LOCUS       sequence-304923-        6740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the medium-affinity adenosine sensor GRAB_Ado1.0med in 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6740)
  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 6740)
  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-hsyn-GRAB_Ado1.0med
FEATURES             Location/Qualifiers
     source          1..6740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     587..646
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    650..674
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1729..1750
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1856..1877)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2117..2136)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2867..3455
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2920..2940)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3338..3349)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3487..3963
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3686..3705)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4003..4143
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4003..4132
                     /label=AAV2 ITR
     rep_origin      4218..4673
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4305..4324)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4515..4536
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4690..4709)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4809..4831
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4869..4887)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4955..5059
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5060..5128,5129..5920)
                     /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(5278..5297)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6091..6679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6580..6599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtatgagg cggggtgggg gtgcctacct gacgaccgac
      181 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      241 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      301 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      361 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      421 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      481 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      541 ggagtcgtgt cgtgcctgag agcgcagtcg agaaaccggt gccaccatgg agacagacac
      601 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca caagtttgta
      661 caaaaaagtt ggcaccggtc ccatcatggg ctcctcggtg tacatcacgg tggagctggc
      721 cattgctgtg ctggccatcc tgggcaatgt gctggtgtgc tgggccgtgt ggctcaacag
      781 caacctgcag aacgtcacca actactttgt ggtgtcactg gcggcggccg acatcgcagt
      841 gggtgtgctc gccatcccct ttgccatcac catcagcacc gggttctgcg ctgcctgcca
      901 cggctgcctc ttcattgcct gcttcgtcct ggtcctcacg cagagctcca tcttcagtct
      961 cctggccatc gccattgacc gctacattgc catccgcatc ccgctccggt acaatggctt
     1021 ggtgaccggc acgagggcta agggcatcat tgccatctgc tgggtgctgt cgtttgccat
     1081 cggcctgact cccatgctag gttggaacaa ctgcggtcag ccaaaggagg gcaagaacca
     1141 ctcccagggc tgcggggagg gccaagtggc ctgtctcttt gaggatgtgg tccccatgaa
     1201 ctacatggtg tacttcaact tctttgcctg tgtgctggtg cccctgctgc tcatgctggg
     1261 tgtctatttg cggatcttcc tggcggcgcg acgacagctg aagcagatgg agagccagcc
     1321 tcccagccgc cggggtccgg acgccgtcgc cgcgccgccg gggggcaccg agcgcaggcc
     1381 caacggtctg ggccccgagc gcagcgcggg cccggggggc gcagaggccg aaccgctgcc
     1441 cacccagctc aacggcgccc ctggcgagcc cgcgccggcc gggccgcgcg acaccgacgc
     1501 gctggacctg gaggagggag gaaacgtcta tatcaaggcc gacaagcaga agaacggcat
     1561 caaggcgaac ttccacatcc gccacaacat cgaggacggc ggcgtgcagc tcgcctacca
     1621 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1681 gagcgtgcag tccaaacttt cgaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1741 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac
     1801 cggagggagc atggtgagaa agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1861 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga
     1921 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1981 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     2041 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg
     2101 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2161 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2221 cctggggcac aagctggagt acaacaccgg agcagcagca cgctggcgcg ggcggcagaa
     2281 ccgcgagctc catgctgcca agtcactggc catcattgtg gggctctttg ccctctgctg
     2341 gctgccccta cacatcatca actgcttcac tttcttctgc cccgactgca gccacgcccc
     2401 tctctggctc cactacctgg ccatcgtcct ctcccacacc aattcggttg tgaatccctt
     2461 catctacgcc taccgtatcc gcgagttccg ccagaccttc cgcaagatca ttcgcagcca
     2521 cgtcctgagg cagcaagaac ctttcaaggc agctggcacc agtgcccggg tcttggcagc
     2581 tcatggcagt gacggagagc aggtcagcct ccgtctcaac ggccacccgc caggagtgtg
     2641 ggccaacggc agtgctcccc accctgagcg gaggcccaat ggctacgccc tggggctggt
     2701 gagtggaggg agtgcccaag agtcccaggg gaacacgggc ctcccagacg tggagctcct
     2761 tagccatgag ctcaagggag tgtgcccaga gccccctggc ctagatgacc ccctggccca
     2821 ggatggagca ggagtgtcct gagaattcga tatcaagctt atcgataatc aacctctgga
     2881 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2941 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3001 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3061 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3121 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3181 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3241 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac
     3301 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3361 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3421 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag
     3481 atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     3541 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     3601 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     3661 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     3721 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     3781 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     3841 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     3901 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     3961 cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     4021 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4081 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     4141 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     4201 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4261 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     4321 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4381 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     4441 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4501 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     4561 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     4621 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     4681 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     4741 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4801 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4861 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4921 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4981 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5041 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5101 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5161 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5221 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5281 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5341 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5401 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5461 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5521 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5581 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5641 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5701 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5761 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5821 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5881 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5941 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6001 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6061 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6121 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6181 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6241 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6301 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6361 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6421 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6481 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6541 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6601 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6661 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6721 ctggcctttt gctcacatgt
//
