pAcGFP1-1

价格:1200元

联系方式:I47-825O-882O

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载体基本信息

出品公司: Clontech
载体名称: pAcGFP1-1
质粒类型: 无启动子载体;荧光报告载体
高拷贝/低拷贝: 高拷贝
克隆方法: 限制性内切酶,多克隆位点
启动子: 无启动子
载体大小: 4150 bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: 绿色荧光蛋白AcGFP
载体抗性: 卡那霉素
筛选标记: Neomycin_1.html' target='_blank'>新霉素Neomycin
克隆菌株: DH5α, HB101
宿主细胞(系): 真核细胞
备注: pAcGFP1-1载体不含启动子,将待研究的目的启动子插入MCS区,用于研究启动子及顺式转录元件在细胞内的作用水平;
AcGFP荧光亮度高。
产品目录号: 632497
稳定性: 瞬表达 或 稳表达
组成型/诱导型: --
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息

pAcGFP1-1载体图谱



pAcGFP1-1 多克隆位点

pAcGFP1-1 载体特征1
pAcGFP1-1 载体特征2

载体简介

pAcGFP1-1载体描述

pAcGFP1-1 encodes the green fluorescent protein AcGFP1, a derivative of AcGFP from Aequorea coerulescens. AcGFP1 has been optimized for brighter fluorescence. (Excitation maximum = 475 nm; emission maximum = 505 nm.) The coding sequence of the AcGFP1 gene contains silent base changes, which correspond to human codon-usage preferences (1). pAcGFP1-1 is a promoterless vector that can be used to monitor transcription from different promoters and promoter/enhancer combinations inserted into the multiple cloning site (MCS). Sequences upstream of AcGFP1 have been converted to a Kozak consensus translation initiation site (2) to enhance translation efficiency in eukaryotic cells. SV40 polyadenylation signals downstream of the AcGFP1 gene direct proper processing of the 3' end of the AcGFP1 mRNA. The vector backbone contains an SV40 origin for replication in mammalian cells expressing the SV40 T antigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production. A neomycin-resistance cassette (Neor) allows stably transfected eukaryotic cells to be selected using G418. This cassette consists of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene. A bacterial promoter upstream of the cassette expresses kanamycin resistance in E. coli.

AcGFP1 can be used as an in vivo reporter of gene expression. Promoters should be cloned into the pAcGFP1-1 MCS upstream from the AcGFP1 coding sequence. Without addition of a functional promoter, this vector will not express AcGFP1. The recombinant AcGFP1 vector can be transfected into mammalian cells using any standard method. If required, stable transfectants can be selected using G418 (3).

pAcGFP1-1载体含有以下元件:
 MCS: 12–83
 Aequorea coerulescens green fluorescent protein (AcGFP1) gene
Kozak consensus translation initiation site: 90–100
Start codon (ATG): 97–99; Stop codon: 814-816
 SV40 early mRNA polyadenylation signal
Polyadenylation signals: 969–974 & 998–1003
mRNA 3' ends: 1007 & 1019
 f1 single-strand DNA origin: 1066–1521
(Packages noncoding strand of AcGFP1-1.)
 Ampicillin resistance (β-lactamase) promoter
–35 region: 1583–1588; –10 region: 1606–1611
Transcription start point: 1618
 SV40 origin of replication: 1862–1997
 SV40 early promoter
Enhancer (72-bp tandem repeats): 1693–1766 & 1767–1838
21-bp repeats: 1842–1862, 1863–1883 & 1885–1905
Early promoter element: 1918–1924
Major transcription start points: 1914, 1952, 1958 & 1963
 Kanamycin/neomycin resistance gene
Neomycin phosphotransferase coding sequences:
Start codon (ATG): 2046–2048; stop codon: 2838–2840
G→A mutation to remove Pst I site: 2228
C→A (Arg→Ser) mutation to remove BssH II site: 2574
 Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signal
Polyadenylation signals: 3076–3081 & 3089–3094
 pUC plasmid replication origin: 3425–4068

Propagation in E. Coli
 Suitable host strains: DH5α, HB101 and other general purpose strains. Single-stranded DNA production requires
a host containing an F plasmid such as JM109 or XL1-Blue.
 Selectable marker: plasmid confers resistance to kanamycin (50 μg/ml) in E. coli hosts.
 E. coli replication origin: pUC
 Copy number: ~500

载体序列

LOCUS       pAcGFP1-1	4150 bp 	DNA	SYN
DEFINITION  pAcGFP1-1
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..4150
                     /organism="pAcGFP1-1"
                     /mol_type="other DNA"
     CDS             97..816
                     /label="ORF frame 1"

     gene            100..813
                     /label="AcGFP1"
                     /gene="AcGFP1"

     misc_feature    1029..1048
                     /label="EBV_rev_primer"

     rep_origin      complement(1198..1504)
                     /label="f1_origin"

     promoter        1583..1611
                     /label="AmpR_promoter"

     misc_feature    complement(1677..1697)
                     /label="pBABE_3_primer"

     misc_feature    complement(1683..1898)
                     /label="SV40_enhancer"

     promoter        1695..1963
                     /label="SV40_promoter"

     rep_origin      1862..1939
                     /label="SV40_origin"

     misc_feature    1924..1943
                     /label="SV40pro_F_primer"

     CDS             2046..2840
                     /label="ORF frame 3"

     gene            2049..2837
                     /label="NeoR/KanR"
                     /gene="NeoR/KanR"

     CDS             complement(2355..2891)
                     /label="ORF frame 3"

     terminator      3015..3284
                     /label="TK_PA_terminator"

     rep_origin      3432..4051
                     /label="pBR322_origin"

ORIGIN
    1 TAGTTATTAC TAGCGCTACC GGACTCAGAT CTCGAGCTCA AGCTTCGAAT TCTGCAGTCG
   61 ACGGTACCGC GGGCCCGGGA TCCACCGGTC GCCACCATGG TGAGCAAGGG CGCCGAGCTG
  121 TTCACCGGCA TCGTGCCCAT CCTGATCGAG CTGAATGGCG ATGTGAATGG CCACAAGTTC
  181 AGCGTGAGCG GCGAGGGCGA GGGCGATGCC ACCTACGGCA AGCTGACCCT GAAGTTCATC
  241 TGCACCACCG GCAAGCTGCC TGTGCCCTGG CCCACCCTGG TGACCACCCT GAGCTACGGC
  301 GTGCAGTGCT TCTCACGCTA CCCCGATCAC ATGAAGCAGC ACGACTTCTT CAAGAGCGCC
  361 ATGCCTGAGG GCTACATCCA GGAGCGCACC ATCTTCTTCG AGGATGACGG CAACTACAAG
  421 TCGCGCGCCG AGGTGAAGTT CGAGGGCGAT ACCCTGGTGA ATCGCATCGA GCTGACCGGC
  481 ACCGATTTCA AGGAGGATGG CAACATCCTG GGCAATAAGA TGGAGTACAA CTACAACGCC
  541 CACAATGTGT ACATCATGAC CGACAAGGCC AAGAATGGCA TCAAGGTGAA CTTCAAGATC
  601 CGCCACAACA TCGAGGATGG CAGCGTGCAG CTGGCCGACC ACTACCAGCA GAATACCCCC
  661 ATCGGCGATG GCCCTGTGCT GCTGCCCGAT AACCACTACC TGTCCACCCA GAGCGCCCTG
  721 TCCAAGGACC CCAACGAGAA GCGCGATCAC ATGATCTACT TCGGCTTCGT GACCGCCGCC
  781 GCCATCACCC ACGGCATGGA TGAGCTGTAC AAGTGAGCGG CCGCGACTCT AGATCATAAT
  841 CAGCCATACC ACATTTGTAG AGGTTTTACT TGCTTTAAAA AACCTCCCAC ACCTCCCCCT
  901 GAACCTGAAA CATAAAATGA ATGCAATTGT TGTTGTTAAC TTGTTTATTG CAGCTTATAA
  961 TGGTTACAAA TAAAGCAATA GCATCACAAA TTTCACAAAT AAAGCATTTT TTTCACTGCA
 1021 TTCTAGTTGT GGTTTGTCCA AACTCATCAA TGTATCTTAA GGCGTAAATT GTAAGCGTTA
 1081 ATATTTTGTT AAAATTCGCG TTAAATTTTT GTTAAATCAG CTCATTTTTT AACCAATAGG
 1141 CCGAAATCGG CAAAATCCCT TATAAATCAA AAGAATAGAC CGAGATAGGG TTGAGTGTTG
 1201 TTCCAGTTTG GAACAAGAGT CCACTATTAA AGAACGTGGA CTCCAACGTC AAAGGGCGAA
 1261 AAACCGTCTA TCAGGGCGAT GGCCCACTAC GTGAACCATC ACCCTAATCA AGTTTTTTGG
 1321 GGTCGAGGTG CCGTAAAGCA CTAAATCGGA ACCCTAAAGG GAGCCCCCGA TTTAGAGCTT
 1381 GACGGGGAAA GCCGGCGAAC GTGGCGAGAA AGGAAGGGAA GAAAGCGAAA GGAGCGGGCG
 1441 CTAGGGCGCT GGCAAGTGTA GCGGTCACGC TGCGCGTAAC CACCACACCC GCCGCGCTTA
 1501 ATGCGCCGCT ACAGGGCGCG TCAGGTGGCA CTTTTCGGGG AAATGTGCGC GGAACCCCTA
 1561 TTTGTTTATT TTTCTAAATA CATTCAAATA TGTATCCGCT CATGAGACAA TAACCCTGAT
 1621 AAATGCTTCA ATAATATTGA AAAAGGAAGA GTCCTGAGGC GGAAAGAACC AGCTGTGGAA
 1681 TGTGTGTCAG TTAGGGTGTG GAAAGTCCCC AGGCTCCCCA GCAGGCAGAA GTATGCAAAG
 1741 CATGCATCTC AATTAGTCAG CAACCAGGTG TGGAAAGTCC CCAGGCTCCC CAGCAGGCAG
 1801 AAGTATGCAA AGCATGCATC TCAATTAGTC AGCAACCATA GTCCCGCCCC TAACTCCGCC
 1861 CATCCCGCCC CTAACTCCGC CCAGTTCCGC CCATTCTCCG CCCCATGGCT GACTAATTTT
 1921 TTTTATTTAT GCAGAGGCCG AGGCCGCCTC GGCCTCTGAG CTATTCCAGA AGTAGTGAGG
 1981 AGGCTTTTTT GGAGGCCTAG GCTTTTGCAA AGATCGATCA AGAGACAGGA TGAGGATCGT
 2041 TTCGCATGAT TGAACAAGAT GGATTGCACG CAGGTTCTCC GGCCGCTTGG GTGGAGAGGC
 2101 TATTCGGCTA TGACTGGGCA CAACAGACAA TCGGCTGCTC TGATGCCGCC GTGTTCCGGC
 2161 TGTCAGCGCA GGGGCGCCCG GTTCTTTTTG TCAAGACCGA CCTGTCCGGT GCCCTGAATG
 2221 AACTGCAAGA CGAGGCAGCG CGGCTATCGT GGCTGGCCAC GACGGGCGTT CCTTGCGCAG
 2281 CTGTGCTCGA CGTTGTCACT GAAGCGGGAA GGGACTGGCT GCTATTGGGC GAAGTGCCGG
 2341 GGCAGGATCT CCTGTCATCT CACCTTGCTC CTGCCGAGAA AGTATCCATC ATGGCTGATG
 2401 CAATGCGGCG GCTGCATACG CTTGATCCGG CTACCTGCCC ATTCGACCAC CAAGCGAAAC
 2461 ATCGCATCGA GCGAGCACGT ACTCGGATGG AAGCCGGTCT TGTCGATCAG GATGATCTGG
 2521 ACGAAGAGCA TCAGGGGCTC GCGCCAGCCG AACTGTTCGC CAGGCTCAAG GCGAGCATGC
 2581 CCGACGGCGA GGATCTCGTC GTGACCCATG GCGATGCCTG CTTGCCGAAT ATCATGGTGG
 2641 AAAATGGCCG CTTTTCTGGA TTCATCGACT GTGGCCGGCT GGGTGTGGCG GACCGCTATC
 2701 AGGACATAGC GTTGGCTACC CGTGATATTG CTGAAGAGCT TGGCGGCGAA TGGGCTGACC
 2761 GCTTCCTCGT GCTTTACGGT ATCGCCGCTC CCGATTCGCA GCGCATCGCC TTCTATCGCC
 2821 TTCTTGACGA GTTCTTCTGA GCGGGACTCT GGGGTTCGAA ATGACCGACC AAGCGACGCC
 2881 CAACCTGCCA TCACGAGATT TCGATTCCAC CGCCGCCTTC TATGAAAGGT TGGGCTTCGG
 2941 AATCGTTTTC CGGGACGCCG GCTGGATGAT CCTCCAGCGC GGGGATCTCA TGCTGGAGTT
 3001 CTTCGCCCAC CCTAGGGGGA GGCTAACTGA AACACGGAAG GAGACAATAC CGGAAGGAAC
 3061 CCGCGCTATG ACGGCAATAA AAAGACAGAA TAAAACGCAC GGTGTTGGGT CGTTTGTTCA
 3121 TAAACGCGGG GTTCGGTCCC AGGGCTGGCA CTCTGTCGAT ACCCCACCGA GACCCCATTG
 3181 GGGCCAATAC GCCCGCGTTT CTTCCTTTTC CCCACCCCAC CCCCCAAGTT CGGGTGAAGG
 3241 CCCAGGGCTC GCAGCCAACG TCGGGGCGGC AGGCCCTGCC ATAGCCTCAG GTTACTCATA
 3301 TATACTTTAG ATTGATTTAA AACTTCATTT TTAATTTAAA AGGATCTAGG TGAAGATCCT
 3361 TTTTGATAAT CTCATGACCA AAATCCCTTA ACGTGAGTTT TCGTTCCACT GAGCGTCAGA
 3421 CCCCGTAGAA AAGATCAAAG GATCTTCTTG AGATCCTTTT TTTCTGCGCG TAATCTGCTG
 3481 CTTGCAAACA AAAAAACCAC CGCTACCAGC GGTGGTTTGT TTGCCGGATC AAGAGCTACC
 3541 AACTCTTTTT CCGAAGGTAA CTGGCTTCAG CAGAGCGCAG ATACCAAATA CTGTCCTTCT
 3601 AGTGTAGCCG TAGTTAGGCC ACCACTTCAA GAACTCTGTA GCACCGCCTA CATACCTCGC
 3661 TCTGCTAATC CTGTTACCAG TGGCTGCTGC CAGTGGCGAT AAGTCGTGTC TTACCGGGTT
 3721 GGACTCAAGA CGATAGTTAC CGGATAAGGC GCAGCGGTCG GGCTGAACGG GGGGTTCGTG
 3781 CACACAGCCC AGCTTGGAGC GAACGACCTA CACCGAACTG AGATACCTAC AGCGTGAGCT
 3841 ATGAGAAAGC GCCACGCTTC CCGAAGGGAG AAAGGCGGAC AGGTATCCGG TAAGCGGCAG
 3901 GGTCGGAACA GGAGAGCGCA CGAGGGAGCT TCCAGGGGGA AACGCCTGGT ATCTTTATAG
 3961 TCCTGTCGGG TTTCGCCACC TCTGACTTGA GCGTCGATTT TTGTGATGCT CGTCAGGGGG
 4021 GCGGAGCCTA TGGAAAAACG CCAGCAACGC GGCCTTTTTA CGGTTCCTGG CCTTTTGCTG
 4081 GCCTTTTGCT CACATGTTCT TTCCTGCGTT ATCCCCTGAT TCTGTGGATA ACCGTATTAC
 4141 CGCCATGCAT
//

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