| 质粒名称: | pAAV-EFS-DIO-GRAB_ATP1.0 |
|---|---|
| 出品公司: | Wekwikgene |
| 目录编号: | 0000137 |
| 产品用途: | Express the ATP sensor GRAB_ATP1.0 in Cre positive cells |
| 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: | EFS |
| 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_ATP1.0 |
| 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: |
A sensitive GRAB sensor for detecting extracellular ATP in vitro and in vivo
Wu Z., He K., Chen Y., Li H., Pan S., Li B., Liu T., Xi F., Deng F., Wang H., Du J., Jing M., Li Y.
Neuron 110, 770-782.e5 (2022)
|
买家导航
pAAV-EFS-DIO-GRAB_ATP1.0载体质粒基本信息
pAAV-EFS-DIO-GRAB_ATP1.0质粒图谱载体图谱和pAAV-EFS-DIO-GRAB_ATP1.0载体序列质粒序列多克隆位点信息
pAAV-EFS-DIO-GRAB_ATP1.0质粒载体简介
pAAV-EFS-DIO-GRAB_ATP1.0质粒序列载体序列
LOCUS pAAV-EFS-DIO-GRAB_ATP1.0 6519 bp DNA circular SYN 21-APR-2023
DEFINITION Express the ATP sensor GRAB_ATP1.0 in Cre positive cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6519)
TITLE An ultrasensitive GRAB sensor for detecting extracellular ATP in
vitro and in vivo
REFERENCE 2 (bases 1 to 6519)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 27, 2026 from SnapGene 6.0.2
https:///www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6519
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 2..131
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 152..363
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
primer_bind 202..228
/label=EFS-F
protein_bind 382..415
/label=lox2272
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GGATACTT). lox2272 sites are compatible with each
other, but incompatible with loxP or loxN sites (Lee and
Saito, 1988)."
protein_bind 466..499
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS complement(510..2405)
/codon_start=1
/label=GRAB_ATP1.0
/translation="TTEVLWPAVPNGTDAAFLAGPGSSWGNSTVASTAAVSSSFKCALT
KTGFQFYYLPAVYILVFIIGFLGNSVAIWMFVFHMKPWSGISVYMFNLALADFLYVLTL
PALIFYYFNKTDWIFGDAMCKLQRFIFHVNLYGSILFLTCISAHRYSGVVYPLKSLGRL
KKKNAICISVLVWLIVVVAISPILFYSGTGVRKNKTITCYDTTSDEYLRSYFIYSMCTT
VAMFCVPLVLILGCYGLIVRALIYPAPAGPRDTDALDLEEGGNVYIKADKQKNGIKANF
HIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFV
TAAGITLGMDELYKGGTGGSMVRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDAT
EGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERT
IFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNTGAAARWRGRSI
YLVIIVLTVFAVSYIPFHVMKTMNLRARLDFQTPAMCAFNDRVYATYQVTRGLASLNSC
VDPILYFLAGDTFRRRLSRATRKASRRSEANLQSKSEDMTLNILPEFKQNGDTSL"
primer_bind complement(557..577)
/label=WKG138-F2
primer_bind 930..951
/label=WKG137-F
primer_bind 1000..1019
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1058..1080
/label=WKG183-R
primer_bind 1065..1086
/label=WKG142-F
primer_bind 1107..1129
/label=WKG146-F
primer_bind 1113..1134
/label=WKG111-F
primer_bind 1259..1280
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1304..1323)
/label=WKG130-F2
primer_bind complement(1386..1407)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1571..1596)
/label=WKG124-F3
primer_bind complement(1578..1597)
/label=WKG148-F3
primer_bind complement(1658..1679)
/label=WKG136-F1
primer_bind complement(1940..1960)
/label=WKG133-F
primer_bind complement(2022..2042)
/label=WKG138-F1
primer_bind complement(2126..2158)
/label=WKG135-F
sig_peptide complement(2430..2492)
/codon_start=1
/product="leader sequence from mouse immunoglobulin kappa
light chain"
/label=Ig-kappa leader
/translation="METDTLLLWVLLLWVPGSTGD"
regulatory 2489..2498
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
protein_bind complement(2505..2538)
/label=lox2272
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GGATACTT). lox2272 sites are compatible with each
other, but incompatible with loxP or loxN sites (Lee and
Saito, 1988)."
protein_bind complement(2589..2622)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 2647..3235
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2700..2720)
/label=WPRE-R
/note="WPRE, reverse primer"
primer_bind complement(2714..2739)
/label=WPRE-Ra
CDS complement(3118..3129)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3267..3743
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3466..3485)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3783..3923
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3783..3912
/label=AAV2 ITR
rep_origin 3998..4453
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4085..4104)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4295..4316
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4470..4489)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4589..4611
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4649..4667)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4735..4839
/gene="bla"
/label=AmpR promoter
CDS 4840..5700
/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(5058..5077)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5871..6459
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6360..6379
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
121 taggggttcc tgcggccgca cgcgtaagct tgggcagagc gcacatcgcc cacagtcccc
181 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
241 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
301 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
361 cagtctagag tcgactccgg aataacttcg tataggatac tttatacgaa gttatgcaga
421 atggtagctg gattgtagct gctattagca atatgaaacc tcttaataac ttcgtatagc
481 atacattata cgaagttatg gcgcatatgt tacaggcttg tatctccatt ctgcttgaac
541 tcaggtaaaa tattgagggt catgtcttca ctcttggatt gcaaatttgc ctcacttctt
601 ctagaagctt tccttgtggc tcgggagagt ctccttctga aagtatctcc cgccaagaaa
661 tagagaatgg ggtccacaca actgttgaga cttgctagac ctcttgtcac ctgatacgtg
721 gcataaaccc tgtcattgaa agcacacatt gctggggtct gaaaatcaag ccgggccctc
781 aagttcatcg ttttcatcac atggaaaggg atgtaagaca cagcaaaaac agtcagtaca
841 atgattacca ggtaaatcga ccgcccgcgc cagcgtgctg ctgctccggt gttgtactcc
901 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
961 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
1021 ttgaagaaga tggtgcgctc ctggatgtag ccttcgggca tggcggactt gaagaagtcg
1081 tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc
1141 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
1201 ttgccctcgg tggcatcgcc ctcaccctcg ccggacacgc tgaacttgtg gccgtttacg
1261 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gccctttctc
1321 accatgctcc ctccggtacc gcccttgtac agctcgtcca tgccgagagt gatcccggcg
1381 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttcgaaagt
1441 ttggactgca cgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
1501 gtgttctgct ggtagtggta ggcgagctgc acgccgccgt cctcgatgtt gtggcggatg
1561 tggaagttcg ccttgatgcc gttcttctgc ttgtcggcct tgatatagac gtttcctccc
1621 tcctccaggt ccagcgcgtc ggtgtcgcgc ggcccggccg gcgcggggta aatcaaagct
1681 ctcacaatta atccgtaaca gcccagaatc agcaccaagg ggacacagaa catggccacg
1741 gtcgtgcaca tgctgtagat gaaataactt cgcaggtact cgtctgaggt ggtgtcgtaa
1801 caggtgatgg ttttgttttt gcggaccccg gtacctgagt agaagaggat gggggagatc
1861 gccaccacca caatgagcca caccagcacg ctgatacaga tcgcattctt ctttttgagc
1921 cggcccaggg acttgagggg gtacaccaca ccgctgtacc ggtgggcact gatgcatgtc
1981 agaaacaaga tgctgccata gaggttcaca tgaaagatga acctctgcag tttacacatg
2041 gcatccccga agatccagtc tgttttattg aagtagtaga agatcagggc tggcagagtc
2101 agcacgtaca agaagtcggc cagagccaaa ttgaacatgt acacggagat gccgctccag
2161 ggcttcatgt ggaagacgaa catccagatg gccacgctgt tgcccaggaa gccgatgatg
2221 aataccaaga tgtagacagc cggcaggtag taaaactgga agcccgtctt ggtcaaggcg
2281 catttgaacg acgaggagac ggcggcagtg gaggcgaccg tgctgttccc ccaggacgaa
2341 cccggaccgg ccaggaaggc agcgtccgtc ccgttgggga cagccggcca cagcacctcg
2401 gtggtgccaa ctttttcgca caaacttgtg tcaccagtgg aacctggaac ccagagcagc
2461 agtacccata gcaggagtgt gtctgtctcc atggtggcgc tagcataact tcgtataaag
2521 tatcctatac gaagttattt gccttaaccc agaaattatc actgttattc tttagaatgg
2581 tgcaaagaat aacttcgtat aatgtatgct atacgaagtt atgaattcga tatcaagctt
2641 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
2701 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
2761 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
2821 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
2881 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
2941 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
3001 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
3061 ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
3121 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
3181 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
3241 taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc
3301 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
3361 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
3421 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
3481 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
3541 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
3601 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
3661 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
3721 ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc
3781 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
3841 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
3901 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
3961 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
4021 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
4081 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
4141 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
4201 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
4261 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
4321 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
4381 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
4441 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
4501 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
4561 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
4621 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
4681 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
4741 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
4801 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
4861 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
4921 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
4981 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
5041 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
5101 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
5161 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
5221 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
5281 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
5341 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
5401 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
5461 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
5521 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
5581 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
5641 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
5701 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
5761 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
5821 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
5881 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
5941 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
6001 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
6061 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
6121 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
6181 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
6241 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
6301 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
6361 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
6421 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
6481 ttacggttcc tggccttttg ctggcctttt gctcacatg
//
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