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:
- pBSc393K
- 载体抗性:
- Chloramphenicol
- 载体长度:
- 4412 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Radeck J, Meyer D, Lautenschlager N, Mascher T.
pBSc393K 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。
pBSc393K 载体序列
LOCUS 40924_7386 4412 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pBSc393K, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4412)
AUTHORS Radeck J, Meyer D, Lautenschlager N, Mascher T.
TITLE Bacillus SEVA siblings: A Golden Gate-based toolbox to create
personalized integrative vectors for Bacillus subtilis
JOURNAL Sci Rep 7 (1), 14134 (2017)
PUBMED 29074996
REFERENCE 2 (bases 1 to 4412)
AUTHORS Radeck J.
TITLE Direct Submission
JOURNAL Submitted (26-APR-2017) Biology, Technische Universitat Dresden,
Zellescher Weg 20b, Dresden 01627, Germany
REFERENCE 3 (bases 1 to 4412)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4412)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep";
date: "2017"; volume: "7"; issue: "1"; pages: "14134"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(26-APR-2017) Biology, Technische Universitat Dresden, Zellescher
Weg 20b, Dresden 01627, Germany"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..4412
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 125..146
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 161..191
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 199..215
/label=lac operator
/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)."
primer_bind 223..239
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 249..305
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(309..325)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator 604..698
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
promoter 793..895
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 896..1552
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
oriT 1715..1823
/label=oriT
/note="incP origin of transfer"
rep_origin 1858..2446
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(2632..2772)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(2877..3065)
/codon_start=1
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
/translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
terminator complement(3347..3374)
/label=T7Te terminator
/note="phage T7 early transcription terminator"
CDS complement(3389..4180)
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
terminator complement(4320..4406)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"