LOCUS       sequence-330880-        6519 bp    DNA     circular SYN 15-DEC-2025
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)
  AUTHORS   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
  TITLE     A sensitive GRAB sensor for detecting extracellular ATP in vitro and
            in vivo.
  JOURNAL   Neuron. 2021 Dec 17. pii: S0896-6273(21)00988-0. doi: 
            10.1016/j.neuron.2021.11.027.
  PUBMED    34942116
REFERENCE   2  (bases 1 to 6519)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neuron. 
            2021 Dec 17. pii: S0896-6273(21)00988-0. doi: 
            10.1016/j.neuron.2021.11.027."
COMMENT     Sequence Label: pAAV-EFS-DIO-GRAB_ATP1.0
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"
     protein_bind    complement(382..415)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(466..499)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1000..1019
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1259..1280
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1386..1407)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2433..2492)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2489..2498
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2505..2538
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2589..2622
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     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"
     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             join(4840..4908,4909..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
//
