LOCUS       sequence-305682-        6743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the adenosine sensor GRAB_Ado1.0 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6743)
  AUTHORS   Peng W, Wu Z, Song K, Zhang S, Li Y, Xu M
  TITLE     Regulation of sleep homeostasis mediator adenosine by basal 
            forebrain glutamatergic neurons.
  JOURNAL   Science. 2020 Sep 4;369(6508). pii: 369/6508/eabb0556. doi: 
            10.1126/science.abb0556.
  PUBMED    32883833
REFERENCE   2  (bases 1 to 6743)
  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: "Science."; 
            date: "2020-09-4"; volume: "369(6508). pii: 369/6508/eabb0556. doi";
            pages: " 10.1126/science.abb0556"
COMMENT     Sequence Label: pAAV-hsyn-GRAB_Ado1.0
FEATURES             Location/Qualifiers
     source          1..6743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      499..508
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     505..564
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    568..592
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1650..1671
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1777..1798)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2038..2057)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2788..3376
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2841..2861)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3259..3270)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3408..3884
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3607..3626)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3924..4064
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3924..4053
                     /label=AAV2 ITR
     rep_origin      4139..4594
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4226..4245)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4436..4457
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4611..4630)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4730..4752
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4790..4808)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4876..4980
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4981..5049,5050..5841)
                     /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(5199..5218)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6012..6600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6501..6520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6662..6743,1..48)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcacgc
       61 gtatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
      121 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
      181 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
      241 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
      301 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
      361 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
      421 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
      481 cgcagtcgag aaaccggtgc caccatggag acagacacac tcctgctatg ggtactgctg
      541 ctctgggttc caggttccac tggtgacaca agtttgtaca aaaaagttgg caccaccggt
      601 cccatcatgg gctcctcggt gtacatcacg gtggagctgg ccattgctgt gctggccatc
      661 ctgggcaatg tgctggtgtg ctgggccgtg tggctcaaca gcaacctgca gaacgtcacc
      721 aactactttg tggtgtcact ggcggcggcc gacatcgcag tgggtgtgct cgccatcccc
      781 tttgccatca ccatcagcac cgggttctgc gctgcctgcc acggctgcct cttcattgcc
      841 tgcttcgtcc tggtcctcac gcagagctcc atcttcagtc tcctggccat cgccattgac
      901 cgctacattg ccatccgcat cccgctccgg tacaatggct tggtgaccgg cacgagggct
      961 aagggcatca ttgccatctg ctgggtgctg tcgtttgcca tcggcctgac tcccatgcta
     1021 ggttggaaca actgcggtca gccaaaggag ggcaagaacc actcccaggg ctgcggggag
     1081 ggccaagtgg cctgtctctt tgaggatgtg gtccccatga actacatggt gtacttcaac
     1141 ttctttgcct gtgtgctggt gcccctgctg ctcatgctgg gtgtctattt gcggatcttc
     1201 ctggcggcgc gacgacagct gaagcagatg gagagccagc ctcccagccg ccggggtccg
     1261 gacgccgtcg ccgcgccgcc ggggggcacc gagcgcaggc ccaacggtct gggccccgag
     1321 cgcagcgcgg gcccgggggg cgcagaggcc gaaccgctgc ccacccagct caacggcgcc
     1381 cctggcgagc ccgcgccggc cgggccgcgc gacaccgacg cgctggacct ggaggaggga
     1441 ggaaacgtct atatcaaggc cgacaagcag aagaacggca tcaaggcgaa cttccacatc
     1501 cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc
     1561 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccaaactt
     1621 tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1681 gggatcactc tcggcatgga cgagctgtac aagggcggta ccggagggag catggtgaga
     1741 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1801 aacggccaca agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg
     1861 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1921 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1981 ttcttcaagt ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaaggac
     2041 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     2101 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     2161 tacaacaccg gagcagcagc acgctggcgc gggcggcaga accgcgagct ccatgctgcc
     2221 aagtcactgg ccatcattgt ggggctcttt gccctctgct ggctgcccct acacatcatc
     2281 aactgcttca ctttcttctg ccccgactgc agccacgccc ctctctggct catgtacctg
     2341 gccatcgtcc tctcccacac caattcggtt gtgaatccct tcatctacgc ctaccgtatc
     2401 cgcgagttcc gccagacctt ccgcaagatc attcgcagcc acgtcctgag gcagcaagaa
     2461 cctttcaagg cagctggcac cagtgcccgg gtcttggcag ctcatggcag tgacggagag
     2521 caggtcagcc tccgtctcaa cggccacccg ccaggagtgt gggccaacgg cagtgctccc
     2581 caccctgagc ggaggcccaa tggctacgcc ctggggctgg tgagtggagg gagtgcccaa
     2641 gagtcccagg ggaacacggg cctcccagac gtggagctcc ttagccatga gctcaaggga
     2701 gtgtgcccag agccccctgg cctagatgac cccctggccc aggatggagc aggagtgtcc
     2761 tgagaattcg atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     2821 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     2881 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     2941 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3001 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3061 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3121 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3181 aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg
     3241 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     3301 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     3361 tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct
     3421 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     3481 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     3541 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     3601 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     3661 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     3721 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     3781 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     3841 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa
     3901 ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     3961 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     4021 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     4081 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     4141 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4201 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4261 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4321 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     4381 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4441 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     4501 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4561 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     4621 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     4681 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4741 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     4801 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4861 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4921 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4981 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5041 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5101 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5161 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5221 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5281 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5341 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5401 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5461 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5521 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5581 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5641 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5701 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5761 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5821 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5881 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5941 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6001 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6061 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6121 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6181 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6241 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6301 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6361 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6421 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6481 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6541 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6601 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6661 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
     6721 tggtcgcccg gcctcagtga gcg
//
