LOCUS       sequence-325521-        6305 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 6305)
  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 6305)
  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..6305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..142
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   13..142
                     /label=AAV2 ITR
     promoter        163..374
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    complement(393..426)
                     /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(477..510)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1011..1030
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1270..1291
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1397..1418)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2444..2503)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2500..2509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2516..2549
                     /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    2600..2633
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2658..3246
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2711..2731)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3129..3140)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3278..3754
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3477..3496)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3794..3934
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3794..3923
                     /label=AAV2 ITR
     rep_origin      4009..4464
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4096..4115)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4306..4327
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4481..4500)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4600..4622
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4660..4678)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4746..4850
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4851..4919,4920..5711)
                     /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(5069..5088)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
       61 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctgcggccgc acgcgtaagc ttgggcagag cgcacatcgc
      181 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
      241 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
      301 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
      361 ccgccagaac acagtctaga gtcgactccg gaataacttc gtataggata ctttatacga
      421 agttatgcag aatggtagct ggattgtagc tgctattagc aatatgaaac ctcttaataa
      481 cttcgtatag catacattat acgaagttat ggcgcatatg ttacaggctt gtatctccat
      541 tctgcttgaa ctcaggtaaa atattgaggg tcatgtcttc actcttggat tgcaaatttg
      601 cctcacttct tctagaagct ttccttgtgg ctcgggagag tctccttctg aaagtatctc
      661 ccgccaagaa atagagaatg gggtccacac aactgttgag acttgctaga cctcttgtca
      721 cctgatacgt ggcataaacc ctgtcattga aagcacacat tgctggggtc tgaaaatcaa
      781 gccgggccct caagttcatc gttttcatca catggaaagg gatgtaagac acagcaaaaa
      841 cagtcagtac aatgattacc aggtaaatcg accgcccgcg ccagcgtgct gctgctccgg
      901 tgttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
      961 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     1021 tgccgtcgtc cttgaagaag atggtgcgct cctggatgta gccttcgggc atggcggact
     1081 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
     1141 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
     1201 tcagggtcag cttgccctcg gtggcatcgc cctcaccctc gccggacacg ctgaacttgt
     1261 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     1321 cgccctttct caccatgctc cctccggtac cgcccttgta cagctcgtcc atgccgagag
     1381 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
     1441 ctttcgaaag tttggactgc acgctcaggt agtggttgtc gggcagcagc acggggccgt
     1501 cgccgatggg ggtgttctgc tggtagtggt aggcgagctg cacgccgccg tcctcgatgt
     1561 tgtggcggat gtggaagttc gccttgatgc cgttcttctg cttgtcggcc ttgatataga
     1621 cgtttcctcc ctcctccagg tccagcgcgt cggtgtcgcg cggcccggcc ggcgcggggt
     1681 aaatcaaagc tctcacaatt aatccgtaac agcccagaat cagcaccaag gggacacaga
     1741 acatggccac ggtcgtgcac atgctgtaga tgaaataact tcgcaggtac tcgtctgagg
     1801 tggtgtcgta acaggtgatg gttttgtttt tgcggacccc ggtacctgag tagaagagga
     1861 tgggggagat cgccaccacc acaatgagcc acaccagcac gctgatacag atcgcattct
     1921 tctttttgag ccggcccagg gacttgaggg ggtacaccac accgctgtac cggtgggcac
     1981 tgatgcatgt cagaaacaag atgctgccat agaggttcac atgaaagatg aacctctgca
     2041 gtttacacat ggcatccccg aagatccagt ctgttttatt gaagtagtag aagatcaggg
     2101 ctggcagagt cagcacgtac aagaagtcgg ccagagccaa attgaacatg tacacggaga
     2161 tgccgctcca gggcttcatg tggaagacga acatccagat ggccacgctg ttgcccagga
     2221 agccgatgat gaataccaag atgtagacag ccggcaggta gtaaaactgg aagcccgtct
     2281 tggtcaaggc gcatttgaac gacgaggaga cggcggcagt ggaggcgacc gtgctgttcc
     2341 cccaggacga acccggaccg gccaggaagg cagcgtccgt cccgttgggg acagccggcc
     2401 acagcacctc ggtggtgcca actttttcgc acaaacttgt gtcaccagtg gaacctggaa
     2461 cccagagcag cagtacccat agcaggagtg tgtctgtctc catggtggcg ctagcataac
     2521 ttcgtataaa gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt
     2581 ctttagaatg gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcg
     2641 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2701 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2761 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2821 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2881 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     2941 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3001 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     3061 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     3121 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3181 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3241 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     3301 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     3361 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     3421 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     3481 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     3541 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     3601 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     3661 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     3721 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     3781 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3841 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3901 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     3961 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4021 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4081 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4141 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4201 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4261 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4321 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     4381 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4441 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4501 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4561 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4621 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4681 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4741 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4801 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4861 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4921 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4981 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5041 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5101 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5161 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5221 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5281 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5341 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5401 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5461 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5521 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5581 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5641 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5701 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5761 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5821 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5881 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5941 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6001 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6061 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6121 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6181 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6241 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6301 ggaga
//
