pCMV-Gaussia Luc

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pCMV-Gaussia Luc载体质粒基本信息

出品公司: Thermofisher
载体名称: pCMV-Gaussia Luc
载体又名: pCMV-Gaussia Luc载体,pCMV-Gaussia Luc质粒,pCMV-Gaussia Luc载体质粒
英文名称: pCMV-Gaussia Luc, pCMV-Gaussia Luc Vector, pCMV-Gaussia Luc Plasmid
质粒类型: 哺乳动物细胞表达载体
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: CMV
载体大小: 5483 bp
5’测序引物及序列: CMV-F: CGCAAATGGGCGGTAGGCGTG
3’测序引物及序列: pBABE 3': ACCCTAACTGACACACATTCC
载体标签: C-Gaussia Luc
载体抗性: 氨苄青霉素
筛选标记: 新霉素Neomycin或者G418
克隆菌株: DH5α,TOP10 等
宿主细胞: 常规细胞系(293细、CHO等)
备注: 用于荧光素酶检测。
产品目录号: 16147
稳定性: 瞬时表达
组成型/诱导型: 组成型
病毒/非病毒: 非病毒
相关文件:  pCMV-Gaussia Luc载体图谱
pCMV-Gaussia Luc载体序列
pCMV-Gaussia Luc载体说明书

pCMV-Gaussia Luc质粒图谱载体图谱和pCMV-Gaussia Luc载体序列质粒序列多克隆位点信息

pCMV-Gaussia Luc载体图谱

pCMV-Gaussia Luc质粒载体简介

pCMV-Gaussia Luc载体,含有受 CMV(巨细胞病毒)启动子控制的 Gaussia 荧光素酶基因。此组成型表达载体可用作归一化对照品,以结合其他报告基因解释实验变化。
 
pCMV-Gaussia Luc载体含有哺乳动物(人)密码子优化的Gaussia荧光素酶,表达N-末端分泌信号。该产品可用作阳性对照载体,在使用Gaussia荧光素酶作为报告载体,评估基因表达效率方面非常有用。该载体同时具有氨苄西林抗性和新霉素抗性。因此,它很容易在大肠杆菌中繁殖,并可用于建立稳定的表达Gaussia荧光素酶的细胞系。
 
另外,有研究使用Gaussia荧光素酶与CRISPR一起用作报告质粒,以研究非编码RNA(ncRNA)的功能:Multiplexable, locus-specific targeting of long RNAs with CRISPR-Display.  Shechner DM, Hacisuleyman E, Younger ST, Rinn JL. July 2015. PubMed
 
pCMV-Gaussia Luc 载体的特点:
 
•用于共转染和归一化的强 (CMV) 组成型启动子
•天然分泌Gaussia 荧光素酶基因,经优化可实现哺乳动物系统中的高表达
•多克隆位点 (MCS) 为调控元件从一个质粒至另一个质粒的转移提供了多功能性
•用于通过降低转录的通读来最小化背景的转录终止位点 (Ter)、Lac 操纵基因 (Lac O1) 和转录中断位点 (TPS)
嘌呤霉素 (Pur) 和氨苄青霉素 (Amp) 标志物分别用于哺乳动物和细菌细胞中的药物选择
• 高拷贝 pUC 细菌 DNA 复制起点
 
Gaussia Luc 载体含有从桡足动物 Gaussia princeps 中克隆的基因。该基因可编码一个自然分泌的生物发光 Gaussia 荧光素酶(约 20 kDa),可测量培养基(实时检测)和细胞裂解物中的报告基因活性。
 
Gaussia荧光素酶(GLuc)是一种由“人源化”序列编码的19kDa蛋白质,它在N端含有天然信号肽,可使其从哺乳动物细胞分泌到细胞培养基中。在SV40启动子控制下的新霉素抗性基因允许使用G418选择质粒稳定整合到哺乳动物细胞基因组中。

pCMV-Gaussia Luc质粒序列载体序列

LOCUS       pCMV-Gaussia Luc        5483 bp ds-DNA     circular SYN 19-OCT-2017
DEFINITION  Control vector for constitutive high-level expression of secreted 
            Gaussia luciferase under control of the CMV promoter.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5483)
  AUTHORS   Thermo Fisher (Pierce)
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 13, 2023 from SnapGene 4.3.6 to Vector NTI(R) 
            format
            https://www.biofeng.com
COMMENT     VNTDATE|666144000|
COMMENT     VNTDBDATE|820022400|
COMMENT     VNTNAME|pCMV-Gaussia Luc|
COMMENT     VNTAUTHORNAME|Thermo Fisher (Pierce)|
FEATURES             Location/Qualifiers
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        615..633
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             647..1204
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
     polyA_signal    1227..1338
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1461..1790
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1641..1776
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        1838..1885
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1904..2503
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
     polyA_signal    2633..2754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2807..3667)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     rep_origin      3931..4519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5261..5277
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     misc_feature    5392..5483
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
ORIGIN
        1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
      601 tggcttatcg aaattaatac gactcactat aggggatcca gccaccatgg gagtcaaagt
      661 tctgtttgcc ctgatctgca tcgctgtggc cgaggccaag cccaccgaga acaacgaaga
      721 cttcaacatc gtggccgtgg ccagcaactt cgcgaccacg gatctcgatg ctgaccgcgg
      781 gaagttgccc ggcaagaagc tgccgctgga ggtgctcaaa gagatggaag ccaatgcccg
      841 gaaagctggc tgcaccaggg gctgtctgat ctgcctgtcc cacatcaagt gcacgcccaa
      901 gatgaagaag ttcatcccag gacgctgcca cacctacgaa ggcgacaaag agtccgcaca
      961 gggcggcata ggcgaggcga tcgtcgacat tcctgagatt cctgggttca aggacttgga
     1021 gcccatggag cagttcatcg cacaggtcga tctgtgtgtg gactgcacaa ctggctgcct
     1081 caaagggctt gccaacgtgc agtgttctga cctgctcaag aagtggctgc cgcaacgctg
     1141 tgcgaccttt gccagcaaga tccagggcca ggtggacaag atcaaggggg ccggtggtga
     1201 ctaagcggcc gcaaaatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1261 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1321 aaatgaggaa attgcatcac aacactcaac cctatctcgg tctattcttt tgatttataa
     1381 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1441 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     1501 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     1561 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     1621 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1681 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     1741 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     1801 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     1861 gtatatcggc atagtataat acgacaaggt gaggaactaa accatgaccg agtacaagcc
     1921 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     1981 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     2041 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     2101 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     2161 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     2221 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     2281 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     2341 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacat ccgcgccccg
     2401 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     2461 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacacgtgc tacgagattt
     2521 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     2581 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     2641 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2701 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     2761 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcattac caatgcttaa
     2821 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     2881 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagc gctgcgatga
     2941 taccgcgaga accacgctca ccggctccgg atttatcagc aataaaccag ccagccggaa
     3001 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3061 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccatcg
     3121 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3181 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3241 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3301 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3361 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3421 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3481 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3541 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3601 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3661 tactcatatt cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3721 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggtcagt gttacaacca
     3781 attaaccaat tctgaacatt atcgcgagcc catttatacc tgaatatggc tcataacacc
     3841 ccttgctcat gaccaaaatc ccttaacgtg agttacgcgc gcgtcgttcc actgagcgtc
     3901 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3961 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4021 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4081 tctagtgtag ccgtagttag cccaccactt caagaactct gtagcaccgc ctacatacct
     4141 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4201 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4261 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4321 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4381 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4441 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4501 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4561 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4621 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4681 agtgagcgag gaagcggaag gcgagagtag ggaactgcca ggcatcaaac taagcagaag
     4741 gcccctgacg gatggccttt ttgcgtttct acaaactctt tctgtgttgt aaaacgacgg
     4801 ccagtcttaa gctcgggccc cctgggcggt tctgataacg agtaatcgtt aatccgcaaa
     4861 taacgtaaaa acccgcttcg gcgggttttt ttatgggggg agtttaggga aagagcattt
     4921 gtcagaatat ttaagggcgc ctgtcacttt gcttgatata tgagaattat ttaaccttat
     4981 aaatgagaaa aaagcaacgc actttaaata agatacgttg ctttttcgat tgatgaacac
     5041 ctataattaa actattcatc tattatttat gattttttgt atatacaata tttctagttt
     5101 gttaaagaga attaagaaaa taaatctcga aaataataaa gggaaaatca gtttttgata
     5161 tcaaaattat acatgtcaac gataatacaa aatataatac aaactataag atgttatcag
     5221 tatttattat catttagaat aaattttgtg tcgcccttaa ttgtgagcgg ataacaatta
     5281 cgagcttcat gcacagtggc gttgacattg attattgact agcatgttct ttcctgcgtt
     5341 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa taacatacgc
     5401 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     5461 tgcaggtgcc agaacatttc tct
//

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