LOCUS       sequence-296618-        7377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the medium-affinity adenosine sensor GRAB_Ado1.0med in 
            astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7377)
  AUTHORS   Wu Z, Cui Y, Wang H, Wu H, Wan Y, Li B, Wang L, Pan S, Peng W, Dong 
            A, Yuan Z, Jing M, Xu M, Luo M, Li Y
  TITLE     Neuronal activity-induced, equilibrative nucleoside 
            transporter-dependent, somatodendritic adenosine release revealed by
            a GRAB sensor.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Apr 4;120(14):e2212387120. doi: 
            10.1073/pnas.2212387120. Epub 2023 Mar 30.
  PUBMED    36996110
REFERENCE   2  (bases 1 to 7377)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-04-4"; volume: "120(14):e2212387120. 
            doi"; pages: " 10.1073/pnas.2212387120. Epub 2023 Mar 30"
COMMENT     Sequence Label: pAAV-GfaABC1D-GRAB_Ado1.0med
FEATURES             Location/Qualifiers
     source          1..7377
                     /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     2366..2387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2493..2514)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2754..2773)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3504..4092
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3557..3577)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3975..3986)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4124..4600
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4323..4342)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4640..4780
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4640..4769
                     /label=AAV2 ITR
     rep_origin      4855..5310
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4942..4961)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5152..5173
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5327..5346)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5446..5468
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5506..5524)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5592..5696
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5697..5765,5766..6557)
                     /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      6728..7316
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7217..7236
                     /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 accggtccca
     1321 tcatgggctc ctcggtgtac atcacggtgg agctggccat tgctgtgctg gccatcctgg
     1381 gcaatgtgct ggtgtgctgg gccgtgtggc tcaacagcaa cctgcagaac gtcaccaact
     1441 actttgtggt gtcactggcg gcggccgaca tcgcagtggg tgtgctcgcc atcccctttg
     1501 ccatcaccat cagcaccggg ttctgcgctg cctgccacgg ctgcctcttc attgcctgct
     1561 tcgtcctggt cctcacgcag agctccatct tcagtctcct ggccatcgcc attgaccgct
     1621 acattgccat ccgcatcccg ctccggtaca atggcttggt gaccggcacg agggctaagg
     1681 gcatcattgc catctgctgg gtgctgtcgt ttgccatcgg cctgactccc atgctaggtt
     1741 ggaacaactg cggtcagcca aaggagggca agaaccactc ccagggctgc ggggagggcc
     1801 aagtggcctg tctctttgag gatgtggtcc ccatgaacta catggtgtac ttcaacttct
     1861 ttgcctgtgt gctggtgccc ctgctgctca tgctgggtgt ctatttgcgg atcttcctgg
     1921 cggcgcgacg acagctgaag cagatggaga gccagcctcc cagccgccgg ggtccggacg
     1981 ccgtcgccgc gccgccgggg ggcaccgagc gcaggcccaa cggtctgggc cccgagcgca
     2041 gcgcgggccc ggggggcgca gaggccgaac cgctgcccac ccagctcaac ggcgcccctg
     2101 gcgagcccgc gccggccggg ccgcgcgaca ccgacgcgct ggacctggag gagggaggaa
     2161 acgtctatat caaggccgac aagcagaaga acggcatcaa ggcgaacttc cacatccgcc
     2221 acaacatcga ggacggcggc gtgcagctcg cctaccacta ccagcagaac acccccatcg
     2281 gcgacggccc cgtgctgctg cccgacaacc actacctgag cgtgcagtcc aaactttcga
     2341 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     2401 tcactctcgg catggacgag ctgtacaagg gcggtaccgg agggagcatg gtgagaaagg
     2461 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2521 gccacaagtt cagcgtgtcc ggcgagggtg agggcgatgc cacctacggc aagctgaccc
     2581 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2641 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     2701 tcaagtccgc catgcccgaa ggctacatcc aggagcgcac catcttcttc aaggacgacg
     2761 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2821 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2881 acaccggagc agcagcacgc tggcgcgggc ggcagaaccg cgagctccat gctgccaagt
     2941 cactggccat cattgtgggg ctctttgccc tctgctggct gcccctacac atcatcaact
     3001 gcttcacttt cttctgcccc gactgcagcc acgcccctct ctggctccac tacctggcca
     3061 tcgtcctctc ccacaccaat tcggttgtga atcccttcat ctacgcctac cgtatccgcg
     3121 agttccgcca gaccttccgc aagatcattc gcagccacgt cctgaggcag caagaacctt
     3181 tcaaggcagc tggcaccagt gcccgggtct tggcagctca tggcagtgac ggagagcagg
     3241 tcagcctccg tctcaacggc cacccgccag gagtgtgggc caacggcagt gctccccacc
     3301 ctgagcggag gcccaatggc tacgccctgg ggctggtgag tggagggagt gcccaagagt
     3361 cccaggggaa cacgggcctc ccagacgtgg agctccttag ccatgagctc aagggagtgt
     3421 gcccagagcc ccctggccta gatgaccccc tggcccagga tggagcagga gtgtcctgag
     3481 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     3541 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3601 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3661 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3721 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3781 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3841 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     3901 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct
     3961 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4021 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4081 ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga
     4141 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     4201 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     4261 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     4321 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     4381 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     4441 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     4501 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     4561 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac
     4621 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4681 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4741 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     4801 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     4861 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4921 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4981 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5041 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     5101 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5161 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     5221 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5281 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     5341 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5401 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5461 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5521 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5581 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5641 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5701 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5761 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5821 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5881 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5941 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6001 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6061 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6121 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6181 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6241 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6301 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6361 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6421 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6481 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6541 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6601 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6661 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6721 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6781 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6841 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     6901 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6961 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7021 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7081 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7141 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7201 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7261 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7321 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
