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:
- pJG4-5 (pB42AD)
- 载体抗性:
- Ampicillin
- 载体长度:
- 6449 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Gyuris J, Golemis E, Chertkov H, Brent R.
- 启动子:
- GAL1
pJG4-5 (pB42AD) 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。
pJG4-5 (pB42AD) 载体序列
LOCUS 40924_26091 6449 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pJG4-5 (pB42AD), complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6449)
AUTHORS Gyuris J, Golemis E, Chertkov H, Brent R.
TITLE Cdi1, a human G1 and S phase protein phosphatase that associates
with Cdk2
JOURNAL Cell 75 (4), 791-803 (1993)
PUBMED 8242750
REFERENCE 2 (bases 1 to 6449)
AUTHORS Gyuris J, Golemis E, Chertkov H, Brent R.
TITLE pB42AD (pJG4-5) complete sequence
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 6449)
AUTHORS Golemis E, Gyuris J, Chertkov H, Brent R.
TITLE Direct Submission
JOURNAL Submitted (19-FEB-1997) CLONTECH Laboratories, Inc., 1020 East
Meadow Circle, Palo Alto, CA 94303-4230, USA
REFERENCE 4 (bases 1 to 6449)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 6449)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell";
date: "1993"; volume: "75"; issue: "4"; pages: "791-803"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(19-FEB-1997) CLONTECH Laboratories, Inc., 1020 East Meadow Circle,
Palo Alto, CA 94303-4230, USA"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT This vector can be obtained from CLONTECH Laboratories, Inc., 1020
East Meadow Circle, Palo Alto, CA 94303-4230, USA. To place an order
call (415) 424-8222 or (800) 662-2566, extension 1. International
customers, please contact your local distributor. For technical
information, call (415) 424-8222 or (800) 662-2566, extension 3.
This sequence has been compiled from information in the sequence
databases, published literature and other sources, together with
partial sequences obtained by CLONTECH. If you suspect there is an
error in this sequence, please contact CLONTECH's Technical Support
Department at (415) 424-8222 or (800) 662-2566, extension 3 or
E-mail TECH@CLONTECH.COM.
FEATURES Location/Qualifiers
source 1..6449
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 81..522
/label=GAL1 promoter
/note="inducible promoter, regulated by Gal4"
CDS 546..566
/codon_start=1
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
CDS 573..809
/codon_start=1
/label=B42 transcriptional activator
/note="yeast transcriptional activator cretaed from E. coli
genomic DNA fragments (Ma and Ptashne, 1987)"
/translation="INKDIEECNAIIEQFIDYLRTGQEMPMEMADQAINVVPGMTPKTI
LHAGPPIQPDWLKSNGFHEIEADVNDTSLLLSGD"
CDS 816..842
/codon_start=1
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
/translation="YPYDVPDYA"
terminator 1004..1191
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
rep_origin 1713..3055
/label=2u ori
/note="yeast 2u plasmid origin of replication"
CDS complement(3419..4090)
/codon_start=1
/label=TRP1
/note="phosphoribosylanthranilate isomerase, required for
tryptophan biosynthesis"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
promoter complement(4091..4192)
/label=TRP1 promoter
promoter 4295..4399
/label=AmpR promoter
CDS 4400..5257
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 5431..6019
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 6307..6328
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6343..6373
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6381..6397
/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 6405..6421
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"