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:
- pBG
- 载体抗性:
- Kanamycin
- 载体长度:
- 3947 bp
- 载体类型:
- Cloning vector
- 复制子:
- R6K γ ori
- 载体来源:
- Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank LM.
- 启动子:
- Pc
pBG 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。
pBG 载体序列
LOCUS 40924_6257 3947 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pBG, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3947)
AUTHORS Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank
LM.
TITLE A Tn7-based device for calibrated heterologous gene expression in
Pseudomonas putida
JOURNAL ACS Synth Biol (2015) In press
PUBMED 26133359
REFERENCE 2 (bases 1 to 3947)
AUTHORS Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank
LM.
TITLE Direct Submission
JOURNAL Submitted (12-JUN-2015) System Biology, Centro Nacional de
Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE 3 (bases 1 to 3947)
AUTHORS Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank
LM.
TITLE Direct Submission
JOURNAL Submitted (13-NOV-2015) System Biology, Centro Nacional de
Biotecnologia, C/Darwin 3, Madrid 28049, Spain
REFERENCE 4 (bases 1 to 3947)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 3947)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol (2015) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(12-JUN-2015) System Biology, Centro Nacional de Biotecnologia,
C/Darwin 3, Madrid 28049, Spain"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(13-NOV-2015) System Biology, Centro Nacional de Biotecnologia,
C/Darwin 3, Madrid 28049, Spain"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
On Nov 13, 2015 this sequence version replaced KT192136.1.
FEATURES Location/Qualifiers
source 1..3947
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 44..120
/label=BCD2 bicistronic linker
/note="BCD2 bicistronic linker"
RBS 112..120
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 139..852
/codon_start=1
/label=superfolder GFP
/note="GFP variant that folds robustly even when fused to
poorly folded proteins (Pedelacq et al., 2006)"
/translation="IHKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
misc_feature 856..912
/label=MCS
/note="pUC18/19 multiple cloning site"
terminator 955..1049
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
misc_feature 1054..1252
/label=Tn7R transposase recognition site
/note="Tn7R transposase recognition site"
CDS 1362..2174
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
oriT 2333..2441
/label=oriT
/note="incP origin of transfer"
rep_origin 2460..2848
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
mobile_element 2860..3025
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
terminator complement(3039..3125)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(3194..3724)
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(3913..3941)
/label=Pc promoter
/note="class 1 integron promoter"