LOCUS       sequence-330732-        7093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the ATP-insensitive control sensor GRAB_ATP1.0mut in 
            astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7093)
  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 7093)
  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-GfaABC1D-GRAB_ATP1.0mut
FEATURES             Location/Qualifiers
     source          1..7093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          995..1127
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1110..1129
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1172..1190
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1207..1216
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1213..1272
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    1276..1300
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2298..2319
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2425..2446)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2686..2705)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3220..3808
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3273..3293)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3691..3702)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3840..4316
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4039..4058)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4356..4496
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4356..4485
                     /label=AAV2 ITR
     rep_origin      4571..5026
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4658..4677)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4868..4889
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5043..5062)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5162..5184
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5222..5240)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5308..5412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5413..5481,5482..6273)
                     /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"
     rep_origin      6444..7032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6933..6952
                     /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 gcgttgtagt taatgattaa cccgccatgc tacttatcta
      181 cgtagccatg ctctaggaag atctaacata tcctggtgtg gagtagggga cgctgctctg
      241 acagaggctc gggggcctga gctggctctg tgagctgggg aggaggcaga cagccaggcc
      301 ttgtctgcaa gcagacctgg cagcattggg ctggccgccc cccagggcct cctcttcatg
      361 cccagtgaat gactcacctt ggcacagaca caatgttcgg ggtgggcaca gtgcctgctt
      421 cccgccgcac cccagccccc ctcaaatgcc ttccgagaag cccattgagc agggggcttg
      481 cattgcaccc cagcctgaca gcctggcatc ttgggataaa agcagcacag ccccctaggg
      541 gctgcccttg ctgtgtggcg ccaccggcgg tggagaacaa ggctctattc agcctgtgcc
      601 caggaaaggg gatcagggga tgcccaggca tggacagtgg gtggcagggg gggagaggag
      661 ggctgtctgc ttcccagaag tccaaggaca caaatgggtg aggggagagc tctccccata
      721 gctgggctgc ggcccaaccc caccccctca ggctatgcca gggggtgttg ccaggggcac
      781 ccgggcatcg ccagtctagc ccactccttc ataaagccct cgcatcccag gagcgagcag
      841 agccagagca ggttggagag gagacgcatc acctccgctg ctcgcaagct ttattgcggt
      901 agtttatcac agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt
      961 aagctgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt
     1021 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     1081 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca
     1141 gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg ctagcctcga
     1201 gaattcgcca ccatggagac agacacactc ctgctatggg tactgctgct ctgggttcca
     1261 ggttccactg gtgacacaag tttgtacaaa aaagttggca ccgccgaggt gctgtggccg
     1321 gctgtcccca acgggacgga cgctgccttc ctggccggtc cgggttcgtc ctgggggaac
     1381 agcacggtcg cctccactgc cgccgtctcc tcgtcgttca aatgcgcctt gaccaagacg
     1441 ggcttccagt tttactacct gccggctgtc tacatcttgg tattcatcat cggcttcctg
     1501 ggcaacagcg tggccatctg gatgttcgtc ttccacatga agccctggag cggcatctcc
     1561 gtgtacatgt tcaatttggc tctggccgac ttcttgtacg tgctgactct gccagccctg
     1621 atcttctact acttcaataa aacagactgg atcttcgggg atgccatgtg taaactgcag
     1681 aggttcatct ttcatgtgaa cctctatggc agcatcttgt ttctgacatg catcagtgcc
     1741 caccggtaca gcggtgtggt gtaccccctc aagtccctgg gccggctcaa aaagaagaat
     1801 gcgatctgta tcagcgtgct ggtgtggctc attgtggtgg tggcgatctc ccccatcctc
     1861 ttctactcag gtaccggggt ccgcaaaaac aaaaccatca cctgttacga caccacctca
     1921 gacgagtacc tgcgaagtta tttcatctac agcatgtgca cgaccgtggc catgttctgt
     1981 gtccccttgg tgctgattct gggctgttac ggattaattg tgagagcttt gatttacccc
     2041 gcgccggccg ggccgcgcga caccgacgcg ctggacctgg aggagggagg aaacgtctat
     2101 atcaaggccg acaagcagaa gaacggcatc aaggcgaact tccacatccg ccacaacatc
     2161 gaggacggcg gcgtgcagct cgcctaccac taccagcaga acacccccat cggcgacggc
     2221 cccgtgctgc tgcccgacaa ccactacctg agcgtgcagt ccaaactttc gaaagacccc
     2281 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2341 ggcatggacg agctgtacaa gggcggtacc ggagggagca tggtgagaaa gggcgaggag
     2401 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2461 ttcagcgtgt ccggcgaggg tgagggcgat gccaccgagg gcaagctgac cctgaagttc
     2521 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     2581 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2641 gccatgcccg aaggctacat ccaggagcgc accatcttct tcaaggacga cggcaactac
     2701 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2761 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacaccgga
     2821 gcagcagcac gctggcgcgg gcggtcgatt tacctggtaa tcattgtact gactgttttt
     2881 gctgtgtctt acatcccttt ccatgtgatg aaaacgatgg ctttgagggc ccggcttgat
     2941 tttcagaccc cagcaatgtg tgctttcaat gacagggttt atgccacgta tcaggtgaca
     3001 agaggtctag caagtctcaa cagttgtgtg gaccccattc tctatttctt ggcgggagat
     3061 actttcagaa ggagactctc ccgagccaca aggaaagctt ctagaagaag tgaggcaaat
     3121 ttgcaatcca agagtgaaga catgaccctc aatattttac ctgagttcaa gcagaatgga
     3181 gatacaagcc tgtaagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
     3241 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3301 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3361 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3421 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3481 tgtcagctcc tttccggaac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3541 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3601 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
     3661 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3721 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3781 cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg
     3841 ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag
     3901 tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct
     3961 tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa
     4021 cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc
     4081 tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt
     4141 tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg
     4201 gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt
     4261 ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga
     4321 ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     4381 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4441 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     4501 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     4561 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4621 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     4681 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4741 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     4801 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4861 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     4921 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     4981 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     5041 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5101 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5161 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5221 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5281 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5341 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5401 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5461 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5521 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5581 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5641 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5701 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5761 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5821 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5881 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5941 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6001 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6061 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6121 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6181 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6241 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6301 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6361 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6421 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6481 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6541 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6601 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6661 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6721 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6781 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6841 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6901 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6961 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7021 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7081 tttgctcaca tgt
//
