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:
- pMQ61
- 载体抗性:
- Kanamycin
- 载体长度:
- 7978 bp
- 载体类型:
- Shuttle vector
- 复制子:
- ori
- 宿主:
- Yeast
- 载体来源:
- Shanks RM, Caiazza NC, Hinsa SM, Toutain CM, O'toole GA.
- 启动子:
- TEF
pMQ61 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。
pMQ61 载体序列
LOCUS 40924_32120 7978 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle vector pMQ61, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7978)
AUTHORS Shanks RM, Caiazza NC, Hinsa SM, Toutain CM, O'toole GA.
TITLE Saccharomyces cerevisiae-Based Molecular Tool Kit for Manipulation
of Genes from Gram-Negative Bacteria
JOURNAL Appl. Environ. Microbiol. 72 (7), 5027-5036 (2006)
PUBMED 16820502
REFERENCE 2 (bases 1 to 7978)
AUTHORS Shanks RMQ., Caiazza NC, Hinsa SM, Toutain CM, O'Toole GA.
TITLE Direct Submission
JOURNAL Submitted (16-SEP-2005) Microbiology and Immunology, Dartmouth
Medical School, Vail Building Room 505, Hanover, NH 03755, USA
REFERENCE 3 (bases 1 to 7978)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7978)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2006"; volume: "72"; issue: "7"; pages:
"5027-5036"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(16-SEP-2005) Microbiology and Immunology, Dartmouth Medical School,
Vail Building Room 505, Hanover, NH 03755, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7978
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1..801
/codon_start=1
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
misc_feature 964..997
/label=loxP
/note="loxP"
protein_bind 964..997
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
gene complement(1060..2416)
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
protein_bind complement(2471..2504)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
rep_origin complement(2555..2906)
/direction=LEFT
/label=pRO1600 oriV
/note="broad-host-range origin of replication from
Pseudomonas aeruginosa plasmid pRO1600; requires the
pRO1600 Rep protein for replication (West et al., 1994)"
CDS 2920..3750
/codon_start=1
/label=pRO1600 Rep
/note="replication protein for the broad-host-range plasmid
pRO1600 from Pseudomonas aeruginosa"
/translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
primer_bind 3914..3930
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 3940..3958
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature complement(3967..4074)
/label=MCS
/note="pBluescript multiple cloning site"
promoter complement(4087..4105)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4126..4142)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4150..4166)
/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(4174..4204)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4219..4240)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4528..5116)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5328..5858)
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(6047..6075)
/label=Pc promoter
/note="class 1 integron promoter"
rep_origin complement(6152..7494)
/direction=LEFT
/label=2u ori
/note="yeast 2u plasmid origin of replication"
promoter 7758..7978
/label=URA3 promoter