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:
- pBBR4pemIK-GW
- 载体抗性:
- Chloramphenicol
- 载体长度:
- 7368 bp
- 载体类型:
- Cloning vector
- 复制子:
- pBBR1 oriV
- 载体来源:
- Burbank L, Wei W.
- 启动子:
- lac UV5
pBBR4pemIK-GW 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。
pBBR4pemIK-GW 载体序列
LOCUS 62056_3730 7368 bp DNA circular SYN 06-DEC-2019
DEFINITION Cloning vector pBBR4pemIK-GW, complete sequence.
ACCESSION MN044102
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7368)
AUTHORS Burbank L, Wei W.
TITLE Broad-Host-Range Plasmids for Constitutive and Inducible Gene
Expression in the Absence of Antibiotic Selection
JOURNAL Microbiol Resour Announc 8 (36), e00769-19 (2019)
PUBMED 31488530
REFERENCE 2 (bases 1 to 7368)
AUTHORS Burbank LP, Wei W.
TITLE Direct Submission
JOURNAL Submitted (06-JUN-2019) Crop Diseases Pests and Genetics Research
Unit, USDA Agricultural Research Service, 9611 S Riverbend Ave,
Parlier, CA 93648, USA
REFERENCE 3 (bases 1 to 7368)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Microbiol
Resour Announc"; date: "2019"; volume: "8"; issue: "36"; pages:
"e00769-19"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-JUN-2019) Crop Diseases Pests and Genetics Research Unit, USDA
Agricultural Research Service, 9611 S Riverbend Ave, Parlier, CA
93648, USA"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7368
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 1..770
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
CDS 771..1430
/label=pBBR1 Rep
/note="replication protein for the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica"
primer_bind 2140..2156
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2166..2184
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 2217..2233
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(2293..2655)
/codon_start=1
/transl_table=11
/product="mRNA interferase PemK"
/label=mRNA interferase PemK
/protein_id="QDJ95328.1"
/translation="MGRCSSSRTRTDMKRGDVYMVDLEPTAGHEQRGHRPVVVISSERF
NRLTSCPVILPITNGGEFASRLGFAVELLGTITTGVVRCDQPRALDLLARNARKVESLP
PNILADVLAKAVTIFQ"
CDS complement(2616..2879)
/codon_start=1
/transl_table=11
/product="antitoxin PemI"
/label=antitoxin PemI
/protein_id="QDJ95329.1"
/translation="MHTTNLRKVGGSIMLAVPPAFLDQLHLEVGATVGLAVTDGRLVIE
PVLHPQYTLDQLLAEAEASGAYPLPPEEREWVDAPAVGRELI"
protein_bind 2965..3089
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 3114..3144
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 3198..3854
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 4199..4501
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(4545..4669)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(4685..4701)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(4731..4749)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4770..4786)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4794..4810)
/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(4818..4848)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4863..4884)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5112..5183
/label=AmpR promoter
CDS 5184..6041
/label=AmpR
/note="beta-lactamase"