LOCUS       sequence-412373-        6623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent dopamine (DA) sensor 
            GRAB_gDA3h in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6623)
  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 6623)
  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-gDA3h
FEATURES             Location/Qualifiers
     source          1..6623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(97..201)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     269..287
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(325..347)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     447..466
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(483..938)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(620..641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     832..851
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1013..1153
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1024..1153
                     /label=AAV2 ITR
     polyA_signal    1193..1669
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1451..1470
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    1701..2289
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1807..1818
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2216..2236
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     3014..3033
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3273..3294
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3400..3421)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4365..4389)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     complement(4393..4452)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      4449..4458
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4909..5038
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(5100..5688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5180..5199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5859..6623,1..27,28..96))
                     /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     6482..6501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
       61 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
      121 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
      181 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
      241 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
      301 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
      361 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      421 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      481 taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      541 attttttaac caatagaccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
      601 gatagagttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      661 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      721 caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      781 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      841 agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      901 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg
      961 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg
     1021 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     1081 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     1141 actaggggtt cctgcggccg ctcggtccgc acgtggttac ctacaaaatc agaaggacag
     1201 ggaagggagc agtggttcac gcctgtaatc ccagcaattt gggaggccaa ggtgggtaga
     1261 tcacctgaga ttaggagttg gagaccagcc tggccaatat ggtgaaaccc cgtctctacc
     1321 aaaaaaacaa aaattagctg agcctggtca tgcatgcctg gaatcccaac aactcgggag
     1381 gctgaggcag gagaatcgct tgaacccagg aggcggagat tgcagtgagc caagattgtg
     1441 ccactgcact ccagcttggt tcccaataga ccccgcaggc cctacaggtt gtcttcccaa
     1501 cttgcccctt gctccatacc acccccctcc accccataat attatagaag gacacctagt
     1561 cagacaaaat gatgcaactt aattttatta ggacaaggct ggtgggcact ggagtggcaa
     1621 cttccagggc caggagaggc actggggagg ggtcacaggg atgccacccg tagatctctc
     1681 gagcagcgct cggtatcgat gcggggaggc ggcccaaagg gagatccgac tcgtctgagg
     1741 gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg aaggaaggtc
     1801 cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc
     1861 aacataggcg agcagccaag gaaaggacga tgatttcccc gacaacacca cggaattgtc
     1921 agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc
     1981 cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg tggtggcaat
     2041 gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac gttgcctgac
     2101 aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg aggagaaaat
     2161 gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag
     2221 cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag
     2281 aggttgatta tcgataagct tgatatcgaa ttctcaggtc gggtgctgcc cgttttgtgt
     2341 gatgggctgg atcttctcaa gagagacgtc agtgtcatag tccaaaatga cagacagggc
     2401 tggggacagc ttctccaagg gcttggctat tcccaccgcc tcctccttct tcaggccctc
     2461 agaggagccc acagcgtgtg ggatcagata caccacattg cagtccttgg agatggaacc
     2521 ccgaggctcg tgatggctgg aaaacaccac agccccattg ttattgatgc taaccgtctc
     2581 tatggcatta ttcgtcgtcg ggcaaagcct gtagcatcct aagagggtgg aaaatgcctt
     2641 ccgaaaatca gcattaaagg cataaatgat ggggttcaag gaggaattag cccacccaaa
     2701 ccacacaaac acgtcaaagg tgatggaatc gatgcagaag ggcttggtct ctccagaccc
     2761 acagaagggc accatgcagt tcaagatgaa gaaagggagc cagcagcaca cgaacacccc
     2821 catgatcacc gacagagtct tcagaacttt agtctctctt ttgaaggaca tcttaaagga
     2881 gctccagcgt gctgctgctc cggtgttgta ctccagcttg tgccccagga tgttgccgtc
     2941 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     3001 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggat
     3061 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     3121 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     3181 cttgccggtg gtgcagatga acttcagggt cagcttgccc tcggtggcat cgccctcacc
     3241 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     3301 caccaccccg gtgaacagct cctcgccctt tctcaccatg ctccctccgg taccgccctt
     3361 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
     3421 gtgatcgcgc ttctcgttgg ggtctttcga aagtttggac tgcacgctca ggtagtggtt
     3481 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtaggcgag
     3541 ctgcacgccg ccgtcctcga tgttgtggcg gatcgtgaag ttcgccttga tgccgttctt
     3601 ctgcttgtcg gccttgatat agacgtttcc tccctcctcc aggtccagcg cgtcggtgtc
     3661 gcgcggcccg gctgagatac gccgtatttg tttctgggcg atcctgtaga tcctggtgta
     3721 ggtgacaatc atgatggcca cagggatgta aaagcttatt agagaggatg aaatggcata
     3781 tgtcctgctc aagctggagt cacagttgtt gatggtcttg cccagggaag tggcattccc
     3841 ctcagaggga cctgtgggtt tcgccttgtg ccagctgagc tgcacgggga tgaaggagat
     3901 aagaaccgac aaagtccatg ccacactgat cagaatgaag gctgccttgg gggtcatctt
     3961 cctctcatac cgaccggggc tagagatggc ccaatacctg tccacactga tcacacagag
     4021 attgaggatg gatgcggtgg agcacatgat gtcaaaggcc acccagatgt tacagaagga
     4081 cccgaagggc cagaagccag cgatctcggc cactgctttc cagggcatga ccaagacggc
     4141 caccaagaga tccgatacag ccaaggagat gacgaagaag ttggtcacct tggatcgcag
     4201 gtggcggaac ctgatcacgg cggcacagac cagcgtgttc cccaggaggg tggacaggat
     4261 aagcagcgac aggaaacagg ctgtgagaat gcggaaggag aagtccctct ccgccaccag
     4321 cccggtgcct tccatggtag acgtgttgag agtcctaccg gtggtgccaa cttttttgta
     4381 caaacttgtg tcaccagtgg aacctggaac ccagagcagc agtacccata gcaggagtgt
     4441 gtctgtctcc atggtggcac cggtttctcg actgcgctct caggcacgac acgactcctc
     4501 cgctgcccac cgcagactga ggcagcgctg agtcgccggc gccgcagcgc agatggtcgc
     4561 gcccgtgccc ccctatctcg cgcctcgcgt ggtgcggtcc ggctgggccg gcggcggcgc
     4621 ggacgcgacc aaggtggccg ggaaggggag tttgcggggg accggcgagt gacgtcagcg
     4681 cgccttcagt gctgaggcgg cggtggcgcg cgccgccagg cgggggcgaa ggcactgtcc
     4741 gcggtgctga agctggcagt gcgcacgcgc ctcgccgcat cctgtttccc ctccccctct
     4801 ctgatagggg atgcgcaatt tggggaatgg gggttgggtg cttgtccagt gggtcggggt
     4861 cggtcgtcag gtaggcaccc ccaccccgcc tcatacgcgt gcggccgcag gaacccctag
     4921 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4981 aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac
     5041 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5101 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5161 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5221 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5281 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5341 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5401 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5461 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5521 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5581 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5641 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5701 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5761 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5821 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5881 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5941 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6001 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6061 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6121 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6181 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6241 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6301 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6361 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6421 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6481 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6541 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6601 gcacccaact gatcttcagc atc
//
