| 出品公司: | -- |
|---|---|
| 载体名称: | pAAV-CAG-Fluc 、pAAV-CAG-luc |
| 质粒类型: | 腺病毒载体；荧光素酶报告载体；荧光蛋白报告载体 |
| 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: | CAG (CMV enhancer/chicken β-actin promoter) |
| 载体大小: | 8323 bp |
| 5' 测序引物及序列: | -- |
| 3' 测序引物及序列: | -- |
| 载体标签: | -- |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | 萤火虫荧光素酶、橙色荧光蛋白 |
| 克隆菌株: | Stbl3 或 NEB Stble |
| 宿主细胞（系）: | 常规哺乳动物细胞系 |
| 备注: | -- |
| 产品目录号: | -- |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 腺病毒 |
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
This AAV is a fLuc construct driven by the ubiquitous CAG promoter. The roughly 1.8Kb CAG promoter (also known as CBA promoter or CAGGS promoter) is a strong synthetic promoter frequently used to drive high levels of gene expression in mammalian cells. The CAG promoter is composed of the following regulatory elements: (C) cytomegalovirus (CMV) early enhancer element; (A) the promoter region, the first exon, and the first intron of chicken beta-Actin gene, and (G) the splice acceptor of the rabbit beta-Globin gene. Like the EF1a promoter, the CAG promoter is commonly used as an alternative for the CMV promoter, from which the expression is decreased due to methylation/silencing. In many cell types tested, the CAG and EF1a promoters give much higher levels of expression than other commonly used cellular promoters such as the UBC and PGK promoters. Firefly luciferase (fLuc) is an oxidative enzyme used in bioluminescence and is distinct from a photoprotein. In the luciferase reaction, light is emitted when luciferase acts on the appropriate luciferin substrate. Photon emission can be detected by a light-sensitive apparatus such as a luminometer or modified optical microscope. Firefly luciferase is commonly used as a reporter to assess the transcriptional activity in cells.
载体序列
LOCUS KC152483 8323 bp DNA circular SYN 03-JUN-2013
DEFINITION Expression vector pAAV-CAG-Fluc, complete sequence.
ACCESSION KC152483
REFERENCE 1 (bases 1 to 8323)
AUTHORS Chen,Y., Cao,L., Luo,C., Ditzel,D.A., Peter,J. and Sprengel,R.
TITLE RANGE: Gene Transfer of Reversibly Controlled Polycistronic Genes
JOURNAL Mol Ther Nucleic Acids 2, E85 (2013)
PUBMED 23571608
JOURNAL Submitted (08-NOV-2012) Molecular Neurobiology, Max Planck
institute for medical research, Jahnstr. 29, Heidelberg, BW 69120,
Germany
FEATURES Location/Qualifiers
source 1..8323
/organism="Expression vector pAAV-CAG-Fluc"
/mol_type="other DNA"
/db_xref="taxon:1337866"
repeat_region 19..162
/note="AAV2 ITR"
/rpt_type=inverted
/rpt_type=terminal
regulatory 200..1140
/regulatory_class="promoter"
/note="CMV earlyenhancer/chicken beta actin promoter: CAG"
misc_feature 1157..2809
/note="firefly luciferase"
misc_feature 2810..2863
/note="2A peptide from Thosea asigna virus"
misc_feature 2876..3724
/note="tTSKid"
misc_feature 2876..3508
/note="Tet repressor: tetR[B/E]"
misc_feature 3509..3526
/note="nuclear localization signal: NLS"
misc_feature 3533..3718
/note="KRAB (Kruppel-associated box) domain; from Rat zinc
finger protein (kid-1)"
misc_feature 3725..3778
/note="2A peptide from Thosea asigna virus"
gene 3791..4447
/gene="mKO"
CDS 3791..4447
/gene="mKO"
/codon_start=1
/transl_table=11
/product="monomeric Kusabira-orange"
/protein_id="AGM53878.1"
/db_xref="GI:507593976"
/translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLR
VTMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGG
SASVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDV
TMYLKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS
"
ncRNA 4478..5074
/ncRNA_class="other"
/note="post-transcriptional regulatory element: WPRE"
misc_feature 5107..5317
/note="bovine growth hormone polyA signal: bGH pA"
repeat_region complement(5327..5470)
/note="AAV2 ITR"
/rpt_type=inverted
/rpt_type=terminal
gene 6659..7321
/gene="Bla"
CDS 6659..7321
/gene="Bla"
/note="ampicillin resistance"
/codon_start=1
/transl_table=11
/product="beta-lactamase"
/protein_id="AGM53877.1"
/db_xref="GI:507593975"
/translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKH
LTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELN
EAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAG
WFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIK
HW"
ORIGIN
1 gggggggggg ggggggggtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
61 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
121 gcgcgcagag agggagtggc caactccatc actaggggtt cctagatcta ggatcacgcg
181 ttctagtcct agggatgtac cattgacgtc aataatgacg tatgttccca tagtaacgcc
241 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
301 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
361 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
421 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
481 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
541 tgcagcgatg ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga
601 ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
661 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
721 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
781 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
841 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
901 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcggggct
961 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
1021 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
1081 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattggatc
1141 tagtcgattc gaattcatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta
1201 tccgctagag gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct
1261 ggttcctgga acaattgctt ttacagatgc acatatcgag gtgaacatca cgtacgcgga
1321 atacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa
1381 tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc
1441 gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct
1501 caacagtatg aacatttcgc agcctaccgt agtgtttgtt tccaaaaagg ggttgcaaaa
1561 aattttgaac gtgcaaaaaa aattaccaat aatccagaaa attattatca tggattctaa
1621 aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg
1681 ttttaatgaa tacgattttg taccagagtc ctttgatcgt gacaaaacaa ttgcactgat
1741 aatgaactcc tctggatcta ctgggttacc taagggtgtg gcccttccgc atagaactgc
1801 ctgcgtcaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac
1861 tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta cactcggata
1921 tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttttacg
1981 atcccttcag gattacaaaa ttcaaagtgc gttgctagta ccaaccctat tttcattctt
2041 cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg
2101 gggcgcacct ctttcgaaag aagtcgggga agcggttgca aaacgcttcc atcttccagg
2161 gatacgacaa ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg
2221 ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga
2281 tctggatacc gggaaaacgc tgggcgttaa tcagagaggc gaattatgtg tcagaggacc
2341 tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga
2401 tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact tcttcatagt
2461 tgaccgcttg aagtctttaa ttaaatacaa aggataccag gtggcccccg ctgaattgga
2521 gtcgatattg ttacaacacc ccaacatctt cgacgcgggc gtggcaggtc ttcccgacga
2581 tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga
2641 aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg
2701 agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat
2761 cagagagatc ctcataaagg ccaagaaggg cggaaagtcc aaattgctcg agggcagagg
2821 aagtcttcta acatgcggtg acgtggagga gaatcccggc cctgcaccgg gatccatgtc
2881 cagattagat aaaagtaaag tgattaacag cgcattagag ctgcttaatg aggtcggaat
2941 cgaaggttta acaacccgta aactcgccca gaagctaggt gtagagcagc ctacattgta
3001 ttggcacgtg cgcaacaagc agactcttat gaacatgctt tcagaggcaa tactggcgaa
3061 gcatcacacc cgttcagcac cgttaccgac tgagagttgg cagcagtttc tccaggaaaa
3121 tgctctgagt ttccgtaaag cattactggt ccatcgtgat ggagcccgat tgcatatagg
3181 gacctctcct acgccccccc agtttgaaca agcagaggcg caactacgct gtctatgcga
3241 tgcagggttt tcggtcgagg aggctctttt cattctgcaa tctatcagcc attttacgtt
3301 gggtgcagta ttagaggagc aagcaacaaa ccagatagaa aataatcatg tgatagacgc
3361 tgcaccacca ttattacaag aggcatttaa tattcaggcg agaacctctg ctgaaatggc
3421 cttccatttc gggctgaaat cattaatatt tggattttct gcacagttag atgaaaaaaa
3481 gcatacaccc attgaggatg gtaataaacc aaaaaagaag agaaagctag cagtgtcagt
3541 gacatttgaa gatgtggctg tgctctttac tcgggacgag tggaagaagc tggatctgtc
3601 tcagagaagc ctgtaccgtg aggtgatgct ggagaattac agcaacctgg cctccatggc
3661 aggattcctg tttaccaaac caaaggtgat ctccctgttg cagcaaggag aggacccgta
3721 tctcgagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctgc
3781 accgggatcc atggtgagtg tgattaaacc agaaatgaag atgaggtact acatggacgg
3841 ctccgtcaat gggcatgagt tcacaattga aggtgaaggc acaggcagac cttacgaggg
3901 acatcaagag atgacactac gcgtcacaat ggccaagggc gggccaatgc ctttcgcgtt
3961 tgacttagtg tcacacgtgt tctgttacgg ccacagacct tttactaaat atccagaaga
4021 gataccagac tatttcaaac aagcatttcc tgaaggcctg tcatgggaaa ggtcgttgga
4081 gttcgaagat ggtgggtccg cttcagtcag tgcgcatata agccttagag gaaacacctt
4141 ctaccacaaa tccaaattta ctggggttaa ctttcctgcc gatggtccta tcatgcaaaa
4201 ccaaagtgtt gattgggagc catcaaccga gaaaattact gccagcgacg gagttctgaa
4261 gggtgatgtt acgatgtacc taaaacttga aggaggcggc aatcacaaat gccaattcaa
4321 gactacttac aaggcggcaa aaaagattct taaaatgcca ggaagccatt acatcagcca
4381 tcgcctcgtc aggaaaaccg aaggcaacat tactgagctg gtagaagatg cagtagctca
4441 ttcctgatgt acaagtaaag cggccgcaag cttatcgata atcaacctct ggattacaaa
4501 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
4561 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
4621 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
4681 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
4741 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
4801 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
4861 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
4921 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
4981 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
5041 cggatctccc tttgggccgc ctccccgcct gatcgatacc gtcgactaga gctcgctgat
5101 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
5161 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
5221 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
5281 gggaggattg ggaagacaat agcaggcatg ctggggagag atctgaggaa cccctagtga
5341 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
5401 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
5461 gagtggccaa cccccccccc cccccccctg cagcctggcg taatagcgaa gaggcccgca
5521 ccgatcgccc ttcccaacag ttgcgtagcc tgaatggcga atggcgcgac gcgccctgta
5581 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
5641 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
5701 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
5761 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
5821 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
5881 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
5941 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
6001 acaaaatatt aacgtttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
6061 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
6121 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
6181 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
6241 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
6301 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
6361 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
6421 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
6481 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
6541 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
6601 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
6661 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
6721 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
6781 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
6841 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
6901 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
6961 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
7021 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
7081 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
7141 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
7201 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
7261 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
7321 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
7381 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
7441 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
7501 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
7561 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
7621 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
7681 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
7741 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
7801 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
7861 actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc
7921 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
7981 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
8041 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
8101 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
8161 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
8221 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
8281 gcctctcccc gcgcgttggc cgattcatta atgcaggctg cag
//
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