LOCUS       sequence-412468-        6533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent dopamine (DA) sensor 
            GRAB_rDA3h in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6533)
  AUTHORS   Zhuo Y, Luo B, Yi X, Dong H, Miao X, Wan J, Williams JT, Campbell 
            MG, Cai R, Qian T, Li F, Weber SJ, Wang L, Li B, Wei Y, Li G, Wang 
            H, Zheng Y, Zhao Y, Wolf ME, Zhu Y, Watabe-Uchida M, Li Y
  TITLE     Improved green and red GRAB sensors for monitoring dopaminergic 
            activity in vivo.
  JOURNAL   Nat Methods. 2023 Nov 30. doi: 10.1038/s41592-023-02100-w.
  PUBMED    38036855
REFERENCE   2  (bases 1 to 6533)
  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: "Nat 
            Methods. 2023 Nov 30. doi: 10.1038/s41592-023-02100-w."
COMMENT     Sequence Label: pAAV-hSyn-GRAB-rDA3h
FEATURES             Location/Qualifiers
     source          1..6533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    2653..3241
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2706..2726)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3124..3135)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3273..3749
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3472..3491)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3789..3929
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3789..3918
                     /label=AAV2 ITR
     rep_origin      4004..4459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4091..4110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4301..4322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4476..4495)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4595..4617
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4655..4673)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4741..4845
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4846..4914,4915..5706)
                     /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(5064..5083)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5877..6465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6366..6385
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6527..6533,1..123)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc
       61 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
      121 cctgcggccg cacgcgtatg aggcggggtg ggggtgccta cctgacgacc gaccccgacc
      181 cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca gagaggggga
      241 ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc
      301 cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt
      361 cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc cgcgccgccg
      421 ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc
      481 tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg
      541 tgtcgtgcct gagagcgcag tcgagaaacc ggtgccacca tgaagacgat catcgccctg
      601 agctacatct tctgcttggt gttcgccgac tacaaggacg atgatgacgc catgaggact
      661 ctgaacacct ctgccatgga cgggactggg ctggtggtgg agagggactt ctctgttcgt
      721 atcctcactg cctgtttcct gtcgctgctc atcctgtcca cgctcctggg gaacacgctg
      781 gtctgtgctg ccgttatcag gttccgacac ctgcggtcca aggtgaccaa cttctttgtc
      841 atctccttgg ctgtgtcaga tctcttgatc gccgtcctgg tcatgccctg gaaggcagtg
      901 gctgagattg ctggcttctg gccctttggg tccttctgta acatctgggt ggcctttgac
      961 atcatgtgct ccactgcatc catcctcaac ctctgtgtga tcagcgtgga caggtattgg
     1021 gctatctcca gccctgagcg gtatgagaga aagatgaccc ccaaggcagc cttcatcctg
     1081 atcagtgtgg catggacctt gtctgtactc atctccttca tcccagtgca gctcagctgg
     1141 cacaaggcaa aacccacaag cccctctgat ggaaatgcca cttccctggc tgagaccata
     1201 gacaactgtg actccagcct cagcaggaca tatgccatct catcctctgt aataagcttt
     1261 tacatccctg tggccatcat gattgtctgc tacaccagga tctacaggat tgctcagaaa
     1321 aagctctcct ccttcatggt ttccgagcgg atgtaccccg aggacggcgc cctgaagagc
     1381 gagatcaaga aggggctgag gctgaaggac ggcggccact acgccgccga ggtcaagacc
     1441 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acatcgtgga catcaagttg
     1501 gacatcgtgt cccacaacga ggactacacc atcgtggaac agtgcgaacg cgccgagggc
     1561 cgccactcca ccggcggcat ggacgagctg tacaagggag gtacaggcgg gagtctggtg
     1621 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1681 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1741 tacgaggcct ttcagaccgc taagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1801 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacattaa gcacccagcc
     1861 gacatccccg actacttcaa gtcgtccttc cccgagggct tcaggtggga gcgcgtgatg
     1921 aacttcgagg acggcggcat tattcacgtg aaccaggact cctccctgca ggacggcgta
     1981 ttcatctaca aggtgaagct gcgcggcacc aacttccccc ccgacggccc cgtaatgcag
     2041 aagaaaacta tggggtggga acatcggaag gacgacctta aaagagaaac taaagtcctg
     2101 aagactctgt cggtgatcat gggtgtgttt gtgtgctgtt ggctaccttt cttcatcttg
     2161 aactgcattt tgcccttctg tgggtctggg gagacgcagc ccttctgcat tgattccaac
     2221 acctttgacg tgtttgtgtg gtttgggtgg gctaattcat ccttgaaccc catcatttat
     2281 gcctttaatg ctgattttcg gaaggcattt tcaaccctct taggatgcta cagactttgc
     2341 cctgcgacga ataatgccat agagacggtg agtatcaata acaatggggc cgcgatgttt
     2401 tccagccatc atgagccacg aggctccatc tccaaggagt gcaatctggt ttacctgatc
     2461 ccacatgctg tgggctcctc tgaggacctg aaaaaggagg aggcagctgg catcgccaga
     2521 cccttggaga agctgtcccc agccctatcg gtcatattgg actatgacac tgacgtctct
     2581 ctggagaaga tccaacccat cacacaaaac ggtcagcacc caacctgaga attcgatatc
     2641 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2701 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2761 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2821 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2881 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2941 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3001 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     3061 ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3121 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3181 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3241 catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag
     3301 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     3361 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     3421 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     3481 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     3541 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     3601 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     3661 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     3721 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga
     3781 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     3841 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     3901 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     3961 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     4021 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4081 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4141 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4201 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4261 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4321 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     4381 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4441 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4501 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4561 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4621 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4681 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4741 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4801 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4861 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4921 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4981 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5041 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5101 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5161 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5221 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5281 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5341 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5401 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5461 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5521 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5581 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5641 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5701 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5761 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5821 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5881 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5941 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6001 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6061 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6121 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6181 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6241 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6301 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6361 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6421 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6481 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg cag
//
