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:
- pDSG398
- 载体抗性:
- Kanamycin
- 载体长度:
- 5975 bp
- 载体类型:
- Cloning vector
- 复制子:
- p15A ori
- 载体来源:
- Glass DS, Riedel-Kruse IH.
pDSG398 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。
pDSG398 载体序列
LOCUS 40924_15500 5975 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector pDSG398, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5975) AUTHORS Glass DS, Riedel-Kruse IH. TITLE A synthetic bacterial cell-cell adhesion toolbox for programming multicellular morphologies and patterns JOURNAL Cell (2018) In press REFERENCE 2 (bases 1 to 5975) AUTHORS Glass DS. TITLE Direct Submission JOURNAL Submitted (14-JUN-2018) Bioengineering, Stanford University, 318 Campus Drive, Clark Center E350, Stanford, CA 94305, USA REFERENCE 3 (bases 1 to 5975) TITLE Direct Submission REFERENCE 4 (bases 1 to 5975) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell (2018) In press" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (14-JUN-2018) Bioengineering, Stanford University, 318 Campus Drive, Clark Center E350, Stanford, CA 94305, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..5975 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 1..36 /label=pLacIQ (BBa_I4032, 1nt deletion) /note="pLacIQ (BBa_I4032, 1nt deletion)" misc_feature 45..56 /label=RBS (BBa_B0034) /note="RBS (BBa_B0034)" misc_feature 45..56 /label=BBa_J04500(1) /note="BBa_J04500(1)" RBS 45..56 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 63..683 /label=TetR /note="tetracycline repressor TetR" terminator 706..777 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 793..820 /label=T7Te terminator /note="phage T7 early transcription terminator" protein_bind 835..853 /label=tet operator /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 860..878 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" misc_feature 897..908 /label=RBS (BBa_B0034)(1) /note="RBS (BBa_B0034)(1)" misc_feature 897..908 /label=BBa_J04500 /note="BBa_J04500" RBS 897..908 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 915..3224 /codon_start=1 /product="mutated Neae" /label=mutated Neae /note="removed restriction sites" /protein_id="AXC07940.1" /translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS AKTAEMTSALNASAVIFFDGATRQVQLQESGGGSVRPGESLRLSCIAAGTLFSGKAVGW YRQAPGKDRELVAALSSGRETYYIGPVKGRFIISRDDNKDTLYLQMNNLKLEDTAIYTC RLGLHWGQGTQVTVSS" CDS 2889..3224 /codon_start=1 /product="N8-1_antiP53TA-R4P43" /label=N8-1_antiP53TA-R4P43 /protein_id="AXC07938.1" /translation="QVQLQESGGGSVRPGESLRLSCIAAGTLFSGKAVGWYRQAPGKDR ELVAALSSGRETYYIGPVKGRFIISRDDNKDTLYLQMNNLKLEDTAIYTCRLGLHWGQG TQVTVSS" misc_feature 3222..3227 /label=stop codons /note="stop codons" misc_feature 3228..3248 /label=BioBrick suffix /note="universal suffix for all parts" terminator 3249..3306 /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." misc_feature complement(3384..3401) /label=VR primer site /note="VR primer site" CDS complement(3467..4279) /label=KanR /note="aminoglycoside phosphotransferase" rep_origin complement(4874..5418) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin." misc_feature 5836..5856 /label=VF2 primer site /note="VF2 primer site" terminator complement(5908..5951) /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 5954..5975 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with 'ATG'"