出品公司: | Clontech |
---|---|
载体名称: | pZsGreen1-1 |
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: | 4126 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ZsGreen1 |
载体抗性: | 卡那霉素 |
筛选标记: | 新霉素(Neomycin) |
克隆菌株: | DH5α, JM109 |
宿主细胞(系): | 真核细胞 |
备注: |
pZsGreen1-1载体不含启动子,用于研究目的启动子及顺式转录元件在细胞内的功能和水平; ZsGreen1是亮度最高的绿色荧光蛋白。 |
产品目录号: | 632473 |
稳定性: | 瞬表达 或 稳表达 |
组成型/诱导型: | -- |
病毒/非病毒: | 非病毒 |
买家导航
pZsGreen1-1载体质粒基本信息
pZsGreen1-1质粒图谱载体图谱和pZsGreen1-1载体序列质粒序列多克隆位点信息
pZsGreen1-1质粒载体简介
pZsGreen1-1载体
pZsGreen1-1 encodes a human codon-optimized variant of wild-type Zoanthus sp. green fluorescent
protein, ZsGreen1 (1). The ZsGreen1 coding sequence contains a series of silent base-pair changes,
which correspond to human codon-usage preferences, for optimal expression in mammalian cells
(2). A single amino acid substitution (Asn-66 to Met) has been made to enhance the emission
characteristics of ZsGreen1 (excitation maximum = 493 nm; emission maximum = 505 nm).
pZsGreen1-1 is a promoterless ZsGreen1 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 ZsGreen1 have been converted to a Kozak consensus translation
initiation site (3) to increase translation efficiency in eukaryotic cells. SV40 polyadenylation signals
downstream of the ZsGreen1 gene direct proper processing of the 3' end of the ZsGreen1 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 singlestranded
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.
载体使用:
ZsGreen1 can be used as an in vivo reporter of gene expression. Promoters should be cloned
into the pZsGreen1-1 MCS upstream from the ZsGreen1 coding sequence. Without addition of a
functional promoter, this vector will not express ZsGreen1. The recombinant ZsGreen1 vector can
be transfected into mammalian cells using any standard transfection method. If required, stable
transfectants can be selected using G418 (4).
载体特征:
MCS: 12–83
Zoanthus sp. human codon-optimized Green Fluorescent Protein (ZsGreen1) gene
Kozak consensus translation initiation site: 90–100
Start codon (ATG): 97–99; Stop codon: 790–792
Asn-66 to Met mutation (A→T, C→G ): 293, 294
SV40 early mRNA polyadenylation signal
Polyadenylation signals: 945–950 & 974–979
mRNA 3' ends: 983 & 995
f1 single-strand DNA origin: 1042–1497
(Packages noncoding strand of ZsGreen1.)
Ampicillin resistance (β-lactamase) promoter
–35 region: 1559–1564; –10 region: 1582–1587
Transcription start point: 1594
SV40 origin of replication: 1838–1973
SV40 early promoter
Enhancer (72-bp tandem repeats): 1669–1742 & 1743–1814
21-bp repeats: 1818–1837, 1839–1859 & 1861–1881
Early promoter element: 1894–1900
Major transcription start points: 1890, 1928, 1934 & 1939
Kanamycin/neomycin resistance gene
Neomycin phosphotransferase coding sequences:
Start codon (ATG): 2022–2024; stop codon: 2814–2816
G→A mutation to remove Pst I site: 2204
C→A (Arg→Ser) mutation to remove BssH II site: 2550
Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signal
Polyadenylation signals: 3052–3057 & 3065–3070
pUC plasmid replication origin: 3401–4044
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
Plasmid incompatibility group: pMB1/Col E1
Green Fluorescent Protein (ZsGreen1)
Excitation/Emission Maxima: 493 nm / 505 nm
pZsGreen1-1质粒序列载体序列
LOCUS pZsGreen1-1 4126 bp DNA SYN DEFINITION pZsGreen1-1 ACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors. FEATURES Location/Qualifiers source 1..4126 /organism="pZsGreen1-1" /mol_type="other DNA" CDS 97..792 /label="ORF frame 1" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH LTEHAIASGSALP*" gene 100..789 /label="Zs_Green" /gene="Zs_Green" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH LTEHAIASGSALP*" misc_feature 1005..1024 /label="EBV_rev_primer" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH LTEHAIASGSALP*" rep_origin complement(1174..1480) /label="f1_origin" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH LTEHAIASGSALP*" promoter 1559..1587 /label="AmpR_promoter" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH LTEHAIASGSALP*" misc_feature complement(1653..1673) /label="pBABE_3_primer" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH LTEHAIASGSALP*" misc_feature complement(1659..1874) /label="SV40_enhancer" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH LTEHAIASGSALP*" promoter 1671..1939 /label="SV40_promoter" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH LTEHAIASGSALP*" rep_origin 1838..1915 /label="SV40_origin" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH LTEHAIASGSALP*" misc_feature 1900..1919 /label="SV40pro_F_primer" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH LTEHAIASGSALP*" CDS 2022..2816 /label="ORF frame 3" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" gene 2025..2813 /label="NeoR/KanR" /gene="NeoR/KanR" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" CDS complement(2331..2867) /label="ORF frame 3" /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ PPHCISHDGYFLGRSKVR*" terminator 2991..3260 /label="TK_PA_terminator" /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ PPHCISHDGYFLGRSKVR*" rep_origin 3408..4027 /label="pBR322_origin" /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ PPHCISHDGYFLGRSKVR*" ORIGIN 1 TAGTTATTAC TAGCGCTACC GGACTCAGAT CTCGAGCTCA AGCTTCGAAT TCTGCAGTCG 61 ACGGTACCGC GGGCCCGGGA TCCACCGGTC GCCACCATGG CCCAGTCCAA GCACGGCCTG 121 ACCAAGGAGA TGACCATGAA GTACCGCATG GAGGGCTGCG TGGACGGCCA CAAGTTCGTG 181 ATCACCGGCG AGGGCATCGG CTACCCCTTC AAGGGCAAGC AGGCCATCAA CCTGTGCGTG 241 GTGGAGGGCG GCCCCTTGCC CTTCGCCGAG GACATCTTGT CCGCCGCCTT CATGTACGGC 301 AACCGCGTGT TCACCGAGTA CCCCCAGGAC ATCGTCGACT ACTTCAAGAA CTCCTGCCCC 361 GCCGGCTACA CCTGGGACCG CTCCTTCCTG TTCGAGGACG GCGCCGTGTG CATCTGCAAC 421 GCCGACATCA CCGTGAGCGT GGAGGAGAAC TGCATGTACC ACGAGTCCAA GTTCTACGGC 481 GTGAACTTCC CCGCCGACGG CCCCGTGATG AAGAAGATGA CCGACAACTG GGAGCCCTCC 541 TGCGAGAAGA TCATCCCCGT GCCCAAGCAG GGCATCTTGA AGGGCGACGT GAGCATGTAC 601 CTGCTGCTGA AGGACGGTGG CCGCTTGCGC TGCCAGTTCG ACACCGTGTA CAAGGCCAAG 661 TCCGTGCCCC GCAAGATGCC CGACTGGCAC TTCATCCAGC ACAAGCTGAC CCGCGAGGAC 721 CGCAGCGACG CCAAGAACCA GAAGTGGCAC CTGACCGAGC ACGCCATCGC CTCCGGCTCC 781 GCCTTGCCCT GAGCGGCCGC GACTCTAGAT CATAATCAGC CATACCACAT TTGTAGAGGT 841 TTTACTTGCT TTAAAAAACC TCCCACACCT CCCCCTGAAC CTGAAACATA AAATGAATGC 901 AATTGTTGTT GTTAACTTGT TTATTGCAGC TTATAATGGT TACAAATAAA GCAATAGCAT 961 CACAAATTTC ACAAATAAAG CATTTTTTTC ACTGCATTCT AGTTGTGGTT TGTCCAAACT 1021 CATCAATGTA TCTTAAGGCG TAAATTGTAA GCGTTAATAT TTTGTTAAAA TTCGCGTTAA 1081 ATTTTTGTTA AATCAGCTCA TTTTTTAACC AATAGGCCGA AATCGGCAAA ATCCCTTATA 1141 AATCAAAAGA ATAGACCGAG ATAGGGTTGA GTGTTGTTCC AGTTTGGAAC AAGAGTCCAC 1201 TATTAAAGAA CGTGGACTCC AACGTCAAAG GGCGAAAAAC CGTCTATCAG GGCGATGGCC 1261 CACTACGTGA ACCATCACCC TAATCAAGTT TTTTGGGGTC GAGGTGCCGT AAAGCACTAA 1321 ATCGGAACCC TAAAGGGAGC CCCCGATTTA GAGCTTGACG GGGAAAGCCG GCGAACGTGG 1381 CGAGAAAGGA AGGGAAGAAA GCGAAAGGAG CGGGCGCTAG GGCGCTGGCA AGTGTAGCGG 1441 TCACGCTGCG CGTAACCACC ACACCCGCCG CGCTTAATGC GCCGCTACAG GGCGCGTCAG 1501 GTGGCACTTT TCGGGGAAAT GTGCGCGGAA CCCCTATTTG TTTATTTTTC TAAATACATT 1561 CAAATATGTA TCCGCTCATG AGACAATAAC CCTGATAAAT GCTTCAATAA TATTGAAAAA 1621 GGAAGAGTCC TGAGGCGGAA AGAACCAGCT GTGGAATGTG TGTCAGTTAG GGTGTGGAAA 1681 GTCCCCAGGC TCCCCAGCAG GCAGAAGTAT GCAAAGCATG CATCTCAATT AGTCAGCAAC 1741 CAGGTGTGGA AAGTCCCCAG GCTCCCCAGC AGGCAGAAGT ATGCAAAGCA TGCATCTCAA 1801 TTAGTCAGCA ACCATAGTCC CGCCCCTAAC TCCGCCCATC CCGCCCCTAA CTCCGCCCAG 1861 TTCCGCCCAT TCTCCGCCCC ATGGCTGACT AATTTTTTTT ATTTATGCAG AGGCCGAGGC 1921 CGCCTCGGCC TCTGAGCTAT TCCAGAAGTA GTGAGGAGGC TTTTTTGGAG GCCTAGGCTT 1981 TTGCAAAGAT CGATCAAGAG ACAGGATGAG GATCGTTTCG CATGATTGAA CAAGATGGAT 2041 TGCACGCAGG TTCTCCGGCC GCTTGGGTGG AGAGGCTATT CGGCTATGAC TGGGCACAAC 2101 AGACAATCGG CTGCTCTGAT GCCGCCGTGT TCCGGCTGTC AGCGCAGGGG CGCCCGGTTC 2161 TTTTTGTCAA GACCGACCTG TCCGGTGCCC TGAATGAACT GCAAGACGAG GCAGCGCGGC 2221 TATCGTGGCT GGCCACGACG GGCGTTCCTT GCGCAGCTGT GCTCGACGTT GTCACTGAAG 2281 CGGGAAGGGA CTGGCTGCTA TTGGGCGAAG TGCCGGGGCA GGATCTCCTG TCATCTCACC 2341 TTGCTCCTGC CGAGAAAGTA TCCATCATGG CTGATGCAAT GCGGCGGCTG CATACGCTTG 2401 ATCCGGCTAC CTGCCCATTC GACCACCAAG CGAAACATCG CATCGAGCGA GCACGTACTC 2461 GGATGGAAGC CGGTCTTGTC GATCAGGATG ATCTGGACGA AGAGCATCAG GGGCTCGCGC 2521 CAGCCGAACT GTTCGCCAGG CTCAAGGCGA GCATGCCCGA CGGCGAGGAT CTCGTCGTGA 2581 CCCATGGCGA TGCCTGCTTG CCGAATATCA TGGTGGAAAA TGGCCGCTTT TCTGGATTCA 2641 TCGACTGTGG CCGGCTGGGT GTGGCGGACC GCTATCAGGA CATAGCGTTG GCTACCCGTG 2701 ATATTGCTGA AGAGCTTGGC GGCGAATGGG CTGACCGCTT CCTCGTGCTT TACGGTATCG 2761 CCGCTCCCGA TTCGCAGCGC ATCGCCTTCT ATCGCCTTCT TGACGAGTTC TTCTGAGCGG 2821 GACTCTGGGG TTCGAAATGA CCGACCAAGC GACGCCCAAC CTGCCATCAC GAGATTTCGA 2881 TTCCACCGCC GCCTTCTATG AAAGGTTGGG CTTCGGAATC GTTTTCCGGG ACGCCGGCTG 2941 GATGATCCTC CAGCGCGGGG ATCTCATGCT GGAGTTCTTC GCCCACCCTA GGGGGAGGCT 3001 AACTGAAACA CGGAAGGAGA CAATACCGGA AGGAACCCGC GCTATGACGG CAATAAAAAG 3061 ACAGAATAAA ACGCACGGTG TTGGGTCGTT TGTTCATAAA CGCGGGGTTC GGTCCCAGGG 3121 CTGGCACTCT GTCGATACCC CACCGAGACC CCATTGGGGC CAATACGCCC GCGTTTCTTC 3181 CTTTTCCCCA CCCCACCCCC CAAGTTCGGG TGAAGGCCCA GGGCTCGCAG CCAACGTCGG 3241 GGCGGCAGGC CCTGCCATAG CCTCAGGTTA CTCATATATA CTTTAGATTG ATTTAAAACT 3301 TCATTTTTAA TTTAAAAGGA TCTAGGTGAA GATCCTTTTT GATAATCTCA TGACCAAAAT 3361 CCCTTAACGT GAGTTTTCGT TCCACTGAGC GTCAGACCCC GTAGAAAAGA TCAAAGGATC 3421 TTCTTGAGAT CCTTTTTTTC TGCGCGTAAT CTGCTGCTTG CAAACAAAAA AACCACCGCT 3481 ACCAGCGGTG GTTTGTTTGC CGGATCAAGA GCTACCAACT CTTTTTCCGA AGGTAACTGG 3541 CTTCAGCAGA GCGCAGATAC CAAATACTGT TCTTCTAGTG TAGCCGTAGT TAGGCCACCA 3601 CTTCAAGAAC TCTGTAGCAC CGCCTACATA CCTCGCTCTG CTAATCCTGT TACCAGTGGC 3661 TGCTGCCAGT GGCGATAAGT CGTGTCTTAC CGGGTTGGAC TCAAGACGAT AGTTACCGGA 3721 TAAGGCGCAG CGGTCGGGCT GAACGGGGGG TTCGTGCACA CAGCCCAGCT TGGAGCGAAC 3781 GACCTACACC GAACTGAGAT ACCTACAGCG TGAGCTATGA GAAAGCGCCA CGCTTCCCGA 3841 AGGGAGAAAG GCGGACAGGT ATCCGGTAAG CGGCAGGGTC GGAACAGGAG AGCGCACGAG 3901 GGAGCTTCCA GGGGGAAACG CCTGGTATCT TTATAGTCCT GTCGGGTTTC GCCACCTCTG 3961 ACTTGAGCGT CGATTTTTGT GATGCTCGTC AGGGGGGCGG AGCCTATGGA AAAACGCCAG 4021 CAACGCGGCC TTTTTACGGT TCCTGGCCTT TTGCTGGCCT TTTGCTCACA TGTTCTTTCC 4081 TGCGTTATCC CCTGATTCTG TGGATAACCG TATTACCGCC ATGCAT //
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pZsGreen1-1质粒载体应用举例
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