| 质粒名称: | pAAV-GFAP-hChR2(H134R)-EYFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 27054 |
| 存储实验室: | Karl Deisseroth |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6290 bp |
| 载体类型: | 哺乳细胞表达,AAV |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | hChR2(H134R),ChR2 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1662 bp |
| 基因突变: | H134R |
| 标签/融合蛋白: | EYFP（插入式C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI, SalI(未破坏) |
| 3' 克隆位点: | EcoRI, HinDIII (未破坏) |
| 5' 测序引物 | ccaaccccaccccctcaggctatg |
| 3' 测序引物 | GCAATAGCATGATACAAAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-GFAP-hChR2(H134R)-EYFP was a gift from Karl Deisseroth (Addgene plasmid # 27054 ; http://n2t.net/addgene:27054 ; RRID:Addgene_27054) |
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
-EYFP质粒图谱.png)
载体简介
载体序列
LOCUS pAAV-GFAP-hChR2(H134R)-EYFP 7952 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pAAV-GFAP-hChR2(H134R)-EYFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7952)
TITLE hChR2 plasmids
REFERENCE 2 (bases 1 to 7952)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7952
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..141
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 12..141
/label=AAV2 ITR
regulatory 2387..2396
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 3329..3338
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3335..4054
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3380..3401)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3641..3660)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3988..4009
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 4079..4667
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4132..4152)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4550..4561)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 4699..5175
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(4898..4917)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 5215..5355
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 5215..5344
/label=AAV2 ITR
rep_origin 5430..5885
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5517..5536)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5727..5748
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5902..5921)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6021..6043
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6081..6099)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6167..6271
/gene="bla"
/label=AmpR promoter
CDS 6272..7132
/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(6490..6509)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7303..7891
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7792..7811
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
121 actccatcac taggggttcc tgcggccgca cgcgtcccac ctccctctct gtgctgggac
181 tcacagaggg agacctcagg aggcagtctg tccatcacat gtccaaatgc agagcatacc
241 ctgggctggg cgcagtggcg cacaactgta attccagcac tttgggaggc tgatgtggaa
301 ggatcacttg agcccagaag ttctagacca gcctgggcaa catggcaaga ccctatctct
361 acaaaaaaag ttaaaaaatc agccacgtgt ggtgacacac acctgtagtc ccagctattc
421 aggaggctga ggtgagggga tcacttaagg ctgggaggtt gaggctgcag tgagtcgtgg
481 ttgcgccact gcactccagc ctgggcaaca gtgagaccct gtctcaaaag acaaaaaaaa
541 aaaaaaaaaa aaaaagaaca tatcctggtg tggagtaggg gacgctgctc tgacagaggc
601 tcgggggcct gagctggctc tgtgagctgg ggaggaggca gacagccagg ccttgtctgc
661 aagcagacct ggcagcattg ggctggccgc cccccagggc ctcctcttca tgcccagtga
721 atgactcacc ttggcacaga cacaatgttc ggggtgggca cagtgcctgc ttcccgccgc
781 accccagccc ccctcaaatg ccttccgaga agcccattga gcagggggct tgcattgcac
841 cccagcctga cagcctggca tcttgggata aaagcagcac agccccctag gggctgccct
901 tgctgtgtgg cgccaccggc ggtggagaac aaggctctat tcagcctgtg cccaggaaag
961 gggatcaggg gatgcccagg catggacagt gggtggcagg gggggagagg agggctgtct
1021 gcttcccaga agtccaagga cacaaatggg tgaggggact gggcagggtt ctgaccctgt
1081 gggaccagag tggagggcgt agatggacct gaagtctcca gggacaacag ggcccaggtc
1141 tcaggctcct agttgggccc agtggctcca gcgtttccaa acccatccat ccccagaggt
1201 tcttcccatc tctccaggct gatgtgtggg aactcgagga aataaatctc cagtgggaga
1261 cggaggggtg gccagggaaa cggggcgctg caggaataaa gacgagccag cacagccagc
1321 tcatgtgtaa cggctttgtg gagctgtcaa ggcctggtct ctgggagaga ggcacaggga
1381 ggccagacaa ggaaggggtg acctggaggg acagatccag gggctaaagt cctgataagg
1441 caagagagtg ccggccccct cttgccctat caggacctcc actgccacat agaggccatg
1501 attgaccctt agacaaaggg ctggtgtcca atcccagccc ccagccccag aactccaggg
1561 aatgaatggg cagagagcag gaatgtggga catctgtgtt caagggaagg actccaggag
1621 tctgctggga atgaggccta gtaggaaatg aggtggccct tgagggtaca gaacaggttc
1681 attcttcgcc aaattcccag caccttgcag gcacttacag ctgagtgaga taatgcctgg
1741 gttatgaaat caaaaagttg gaaagcaggt cagaggtcat ctggtacagc ccttccttcc
1801 cttttttttt tttttttttt gtgagacaag gtctctctct gttgcccagg ctggagtggc
1861 gcaaacacag ctcactgcag cctcaaccta ctgggctcaa gcaatcctcc agcctcagcc
1921 tcccaaagtg ctgggattac aagcatgagc caccccactc agccctttcc ttccttttta
1981 attgatgcat aataattgta agtattcatc atggtccaac caaccctttc ttgacccacc
2041 ttcctagaga gagggtcctc ttgcttcagc ggtcagggcc ccagacccat ggtctggctc
2101 caggtaccac ctgcctcatg caggagttgg cgtgcccagg aagctctgcc tctgggcaca
2161 gtgacctcag tggggtgagg ggagctctcc ccatagctgg gctgcggccc aaccccaccc
2221 cctcaggcta tgccaggggg tgttgccagg ggcacccggg catcgccagt ctagcccact
2281 ccttcataaa gccctcgcat cccaggagcg agcagagcca gagcaggttg gagaggagac
2341 gcatcacctc cgctgctcgc cgggatcctc tagagtcgac tccggagcca ccatggacta
2401 tggcggcgct ttgtctgccg tcggacgcga acttttgttc gttactaatc ctgtggtggt
2461 gaacgggtcc gtcctggtcc ctgaggatca atgttactgt gccggatgga ttgaatctcg
2521 cggcacgaac ggcgctcaga ccgcgtcaaa tgtcctgcag tggcttgcag caggattcag
2581 cattttgctg ctgatgttct atgcctacca aacctggaaa tctacatgcg gctgggagga
2641 gatctatgtg tgcgccattg aaatggttaa ggtgattctc gagttctttt ttgagtttaa
2701 gaatccctct atgctctacc ttgccacagg acaccgggtg cagtggctgc gctatgcaga
2761 gtggctgctc acttgtcctg tcatccttat ccgcctgagc aacctcaccg gcctgagcaa
2821 cgactacagc aggagaacca tgggactcct tgtctcagac atcgggacta tcgtgtgggg
2881 ggctaccagc gccatggcaa ccggctatgt taaagtcatc ttcttttgtc ttggattgtg
2941 ctatggcgcg aacacatttt ttcacgccgc caaagcatat atcgagggtt atcatactgt
3001 gccaaagggt cggtgccgcc aggtcgtgac cggcatggca tggctgtttt tcgtgagctg
3061 gggtatgttc ccaattctct tcattttggg gcccgaaggt tttggcgtcc tgagcgtcta
3121 tggctccacc gtaggtcaca cgattattga tctgatgagt aaaaattgtt gggggttgtt
3181 gggacactac ctgcgcgtcc tgatccacga gcacatattg attcacggag atatccgcaa
3241 aaccaccaaa ctgaacatcg gcggaacgga gatcgaggtc gagactctcg tcgaagacga
3301 agccgaggcc ggagccgtgc cagcggccgc caccatggtg agcaagggcg aggagctgtt
3361 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
3421 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
3481 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct
3541 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
3601 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
3661 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
3721 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
3781 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
3841 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
3901 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag
3961 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
4021 gatcactctc ggcatggacg agctgtacaa gtaagaattc gatatcaagc ttatcgataa
4081 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
4141 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
4201 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
4261 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
4321 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
4381 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
4441 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
4501 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
4561 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
4621 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc
4681 gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
4741 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
4801 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
4861 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
4921 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
4981 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
5041 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
5101 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
5161 cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac
5221 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
5281 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
5341 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
5401 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
5461 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
5521 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
5581 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
5641 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
5701 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
5761 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
5821 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
5881 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
5941 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
6001 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
6061 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
6121 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
6181 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
6241 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
6301 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
6361 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
6421 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
6481 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
6541 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
6601 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
6661 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
6721 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
6781 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
6841 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
6901 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
6961 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
7021 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
7081 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
7141 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
7201 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
7261 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
7321 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
7381 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
7441 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
7501 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
7561 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
7621 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
7681 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
7741 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
7801 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
7861 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
7921 cctggccttt tgctggcctt ttgctcacat gt
//
