| 质粒名称: | CAG-NLS-GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 104061 |
| 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: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | AAV |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 30°C |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | GFP |
| 物种来源: | -- |
| 插入大小: | 720 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CAG |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | CAG-NLS-GFP was a gift from Viviana Gradinaru (Addgene plasmid # 104061 ; http://n2t.net/addgene:104061 ; RRID:Addgene_104061) |
| 参考文献部分: | Systemic AAV vectors for widespread and targeted gene delivery in rodents. Challis RC, Ravindra Kumar S, Chan KY, Challis C, Beadle K, Jang MJ, Kim HM, Rajendran PS, Tompkins JD, Shivkumar K, Deverman BE, Gradinaru V. Nat Protoc. 2019 Feb;14(2):379-414. doi: 10.1038/s41596-018-0097-3. 10.1038/s41596-018-0097-3 PubMed 30626963 |
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 CAG-NLS-GFP 5829 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION AAV expression of nuclear localized GFP with SV40 NLS.
ACCESSION .
VERSION .
KEYWORDS CAG-NLS-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5829)
AUTHORS Challis RC, Ravindra Kumar S, Chan KY, Challis C, Beadle K, Jang MJ,
Kim HM, Rajendran PS, Tompkins JD, Shivkumar K, Deverman BE,
Gradinaru V
TITLE Systemic AAV vectors for widespread and targeted gene delivery in
rodents.
JOURNAL Nat Protoc. 2019 Feb;14(2):379-414. doi: 10.1038/s41596-018-0097-3.
PUBMED 30626963
REFERENCE 2 (bases 1 to 5829)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5829
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..130
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
enhancer 286..589
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 982..1004
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1066..1085
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1182..1202
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1212..1931
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1212..1928
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1257..1278)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1518..1537)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1865..1886
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 1956..2544
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2009..2029)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2427..2438)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 2576..3052
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2775..2794)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3092..3232
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3092..3221
/label=AAV2 ITR
rep_origin 3307..3762
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3394..3413)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3604..3625
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3779..3798)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3898..3920
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3958..3976)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4044..4148
/gene="bla"
/label=AmpR promoter
CDS 4149..5009
/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(4367..4386)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5180..5768
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5669..5688
/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 tagagggagt
181 ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt aacactagtt attaatagta
241 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
301 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
361 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
421 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
481 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
541 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
601 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
661 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc
721 gggggggggg gggggggggg gggggggggg gggggcgggg cggcgcggtg cggcgggctg
781 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga
841 ggggctccgg gagcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
901 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
961 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
1021 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
1081 ttgtgctgtc tcatcatttt ggcaaagaat tgcggcccaa cggtaccgcc accatgtcta
1141 gtgatgatga ggctactgct gactctcaac attctactcc tccaaaaaag aagagaaagg
1201 tagaagaccc catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
1261 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
1321 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
1381 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
1441 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
1501 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
1561 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
1621 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
1681 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
1741 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
1801 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
1861 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
1921 tgtacaagta agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
1981 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
2041 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
2101 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
2161 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
2221 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
2281 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
2341 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
2401 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
2461 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
2521 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
2581 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
2641 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
2701 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
2761 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
2821 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
2881 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
2941 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
3001 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
3061 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
3121 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
3181 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
3241 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
3301 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
3361 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
3421 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
3481 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
3541 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
3601 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
3661 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
3721 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
3781 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
3841 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
3901 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
3961 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
4021 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
4081 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
4141 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
4201 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
4261 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
4321 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
4381 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
4441 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
4501 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
4561 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
4621 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
4681 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
4741 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
4801 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
4861 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
4921 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
4981 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
5041 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
5101 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
5161 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
5221 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
5281 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
5341 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
5401 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
5461 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
5521 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
5581 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
5641 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
5701 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
5761 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
5821 ctcacatgt
//
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