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:
- pSH63
- 载体抗性:
- Ampicillin
- 载体长度:
- 6869 bp
- 载体类型:
- Recombinase expression vector
- 复制子:
- ori
- 载体来源:
- Gueldener U, Heinisch J, Koehler GJ, Voss D, Hegemann JH.
- 筛选标记:
- TRP1
- 启动子:
- GAL1
pSH63 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。
pSH63 载体序列
LOCUS 40924_40082 6869 bp DNA circular SYN 18-DEC-2018
DEFINITION Recombinase expression vector pSH63, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6869)
AUTHORS Gueldener U, Heinisch J, Koehler GJ, Voss D, Hegemann JH.
TITLE A second set of loxP marker cassettes for Cre-mediated multiple gene
knockouts in budding yeast
JOURNAL Nucleic Acids Res. 30 (6), E23 (2002)
PUBMED 11884642
REFERENCE 2 (bases 1 to 6869)
AUTHORS Gueldener U, Heinisch JH, Voss D, Koehler G, Hegemann JH.
TITLE Direct Submission
JOURNAL Submitted (23-AUG-2000) Institut fuer Mikrobiologie,
Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf
40225, Germany
REFERENCE 3 (bases 1 to 6869)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6869)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "2002"; volume: "30"; issue: "6"; pages: "E23"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(23-AUG-2000) Institut fuer Mikrobiologie,
Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf
40225, Germany"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6869
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 187..467
/label=TRP1 promoter
CDS 468..1139
/codon_start=1
/label=TRP1
/note="phosphoribosylanthranilate isomerase, required for
tryptophan biosynthesis"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
rep_origin complement(1241..1696)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 1841..1857
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 1867..1885
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
terminator complement(1904..2151)
/label=CYC1 terminator
/note="transcription terminator for CYC1"
CDS complement(2473..3501)
/codon_start=1
/label=Cre
/note="site-specific recombinase"
/translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRTLDSETGAMVRLLE
DGD"
primer_bind complement(3599..3615)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3630..4071)
/label=GAL1 promoter
/note="inducible promoter, regulated by Gal4"
promoter complement(4095..4113)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4134..4150)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4158..4174)
/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(4182..4212)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4227..4248)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4536..5124)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5298..6155)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6156..6260)
/label=AmpR promoter
misc_feature 6297..6800
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"