| 质粒名称: | pAAV-CAG-Fluc |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 83281 |
| 存储实验室: | Mark Kay |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV with CAG promoter |
| 空载体大小: | 4721 bp |
| 完整质粒大小: | 6374 bp |
| 载体类型: | 哺乳细胞表达，腺病毒，AAV |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Firefly Luciferase |
| 物种来源: | Photinus pyralis (Firefly) |
| 插入大小: | 1653 bp |
| 启动子: | CAG |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamH1 (not destroyed) |
| 3’克隆位点: | EcoR1 (not destroyed) |
| 5’测序引物 | GTTATTGTGCTGTCTCATC |
| 3’测序引物 | CATTAAAGCAGCGTATCC |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-CAG-FLuc was a gift from Mark Kay (Addgene plasmid # 83281 ; http://n2t.net/addgene:83281 ; RRID:Addgene_83281) |
| 参考文献部分: | Using a barcoded AAV capsid library to select for clinically relevant gene therapy vectors. Pekrun K, De Alencastro G, Luo QJ, Liu J, Kim Y, Nygaard S, Galivo F, Zhang F, Song R, Tiffany MR, Xu J, Hebrok M, Grompe M, Kay MA. JCI Insight. 2019 Nov 14;4(22). pii: 131610. doi: 10.1172/jci.insight.131610. 10.1172/jci.insight.131610 PubMed 31723052 |
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
载体序列
LOCUS pAAV-CAG-FLuc 6374 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION recombinant AAV vector packaging single-stranded Firefly Luciferase
under the CAG promoter (includes WPRE element).
ACCESSION .
VERSION .
KEYWORDS pAAV-CAG-FLuc
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6374)
AUTHORS Pekrun K, De Alencastro G, Luo QJ, Liu J, Kim Y, Nygaard S, Galivo
F, Zhang F, Song R, Tiffany MR, Xu J, Hebrok M, Grompe M, Kay MA
TITLE Using a barcoded AAV capsid library to select for clinically
relevant gene therapy vectors.
JOURNAL JCI Insight. 2019 Nov 14;4(22). pii: 131610. doi:
10.1172/jci.insight.131610.
PUBMED 31723052
REFERENCE 2 (bases 1 to 6374)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6374
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(11..28)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(182..770)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(262..281)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(941..1801)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 1564..1583
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1802..1906)
/gene="bla"
/label=AmpR promoter
rep_origin complement(1932..2387)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2069..2090)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2281..2300
/label=F1ori-R
/note="F1 origin, reverse primer"
repeat_region 2459..2588
/label=AAV2 ITR
promoter 2899..3176
/label=chicken beta-actin promoter
primer_bind 3455..3477
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 3539..3558
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 3591..3600
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3597..5249
/codon_start=1
/product="firefly luciferase"
/label=luciferase
/note="synthetic luc2 version of the luciferase gene"
/translation="MEDAKNIRKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGETLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
misc_feature 5276..5864
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5329..5349)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5747..5758)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 5907..6028
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5918..5937)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5972..5991
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
repeat_region 6190..6319
/label=AAV2 ITR
ORIGIN
1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
2641 gaagagtacc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
2701 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
2761 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
2821 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
2881 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
2941 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
3001 gggcgggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg
3061 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
3121 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
3181 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
3241 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
3301 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
3361 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
3421 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
3481 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
3541 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggatcc gccaccatgg
3601 aagatgccaa aaacattagg aagggcccag cgccattcta cccactcgaa gacgggaccg
3661 ccggcgagca gctgcacaaa gccatgaagc gctacgccct ggtgcccggc accatcgcct
3721 ttaccgacgc acatatcgag gtggacatta cctacgccga gtacttcgag atgagcgttc
3781 ggctggcaga agctatgaag cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca
3841 gcgagaatag cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg
3901 tggccccagc taacgacatc tacaacgagc gcgagctgct gaacagcatg ggcatcagcc
3961 agcccaccgt cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga
4021 agctaccgat catacaaaag atcatcatca tggatagcaa gaccgactac cagggcttcc
4081 aaagcatgta caccttcgtg acttcccatt tgccacccgg cttcaacgag tacgacttcg
4141 tgcccgagag cttcgaccgg gacaaaacca tcgccctgat catgaacagt agtggcagta
4201 ccggattgcc caagggcgta gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg
4261 cccgcgaccc catcttcggc aaccagatca tccccgacac cgctatcctc agcgtggtgc
4321 catttcacca cggcttcggc atgttcacca cgctgggcta cttgatctgc ggctttcggg
4381 tcgtgctcat gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa gactataaga
4441 ttcaatctgc cctgctggtg cccacactat ttagcttctt cgctaagagc actctcatcg
4501 acaagtacga cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg
4561 aggtaggtga ggccgtggcc aaacgcttcc acctaccagg catccgccag ggctacggcc
4621 tgacagaaac aaccagcgcc attctgatca cccccgaagg ggacgacaag cctggcgcag
4681 taggcaaggt ggtgcccttc ttcgaggcta aggtggtgga cttggacacc ggtgagacac
4741 tgggtgtgaa ccagcgcggc gagctgtgcg tccgtggccc catgatcatg agcggctacg
4801 ttaacaaccc cgaggctaca aacgctctca tcgacaagga cggctggctg cacagcggcg
4861 acatcgccta ctgggacgag gacgagcact tcttcatcgt ggaccggctg aagagcctga
4921 tcaaatacaa gggctaccag gtagccccag ccgaactgga gagcatcctg ctgcaacacc
4981 ccaacatctt cgacgccggg gtcgccggcc tgcccgacga cgatgccggc gagctgcccg
5041 ccgcagtcgt cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc gtggactatg
5101 tggccagcca ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg
5161 tgcctaaagg actgaccggc aagttggacg cccgcaagat ccgcgagatt ctcattaagg
5221 ccaagaaggg cggcaagatc gccgtgtaag aattcgatat atcaagctta tcgataatca
5281 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
5341 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
5401 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
5461 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
5521 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
5581 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
5641 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
5701 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
5761 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
5821 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
5881 cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
5941 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
6001 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
6061 ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
6121 aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca
6181 ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
6241 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
6301 tgagcgagcg agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
6361 gcgtattggg cgct
//
