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:
- pSI-DGB2
- 载体抗性:
- Gentamycin
- 载体长度:
- 4312 bp
- 载体类型:
- Cloning vector
- 复制子:
- R6K γ ori
- 载体来源:
- Kriz A, Schmid K, Baumgartner N, Ziegler U, Berger I, Ballmer-Hofer K, Berger P.
- 启动子:
- CMV
pSI-DGB2 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。
pSI-DGB2 载体序列
LOCUS 40924_40247 4312 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pSI-DGB2, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4312)
AUTHORS Kriz A, Schmid K, Baumgartner N, Ziegler U, Berger I, Ballmer-Hofer
K, Berger P.
TITLE A plasmid-based multigene expression system for mammalian cells
JOURNAL Nat Commun 1, 120 (2010)
PUBMED 21081918
REFERENCE 2 (bases 1 to 4312)
AUTHORS Kriz A, Schmid K, Baumgartner N, Ziegler U, Berger I, Ballmer-Hofer
K, Berger P.
TITLE Direct Submission
JOURNAL Submitted (11-OCT-2010) Laboratory of Biomolecular Research, Paul
Scherrer Institut, OFLC / 101, Villigen PSI CH-5232, Switzerland
REFERENCE 3 (bases 1 to 4312)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4312)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun
1, 120 (2010)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(11-OCT-2010) Laboratory of Biomolecular Research, Paul Scherrer
Institut, OFLC / 101, Villigen PSI CH-5232, Switzerland"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4312
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 71..374
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 375..578
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 623..641
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 703..1419
/codon_start=1
/label=EBFP2
/note="enhanced blue variant of GFP (Ai et al., 2007)"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
EFRTAAGITLGMDELYK"
protein_bind 1478..1499
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1514..1544
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1552..1568
/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 1576..1592
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
RBS 1619..1641
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1766..2473
/codon_start=1
/label=GFP
/note="Aequorea victoria green fluorescent protein"
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
ICTTGKLPVPWPTLVTTLCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
FKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
VTAAGITHGMDELY"
polyA_signal 2666..2890
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
CDS complement(3120..3650)
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
rep_origin complement(3848..4229)
/direction=LEFT
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
protein_bind complement(4271..4304)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."