LOCUS       sequence-304900-        6743 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 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_Ado1.0mut
FEATURES             Location/Qualifiers
     source          1..6743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      553..562
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     559..618
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    622..646
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1704..1725
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1831..1852)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2092..2111)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2842..3430
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2895..2915)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3313..3324)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3462..3938
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3661..3680)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3978..4118
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3978..4107
                     /label=AAV2 ITR
     rep_origin      4193..4648
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4280..4299)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4490..4511
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4665..4684)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4784..4806
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4844..4862)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4930..5034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5035..5103,5104..5895)
                     /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(5253..5272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6066..6654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6555..6574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6716..6743,1..102)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
       61 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtatga
      121 ggcggggtgg gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc
      181 ccattcccca aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag
      241 gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg
      301 cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa
      361 ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc
      421 acgcgaggcg cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact
      481 cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt
      541 cgagaaaccg gtgccaccat ggagacagac acactcctgc tatgggtact gctgctctgg
      601 gttccaggtt ccactggtga cacaagtttg tacaaaaaag ttggcaccac cggtcccatc
      661 atgggctcct cggtgtacat cacggtggag ctggccattg ctgtgctggc catcctgggc
      721 aatgtgctgg tgtgctgggc cgtgtggctc aacagcaacc tgcagaacgt caccaactac
      781 tttgtggtgt cactggcggc ggccgacatc gcagtgggtg tgctcgccat cccctttgcc
      841 atcaccatca gcaccgggtt ctgcgctgcc tgccacggct gcctcttcat tgcctgcttc
      901 gtcctggtcc tcacgcagag ctccatcttc agtctcctgg ccatcgccat tgaccgctac
      961 attgccatcc gcatcccgct ccggtacaat ggcttggtga ccggcacgag ggctaagggc
     1021 atcattgcca tctgctgggt gctgtcgttt gccatcggcc tgactcccat gctaggttgg
     1081 aacaactgcg gtcagccaaa ggagggcaag aaccactccc agggctgcgg ggagggccaa
     1141 gtggcctgtc tcgctgagga tgtggtcccc atgaactaca tggtgtactt caacttcttt
     1201 gcctgtgtgc tggtgcccct gctgctcatg ctgggtgtct atttgcggat cttcctggcg
     1261 gcgcgacgac agctgaagca gatggagagc cagcctccca gccgccgggg tccggacgcc
     1321 gtcgccgcgc cgccgggggg caccgagcgc aggcccaacg gtctgggccc cgagcgcagc
     1381 gcgggcccgg ggggcgcaga ggccgaaccg ctgcccaccc agctcaacgg cgcccctggc
     1441 gagcccgcgc cggccgggcc gcgcgacacc gacgcgctgg acctggagga gggaggaaac
     1501 gtctatatca aggccgacaa gcagaagaac ggcatcaagg cgaacttcca catccgccac
     1561 aacatcgagg acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc
     1621 gacggccccg tgctgctgcc cgacaaccac tacctgagcg tgcagtccaa actttcgaaa
     1681 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1741 actctcggca tggacgagct gtacaagggc ggtaccggag ggagcatggt gagaaagggc
     1801 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1861 cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca cctacggcaa gctgaccctg
     1921 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1981 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2041 aagtccgcca tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc
     2101 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2161 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2221 accggagcag cagcacgctg gcgcgggcgg cagaaccgcg agctccatgc tgccaagtca
     2281 ctggccatca ttgtggggct ctttgccctc tgctggctgc ccctacacat catcaactgc
     2341 ttcactttct tctgccccga ctgcagccac gcccctctct ggctcatgta cctggccatc
     2401 gtcctctccc acaccaattc ggttgtgaat cccttcatct acgcctaccg tatccgcgag
     2461 ttccgccaga ccttccgcaa gatcattcgc agccacgtcc tgaggcagca agaacctttc
     2521 aaggcagctg gcaccagtgc ccgggtcttg gcagctcatg gcagtgacgg agagcaggtc
     2581 agcctccgtc tcaacggcca cccgccagga gtgtgggcca acggcagtgc tccccaccct
     2641 gagcggaggc ccaatggcta cgccctgggg ctggtgagtg gagggagtgc ccaagagtcc
     2701 caggggaaca cgggcctccc agacgtggag ctccttagcc atgagctcaa gggagtgtgc
     2761 ccagagcccc ctggcctaga tgaccccctg gcccaggatg gagcaggagt gtcctgagaa
     2821 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     2881 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2941 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3001 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3061 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3121 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3181 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3241 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     3301 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3361 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3421 gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc
     3481 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     3541 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     3601 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     3661 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     3721 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     3781 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     3841 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     3901 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     3961 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     4021 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     4081 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     4141 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4201 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4261 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4321 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4381 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4441 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4501 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     4561 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4621 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     4681 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4741 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4801 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4861 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4921 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4981 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5041 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5101 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5161 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5221 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5281 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5341 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5401 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5461 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5521 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5581 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5641 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5701 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5761 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5821 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5881 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5941 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6001 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc
     6721 aggcagctgc gcgctcgctc gct
//
