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。
The pCAMBIA1302 is a Agrobacterium binary vector for plant transformation, with hygromycin- and kanamycin-resistance and GFP genes.
- 载体名称:
- pCAMBIA1302
- 载体抗性:
- Kanamycin
- 载体长度:
- 10549 bp
- 载体类型:
- Plant Vectors
- 复制子:
- ori
- 宿主:
- Plants
- 载体来源:
- Cambia
- 拷贝数:
- High copy number
- 启动子:
- CaMV 35S (enhanced)
- 5'引物:
- M13 fwd
- 3'引物:
- M13 rev
- 感受态:
- Stbl3
- 培养温度:
- 37℃
pCAMBIA1302 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。
pCAMBIA1302 载体序列
LOCUS Exported 10549 bp DNA circular SYN 03-SEP-2024
DEFINITION Agrobacterium binary vector for plant transformation, with
hygromycin- and kanamycin-resistance and GFP genes.
ACCESSION AF234298
VERSION .
KEYWORDS pCAMBIA1302
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10549)
AUTHORS Cambia
TITLE Direct Submission
REFERENCE 2 (bases 1 to 10549)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT The GenBank record was corrected by inserting a G at position 3443.
FEATURES Location/Qualifiers
source 1..10549
/lab_host="Plant Cells"
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 17..727
/codon_start=1
/gene="mgfp5"
/product="GFP with folding enhancement mutations"
/label=mgfp5
/note="mgfp5"
/note="suitable for expression in plants due to removal of
a cryptic intron"
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
FKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
CDS 734..751
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis affinity tag
/note="6xHis"
/translation="HHHHHH"
terminator 783..1035
/note="NOS terminator"
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 1057..1081
/label=RB T-DNA repeat
/note="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
CDS 2382..3011
/codon_start=1
/product="stability protein from Pseudomonas plasmid pVS1"
/label=stability protein from Pseudomonas plasmid pVS1
/note="pVS1 StaA"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 3440..4513
/codon_start=1
/product="replication protein from Pseudomonas plasmid
pVS1"
/label=replication protein from Pseudomonas plasmid
pVS1
/note="pVS1 RepA"
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 4579..4773
/label=pVS1 oriV
/note="pVS1 oriV"
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 5117..5257
/label=bom
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(5443..6031)
/direction=LEFT
/label=ori
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6118..6912)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=aphA-3
/note="KanR"
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 7337..7361
/label=LB T-DNA repeat
/note="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 7439..7613
/label=CaMV poly(A) signal
/note="CaMV poly(A) signal"
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(7654..8679)
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=hygromycin B phosphotransferase
/note="HygR"
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(8746..9422)
/note="CaMV 35S promoter (enhanced)"
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
promoter 9649..9679
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 9687..9703
/label=lac repressor encoded by lacI binding site
/bound_moiety="lac repressor encoded by lacI"
/note="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)."
primer_bind 9711..9727
/label=M13 rev
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 9723..9955
/codon_start=1
/gene="lacZ (truncated)"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ (truncated)
/note="lacZ-alpha"
/translation="MT*LRIRARYPGIL*SRPAGMQAWHWPSFYNVVTGKTLALPNLIA
LQHIPLSPAGVIAKRPAPIALPNSCAA*MANA"
misc_feature 9736..9792
/label=MCS
/note="MCS"
/note="pUC18/19 multiple cloning site"
primer_bind complement(9796..9812)
/label=M13 fwd
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 10189..10534
/note="CaMV 35S promoter"
/note="strong constitutive promoter from cauliflower mosaic
virus"