| 质粒名称: | pAAV-CAG-mTurquoise2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 99122 |
| 存储实验室: | Viviana Gradinaru |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 5891bp |
| 载体类型: | 哺乳细胞表达, AAV |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Turquoise2 |
| 插入大小: | 3305bp |
| 启动子: | CAG |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | KpnI (not destroyed) |
| 3’克隆位点: | EcoRI/HindIII (not destroyed) |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-CAG-mTurquoise2 was a gift from Viviana Gradinaru (Addgene plasmid # 99122 ; http://n2t.net/addgene:99122 ; RRID:Addgene_99122) |
| 参考文献部分: | Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V. Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub 2017 Jun 26. 10.1038/nn.4593 PubMed 28671695 |
买家导航
pAAV-CAG-mTurquoise2载体质粒基本信息
pAAV-CAG-mTurquoise2质粒图谱载体图谱和pAAV-CAG-mTurquoise2载体序列质粒序列多克隆位点信息
pAAV-CAG-mTurquoise2质粒载体简介
pAAV-CAG-mTurquoise2质粒序列载体序列
LOCUS pAAV-CAG-mTurquoise2 5891 bp DNA circular SYN 25-AUG-2024
DEFINITION An AAV genome that ubiquitously expresses the fluorescent protein
mTurquoise2 from the CAG promoter.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5891)
AUTHORS Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL,
Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V
TITLE Engineered AAVs for efficient noninvasive gene delivery to the
central and peripheral nervous systems.
JOURNAL Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub
2017 Jun 26.
PUBMED 28671695
REFERENCE 2 (bases 1 to 5891)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nn.4593";
journalName: "Nat Neurosci"; date: "2017-08"; volume: "20"; issue:
"8"; pages: "1172-1179"
FEATURES Location/Qualifiers
source 1..5891
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 1..130
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
enhancer 210..513
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 515..792
/label=chicken beta-actin promoter
primer_bind 1122..1144
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1206..1225
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1268..1277
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1274..1993
/codon_start=1
/product="enhanced monomeric variant of CFP (Goedhart et
al., 2012)"
/label=mTurquoise2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1274..1993
/codon_start=1
/product="enhanced monomeric variant of CFP (Goedhart et
al., 2010)"
/label=mTurquoise
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1319..1340)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1580..1599)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1927..1948
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 2018..2606
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2071..2091)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2489..2500)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 2638..3114
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2837..2856)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3154..3294
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3154..3283
/label=AAV2 ITR
rep_origin 3369..3824
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3456..3475)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3666..3687
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3841..3860)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3960..3982
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4020..4038)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4106..4210
/gene="bla"
/label=AmpR promoter
CDS 4211..5071
/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 complement(4429..4448)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5242..5830
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5731..5750
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
121 aggggttcct gcggccgcac gcgtaacact agttattaat agtaatcaat tacggggtca
181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
481 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
541 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
601 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg
661 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
721 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
781 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
841 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
901 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
961 tggctgcgtg aaagccttga ggggctccgg gagcgccggc aggaaggaaa tgggcgggga
1021 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
1081 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
1141 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
1201 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgcggcccaa
1261 cggtaccgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
1321 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
1381 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
1441 gccctggccc accctcgtga ccaccctgtc ctggggcgtg cagtgcttcg cccgctaccc
1501 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
1561 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
1621 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
1681 catcctgggg cacaagctgg agtacaacta cttcagcgac aacgtctata tcaccgccga
1741 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg
1801 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
1861 gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg
1921 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
1981 gctgtacaag tgagaattcg atatcaagct tatcgataat caacctctgg attacaaaat
2041 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
2101 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
2161 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
2221 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
2281 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
2341 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
2401 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
2461 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
2521 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
2581 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg
2641 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
2701 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
2761 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
2821 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
2881 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
2941 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
3001 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
3061 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
3121 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
3181 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
3241 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
3301 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
3361 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
3421 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
3481 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
3541 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
3601 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
3661 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
3721 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
3781 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
3841 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
3901 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
3961 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
4021 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
4081 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
4141 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
4201 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
4261 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
4321 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
4381 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
4441 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
4501 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
4561 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
4621 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
4681 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
4741 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
4801 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
4861 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
4921 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
4981 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
5041 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
5101 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
5161 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
5221 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
5281 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
5341 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
5401 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
5461 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
5521 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
5581 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
5641 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
5701 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
5761 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
5821 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
5881 tgctcacatg t
//
