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:
- pRED56-MD8-Pnos-55
- 载体抗性:
- Ampicillin
- 载体长度:
- 8695 bp
- 载体类型:
- Gateway recycling vector
- 复制子:
- ori
- 宿主:
- Plants
- 载体来源:
- Saisho T, Shibahara K, Nakao A, Tanaka Y, Nishimura K, Kimura T, Nakagawa T.
- 启动子:
- NOS
pRED56-MD8-Pnos-55 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。
pRED56-MD8-Pnos-55 载体序列
LOCUS 40924_36698 8695 bp DNA circular SYN 18-DEC-2018
DEFINITION Gateway recycling vector pRED56-MD8-Pnos-55 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8695)
AUTHORS Saisho T, Shibahara K, Nakao A, Tanaka Y, Nishimura K, Kimura T,
Nakagawa T.
TITLE Improved Gateway Recycling Cloning System for Multigene Construction
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8695)
AUTHORS Saisho T, Shibahara K, Nakao A, Tanaka Y, Nishimura K, Kimura T,
Nakagawa T.
TITLE Direct Submission
JOURNAL Submitted (02-NOV-2015) Contact:Tsuyoshi Nakagawa Shimane
University, Interdisciplinary Center for Science Research; 1060
Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE 3 (bases 1 to 8695)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8695)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-NOV-2015) Contact:Tsuyoshi Nakagawa Shimane University,
Interdisciplinary Center for Science Research; 1060 Nishikawatsu,
Matsue, Shimane 690-8504, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT constructed using pDONR201.
FEATURES Location/Qualifiers
source 1..8695
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 28..127
/label=attL1
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 674..1071
/label=MD8
/note="MD8"
promoter 1192..1375
/label=NOS promoter
/note="nopaline synthase promoter"
CDS 1393..2067
/codon_start=1
/label=mRFP1
/note="monomeric derivative of DsRed (Campbell et al.,
2002)"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
protein_bind 2077..2201
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 2238..2268
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 2322..2978
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 3323..3625
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(3669..3793)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
terminator 3819..4071
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
protein_bind 4280..4404
/label=attR4
/note="recombination site for the Gateway(R) LR reaction"
promoter 4429..4459
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
misc_feature 4731..4771
/label=rare cutter sites
/note="rare cutter sites"
CDS complement(4863..5720)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5721..5825)
/label=AmpR promoter
protein_bind complement(6122..6245)
/label=attR3
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(6571..6666)
/label=attL6
/note="recombination site for the Gateway(R) LR reaction"
CDS 6790..7596
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 7742..8330
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator complement(8465..8492)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(8584..8670)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"