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:
- pMRE105_PnptII_eYFP
- 载体抗性:
- Ampicillin
- 载体长度:
- 13484 bp
- 载体类型:
- Tn7 delivery vector
- 复制子:
- pSC101 ori
- 载体来源:
- Remus-Emsermann MNP., Gisler P, Drissner D.
- 启动子:
- araBAD
pMRE105_PnptII_eYFP 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。
pMRE105_PnptII_eYFP 载体序列
LOCUS V004476 13484 bp DNA circular SYN 18-DEC-2018 DEFINITION Exported. ACCESSION V004476 VERSION V004476 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 13484) AUTHORS Remus-Emsermann MNP., Gisler P, Drissner D. TITLE MiniTn7-transposon delivery vectors for inducible or constitutive fluorescent protein expression in Enterobacteriaceae JOURNAL Unpublished REFERENCE 2 (bases 1 to 13484) AUTHORS Remus-Emsermann MNP., Drissner D. TITLE Direct Submission JOURNAL Submitted (23-MAR-2016) Institute for Food Sciences, Agroscope, Schloss 1, Waedenswil, Zurich 8820, Switzerland REFERENCE 3 (bases 1 to 13484) TITLE Direct Submission REFERENCE 4 (bases 1 to 13484) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (23-MAR-2016) Institute for Food Sciences, Agroscope, Schloss 1, Waedenswil, Zurich 8820, Switzerland" SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..13484 /mol_type="other DNA" /organism="synthetic DNA construct" CDS complement(149..1024) /label="araC" /note="L-arabinose regulatory protein" promoter 1051..1335 /label="araBAD promoter" /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" CDS 1453..2271 /gene="tnsA" /label="Transposon Tn7 transposition protein TnsA" /note="Transposon Tn7 transposition protein TnsA from Escherichia coli. Accession#: P13988" CDS 2261..4366 /gene="tnsB" /label="Transposon Tn7 transposition protein TnsB" /note="Transposon Tn7 transposition protein TnsB from Escherichia coli. Accession#: P13989" CDS 4366..6030 /gene="tnsC" /label="Transposon Tn7 transposition protein TnsC" /note="Transposon Tn7 transposition protein TnsC from Escherichia coli. Accession#: P05846" CDS 6036..7562 /codon_start=1 /gene="tnsD" /product="TnsD" /label="tnsD" /note="Tn7 transposition site-selector" /protein_id="AMZ00009.1" /translation="MRNFPVPYSNELIYSTIARAGVYQGIVSPKQLLDEVYGNRKVVAT LGLPSHLGVIARHLHQTGRYAVQQLIYEHTLFPLYAPFVGKERRDEAIRLMEYQAQGAV HLMLGVAASRVKSDNRFRYCPDCVALQLNRYGEAFWQRDWYLPALPYCPKHGALVFFDR AVDDHRHQFWALGHTELLSDYPKDSLSQLTALAAYIAPLLDAPRAQELSPSLEQWTLFY QRLAQDLGLTKSKHIRHDLVAERVRQTFSDEALEKLDLKLAENKDTCWLKSIFRKHRKA FSYLQHSIVWQALLPKLTVIEALQQASALTEHSITTRPVSQSVQPNSEDLSVKHKDWQQ LVHKYQGIKAARQSLEGGVLYAWLYRHDRDWLVHWNQQHQQERLAPAPRVDWNQRDRIA VRQLLRIIKRLDSSLDHPRATSSWLLKQTPNGTSLAKNLQKLPLVALCLKRYSESVEDY QIRRISQAFIKLKQEDVELRRWRLLRSATLSKERITEEAQRFLEMVYGEE" gene 6036..7562 /gene="tnsD" /label="tnsD" terminator 8196..8282 /label="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" terminator 8374..8401 /label="rrnB T2 terminator" /note="transcription terminator T2 from the E. coli rrnB gene" promoter 8420..8511 /label="AmpR promoter" CDS 8512..9369 /label="AmpR" /note="beta-lactamase" oriT complement(9543..9652) /direction=LEFT /label="oriT" /note="incP origin of transfer" rep_origin 10392..10614 /label="pSC101 ori" /note="low-copy replication origin that requires the Rep101 protein" CDS 10662..11609 /label="Rep101(Ts)" /note="temperature-sensitive version of the RepA protein needed for replication with the pSC101 origin (Armstrong et al., 1984)" mobile_element 12045..12210 /label="Tn7L" /note="mini-Tn7 element (left end of the Tn7 transposon)" CDS 12509..13225 /label="EYFP" /note="enhanced YFP" repeat_region 13273..13484 /label="Tn7 right end" /note="Tn7 right end"