| 质粒名称: | pAAV-hSyn-GRAB_ACh3.0 |
|---|---|
| 出品公司: | Wekwikgene |
| 目录编号: | 0000110 |
| 产品用途: | Expresses the ACh sensor GRAB_ACh3.0 in neurons |
| 存储实验室: | Yulong Li Lab |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 表达系统: | 腺病毒，哺乳动物表达 |
| 启动子: | hSyn |
| 拷贝数: | 低拷贝 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a，TOP10 |
|
插入基因信息 |
|
| 插入基因名称: | GRAB_ACh3.0 |
| 物种来源: | Synthetic |
| 密码子优化: | 哺乳细胞表达优化 |
|
文献引用方法 |
|
| 参考文献部分: |
An optimized acetylcholine sensor for monitoring in vivo cholinergic activity
Jing M., Li Y., Zeng J., Huang P., Skirzewski M., Kljakic O., Peng W, Qian T., Tan K., Zou J., Trinh S., Wu R., Zhang S., Pan, Hires S.A., Xu M., Li H., Saksida L.M., Prado V.F., Bussey T.J., Prado M.A.M., Chen L., Cheng H., Li Y.
Nat. Methods 2020 1711 17, 1139–1146 (2020)
|
买家导航
pAAV-hSyn-GRAB_ACh3.0载体质粒基本信息
pAAV-hSyn-GRAB_ACh3.0质粒图谱载体图谱和pAAV-hSyn-GRAB_ACh3.0载体序列质粒序列多克隆位点信息
pAAV-hSyn-GRAB_ACh3.0质粒载体简介
pAAV-hSyn-GRAB_ACh3.0质粒序列载体序列
LOCUS pAAV-hSyn-GRAB_ACh3.0 6389 bp DNA circular SYN 21-APR-2023
DEFINITION express the genetically-encoded fluorescent acethycholine(ACh)
sensor GRAB-ACh3.0.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6389)
AUTHORS Jing M, Li Y, Zeng J, Huang P, Skirzewski M, Kljakic O, Peng W, Qian
T, Tan K, Zou J, Trinh S, Wu R, Zhang S, Pan S, Hires SA, Xu M, Li
H, Saksida LM, Prado VF, Bussey TJ, Prado MAM, Chen L, Cheng H, Li Y
TITLE An optimized acetylcholine sensor for monitoring in vivo cholinergic
activity.
JOURNAL Nat Methods. 2020 Sep 28. pii: 10.1038/s41592-020-0953-2. doi:
10.1038/s41592-020-0953-2.
PUBMED 32989318
REFERENCE 2 (bases 1 to 6389)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 24, 2026 from SnapGene 6.0.2
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods. 2020 Sep 28. pii: 10.1038/s41592-020-0953-2. doi:
10.1038/s41592-020-0953-2."
FEATURES Location/Qualifiers
source 1..6389
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 103..546
/label=hSyn
regulatory 553..562
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
sig_peptide 559..621
/codon_start=1
/product="leader sequence from mouse immunoglobulin kappa
light chain"
/label=Ig-kappa leader
/translation="METDTLLLWVLLLWVPGSTGD"
protein_bind 622..646
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 646..2463
/codon_start=1
/label=GRAB_Ach3.0
/translation="TTLHNNSTTSPLFPNISSSWIHSPSDAGLPPGTVTHFGSYNVSRA
AGNFSSPDGTTDDPLGGHTVWQVVFIAFLTGILALVTIIGNILVIVSFKVNKQLKTVNN
YFLLSLACADLIIGVISMNLFTTYIIMNRWALGNLACDLWLAIDYVASNASVMNLLVIS
FDRYFSITRPLTYRAKRTTKRAGVMIGLAWVISFVLWAPAILFWQYFVGKRTVPPGECF
IQFLSEPTITFGTAIAAFYMPVTIMTILYWRIYKETEGGNVYIKADKQKNGIKANFHIR
HNIEDGGVQLAYHYQQNTPIGDGPVLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAA
GITNGMDELYKGGTGGSMVRKGEELFTGVVPILVELDGDVNGHKFSVTGEGEGDATIGK
LTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFF
KGDGNYKTRAIVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNHNAKTLSAILLAFII
TWTPYNIMVLVNTFCDSCIPKTFWNLGYWLCYINSTVNPVCYALCNKTFRTTFKMLLLC
QCDKKKRRKQQYQQRQSVIFHKRAPEQAL"
primer_bind 1153..1162
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind 1201..1224
/label=WKG110-F
primer_bind 1635..1656
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1762..1783)
/label=EGFP-N
/note="EGFP, reverse primer"
misc_feature 2488..3076
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2541..2561)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2959..2970)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3108..3584
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3307..3326)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3624..3764
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3624..3753
/label=AAV2 ITR
rep_origin 3839..4294
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3926..3945)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4136..4157
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4311..4330)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4430..4452
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4490..4508)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4576..4680
/gene="bla"
/label=AmpR promoter
CDS 4681..5541
/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(4899..4918)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5712..6300
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6201..6220
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
repeat_region join(6362..6389,1..102)
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
1 cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
61 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtatga
121 ggcggggtgg gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc
181 ccattcccca aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag
241 gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg
301 cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa
361 ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc
421 acgcgaggcg cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact
481 cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt
541 cgagaaaccg gtgccaccat ggagacagac acactcctgc tatgggtact gctgctctgg
601 gttccaggtt ccactggtga cacaagtttg tacaaaaaag ttggcaccac cttgcacaat
661 aacagtacaa cctcgccttt gtttccaaac atcagctcct cctggataca cagcccctcc
721 gatgcagggc tgcccccggg aaccgtcact catttcggca gctacaatgt ttctcgagca
781 gctggcaatt tctcctctcc agacggtacc accgatgacc ctctgggagg tcataccgtc
841 tggcaagtgg tcttcatcgc tttcttaacg ggcatcctgg ccttggtgac catcatcggc
901 aacatcctgg taattgtgtc atttaaggtc aacaagcagc tgaagacagt caacaactac
961 ttcctcttaa gcctggcctg tgccgatctg attatcgggg tcatttcaat gaatctgttt
1021 acgacctaca tcatcatgaa tcgatgggcc ttagggaact tggcctgtga cctctggctt
1081 gccattgact acgtagccag caatgcctct gttatgaatc ttctggtcat cagctttgac
1141 agatactttt ccatcacgag gccgctcacg taccgagcca aacgaacaac aaagagagcc
1201 ggtgtgatga tcggtctggc ttgggtcatc tcctttgtcc tttgggctcc tgccatcttg
1261 ttctggcaat actttgttgg aaagagaact gtgcctccgg gagagtgctt cattcagttc
1321 ctcagtgagc ccaccattac ttttggcaca gccatcgctg ctttttatat gcctgtcacc
1381 attatgacta ttttatactg gaggatctat aaggaaactg aaggaggaaa cgtctatatc
1441 aaggccgaca agcagaagaa cggcatcaag gcgaacttcc acatccgcca caacatcgag
1501 gacggcggcg tgcagctcgc ctaccactac cagcagaaca cccccatcgg cgacggcccc
1561 gtgctgctgc ccgacaacca ctacctgagc gtgcagtcca aactttcgaa agaccccaac
1621 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactaatggc
1681 atggacgagc tgtacaaggg cggtaccgga gggagcatgg tgagaaaggg cgaggagctg
1741 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
1801 agcgtgaccg gcgagggtga gggcgatgcc accatcggca agctgaccct gaagttcatc
1861 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
1921 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
1981 atgcccgaag gctacatcca ggagcgcacc atcttcttca agggcgacgg caattacaag
2041 acccgcgcca ttgtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
2101 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ccataatgcc
2161 aagaccctca gtgcgatctt gcttgccttc atcatcactt ggaccccata caacatcatg
2221 gttctggtga acaccttttg tgacagctgc atacccaaaa ccttttggaa tctgggctac
2281 tggctgtgct acatcaacag caccgtgaac cccgtgtgct atgctctgtg caacaaaaca
2341 ttcagaacca ctttcaagat gctgctgctg tgccagtgtg acaaaaaaaa gaggcgcaag
2401 cagcagtacc agcagagaca gtcggtcatt tttcacaagc gcgcacccga gcaggccttg
2461 taggaattcg atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
2521 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
2581 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
2641 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
2701 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
2761 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
2821 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
2881 aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg
2941 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
3001 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
3061 tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct
3121 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
3181 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
3241 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
3301 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
3361 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
3421 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
3481 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
3541 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa
3601 ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
3661 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
3721 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
3781 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
3841 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
3901 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
3961 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
4021 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
4081 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
4141 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
4201 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
4261 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
4321 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
4381 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
4441 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
4501 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
4561 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
4621 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
4681 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
4741 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
4801 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
4861 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
4921 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
4981 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
5041 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
5101 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
5161 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
5221 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
5281 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
5341 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
5401 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
5461 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
5521 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
5581 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
5641 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
5701 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
5761 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
5821 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
5881 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
5941 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
6001 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
6061 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
6121 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
6181 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
6241 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
6301 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
6361 tcctgcaggc agctgcgcgc tcgctcgct
//
