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:
- pN-TAPa
- 载体抗性:
- Streptomycin
- 载体长度:
- 12537 bp
- 载体类型:
- N-terminal TAPa T-DNA vector
- 复制子:
- ori
- 载体来源:
- Rubio V, Shen Y, Saijo Y, Liu Y, Gusmaroli G, Dinesh-Kumar SP, Deng XW.
- 启动子:
- CaMV35S(enhanced)
pN-TAPa 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。
pN-TAPa 载体序列
LOCUS 40924_32968 12537 bp DNA circular SYN 18-DEC-2018
DEFINITION N-terminal TAPa T-DNA vector pN-TAPa, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12537)
AUTHORS Rubio V, Shen Y, Saijo Y, Liu Y, Gusmaroli G, Dinesh-Kumar SP, Deng
XW.
TITLE An alternative tandem affinity purification strategy applied to
Arabidopsis protein complex isolation
JOURNAL Plant J. 41 (5), 767-778 (2005)
PUBMED 15703063
REFERENCE 2 (bases 1 to 12537)
AUTHORS Rubio V, Deng XW.
TITLE Direct Submission
JOURNAL Submitted (21-OCT-2004) MCDB, Yale University, 165, Prospect St.,
New Haven, CT 06511, USA
REFERENCE 3 (bases 1 to 12537)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 12537)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant J.";
date: "2005"; volume: "41"; issue: "5"; pages: "767-778"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-OCT-2004) MCDB, Yale University, 165, Prospect St., New Haven,
CT 06511, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12537
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 96..769
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
misc_feature 791..846
/label=TMV Omega
/note="translational enhancer from the tobacco mosaic virus
5'-leader sequence (Gallie et al., 1988)"
regulatory 857..866
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 884..1057
/label=ProtA
/note="IgG-binding unit of Staphylococcus aureus protein A"
CDS 1061..1231
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
EAKKLNDAQAPK"
CDS 1256..1279
/label=HRV 3C site
/note="recognition and cleavage site for human rhinovirus
3C and PreScission proteases"
CDS 1286..1303
/label=6xHis
/note="6xHis affinity tag"
CDS 1328..1357
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
CDS 1364..1393
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1400..1429
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1448..1477
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1484..1513
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1520..1549
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1568..1597
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1604..1633
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1640..1669
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
protein_bind 1704..1828
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 2002..2104
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 2105..2761
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 3106..3408
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(3452..3576)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
terminator 3605..3857
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3873..3889)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3897..3913
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(3921..3951)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3966..3987)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 4152..4176
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 5476..6102
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
CDS 6539..7603
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
rep_origin 7672..7866
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 8210..8350
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(8536..9124)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9371..10159)
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
misc_feature 10687..10711
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal complement(10809..10983)
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(11021..11551)
/label=GmR
/note="gentamycin acetyltransferase"
promoter complement(11611..12289)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
primer_bind 12518..12534
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"