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:
- Master_Architecture_RbsR_KSL
- 载体抗性:
- Kanamycin
- 载体长度:
- 5228 bp
- 载体类型:
- Cloning vector
- 复制子:
- pSC101 ori
- 载体来源:
- Rondon RE, Groseclose TM, Short AE, Wilson CJ.
Master_Architecture_RbsR_KSL 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。
Master_Architecture_RbsR_KSL 载体序列
LOCUS 62056_1545 5228 bp DNA circular SYN 03-DEC-2019
DEFINITION Cloning vector Master_Architecture_RbsR_KSL, complete sequence.
ACCESSION MN207946
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5228)
AUTHORS Rondon RE, Groseclose TM, Short AE, Wilson CJ.
TITLE Transcriptional programming using engineered systems of
transcription factors and genetic architectures
JOURNAL Nat Commun 10 (1), 4784 (2019)
PUBMED 31636266
REFERENCE 2 (bases 1 to 5228)
AUTHORS Rondon RE, Groseclose T, Short A, Wilson CJ.
TITLE Direct Submission
JOURNAL Submitted (21-JUL-2019) Chemical and Biomolecular Engineering,
Georgia Institute of Technology, 950 Atlantic dr. NW, 5110E,
Atlanta, GA 30332, United States
REFERENCE 3 (bases 1 to 5228)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun"; date: "2019"; volume: "10"; issue: "1"; pages: "4784"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-JUL-2019) Chemical and Biomolecular Engineering, Georgia
Institute of Technology, 950 Atlantic dr. NW, 5110E, Atlanta, GA
30332, United States"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..5228
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 51..56
/label=trc promoter -35 hexamer TTGACA
/note="trc promoter -35 hexamer TTGACA"
/regulatory_class="promoter"
regulatory 57..73
/label=operator Ottg
/note="operator Ottg"
/regulatory_class="other"
regulatory 74..79
/label=trc promoter -10 hexamer TATAAT
/note="trc promoter -10 hexamer TATAAT"
/regulatory_class="promoter"
regulatory 95..114
/label=operator Oagg
/note="operator Oagg"
/regulatory_class="other"
regulatory 133..182
/label=RiboJ10
/note="RiboJ10"
/regulatory_class="insulator"
regulatory 224..235
/label=strong bacterial RBS
/note="strong bacterial RBS"
/regulatory_class="ribosome_binding_site"
CDS 242..955
/label=superfolder GFP
/note="GFP variant that folds robustly even when fused to
poorly folded proteins (Nager et al., 2011)"
terminator 970..1056
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(1261..2208)
/label=Rep101
/note="RepA protein needed for replication with the pSC101
origin"
rep_origin complement(2256..2478)
/direction=LEFT
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
regulatory 2565..2570
/label=pLL promoter -35 hexamer TTGACA
/note="pLL promoter -35 hexamer TTGACA"
/regulatory_class="promoter"
regulatory 2571..2587
/label=operator Osym
/note="operator Osym"
/regulatory_class="other"
regulatory 2588..2593
/label=pLL promoter -10 hexamer GATACT
/note="pLL promoter -10 hexamer GATACT"
/regulatory_class="promoter"
regulatory 2609..2628
/label=operator Otta
/note="operator Otta"
/regulatory_class="other"
misc_RNA 2646..2696
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
regulatory 2698..2719
/label=RiboJ
/note="RiboJ"
/regulatory_class="insulator"
regulatory 2724..2735
/label=strong bacterial RBS
/note="strong bacterial RBS"
/regulatory_class="ribosome_binding_site"
CDS 2742..3740
/codon_start=1
/transl_table=11
/product="RbsR KSL"
/label=RbsR KSL
/protein_id="QFU95561.1"
/translation="MKPVTLYDVAEYAGVSKSTVSLVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKASHTIGMLITASTNPFYSELVRGVERSCFERGYSLVLCNTEGDEQ
RMNRNLETLMQKRVDGLLLLCTETHQPSREIMQRYPTVPTVMMDWAPFDGDSDLIQDNS
LLGGDLATQYLIDKGHTRIACITGPLDKTPARLRLEGYRAAMKRAGLNIPDGYEVTGDF
EFNGGFDAMRQLLSHPLRPQAVFTGNDAMAVGVYQALYQAELQVPQDIAVIGYDDIELA
SFMTPPLTTIHQPKDELGELAIDVLIHRITQPTLQQQRLQLTPILMERGSA"
terminator 3741..3772
/label=tonB terminator
/note="bidirectional E. coli tonB-P14 transcription
terminator"
terminator complement(4173..4267)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(4301..5092)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"