA
FSYLQHSIVWQALLPKLTVIEALQQASALTEHSITTRPVSQSVQPNSEDLSVKHKDWQQ
LVHKYQGIKAARQSLEGGVLYAWLYRHDRDWLVHWNQQHQQERLAPAPRVDWNQRDRIA
VRQLLRIIKRLDSSLDHPRATSSWLLKQTPNGTSLAKNLQKLPLVALCLKRYSESVEDY
QIRRISQAFIKLKQEDVELRRWRLLRSATLSKERITEEAQRFLEMVYGEE"
gene complement(690..2216)
/gene="tnsD"
/label="tnsD"
CDS complement(2222..3886)
/gene="tnsC"
/label="Transposon Tn7 transposition protein TnsC"
/note="Transposon Tn7 transposition protein TnsC from
Escherichia coli. Accession#: P05846"
CDS complement(3886..5991)
/gene="tnsB"
/label="Transposon Tn7 transposition protein TnsB"
/note="Transposon Tn7 transposition protein TnsB from
Escherichia coli. Accession#: P13989"
mobile_element 6709..6933
/label="Tn7R"
/note="mini-Tn7 element (right end of the Tn7 transposon)"
rep_origin complement(7197..7785)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7971..7987)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 7995..8011
/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(8019..8049)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(8064..8085)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
oriT 8461..8569
/label="oriT"
/note="incP origin of transfer"
rep_origin 8592..8976
/label="R6K gamma ori"
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS complement(9388..10245)
/label="AmpR"
/note="beta-lactamase"
promoter complement(10246..10350)
/label="AmpR promoter"