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:
- pBS[NBL KO 1]
- 载体抗性:
- Ampicillin
- 载体长度:
- 10843 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Davis RJ, Belikoff EJ, Scholl EH, Li F, Scott MJ.
pBS[NBL KO 1] 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。
pBS[NBL KO 1] 载体序列
LOCUS 40924_7481 10843 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pBS[NBL KO 1], complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10843)
AUTHORS Davis RJ, Belikoff EJ, Scholl EH, Li F, Scott MJ.
TITLE no blokes Is Essential for Male Viability and X Chromosome Gene
Expression in the Australian Sheep Blowfly
JOURNAL Curr. Biol. (2018) In press
PUBMED 29887311
REFERENCE 2 (bases 1 to 10843)
AUTHORS Davis RJ, Belikoff EJ, Scholl EH, Li F, Scott MJ.
TITLE Direct Submission
JOURNAL Submitted (01-APR-2018) Entomology and Plant Pathology, North
Carolina State University, 112 Derieux Place, 1546 Thomas Hall,
Raleigh, NC 27607, USA
REFERENCE 3 (bases 1 to 10843)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10843)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Curr. Biol.
(2018) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(01-APR-2018) Entomology and Plant Pathology, North Carolina State
University, 112 Derieux Place, 1546 Thomas Hall, Raleigh, NC 27607,
USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..10843
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(1..1133)
/label=homology arm derived from Lucilia cuprina NBL
/note="homology arm derived from Lucilia cuprina NBL"
promoter complement(1164..1182)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1203..1219)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1227..1243
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(1251..1281)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1296..1317)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1605..2193)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2367..3224)
/label=AmpR
/note="beta-lactamase"
promoter complement(3225..3329)
/label=AmpR promoter
rep_origin complement(3355..3810)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 3952..3968
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 3978..3996
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature complement(4008..5073)
/label=homology arm derived from Lucilia cuprina NBL
/note="homology arm derived from Lucilia cuprina NBL"
regulatory 5074..8495
/label=derived from Lucilia cuprina h83
/note="derived from Lucilia cuprina h83"
/regulatory_class="promoter"
CDS 8496..9170
/label=DsRed-Express2
/note="noncytotoxic tetrameric variant of DsRed fluorescent
protein (Strack et al., 2008)"
misc_feature 9174..10599
/note="derived from Lucilia cuprina atub UTR and
polyadenylation signal sequence"
protein_bind 10663..10762
/label=phage phi-C31 attP
/note="attachment site of phage phi-C31"