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:
- pCS2+MT-Gli3-6xHIS
- 载体抗性:
- Ampicillin
- 载体长度:
- 9093 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Vega Lopez GA, Cerrizuela S, Aguero TH, Palacio MB, Aybar MJ.
- 启动子:
- sCMV
pCS2+MT-Gli3-6xHIS 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。
pCS2+MT-Gli3-6xHIS 载体序列
LOCUS V008148 9093 bp DNA circular SYN 17-DEC-2018
DEFINITION Exported.
ACCESSION V008148
VERSION V008148
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9093)
AUTHORS Vega Lopez GA, Cerrizuela S, Aguero TH, Palacio MB, Aybar MJ.
TITLE The gli3 transcriptional activator is a key effector of Indian
hedgehog signaling during Xenopus neural crest development
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9093)
AUTHORS Vega Lopez GA, Cerrizuela S, Aguero TH, Palacio MB, Aybar MJ.
TITLE Direct Submission
JOURNAL Submitted (03-JAN-2017) Developmental Biology, Instituto Superior de
Investigaciones Biologicas (INSIBIO, CONICET-UNT), Chacabuco 461,
Tucuman, Tucuman T4000ILI, Argentina
REFERENCE 3 (bases 1 to 9093)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9093)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Assembly Method :: vector nti v. 11
Sequencing Technology :: Applied Biosystems 3130 Genetic Analyzer
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(03-JAN-2017) Developmental Biology, Instituto Superior de
Investigaciones Biologicas (INSIBIO, CONICET-UNT), Chacabuco 461,
Tucuman, Tucuman T4000ILI, Argentina"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9093
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 35..53
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
CDS 103..132
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label="Myc"
/translation="EQKLISEEDL"
CDS 142..171
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label="Myc"
/translation="EQKLISEEDL"
CDS 181..210
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label="Myc"
/translation="EQKLISEEDL"
CDS 220..249
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label="Myc"
/translation="EQKLISEEDL"
CDS 259..288
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label="Myc"
/translation="EQKLISEEDL"
CDS 322..351
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label="Myc"
/translation="EQKLISEEDL"
CDS 379..5085
/gene="gli3"
/label="Zinc finger protein GLI3"
/note="Zinc finger protein GLI3 from Xenopus laevis.
Accession#: Q91660"
CDS 5092..5109
/label="6xHis"
/note="6xHis affinity tag"
promoter complement(5120..5137)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal complement(5142..5276)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(5389..5407)
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5428..5444)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5452..5468)
/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(5476..5506)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(5521..5542)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5830..6418)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6592..7449)
/label="AmpR"
/note="beta-lactamase"
promoter complement(7450..7554)
/label="AmpR promoter"
rep_origin complement(7580..8035)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 8109..9093
/label="CMV IE94 promoter"
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"