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:
- pMG_DV
- 载体长度:
- 6435 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Stach C, McCann M, O'Brien C, Le TS
- 启动子:
- UbC
pMG_DV 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。
pMG_DV 载体序列
LOCUS V015655 6435 bp DNA circular SYN 23-OCT-2019
DEFINITION Exported.
ACCESSION V015655
VERSION V015655
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6435)
AUTHORS Stach C, McCann M, O'Brien C, Le TS, Somia N, Chen X, Lee K, Fu HY,
Daoutidis P, Zhao L, Hu WS, Smanski MJ.
TITLE Model-driven engineering of N-linked glycosylation in Chinese
Hamster Ovary cells
JOURNAL ACS Synth Biol (2019) In press
PUBMED 31596566
REFERENCE 2 (bases 1 to 6435)
AUTHORS Stach CS.
TITLE Direct Submission
JOURNAL Submitted (28-AUG-2019) Biochemistry, Molecular Biology, and
Biophysics; Chemical Engineering and Materials Science, University
of Minnesota, 1479 Gortner Ave, Suite 344, Saint Paul, Minnesota
55108, United States
REFERENCE 3 (bases 1 to 6435)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol (2019) In press"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(28-AUG-2019) Biochemistry, Molecular Biology, and Biophysics;
Chemical Engineering and Materials Science, University of Minnesota,
1479 Gortner Ave, Suite 344, Saint Paul, Minnesota 55108, United
States"
FEATURES Location/Qualifiers
source 1..6435
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind complement(21..54)
/label="FRT (minimal)"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
misc_feature 55..65
/label="AarI site"
/note="AarI site"
misc_feature 66..69
/label="Golden gate cloning scar"
/note="Golden gate cloning scar"
misc_feature 70..77
/label="BbsI site"
/note="BbsI site"
primer_bind 279..295
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 296..352
/label="MCS"
/note="pUC18/19 multiple cloning site"
primer_bind complement(365..381)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(389..405)
/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(413..443)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(458..479)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 560..567
/label="BbsI site"
/note="BbsI site"
misc_feature 568..571
/label="Golden gate cloning scar"
/note="Golden gate cloning scar"
misc_feature 572..582
/label="AarI site"
/note="AarI site"
promoter 588..1799
/label="UbC promoter"
/note="human ubiquitin C promoter"
CDS 1831..2250
/gene="bsr"
/label="Blasticidin-S deaminase"
/note="Blasticidin-S deaminase from Bacillus cereus.
Accession#: P33967"
polyA_signal 2257..2305
/label="poly(A) signal"
/note="synthetic polyadenylation signal"
protein_bind complement(2306..2339)
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 2358..2396
/label="PhiC31 attP"
/note="PhiC31 attP"
CDS complement(2719..3279)
/codon_start=1
/transl_table=11
/product="kanamycin resistance protein"
/label="kanamycin resistance protein"
/protein_id="QFQ66234.1"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLHV
IYTLLLPRKYPSWLMQCGGCIRLIRLPAHSTTKRNIASSEHVLGWKPVLSIRMIWTKSI
RGSRQPNCSPGSRRACPTARISS"
terminator 3812..3898
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3990..4017
/label="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
rep_origin 4151..4739
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(4925..5065)
/label="bom"
/note="basis of mobility region from pBR322"
CDS complement(6362..6379)
/label="6xHis"
/note="6xHis affinity tag"
protein_bind 6400..6433
/label="attB"
/note="minimal attB site for the phi-C31 integrase (Groth
et al., 2000)"