首页技术支持质粒载体

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。

p35S-GFP carries the gene for the green fluorescent protein (GFP) from the bioluminescent jellyfish Aequorea victoria. p35S-GFP was constructed by replacing the GUS coding sequence of pBI221 with a functional GFP gene, thereby placing the GFP gene under the control of the CaMV 35S promoter. Protoplasts were viewed by incident-light fluorescence microscopy 24h after electroporation. 20-60% of the protoplasts emitted an intense green light when illuminated with blue (450-490 nm) light.

载体名称:
p35S-GFP
载体抗性:
Ampicillin
载体长度:
4518 bp
载体类型:
Plant GFP
复制子:
ori
宿主:
Plants
载体来源:
Niedz RP, Sussman MR, Satterlee JS.
启动子:
CaMV35S(long)
感受态:
stbl3
培养温度:
37℃

p35S-GFP 载体图谱

p35S-GFP4518 bp600120018002400300036004200CaMV 35S promoterGFPNOS terminatorM13 fwdAmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 rev

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。

p35S-GFP 载体序列

LOCUS       40924_2622        4518 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p35S-GFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4518)
  AUTHORS   Niedz RP, Sussman MR, Satterlee JS.
  TITLE     Green fluorescent protein: an in vivo reporter of plant gene 
            expression
  JOURNAL   Plant Cell Rep. 14 (7), 403-406 (1995)
  PUBMED    24185445
REFERENCE   2  (bases 1 to 4518)
  AUTHORS   Kitts PA.
  TITLE     CLONTECH Vectors On Disk version 1.3
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 4518)
  AUTHORS   Kitts PA.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-JUN-1995) Paul A. Kitts, CLONTECH Laboratories, Inc., 
            4030 Fabian Way, Palo Alto, CA 94303, USA
REFERENCE   4  (bases 1 to 4518)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4518)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell 
            Rep."; date: "1995"; volume: "14"; issue: "7"; pages: "403-406"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (05-JUN-1995) Paul A. Kitts, CLONTECH Laboratories, Inc., 4030 
            Fabian Way, Palo Alto, CA 94303, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     This vector can be obtained from CLONTECH Laboratories, Inc., 4030 
            Fabian Way, Palo Alto, CA 94303, USA. To place an order call (415) 
            424-8222 or (800) 662-2566, extension 1. International customers, 
            please contact your local distributor. For technical information, 
            call (415) 424-8222 or (800) 662-2566, extension 3. This sequence 
            has been compiled from information in the sequence databases, 
            published literature and other sources, together with partial 
            sequences obtained by CLONTECH. If you suspect there is an error in 
            this sequence, please contact CLONTECH's Technical Service 
            Department at (415) 424-8222 or (800) 662-2566, extension 3 or 
            E-mail TECH@CLONTECH.COM.
FEATURES             Location/Qualifiers
     source          1..4518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        512..857
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             896..1609
                     /codon_start=1
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1629..1881
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1890..1906)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2380..2484
                     /label=AmpR promoter
     CDS             2485..3342
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3516..4104
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4392..4413
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4428..4458
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4466..4482
                     /label=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     4490..4506
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"