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:
- pcDNA3.1/His C
- 载体抗性:
- Ampicillin
- 载体长度:
- 5513 bp
- 载体类型:
- Mammalian Expression Vectors
- 复制子:
- ori
- 载体来源:
- Invitrogen (Life Technologies)
- 筛选标记:
- Neomycin/G418(Geneticin)
- 拷贝数:
- High copy number
pcDNA3.1/His C 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。
pcDNA3.1/His C 载体序列
LOCUS pcDNA3.1_His_C. 5513 bp DNA circular SYN 01-JAN-1980
DEFINITION Mammalian vector for expressing proteins with a cleavable N-terminal
6xHis tag. For other reading frames, use pcDNA(TM)3.1/His A or
pcDNA(TM)3.1/His B.
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1 His C
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5513)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5513)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5513
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 920..922
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 932..949
/label=6xHis
/note="6xHis affinity tag"
CDS 953..985
/label=T7 tag (gene 10 leader)
/note="leader peptide from bacteriophage T7 gene 10"
CDS 989..1012
/label=Xpress(TM) tag
/note="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
misc_feature 1012..1084
/label=MCS
/note="MCS"
/note="multiple cloning site"
polyA_signal 1110..1334
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1380..1808
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1822..2151
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 2218..3009
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
polyA_signal 3186..3319
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3356..3372)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3380..3396)
/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)."
promoter complement(3404..3434)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3449..3470)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3758..4346)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4520..5377)
/label=AmpR
/note="beta-lactamase"
promoter complement(5378..5482)
/label=AmpR promoter