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:
- pKSS
- 载体抗性:
- Ampicillin
- 载体长度:
- 4133 bp
- 载体类型:
- Phagemid cloning vector
- 复制子:
- ori
- 载体来源:
- Selzer G, Som T, Itoh T, Tomizawa J.
- 启动子:
- T3
pKSS 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。
pKSS 载体序列
LOCUS V005182 4133 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V005182
VERSION V005182
KEYWORDS direct-selection cloning vehicle; positive-selection; pheS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 4133)
AUTHORS Selzer G, Som T, Itoh T, Tomizawa J.
TITLE The origin of replication of plasmid p15A and comparative studies on
the nucleotide sequences around the origin of related plasmids
JOURNAL Cell 32 (1), 119-129 (1983)
PUBMED 6186390
REFERENCE 2 (bases 1 to 4133)
AUTHORS Fayat G, Mayaux JF, Sacerdot C, Fromant M, Springer M,
Grunberg-Manago M, Blanquet S.
TITLE Escherichia coli phenylalanyl-tRNA synthetase operon region.
Evidence for an attenuation mechanism. Identification of the gene
for the ribosomal protein L20
JOURNAL J. Mol. Biol. 171 (3), 239-261 (1983)
PUBMED 6317865
REFERENCE 3 (bases 3864 to 3956)
AUTHORS Yanisch-Perron C, Vieira J, Messing J.
TITLE Improved M13 phage cloning vectors and host strains: nucleotide
sequences of the M13mp18 and pUC19 vectors
JOURNAL Gene 33 (1), 103-119 (1985)
PUBMED 2985470
REFERENCE 4 (bases 1 to 4133)
AUTHORS Balbas P, Soberon X, Merino E, Zurita M, Lomeli H, Valle F, Flores
N, Bolivar F.
TITLE Plasmid vector pBR322 and its special-purpose derivatives--a review
JOURNAL Gene 50 (1-3), 3-40 (1986)
PUBMED 3034735
REFERENCE 5 (bases 4133 to 4133)
AUTHORS Short JM, Fernandez JM, Sorge JA, Huse WD.
TITLE Lambda ZAP: a bacteriophage lambda expression vector with in vivo
excision properties
JOURNAL Nucleic Acids Res. 16 (15), 7583-7600 (1988)
PUBMED 2970625
REFERENCE 6 (bases 1 to 4133)
AUTHORS Kast P, Hennecke H.
TITLE Amino acid substrate specificity of Escherichia coli
phenylalanyl-tRNA synthetase altered by distinct mutations
JOURNAL J. Mol. Biol. 222 (1), 99-124 (1991)
PUBMED 1942071
REFERENCE 7 (bases 1 to 4133)
AUTHORS Alting-Mees MA, Sorge JA, Short JM.
TITLE pBluescriptII: multifunctional cloning and mapping vectors
JOURNAL Meth. Enzymol. 216, 483-495 (1992)
PUBMED 1362236
REFERENCE 8 (bases 1 to 4133)
AUTHORS Lin-Chao S, Chen WT, Wong TT.
TITLE High copy number of the pUC plasmid results from a
Rom/Rop-suppressible point mutation in RNA II
JOURNAL Mol. Microbiol. 6 (22), 3385-3393 (1992)
PUBMED 1283002
REFERENCE 9 (bases 4133 to 4133)
AUTHORS Kast P.
TITLE pKSS--a second-generation general purpose cloning vector for
efficient positive selection of recombinant clones
JOURNAL Gene 138 (1-2), 109-114 (1994)
PUBMED 8125286
REFERENCE 10 (bases 1 to 4133)
AUTHORS Kast P.
TITLE Direct Submission
JOURNAL Submitted (10-SEP-1993) Dept. of Chemistry and Molecular Biology,
The Scripps Research Institute, 10550 North Torrey Pines Road, La
Jolla, CA 92037, USA
REFERENCE 11 (bases 1 to 4133)
TITLE Direct Submission
REFERENCE 12 (bases 1 to 4133)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell";
date: "1983"; volume: "32"; issue: "1"; pages: "119-129"
SGRef: number: 2; type: "Journal Article"; journalName: "J. Mol.
Biol."; date: "1983"; volume: "171"; issue: "3"; pages: "239-261"
SGRef: number: 3; type: "Journal Article"; journalName: "Gene";
date: "1985"; volume: "33"; issue: "1"; pages: "103-119"
SGRef: number: 4; type: "Journal Article"; journalName: "Gene 50
(1-3), 3-40 (1986)"
SGRef: number: 5; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "1988"; volume: "16"; issue: "15"; pages:
"7583-7600"
SGRef: number: 6; type: "Journal Article"; journalName: "J. Mol.
Biol."; date: "1991"; volume: "222"; issue: "1"; pages: "99-124"
SGRef: number: 7; type: "Journal Article"; journalName: "Meth.
Enzymol. 216, 483-495 (1992)"
SGRef: number: 8; type: "Journal Article"; journalName: "Mol.
Microbiol."; date: "1992"; volume: "6"; issue: "22"; pages:
"3385-3393"
SGRef: number: 9; type: "Journal Article"; journalName: "Gene 138
(1-2), 109-114 (1994)"
SGRef: number: 10; type: "Journal Article"; journalName: "Submitted
(10-SEP-1993) Dept. of Chemistry and Molecular Biology, The Scripps
Research Institute, 10550 North Torrey Pines Road, La Jolla, CA
92037, USA"
SGRef: number: 11; type: "Journal Article"
On Oct 6, 1993 this sequence version replaced gi:403935.
FEATURES Location/Qualifiers
source 1..4133
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 18..34
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
CDS 146..1126
/gene="pheS"
/label="Phenylalanine--tRNA ligase alpha subunit"
/note="Phenylalanine--tRNA ligase alpha subunit from
Escherichia coli O139:H28 (strain E24377A / ETEC).
Accession#: A7ZMI2"
primer_bind complement(1244..1260)
/label="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(1293..1311)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1318..1334)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 1476..1931
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1957..2061
/label="AmpR promoter"
CDS 2062..2919
/label="AmpR"
/note="beta-lactamase"
rep_origin 3093..3681
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 3864..3956
/codon_start=1
/gene="'lacI"
/product="lac repressor fragment"
/label="'lacI"
/note="vestigal C-terminal portion of lac repressor; this
3' portion of lacI is not functional"
/citation=[3]
/citation=[7]
/protein_id="AAB61949.1"
/translation="APNTQTASPRALADSLMQLARQVSRLESGQ"
gene 3864..3956
/gene="'lacI"
/label="'lacI"
protein_bind 3969..3990
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4005..4035
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4043..4059
/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 4067..4083
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 4104..4122
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
old_sequence 4133
/replace="tg"
/note="all pBluescript I KS (+/-) vectors (and pKSS)
exhibit the deletion of the G nucleotide 5' to the KpnI
site [8,9]"
/citation=[5]
/citation=[7]
/citation=[9]