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pCAMBIA1302 质粒 (编号: V010892)

Note: The pCAMBIA1302 is a Agrobacterium binary vector for plant transformation, with hygromycin- and kanamycin-resistance and GFP genes.

pCAMBIA130210549 bp5001000150020002500300035004000450050005500600065007000750080008500900095001000010500lac promoterlac operatorlacZ (truncated)CaMV 35S promotermgfp56xHis affinity tagNOS terminatorRB T-DNA repeatpVS1 StaApVS1 RepApVS1 oriVbomoriaphA-3LB T-DNA repeatCaMV poly(A) signalHygRCaMV 35S promoter (enhanced)
复制序列 NovoBuilder中查看图谱

基本信息

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:
DH10B
培养温度:
37℃

产品信息

质粒编号:V010892
质粒名称:pCAMBIA1302
规格:5 μg (冻干粉)

下载资源

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。

确保质粒的关键元件正确，但是我们并不能保证实验效果。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。

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。

参考文献

  • Lim HS, Ko TS, Lambert KN, Kim HG, Korban SS, Hartman GL, Domier LL. Soybean mosaic virus helper component-protease enhances somatic embryo production and stabilizes transgene expression in soybean. Plant Physiol Biochem. 2005 Oct-Nov;43(10-11):1014-21. doi: 10.1016/j.plaphy.2005.08.012. Epub 2005 Oct 19. PMID: 16316753.

  • Rizwan HM, Yang Q, Yousef AF, Zhang X, Sharif Y, Kaijie J, Shi M, Li H, Munir N, Yang X, Wei X, Oelmüller R, Cheng C, Chen F. Establishment of a Novel and Efficient Agrobacterium-Mediated in Planta Transformation System for Passion Fruit (Passiflora edulis). Plants (Basel). 2021 Nov 15;10(11):2459. doi: 10.3390/plants10112459. PMID: 34834821; PMCID: PMC8621743.

  • Gupta T, Singhal T, Dhir S, Chandel V, Rishi N, Raj SK, Srivastava A. Demonstration of seed inoculation of dimer infectious clones of mungbean yellow mosaic India virus-soybean isolate. 3 Biotech. 2023 Nov;13(11):360. doi: 10.1007/s13205-023-03778-7. Epub 2023 Oct 11. PMID: 37840874; PMCID: PMC10567615.

pCAMBIA1302 质粒 (编号: V010892)序列

LOCUS       Exported               10549 bp DNA     circular SYN 10-SEP-2025
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)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 10549)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; 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"
     source          join(1090..10549,1..1089)
                     /lab_host="Plant Cells"
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        189..219
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    227..243
                     /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     251..267
                     /label=M13 rev
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             263..495
                     /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    276..332
                     /label=MCS
                     /note="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(336..352)
                     /label=M13 fwd
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        729..1074
                     /note="CaMV 35S promoter"
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1106..1816
                     /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             1823..1840
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis affinity tag
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      1872..2124
                     /note="NOS terminator"
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2146..2170
                     /label=RB T-DNA repeat
                     /note="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3471..4100
                     /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             4529..5602
                     /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      5668..5862
                     /label=pVS1 oriV
                     /note="pVS1 oriV"
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    6206..6346
                     /label=bom
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6532..7120)
                     /direction=LEFT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7207..8001)
                     /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    8426..8450
                     /label=LB T-DNA repeat
                     /note="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    8528..8702
                     /label=CaMV poly(A) signal
                     /note="CaMV poly(A) signal"
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(8743..9768)
                     /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(9835..10511)
                     /note="CaMV 35S promoter (enhanced)"
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"