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:
- pDSG402
- 载体抗性:
- Kanamycin
- 载体长度:
- 6005 bp
- 载体类型:
- Cloning vector
- 复制子:
- p15A ori
- 载体来源:
- Glass DS, Riedel-Kruse IH.
pDSG402 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。
pDSG402 载体序列
LOCUS 40924_15520 6005 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pDSG402, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6005)
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 6005)
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 6005)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6005)
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..6005
/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..3254
/codon_start=1
/product="mutated Neae"
/label=mutated Neae
/note="removed restriction sites"
/protein_id="AXC07764.1"
/translation="MITHGCYTRTRHKHKLKKTLIMLSAGLGLFFYVNQNSFANGENYF
KLGSDSKLLTHDSYQNRLFYTLKTGETVADLSKSQDINLSTIWSLNKHLYSSESEMMKA
APGQQIILPLKKLPFEYSALPLLGSAPLVAAGGVAGHTNKLTKMSPDVTKSNMTDDKAL
NYAAQQAASLGSQLQSRSLNGDYAKDTALGIAGNQASSQLQAWLQHYGTAEVNLQSGNN
FDGSSLDFLLPFYDSEKMLAFGQVGARYIDSRFTANLGAGQRFFLPANMLGYNVFIDQD
FSGDNTRLGIGGEYWRDYFKSSVNGYFRMSGWHESYNKKDYDERPANGFDIRFNGYLPS
YPALGAKLIYEQYYGDNVALFNSDKLQSNPGAATVGVNYTPIPLVTMGIDYRHGTGNEN
DLLYSMQFRYQFDKSWSQQIEPQYVNELRTLSGSRYDLVQRNNNIILEYKKQDILSLNI
PHDINGTEHSTQKIQLIVKSKYGLDRIVWDDSALRSQGGQIQHSGSQSAQDYQAILPAY
VQGGSNIYKVTARAYDRNGNSSNNVQLTITVLSNGQVVDQVGVTDFTADKTSAKADNAD
TITYTATVKKNGVAQANVPVSFNIVSGTATLGANSAKTDANGKATVTLKSSTPGQVVVS
AKTAEMTSALNASAVIFFDGATRQVQLQESGGGLVQAGGSLRLSCAASGRAVSMYNMGW
FRQAPGKEREFVAVISTASTYYVDSVKGRFTISRDYAKNAVYLQMNSLKPEDTAVYYCA
ATLKTYRWPPTTYDYWGRGTQVTVSS"
CDS 2889..3254
/codon_start=1
/product="N8-5_antiP53TA-R3P3"
/label=N8-5_antiP53TA-R3P3
/protein_id="AXC07762.1"
/translation="QVQLQESGGGLVQAGGSLRLSCAASGRAVSMYNMGWFRQAPGKER
EFVAVISTASTYYVDSVKGRFTISRDYAKNAVYLQMNSLKPEDTAVYYCAATLKTYRWP
PTTYDYWGRGTQVTVSS"
misc_feature 3252..3257
/label=stop codons
/note="stop codons"
misc_feature 3258..3278
/label=BioBrick suffix
/note="universal suffix for all parts"
terminator 3279..3336
/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(3414..3431)
/label=VR primer site
/note="VR primer site"
CDS complement(3497..4309)
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin complement(4904..5448)
/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 5866..5886
/label=VF2 primer site
/note="VF2 primer site"
terminator complement(5938..5981)
/label=bacterial terminator
/note="putative bacterial transcription terminator"
misc_feature 5984..6005
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
'ATG'"