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:
- pYZ3N-GFP
- 载体抗性:
- Ampicillin
- 载体长度:
- 9956 bp
- 载体类型:
- Expression vector
- 复制子:
- ori
- 载体来源:
- Zhao Y, Elder RT, Chen M.
- 启动子:
- nmt1
pYZ3N-GFP 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。
pYZ3N-GFP 载体序列
LOCUS 40924_48308 9956 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pYZ3N-GFP, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9956)
AUTHORS Zhao Y, Elder RT, Chen M.
TITLE Fission yeast expression vector with GFP tag and positive selection
for insert
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9956)
AUTHORS Zhao Y, Chen M, Elder RT, Cao J.
TITLE Direct Submission
JOURNAL Submitted (11-FEB-1998) Departments of Pediatrics and
Microbiology-Immunology, Northwestern University Medical School,
2430 N Halsted Street, #218, Chicago, IL 60614, USA
REFERENCE 3 (bases 1 to 9956)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9956)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(11-FEB-1998) Departments of Pediatrics and Microbiology-Immunology,
Northwestern University Medical School, 2430 N Halsted Street, #218,
Chicago, IL 60614, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9956
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 48..1210
/label=nmt1 promoter
/note="wild-type nmt1 promoter from Schizosaccharomyces
pombe, conferring strong thiamine-repressible expression"
CDS 1225..1938
/label=GFP
/note="Aequorea victoria green fluorescent protein"
primer_bind 2094..2110
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 2111..2167
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(2180..2196)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2204..2220
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(2228..2258)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2273..2294)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
regulatory 2394..3403
/label=from Schizosaccharomyces pombe nmt1
/note="from Schizosaccharomyces pombe nmt1"
/regulatory_class="terminator"
rep_origin complement(3477..4260)
/direction=LEFT
/label=ars1
/note="Schizosaccharomyces pombe autonomously replicating
sequence ars1"
primer_bind complement(4610..4626)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4839..5294
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5576..5680
/label=AmpR promoter
CDS 5681..6538
/label=AmpR
/note="beta-lactamase"
rep_origin 6712..7300
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 7588..7609
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7624..7654
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7662..7678
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 7686..7702
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(8206..9297)
/label=LEU2
/note="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
promoter complement(9310..9714)
/label=LEU2 promoter