| 质粒名称: | pAAV-CaMKIIa-eNpHR 3.0-EYFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 26971 |
| 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: | 5332 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 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: | eNpHR 3.0 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1683 bp |
| 基因突变: | Trafficking Signal (TS) ER Export Signal |
| GenBank编号 | -- |
| 启动子 | -- |
| 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, AgeI (未破坏) |
| 3' 克隆位点: | EcoRI, HinDIII (未破坏) |
| 5' 测序引物 | ctcagaagccccaagctcgtc |
| 3' 测序引物 | GCAATAGCATGATACAAAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-CaMKIIa-eNpHR 3.0-EYFP was a gift from Karl Deisseroth (Addgene plasmid # 26971 ; http://n2t.net/addgene:26971 ; RRID:Addgene_26971) |
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-CaMKIIa-eNpHR 3.0-EYFP 7058 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION AAV expression of eNpHR 3.0 fused to EYFP driven by CaMKIIa promoter
for optogenetic inhibition.
ACCESSION .
VERSION .
KEYWORDS pAAV-CaMKIIa-eNpHR 3.0-EYFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7058)
TITLE eNpHR3.0-EYFP
REFERENCE 2 (bases 1 to 7058)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7058
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 54..183
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
CDS 2473..3186
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(2515..2536)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2776..2795)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3123..3144
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 3238..3826
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3291..3311)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3709..3720)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3858..4334
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(4057..4076)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 4374..4514
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4374..4503
/label=AAV2 ITR
rep_origin 4589..5044
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4676..4695)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4886..4907
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5061..5080)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5180..5202
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5240..5258)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5326..5430
/gene="bla"
/label=AmpR promoter
CDS 5431..6291
/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(5649..5668)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6462..7050
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6951..6970
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag
61 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc
121 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
181 cctgcggccg cacgcgttta acattatggc cttaggtcac ttcatctcca tggggttctt
241 cttctgattt tctagaaaat gagatggggg tgcagagagc ttcctcagtg acctgcccag
301 ggtcacatca gaaatgtcag agctagaact tgaactcaga ttactaatct taaattccat
361 gccttggggg catgcaagta cgatatacag aaggagtgaa ctcattaggg cagatgacca
421 atgagtttag gaaagaagag tccagggcag ggtacatcta caccacccgc ccagccctgg
481 gtgagtccag ccacgttcac ctcattatag ttgcctctct ccagtcctac cttgacggga
541 agcacaagca gaaactggga caggagcccc aggagaccaa atcttcatgg tccctctggg
601 aggatgggtg gggagagctg tggcagaggc ctcaggaggg gccctgctgc tcagtggtga
661 cagatagggg tgagaaagca gacagagtca ttccgtcagc attctgggtc tgtttggtac
721 ttcttctcac gctaaggtgg cggtgtgata tgcacaatgg ctaaaaagca gggagagctg
781 gaaagaaaca aggacagaga cagaggccaa gtcaaccaga ccaattccca gaggaagcaa
841 agaaaccatt acagagacta caagggggaa gggaaggaga gatgaattag cttcccctgt
901 aaaccttaga acccagctgt tgccagggca acggggcaat acctgtctct tcagaggaga
961 tgaagttgcc agggtaacta catcctgtct ttctcaagga ccatcccaga atgtggcacc
1021 cactagccgt taccatagca actgcctctt tgccccactt aatcccatcc cgtctgttaa
1081 aagggcccta tagttggagg tgggggaggt aggaagagcg atgatcactt gtggactaag
1141 tttgttcgca tccccttctc caaccccctc agtacatcac cctgggggaa cagggtccac
1201 ttgctcctgg gcccacacag tcctgcagta ttgtgtatat aaggccaggg caaagaggag
1261 caggttttaa agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac
1321 agggcgtttc ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg
1381 tgagcagcca cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt
1441 ttgcactcag gagcacgggc aggcgagtgg cccctagttc tgggggcagc tctagagcgg
1501 taccggatcc ccgggtaccg gtaggccacc atgacagaga ccctgcctcc cgtgaccgag
1561 agtgccgtgg cccttcaagc cgaggttacc caaagggagt tgttcgagtt cgtgctgaac
1621 gaccctttgc ttgcaagcag tctctatatc aacatcgcac ttgcaggact gagtatactg
1681 ctgttcgttt ttatgacccg aggactcgat gatccacggg caaaacttat tgctgtgtca
1741 accatccttg tgcctgtcgt cagcattgcc tcctacactg gattggcgag cggcctgaca
1801 atttccgttc ttgaaatgcc agcgggccat tttgcagaag gcagctcagt gatgctggga
1861 ggagaagagg tagatggtgt agtcaccatg tggggacggt atctcacctg ggcactttcc
1921 acgcccatga ttctcctcgc tctgggtctc ctggccggaa gcaatgctac aaagctcttc
1981 acagctatca ctttcgatat cgctatgtgc gtgactggcc ttgccgcggc cctgactacc
2041 tcctcccacc tcatgagatg gttctggtac gctatcagtt gtgcatgctt tctggtggtc
2101 ttgtatatcc tgctggtgga gtgggcacag gacgccaaag ccgcgggaac cgctgacatg
2161 ttcaataccc tgaagctgtt gacagtagtg atgtggctgg ggtatccaat tgtgtgggct
2221 cttggagtcg agggtatcgc ggtgttgccc gttggggtga cgagctgggg atattctttc
2281 ctggatatcg tggcaaagta cattttcgca ttcttgctcc tgaactatct gacgtcaaac
2341 gaatctgtcg tgtccggcag cattttggat gttccatctg cttctgggac cccggctgat
2401 gatgcggccg ccaagagcag gatcaccagc gagggcgagt acatccccct ggaccagatc
2461 gacatcaacg tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
2521 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
2581 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
2641 tggcccaccc tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac
2701 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
2761 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
2821 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
2881 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
2941 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
3001 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
3061 gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
3121 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
3181 tacaagttct gctacgagaa cgaggtgtaa tgagaattcg atatcaagct tatcgataat
3241 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
3301 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
3361 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
3421 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
3481 tggggcattg ccaccacctg tcagctcctt tccggaactt tcgctttccc cctccctatt
3541 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
3601 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
3661 tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
3721 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
3781 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg
3841 ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc
3901 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt
3961 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg
4021 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg
4081 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc
4141 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt
4201 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc
4261 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc
4321 ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc
4381 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
4441 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
4501 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
4561 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
4621 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
4681 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
4741 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
4801 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
4861 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
4921 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
4981 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
5041 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
5101 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
5161 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
5221 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
5281 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
5341 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
5401 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
5461 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
5521 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
5581 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
5641 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
5701 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
5761 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
5821 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
5881 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
5941 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
6001 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
6061 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
6121 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
6181 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
6241 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
6301 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
6361 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
6421 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
6481 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
6541 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
6601 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
6661 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
6721 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
6781 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
6841 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
6901 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
6961 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
7021 tgctcgtcag gggggcggag cctatggaaa aacgccag
//
