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:
- pLYS1B
- 载体抗性:
- Ampicillin
- 载体长度:
- 5347 bp
- 载体类型:
- S. pombe expression vector
- 复制子:
- ori
- 宿主:
- Yeast
- 载体来源:
- Matsuyama A, Shirai A, Yoshida M.
- 启动子:
- TEF1
pLYS1B 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。
pLYS1B 载体序列
LOCUS 40924_29511 5347 bp DNA circular SYN 18-DEC-2018
DEFINITION S. pombe expression vector pLYS1B DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5347)
AUTHORS Matsuyama A, Shirai A, Yoshida M.
TITLE A novel series of vectors for chromosomal integration in fission
yeast
JOURNAL Biochem. Biophys. Res. Commun. 374 (2), 315-319 (2008)
PUBMED 18634753
REFERENCE 2 (bases 1 to 5347)
AUTHORS Matsuyama A, Shirai A, Yoshida M.
TITLE Fission yeast integration vector pLYS1B
JOURNAL Published Only in Database (2007)
REFERENCE 3 (bases 1 to 5347)
AUTHORS Matsuyama A, Shirai A, Yoshida M.
TITLE Direct Submission
JOURNAL Submitted (15-OCT-2007) Contact:Akihisa Matsuyama RIKEN, Chemical
Genetics Laboratory; Hirosawa 2-1, Wako, Saitama 351-0198, Japan URL
:http://cgl.riken.go.jp/
REFERENCE 4 (bases 1 to 5347)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 5347)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biochem.
Biophys. Res. Commun."; date: "2008"; volume: "374"; issue: "2";
pages: "315-319"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Published
Only in Database (2007)"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(15-OCT-2007) Contact:Akihisa Matsuyama RIKEN, Chemical Genetics
Laboratory; Hirosawa 2-1, Wako, Saitama 351-0198, Japan URL
:http://cgl.riken.go.jp/"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT Fission yeast expression vector.
FEATURES Location/Qualifiers
source 1..5347
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 21..208
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
misc_feature 242..956
/pseudo
/gene="lys1"
/label=derived from Schizosaccharomyces pombe
/note="derived from Schizosaccharomyces pombe"
CDS 242..956
/pseudo
/codon_start=1
/gene="lys1"
/label=lys1
/note="NotI site is created at the middle of this fragment
(613-620)."
/translation="VLCSTCAVSRSNIFGIAKRCHSSRTWHEECPTFGDQSF*Y*QDMR
YRRSW*NISSCWWIG*GLPWK**ID*QKILKELVC*SLQVCRPNSRKCALETLLVRNTR
SYVSFW*LGPLSPNW*CRM*RPPTIKLKFVVSELS*VKLIPIFLVILMSVKILPLFDVI
RMKNPR*LHTLYLKV*IKMTLIALLSRKILSSMV*RSTGSLYMIYGNILRLNFLVMLFL
L*LFPFIKCLSILMER"
gene 242..956
/pseudo
/gene="lys1"
/label=lys1
promoter 962..1369
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
promoter 1376..1423
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 1442..1837
/codon_start=1
/label=BSD
/note="blasticidin S deaminase"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
terminator 1934..2181
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(2207..2223)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 2436..2891
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3173..3277
/label=AmpR promoter
CDS 3278..4135
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4309..4897
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 5185..5206
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5221..5251
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5259..5275
/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 5283..5299
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"