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首页    载体及质粒    pZsGreen1-1
pZsGreen1-1vectormap

pZsGreen1-1

pZsGreen1-1

pZsGreen1-1

 



编号

载体名称

北京华越洋VECT75376

pZsGreen1-1

 

pZsGreen1-1载体基本信息:

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:

Neomycin_1.html'   target='_blank'>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。

稳定性:

瞬表达 稳表达

组成型/诱导型:

--

病毒/非病毒:

非病毒

 

pZsGreen1-1载体质粒图谱和多克隆位点信息

pZsGreen1-1载体简介:

 

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.

 

 

pZsGreen1-1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).

 

 

pZsGreen1-1载体特征:

 

 MCS: 1283

 Zoanthus sp. human codon-optimized Green Fluorescent Protein (ZsGreen1) gene

Kozak consensus translation initiation site: 90100

Start codon (ATG): 9799; Stop codon: 790792

Asn-66 to Met mutation (AT, CG ): 293, 294

 SV40 early mRNA polyadenylation signal

Polyadenylation signals: 945950 & 974979

mRNA 3' ends: 983 & 995

 f1 single-strand DNA origin: 10421497

(Packages noncoding strand of ZsGreen1.)

 Ampicillin resistance (β-lactamase) promoter

35 region: 15591564; 10 region: 15821587

Transcription start point: 1594

 SV40 origin of replication: 18381973

 SV40 early promoter

Enhancer (72-bp tandem repeats): 16691742 & 17431814

21-bp repeats: 18181837, 18391859 & 18611881

Early promoter element: 18941900

Major transcription start points: 1890, 1928, 1934 & 1939

 Kanamycin/neomycin resistance geneNeomycin phosphotransferase coding sequences:

Start codon (ATG): 20222024; stop codon: 28142816

GA mutation to remove Pst I site: 2204

CA (ArgSer) mutation to remove BssH II site: 2550

 Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signal

Polyadenylation signals: 30523057 & 30653070

 pUC plasmid replication origin: 34014044

 

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载体序列:

 

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
//


 

 

pZsGreen1-1载体其他相关的其他类型载体:


TOPFlash

pISRE-TA-Luc

TEV protease

pIL253

SuperTopFlash

pIJ8860

pZsGreen1-DR

pHY300PLK

pZsGreen1-1

pHY300

pZL507

pHT304

pYCT

pHT43

pXMJ19

pHT01

pVLT33

pHSE-Luc

pUG6

pHIS1525

pTimer-1

pHGF9050

pTimer

pHD_v2

pTA-Luc

pHcRed1-DR

pTALETF_v2 (NN)

pHcRed1-1

pTALETF_v2 (NI)

pHCMC05

pTALETF_v2 (NG)

pHCMC04

pTALETF_v2 (HD)

pGRE-luc

pTALEN_v2 (NN)

pGFP315

pTALEN_v2 (NI)

pGFP22

pTALEN_v2 (NG)

pGEM-3

pTALEN_v2 (HD)

pGAS-TA-Luc

pSRE-Luc

pE2F-TA-Luc

pSH65

pDsRed-Express-DR

pSH63

pDsRed-Express2-1

pSH47

pDsRed-Express-1

pSG1154

pDsRed2-1

pset152

pDG1363

pRK2013

pCSN44

pQE-TriSystem

pDD-tdTomato Reporter

pOS7001

pCRE-Luc

pNZ8048

pCR-Blunt

pNN_v2

pCMV-Raf-1

pNI v2

pCMV-p53

pNG_v2

pCMV-IκBα

pMUTIN4

pCMV-CREB

pMG36e

pCaspase3-sensor

pMetLuc2-Reporter

pCantab 5E

pMetLuc2-Control

pBT2

pmCherry-1

pBR322

pMA09-J

pBIG2R-HPH2

pMA09-H

pAP-1-Luc

pMA5MCS

pAN8-1

pKOR1

pAD43-25

pkc1139

pAD43-25

pK18mobSacB

pAcGFP1-1

pJLA502

HRV 3C protease

pIκB-EGFP

GG499

pAN7-1

p53-Luc

pAP1(PMA)-TA-Luc