LOCUS       sequence-330263-        6467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the ATP-insensitive control sensor GRAB_ATP1.0mut in 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6467)
  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 6467)
  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-hsyn-GRAB_ATP1.0mut
FEATURES             Location/Qualifiers
     source          1..6467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     587..646
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    650..674
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1672..1693
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1799..1820)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2060..2079)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2594..3182
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2647..2667)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3065..3076)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3214..3690
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3413..3432)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3730..3870
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3730..3859
                     /label=AAV2 ITR
     rep_origin      3945..4400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4032..4051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4242..4263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4417..4436)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4536..4558
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4596..4614)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4682..4786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4787..4855,4856..5647)
                     /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(5005..5024)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5818..6406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6307..6326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtatgagg cggggtgggg gtgcctacct gacgaccgac
      181 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      241 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      301 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      361 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      421 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      481 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      541 ggagtcgtgt cgtgcctgag agcgcagtcg agaaaccggt gccaccatgg agacagacac
      601 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca caagtttgta
      661 caaaaaagtt ggcaccgccg aggtgctgtg gccggctgtc cccaacggga cggacgctgc
      721 cttcctggcc ggtccgggtt cgtcctgggg gaacagcacg gtcgcctcca ctgccgccgt
      781 ctcctcgtcg ttcaaatgcg ccttgaccaa gacgggcttc cagttttact acctgccggc
      841 tgtctacatc ttggtattca tcatcggctt cctgggcaac agcgtggcca tctggatgtt
      901 cgtcttccac atgaagccct ggagcggcat ctccgtgtac atgttcaatt tggctctggc
      961 cgacttcttg tacgtgctga ctctgccagc cctgatcttc tactacttca ataaaacaga
     1021 ctggatcttc ggggatgcca tgtgtaaact gcagaggttc atctttcatg tgaacctcta
     1081 tggcagcatc ttgtttctga catgcatcag tgcccaccgg tacagcggtg tggtgtaccc
     1141 cctcaagtcc ctgggccggc tcaaaaagaa gaatgcgatc tgtatcagcg tgctggtgtg
     1201 gctcattgtg gtggtggcga tctcccccat cctcttctac tcaggtaccg gggtccgcaa
     1261 aaacaaaacc atcacctgtt acgacaccac ctcagacgag tacctgcgaa gttatttcat
     1321 ctacagcatg tgcacgaccg tggccatgtt ctgtgtcccc ttggtgctga ttctgggctg
     1381 ttacggatta attgtgagag ctttgattta ccccgcgccg gccgggccgc gcgacaccga
     1441 cgcgctggac ctggaggagg gaggaaacgt ctatatcaag gccgacaagc agaagaacgg
     1501 catcaaggcg aacttccaca tccgccacaa catcgaggac ggcggcgtgc agctcgccta
     1561 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1621 cctgagcgtg cagtccaaac tttcgaaaga ccccaacgag aagcgcgatc acatggtcct
     1681 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggcgg
     1741 taccggaggg agcatggtga gaaagggcga ggagctgttc accggggtgg tgcccatcct
     1801 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggtgaggg
     1861 cgatgccacc gagggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1921 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1981 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acatccagga
     2041 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2101 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2161 catcctgggg cacaagctgg agtacaacac cggagcagca gcacgctggc gcgggcggtc
     2221 gatttacctg gtaatcattg tactgactgt ttttgctgtg tcttacatcc ctttccatgt
     2281 gatgaaaacg atggctttga gggcccggct tgattttcag accccagcaa tgtgtgcttt
     2341 caatgacagg gtttatgcca cgtatcaggt gacaagaggt ctagcaagtc tcaacagttg
     2401 tgtggacccc attctctatt tcttggcggg agatactttc agaaggagac tctcccgagc
     2461 cacaaggaaa gcttctagaa gaagtgaggc aaatttgcaa tccaagagtg aagacatgac
     2521 cctcaatatt ttacctgagt tcaagcagaa tggagataca agcctgtaag aattcgatat
     2581 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2641 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2701 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2761 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2821 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2881 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2941 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     3001 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3061 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3121 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3181 gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca
     3241 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     3301 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     3361 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     3421 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     3481 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     3541 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     3601 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     3661 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg
     3721 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3781 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     3841 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     3901 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     3961 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4021 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4081 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4141 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4201 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4261 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     4321 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4381 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     4441 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4501 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4561 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     4621 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     4681 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4741 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4801 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4861 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4921 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4981 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5041 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5101 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5161 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5221 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5281 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5341 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5401 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5461 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5521 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5581 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5641 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5701 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5761 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5821 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5881 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5941 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6001 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6061 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6121 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6181 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6241 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6301 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6361 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6421 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
