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。
The pLac-GFP construct constitutively expresses GFP and serves as a control throughout
- 载体名称:
- pLac-GFP
- 载体抗性:
- Ampicillin
- 载体长度:
- 6465 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Swofford CA, Van Dessel N, Forbes NS.
- 启动子:
- lac
- 感受态:
- Top 10
- 培养温度:
- 37℃
pLac-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。
pLac-GFP 载体序列
LOCUS V005086 6465 bp DNA circular SYN 31-AUG-2023
DEFINITION Exported.
ACCESSION V005086
VERSION V005086
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6465)
AUTHORS Swofford CA, Van Dessel N, Forbes NS.
TITLE Quorum-sensing Salmonella selectively trigger protein expression
within tumors
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 112 (11), 3457-3462 (2015)
PUBMED 25737556
REFERENCE 2 (bases 1 to 6465)
AUTHORS Swofford CA, Van Dessel N, Forbes NS.
TITLE Direct Submission
JOURNAL Submitted (18-DEC-2014) Chemical Engineering, University of
Massachusetts Amherst, 686 North Pleasant Street, Amherst, MA
01003-9303, USA
REFERENCE 3 (bases 1 to 6465)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6465)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
Acad. Sci. U.S.A."; date: "2015"; volume: "112"; issue: "11"; pages:
"3457-3462"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(18-DEC-2014) Chemical Engineering, University of Massachusetts
Amherst, 686 North Pleasant Street, Amherst, MA 01003-9303, USA"
SGRef: number: 3; type: "Journal Article"
SGRef: number: 1; type: ""Journal Article""; journalName: ""Proc.
Natl. Acad. Sci. U.S.A.""; date: ""2015""; volume: ""112""; issue:
""11""; pages: ""3457-3462""
SGRef: number: 2; type: ""Journal Article""; journalName:
""Submitted (18-DEC-2014) Chemical Engineering, University of
Massachusetts Amherst, 686 North Pleasant Street, Amherst, MA
01003-9303, USA"" SGRef: number: 3; type: ""Journal Article""
FEATURES Location/Qualifiers
source 1..6465
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 133..154
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 169..199
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 207..223
/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 231..247
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 275..988
/label="yeGFP"
/note="yeast-enhanced green fluorescent protein"
oriT 1615..1702
/label="RSF1010 oriT"
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
CDS 1783..3132
/codon_start=1
/label="DNA primase"
/translation="MAIYHLTAKTGSRSGGQSARAKADYIQREGKYARDMDEVLHAESG
HMPEFVERPADYWDAADLYERANGRLFKEVEFALPVELTLDQQKALASEFAQHLTGAER
LPYTLAIHAGGGENPHCHLMISERINDGIERPAAQWFKRYNGKTPEKGGAQKTEALKPK
AWLEQTREAWADHANRALERAGHDARIDHRTLEAQGIERLPGVHLGPNVVEMEGRGIRT
DRADVALNIDTANAQIIDLQEYREAIDHERNRQSEEIQRHQRVSGADRTAGPEHGDTGR
RSPAGHEPDPAGQRGAGGGVAESPAPDRGGMGGAGQRVAGGSRRGEQRRAERPERVAGV
ALEAMANRDAGFHDAYGGAADRIVALARPDATDNRGRLDLAALGGPMKNDRTLQAIGRQ
LKAMGCERSSASSLTDSLRSVVRLRRAVSAHSKAVIRLSTESGDNAGKNM"
rep_origin complement(3186..3774)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3948..4805)
/label="AmpR"
/note="beta-lactamase"
promoter complement(4806..4910)
/label="AmpR promoter"
CDS complement(5090..6193)
/gene="asd"
/label="Aspartate-semialdehyde dehydrogenase"
/note="Aspartate-semialdehyde dehydrogenase from Salmonella
typhimurium (strain LT2 / SGSC1412 / ATCC 700720).
Accession#: P0A1F8"