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。
Mobilisable shuttle and expression vector, pBBR1MCS series. Vector is mobilisable via RP4/RK2 mating system, e.g. using E.coli strains S17 or SM10. Note: one base pair deletion mutation near C-terminal mob protein
- 载体名称:
- pBBR1MCS-5
- 载体抗性:
- Gentamicin
- 载体长度:
- 4771 bp
- 载体类型:
- Broad Host Shuttle Vectors
- 复制子:
- pBBR1 oriV
- 拷贝数:
- Low copy number
- 启动子:
- Pc
- 感受态:
- DH5alpha
- 培养温度:
- 37℃
pBBR1MCS-5 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。
pBBR1MCS-5 载体序列
LOCUS Exported 4771 bp DNA circular SYN 31-AUG-2024
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4771)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 4771)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 4771)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4771)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4771
/mol_type="other DNA"
/organism="synthetic DNA construct"
source join(229..4771,1..228)
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(24..1027)
/codon_start=1
/product="mob encodes the plasmid mobilization functions
that allow conjugal delivery of the plasmids into a variety
of bacteria from E. coli strains harboring the RK2 conjugal
transfer functions."
/label=mob
/translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLRRC"
promoter complement(1095..1146)
/label=promoter for mob
misc_feature 1106..1128
/label=RSA
/note="transfer origins (also called recombination site A
[RSA]), is known as the specific site necessary for
mobilization and recombination mediated by a Mob/Pre
protein. "
rep_origin 1251..2022
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
CDS 2023..2682
/codon_start=1
/label=pBBR1 Rep
/note="replication protein for the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica"
/translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
primer_bind 3394..3410
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 3420..3438
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 3447..3554
/label=MCS
/note="pBluescript multiple cloning site"
promoter complement(3567..3585)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3606..3622)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3630..3646)
/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(3654..3684)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3699..3720)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3990..4018
/label=Pc promoter
/note="class 1 integron promoter"
CDS 4207..4737
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"