LOCUS       sequence-296616-        6754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the adenosine-insensitive control sensor GRAB_Ado1.0mut in 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6754)
  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 6754)
  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_Ado1.0mut
FEATURES             Location/Qualifiers
     source          1..6754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      592..601
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     598..657
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    661..685
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1743..1764
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1870..1891)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2131..2150)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2881..3469
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2934..2954)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3352..3363)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3501..3977
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3700..3719)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4017..4157
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4017..4146
                     /label=AAV2 ITR
     rep_origin      4232..4687
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4319..4338)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4529..4550
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4704..4723)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4823..4845
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4883..4901)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4969..5073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5074..5142,5143..5934)
                     /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(5292..5311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6105..6693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6594..6613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtatgag gcggggtggg ggtgcctacc
      181 tgacgaccga ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc
      241 ccctatcaga gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt
      301 cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca
      361 ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg
      421 gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg
      481 ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg
      541 tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc gagaaaccgg tgccaccatg
      601 gagacagaca cactcctgct atgggtactg ctgctctggg ttccaggttc cactggtgac
      661 acaagtttgt acaaaaaagt tggcaccacc ggtcccatca tgggctcctc ggtgtacatc
      721 acggtggagc tggccattgc tgtgctggcc atcctgggca atgtgctggt gtgctgggcc
      781 gtgtggctca acagcaacct gcagaacgtc accaactact ttgtggtgtc actggcggcg
      841 gccgacatcg cagtgggtgt gctcgccatc ccctttgcca tcaccatcag caccgggttc
      901 tgcgctgcct gccacggctg cctcttcatt gcctgcttcg tcctggtcct cacgcagagc
      961 tccatcttca gtctcctggc catcgccatt gaccgctaca ttgccatccg catcccgctc
     1021 cggtacaatg gcttggtgac cggcacgagg gctaagggca tcattgccat ctgctgggtg
     1081 ctgtcgtttg ccatcggcct gactcccatg ctaggttgga acaactgcgg tcagccaaag
     1141 gagggcaaga accactccca gggctgcggg gagggccaag tggcctgtct cgctgaggat
     1201 gtggtcccca tgaactacat ggtgtacttc aacttctttg cctgtgtgct ggtgcccctg
     1261 ctgctcatgc tgggtgtcta tttgcggatc ttcctggcgg cgcgacgaca gctgaagcag
     1321 atggagagcc agcctcccag ccgccggggt ccggacgccg tcgccgcgcc gccggggggc
     1381 accgagcgca ggcccaacgg tctgggcccc gagcgcagcg cgggcccggg gggcgcagag
     1441 gccgaaccgc tgcccaccca gctcaacggc gcccctggcg agcccgcgcc ggccgggccg
     1501 cgcgacaccg acgcgctgga cctggaggag ggaggaaacg tctatatcaa ggccgacaag
     1561 cagaagaacg gcatcaaggc gaacttccac atccgccaca acatcgagga cggcggcgtg
     1621 cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1681 gacaaccact acctgagcgt gcagtccaaa ctttcgaaag accccaacga gaagcgcgat
     1741 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1801 tacaagggcg gtaccggagg gagcatggtg agaaagggcg aggagctgtt caccggggtg
     1861 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1921 gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1981 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2041 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2101 tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2161 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2221 gaggacggca acatcctggg gcacaagctg gagtacaaca ccggagcagc agcacgctgg
     2281 cgcgggcggc agaaccgcga gctccatgct gccaagtcac tggccatcat tgtggggctc
     2341 tttgccctct gctggctgcc cctacacatc atcaactgct tcactttctt ctgccccgac
     2401 tgcagccacg cccctctctg gctcatgtac ctggccatcg tcctctccca caccaattcg
     2461 gttgtgaatc ccttcatcta cgcctaccgt atccgcgagt tccgccagac cttccgcaag
     2521 atcattcgca gccacgtcct gaggcagcaa gaacctttca aggcagctgg caccagtgcc
     2581 cgggtcttgg cagctcatgg cagtgacgga gagcaggtca gcctccgtct caacggccac
     2641 ccgccaggag tgtgggccaa cggcagtgct ccccaccctg agcggaggcc caatggctac
     2701 gccctggggc tggtgagtgg agggagtgcc caagagtccc aggggaacac gggcctccca
     2761 gacgtggagc tccttagcca tgagctcaag ggagtgtgcc cagagccccc tggcctagat
     2821 gaccccctgg cccaggatgg agcaggagtg tcctgagaat tcgatatcaa gcttatcgat
     2881 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2941 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3001 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3061 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3121 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3181 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3241 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     3301 gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3361 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3421 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga
     3481 gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     3541 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     3601 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     3661 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     3721 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     3781 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     3841 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     3901 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     3961 ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga
     4021 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4081 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4141 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4201 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     4261 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4321 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4381 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4441 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4501 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4561 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4621 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     4681 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4741 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4801 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4861 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4921 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4981 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5041 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5101 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5161 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5221 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5281 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5341 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5401 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5461 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5521 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5581 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5641 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5701 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5761 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5821 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5881 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5941 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6001 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6061 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6121 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6181 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6241 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6301 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6361 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6421 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6481 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6541 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6601 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6661 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6721 ttcctggcct tttgctggcc ttttgctcac atgt
//
