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:
- pGREG505-C
- 载体抗性:
- Ampicillin
- 载体长度:
- 9261 bp
- 载体类型:
- PGREG expression vector
- 复制子:
- ori
- 宿主:
- Yeast
- 载体来源:
- Karreman RJ, Morrissey JP.
- 启动子:
- LEU2
pGREG505-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。
pGREG505-C 载体序列
LOCUS 40924_22651 9261 bp DNA circular SYN 18-DEC-2018
DEFINITION PGREG expression vector pGREG505-C, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9261)
AUTHORS Karreman RJ, Morrissey JP.
TITLE Controlling expression of genes in Saccharomyces cerevisiae using
variants of the pGREG recombinational vectors
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9261)
AUTHORS Karreman RJ, Morrissey JP.
TITLE Direct Submission
JOURNAL Submitted (28-JUN-2010) Microbiology Department, University College
Cork, College Rd, Cork City, Cork 0000, Ireland
REFERENCE 3 (bases 1 to 9261)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9261)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(28-JUN-2010) Microbiology Department, University College Cork,
College Rd, Cork City, Cork 0000, Ireland"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9261
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 7..462
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 762..1166
/label=LEU2 promoter
CDS 1179..2270
/label=LEU2
/note="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
misc_feature complement(3005..3508)
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 3545..3649
/label=AmpR promoter
CDS 3650..4507
/label=AmpR
/note="beta-lactamase"
rep_origin 4681..5269
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 5557..5578
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5593..5623
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5631..5647
/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 5655..5671
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 5692..5710
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
regulatory 5729..6015
/label=Saccharomyces cerevisiae CYC1 promoter (weak)
/note="Saccharomyces cerevisiae CYC1 promoter (weak)"
/regulatory_class="promoter"
misc_feature 6040..6073
/note="Rec1 site; pGREG recombinational cloning site I"
misc_feature 6067..6072
/label=SalI restriction enzyme cut site
/note="SalI restriction enzyme cut site"
promoter 6362..6549
/label=HIS3 promoter
CDS 6550..7206
/label=HIS3
/note="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
misc_feature 7235..7240
/label=SalI restriction enzyme cut site
/note="SalI restriction enzyme cut site"
terminator 7247..7494
/label=CYC1 terminator
/note="transcription terminator for CYC1"
misc_feature 7507..7540
/note="LoxP site (cre/lox KanMX marker rescue); Guldener,
et. al. 1996, Nucleic Acids Research 24, 2519-2524"
protein_bind 7507..7540
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
gene complement(7598..8954)
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
protein_bind complement(9009..9042)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
promoter complement(9079..9097)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(9107..9123)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"