LOCUS       sequence-296619-        7380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the adenosine-insensitive control sensor GRAB_Ado1.0mut in 
            astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7380)
  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 7380)
  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-GfaABC1D-GRAB_Ado1.0mut
FEATURES             Location/Qualifiers
     source          1..7380
                     /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
     intron          1006..1138
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1121..1140
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1183..1201
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1218..1227
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1224..1283
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    1287..1311
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2369..2390
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2496..2517)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2757..2776)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3507..4095
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3560..3580)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3978..3989)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4127..4603
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4326..4345)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4643..4783
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4643..4772
                     /label=AAV2 ITR
     rep_origin      4858..5313
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4945..4964)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5155..5176
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5330..5349)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5449..5471
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5509..5527)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5595..5699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5700..5768,5769..6560)
                     /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      6731..7319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7220..7239
                     /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 cgcgttgtag ttaatgatta acccgccatg
      181 ctacttatct acgtagccat gctctaggaa gatctaacat atcctggtgt ggagtagggg
      241 acgctgctct gacagaggct cgggggcctg agctggctct gtgagctggg gaggaggcag
      301 acagccaggc cttgtctgca agcagacctg gcagcattgg gctggccgcc ccccagggcc
      361 tcctcttcat gcccagtgaa tgactcacct tggcacagac acaatgttcg gggtgggcac
      421 agtgcctgct tcccgccgca ccccagcccc cctcaaatgc cttccgagaa gcccattgag
      481 cagggggctt gcattgcacc ccagcctgac agcctggcat cttgggataa aagcagcaca
      541 gccccctagg ggctgccctt gctgtgtggc gccaccggcg gtggagaaca aggctctatt
      601 cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc atggacagtg ggtggcaggg
      661 ggggagagga gggctgtctg cttcccagaa gtccaaggac acaaatgggt gaggggagag
      721 ctctccccat agctgggctg cggcccaacc ccaccccctc aggctatgcc agggggtgtt
      781 gccaggggca cccgggcatc gccagtctag cccactcctt cataaagccc tcgcatccca
      841 ggagcgagca gagccagagc aggttggaga ggagacgcat cacctccgct gctcgcaagc
      901 tttattgcgg tagtttatca cagttaaatt gctaacgcag tcagtgcttc tgacacaaca
      961 gtctcgaact taagctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt
     1021 acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg
     1081 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt
     1141 gtccactccc agttcaatta cagctcttaa ggctagagta cttaatacga ctcactatag
     1201 gctagcctcg agaattcgcc accatggaga cagacacact cctgctatgg gtactgctgc
     1261 tctgggttcc aggttccact ggtgacacaa gtttgtacaa aaaagttggc accaccggtc
     1321 ccatcatggg ctcctcggtg tacatcacgg tggagctggc cattgctgtg ctggccatcc
     1381 tgggcaatgt gctggtgtgc tgggccgtgt ggctcaacag caacctgcag aacgtcacca
     1441 actactttgt ggtgtcactg gcggcggccg acatcgcagt gggtgtgctc gccatcccct
     1501 ttgccatcac catcagcacc gggttctgcg ctgcctgcca cggctgcctc ttcattgcct
     1561 gcttcgtcct ggtcctcacg cagagctcca tcttcagtct cctggccatc gccattgacc
     1621 gctacattgc catccgcatc ccgctccggt acaatggctt ggtgaccggc acgagggcta
     1681 agggcatcat tgccatctgc tgggtgctgt cgtttgccat cggcctgact cccatgctag
     1741 gttggaacaa ctgcggtcag ccaaaggagg gcaagaacca ctcccagggc tgcggggagg
     1801 gccaagtggc ctgtctcgct gaggatgtgg tccccatgaa ctacatggtg tacttcaact
     1861 tctttgcctg tgtgctggtg cccctgctgc tcatgctggg tgtctatttg cggatcttcc
     1921 tggcggcgcg acgacagctg aagcagatgg agagccagcc tcccagccgc cggggtccgg
     1981 acgccgtcgc cgcgccgccg gggggcaccg agcgcaggcc caacggtctg ggccccgagc
     2041 gcagcgcggg cccggggggc gcagaggccg aaccgctgcc cacccagctc aacggcgccc
     2101 ctggcgagcc cgcgccggcc gggccgcgcg acaccgacgc gctggacctg gaggagggag
     2161 gaaacgtcta tatcaaggcc gacaagcaga agaacggcat caaggcgaac ttccacatcc
     2221 gccacaacat cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag aacaccccca
     2281 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcgtgcag tccaaacttt
     2341 cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2401 ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc atggtgagaa
     2461 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2521 acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac ggcaagctga
     2581 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2641 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2701 tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc ttcaaggacg
     2761 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2821 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2881 acaacaccgg agcagcagca cgctggcgcg ggcggcagaa ccgcgagctc catgctgcca
     2941 agtcactggc catcattgtg gggctctttg ccctctgctg gctgccccta cacatcatca
     3001 actgcttcac tttcttctgc cccgactgca gccacgcccc tctctggctc atgtacctgg
     3061 ccatcgtcct ctcccacacc aattcggttg tgaatccctt catctacgcc taccgtatcc
     3121 gcgagttccg ccagaccttc cgcaagatca ttcgcagcca cgtcctgagg cagcaagaac
     3181 ctttcaaggc agctggcacc agtgcccggg tcttggcagc tcatggcagt gacggagagc
     3241 aggtcagcct ccgtctcaac ggccacccgc caggagtgtg ggccaacggc agtgctcccc
     3301 accctgagcg gaggcccaat ggctacgccc tggggctggt gagtggaggg agtgcccaag
     3361 agtcccaggg gaacacgggc ctcccagacg tggagctcct tagccatgag ctcaagggag
     3421 tgtgcccaga gccccctggc ctagatgacc ccctggccca ggatggagca ggagtgtcct
     3481 gagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     3541 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3601 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3661 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3721 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3781 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3841 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3901 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     3961 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     4021 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     4081 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
     4141 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     4201 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     4261 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     4321 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     4381 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     4441 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     4501 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     4561 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     4621 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     4681 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4741 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     4801 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4861 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4921 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4981 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5041 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     5101 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5161 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     5221 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5281 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     5341 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     5401 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     5461 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     5521 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5581 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5641 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5701 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5761 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5821 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5881 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5941 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6001 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6061 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6121 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6181 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6241 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6301 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6361 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6421 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6481 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6541 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6601 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6661 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6721 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6781 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6841 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     6901 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6961 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7021 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7081 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7141 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7201 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7261 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7321 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
