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:
- pCVD047
- 载体抗性:
- Ampicillin
- 载体长度:
- 7140 bp
- 载体类型:
- Broad host range vector
- 复制子:
- RSF1010 oriV
- 载体来源:
- Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
pCVD047 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。
pCVD047 载体序列
LOCUS 40924_49217 7140 bp DNA circular SYN 18-DEC-2018
DEFINITION Broad host range vector vector pCVD047, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7140)
AUTHORS Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B,
Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
TITLE Broad-host-range vector system for synthetic biology and
biotechnology in cyanobacteria
JOURNAL Nucleic Acids Res. (2014) In press
PUBMED 25074377
REFERENCE 2 (bases 1 to 7140)
AUTHORS Taton A, Unglaub F, Wright N, Zeng WY, Paz Yepes J, Brahamsha B,
Palenik B, Peterson T, Haerizadeh F, Golden SS, Golden JW.
TITLE Direct Submission
JOURNAL Submitted (17-JUN-2014) Division of Biological Sciences, University
of California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
REFERENCE 3 (bases 1 to 7140)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7140)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res. (2014) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(17-JUN-2014) Division of Biological Sciences, University of
California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7140
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(124..981)
/label=AmpR
/note="beta-lactamase"
promoter complement(982..1086)
/label=AmpR promoter
misc_feature 1112..1117
/label=ZraI
/note="ZraI"
misc_feature 1227..1232
/label=ZraI
/note="ZraI"
misc_feature 1230..1250
/note="G5C5; GC-adaptor for seamless assembly, flanking a
BHR replicons, neutral sites, or cyanobacterial replicons;
synthetic"
misc_feature 1251..1256
/label=XbaI
/note="XbaI"
regulatory 1288..1314
/note="trpA T; trpA terminator from Lorist6 INSD accession
X98450, one nt shorter than reference; Lorist6"
/regulatory_class="terminator"
CDS complement(1650..2498)
/label=RSF1010 RepC
/note="replication protein C of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS complement(2488..3324)
/label=RSF1010 RepA
/note="replication protein A of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS complement(3354..3560)
/codon_start=1
/product="repF"
/label=repF
/note="repF-RSF1010; replication gene; derived from
pRL1383a in INSD accession AF403426, a derivative of
RSF1010 INSD accession M28829"
/protein_id="AII99706.1"
/translation="MKDQKDKQTGDLLASPDAVRQARYAERMKAKGMRQRKFWLTDDEY
EALRECLEELRAAQGGGSDPASA"
CDS complement(3562..3774)
/codon_start=1
/product="repE"
/label=repE
/note="repE-RSF1010; replication gene; derived from
pRL1383a in INSD accession AF403426, a derivative of
RSF1010 INSD accession M28829"
/protein_id="AII99707.1"
/translation="MEYEKSASGSVYLIKSDKGYWLPGGFGYTSNKAEAGRFSVADMAS
LNLDGCTLSLFREDKPFGPGKFLGD"
regulatory complement(3804..3831)
/note="P4; derived from pRL1383a in INSD accession
AF403426, a derivative of RSF1010 INSD accession M28829;
the P4 promoter is under the auto-regulated control of the
F repressor, whose gene is located in the same operon,
providing transcription of E-F-repA-repC-operon"
/regulatory_class="promoter"
CDS complement(3838..4806)
/label=RSF1010 RepB
/note="replication protein B of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS complement(4803..5216)
/codon_start=1
/product="mobB"
/label=mobB
/note="mobilization gene; derived from pRL1383a in INSD
accession AF403426, a derivative of RSF1010 INSD accession
M28829"
/protein_id="AII99704.1"
/translation="MNAIDRVKKSRGINELAEQIEPLAQSMATLADEARQVMSQTKQAS
EAQAAEWLKAQRQTGAAWVELAKELREVAAEVSSAAQSARSASRGWHWKLWLTVMLASM
MPTVVLLIASLLLLDLTPLTTEDGSIWLRLVAR"
misc_feature 5662..5667
/label=AgeI
/note="AgeI"
oriT complement(6045..6132)
/direction=LEFT
/label=RSF1010 oriT
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
regulatory complement(6141..6172)
/note="P1; derived from pRL1383a in INSD accession
AF403426, a derivative of RSF1010 INSD accession M28829;
the P1 promoter is controlled by MobC and MobA provide
transcription of mobA/repB, mobB, and, probably,
E-F-repA-repC-operon"
/regulatory_class="promoter"
CDS 6163..6447
/codon_start=1
/product="mobC"
/label=mobC
/note="mobilization gene; derived from pRL1383a in INSD
accession AF403426, a derivative of RSF1010 INSD accession
M28829"
/protein_id="AII99703.1"
/translation="MVKGSNKAADRLAKLEEQRARINAEIQRERAREQQQERKNETRRK
VLVGAMILAKVNSSEWPEDRLMAAMDAYLERDHDRALFGLPPRQKDEPG"
rep_origin 6474..6868
/label=RSF1010 oriV
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
terminator complement(7035..7078)
/label=bacterial terminator
/note="putative bacterial transcription terminator"
regulatory complement(7044..7072)
/note="TrrnC; rrnC terminator from Lorist6 INSD accession
X98450; Lorist6"
/regulatory_class="terminator"
misc_feature 7111..7116
/label=NheI
/note="NheI"
misc_feature 7117..7137
/note="GC; GC-adaptor for seamless assembly, flanking BHR
replicons or neutral sites; synthetic"
misc_feature 7135..7140
/label=ZraI
/note="ZraI"