| 质粒名称: | pAAV-IRES-hrGFP-mWnt3a |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 89771 |
| 存储实验室: | Ken-Ichi Takemaru |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV-IRES-hrGFP |
| 载体出品公司: | Agilent Technologies |
| 空载体大小: | 6100bp |
| 完整质粒大小: | 7177bp |
| 载体类型: | 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: | mouse Wnt3a |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 1060bp |
| 启动子: | CMV |
| 标签/融合蛋白: | 无 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI(未破坏) |
| 3’克隆位点: | XhoI (未破坏) |
| 5’测序引物 | ATTCTGAGTCCAAGCTAGGC (beta-globin) |
| 3’测序引物 | TAGAAGGACACCTAGTCAGA (hGH polyA) |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-IRES-hrGFP-mWnt3a was a gift from Ken-Ichi Takemaru (Addgene plasmid # 89771 ; http://n2t.net/addgene:89771 ; RRID:Addgene_89771) |
| 参考文献部分: | Abrogation of beta-catenin signaling in oligodendrocyte precursor cells reduces glial scarring and promotes axon regeneration after CNS injury. Rodriguez JP, Coulter M, Miotke J, Meyer RL, Takemaru K, Levine JM. J Neurosci. 2014 Jul 30;34(31):10285-97. doi: 10.1523/JNEUROSCI.4915-13.2014. 10.1523/JNEUROSCI.4915-13.2014 PubMed 25080590 |
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-IRES-hrGFP-mWnt3a 7177 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses mouse Wnt3a in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pAAV-IRES-hrGFP-mWnt3a
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7177)
AUTHORS Rodriguez JP, Coulter M, Miotke J, Meyer RL, Takemaru K, Levine JM
TITLE Abrogation of beta-catenin signaling in oligodendrocyte precursor
cells reduces glial scarring and promotes axon regeneration after
CNS injury.
JOURNAL J Neurosci. 2014 Jul 30;34(31):10285-97. doi:
10.1523/JNEUROSCI.4915-13.2014.
PUBMED 25080590
REFERENCE 2 (bases 1 to 7177)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7177
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 305..508
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 459..479
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 505..529
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 1106..1124
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
CDS 2253..2324
/codon_start=1
/product="three tandem FLAG(R) epitope tags"
/label=3xFLAG
/translation="DYKDDDDKDYKDDDDKDYKDDDDK"
misc_feature 2360..2942
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2526..2543)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2753..2772
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 2943..3662
/codon_start=1
/product="humanized Renilla green fluorescent protein"
/label=hrGFP
/note="mammalian codon-optimized"
/translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
AQLTSLGKPLGSLHEWV"
primer_bind complement(3692..3711)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3693..3711)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 3719..4195
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3918..3937)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 4235..4375
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4235..4364
/label=AAV2 ITR
rep_origin 4450..4905
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4537..4556)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4747..4768
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4922..4941)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5041..5063
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5101..5119)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5187..5291
/gene="bla"
/label=AmpR promoter
CDS 5292..6152
/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(5510..5529)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6323..6911
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6812..6831
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
repeat_region 6973..7102
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggatt
601 cgaatcccgg ccgggaacgg tgcattggaa cgcggattcc ccgtgccaag agtgacgtaa
661 gtaccgccta tagagtctat aggcccacaa aaaatgcttt cttcttttaa tatacttttt
721 tgtttatctt atttctaata ctttccctaa tctctttctt tcagggcaat aatgatacaa
781 tgtatcatgc ctctttgcac cattctaaag aataacagtg ataatttctg ggttaaggca
841 atagcaatat ttctgcatat aaatatttct gcatataaat tgtaactgat gtaagaggtt
901 tcatattgct aatagcagct acaatccagc taccattctg cttttatttt atggttggga
961 taaggctgga ttattctgag tccaagctag gcccttttgc taatcatgtt catacctctt
1021 atcttcctcc cacagctcct gggcaacgtg ctggtctgtg tgctggccca tcactttggc
1081 aaagaattgg gattcgaaca tcgataatta accctcacta aagggaacaa aagctggagc
1141 tccaccgcgg tggcggccgc tctagcccgg gcggatccga attcaccatg gctcctctcg
1201 gatacctctt agtgctctgc agcctgaagc aggctctggg cagctacccg atctggtggt
1261 ccttggctgt gggaccccag tactcctctc tgagcactca gcccattctc tgtgccagca
1321 tcccaggcct ggtaccgaag cagctgcgct tctgcaggaa ctacgtggag atcatgccca
1381 gcgtggctga gggtgtcaaa gcgggcatcc aggagtgcca gcaccagttc cgaggccggc
1441 gttggaactg caccaccgtc agcaacagcc tggccatctt tggccctgtt ctggacaaag
1501 ccacccggga gtcagccttt gtccatgcca tcgcctccgc tggagtagct ttcgcagtga
1561 cacgctcctg tgcagaggga tcagctgcta tctgtgggtg cagcagccgc ctccagggct
1621 ccccaggcga gggctggaag tggggcggct gtagtgagga cattgaattt ggaggaatgg
1681 tctctcggga gtttgccgat gccagggaga accggccgga tgcccgctct gccatgaacc
1741 gtcacaacaa tgaggctggg cgccaggcca tcgccagtca catgcacctc aagtgcaaat
1801 gccacgggct atctggcagc tgtgaagtga agacctgctg gtggtcgcag ccggacttcc
1861 gcaccatcgg ggatttcctc aaggacaagt atgacagtgc ctcggagatg gtggtagaga
1921 aacaccgaga gtctcgtggc tgggtggaga ccctgaggcc acgttacacg tacttcaagg
1981 tgccgacaga acgcgacctg gtctactacg aggcctcacc caacttctgc gaacctaacc
2041 ccgaaaccgg ctccttcggg acgcgtgacc gcacctgcaa tgtgagctcg catggcatag
2101 atgggtgcga cctgttgtgc tgcgggcgcg ggcataacgc gcgcactgag cgacggaggg
2161 agaaatgcca ctgtgttttc cattggtgct gctacgtcag ctgccaggag tgcacacgtg
2221 tctatgacgt gcacacctgc aagtagctcg aggactacaa ggatgacgat gacaaggatt
2281 acaaagacga cgatgataag gactataagg atgatgacga caaataatag caattcctcg
2341 acgactgcat agggttaccc ccctctccct ccccccccnt aacgttactg gccgaagccg
2401 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
2461 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
2521 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
2581 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
2641 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
2701 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
2761 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
2821 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaacgtcta
2881 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atggccacaa
2941 ccatggtgag caagcagatc ctgaagaaca ccggcctgca ggagatcatg agcttcaagg
3001 tgaacctgga gggcgtggtg aacaaccacg tgttcaccat ggagggctgc ggcaagggca
3061 acatcctgtt cggcaaccag ctggtgcaga tccgcgtgac caagggcgcc cccctgccct
3121 tcgccttcga catcctgagc cccgccttcc agtacggcaa ccgcaccttc accaagtacc
3181 ccgaggacat cagcgacttc ttcatccaga gcttccccgc cggcttcgtg tacgagcgca
3241 ccctgcgcta cgaggacggc ggcctggtgg agatccgcag cgacatcaac ctgatcgagg
3301 agatgttcgt gtaccgcgtg gagtacaagg gccgcaactt ccccaacgac ggccccgtga
3361 tgaagaagac catcaccggc ctgcagccca gcttcgaggt ggtgtacatg aacgacggcg
3421 tgctggtggg ccaggtgatc ctggtgtacc gcctgaacag cggcaagttc tacagctgcc
3481 acatgcgcac cctgatgaag agcaagggcg tggtgaagga cttccccgag taccacttca
3541 tccagcaccg cctggagaag acctacgtgg aggacggcgg cttcgtggag cagcacgaga
3601 ccgccatcgc ccagctgacc agcctgggca agcccctggg cagcctgcac gagtgggtgt
3661 aatagggtac caggtaagtg tacccaattc gccctatagt gagtcgtatt agatctacgg
3721 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
3781 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
3841 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
3901 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
3961 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
4021 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
4081 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
4141 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
4201 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
4261 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
4321 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
4381 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
4441 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
4501 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
4561 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
4621 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
4681 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
4741 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
4801 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
4861 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
4921 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
4981 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
5041 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
5101 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
5161 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
5221 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
5281 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
5341 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
5401 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
5461 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
5521 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
5581 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
5641 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
5701 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
5761 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
5821 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
5881 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
5941 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
6001 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
6061 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
6121 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
6181 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
6241 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
6301 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
6361 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
6421 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
6481 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
6541 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
6601 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
6661 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
6721 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
6781 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
6841 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
6901 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
6961 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
7021 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
7081 aactccatca ctaggggttc ctgcggccgc acgcgtctag ttattaatag taatcaatta
7141 cggggtcatt agttcatagc ccatatatgg agttccg
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：HA-Rab8a-WT
- 下一篇：没有了
