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 |
买家导航
pCMV-Gaussia Luc载体质粒基本信息
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)
• 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 //