| 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
载体基本信息
载体图谱质粒图谱和多克隆位点信息
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。
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
//
